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Lewin γονίδια χ ( μέρος iii)

The fields of molecular biology and molecular genetics is rapidly changing with new data acquired daily and new insights into well-studied processes presented on a scale of weeks or months rather than years.

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Meroypoqn Kol q;" r q "4 't{ n MeroypoqlKoi Mnxovlopoi 25 3u r? OoToypoQ(o ;le tqv o6eto rov Jie Ren rot Jun Ma, U- niversity of Cincinnati College oI Medicine rot Cincin- nati Children's HosPital. Metoypoqrl otouq [lPoxoPutireq Metoyporpn orouq Europudrreq Iuppoqn rt EneEepYooio rou RNA Iro0eporrlro rL Evtontopdq rou mRNA Kototrurtro RNA Meroqpoorl XprlotgonottitvToq T0 [evertro K66tro KEOAAAIO 19 KEOAAAIO 20 KEOAAAIO 21 KEOAAAIO 22 KEOAAAIO 23 KEOAAAIO 24 KEOAAAIO 25 509 III
O Phantatomix/Photo Researchers. lnc ffi * K0T0 rIV ertqrlKuvon, I UeToypoqxn Quo0tr[60 UcToKI- veiror roro prlrog rou DNA rr r; otruoOo rou RNA enr- prlruveror oe roreuOuvorl 5'-+3' pe npooOflr4 vourtre- ond[olv oro 3' oxpo. * H Ueroypoqrl oropord ror ro d[r],trrvo rou DNA [ovo- oXrlporl(erot g6trrq q RNA notrupepdorl onoouv6e0e[ ond ro DNA erpoye(o oe Uro 0eon repponopou. H Borqprorrl RNA l-lotrupepooq Anoretre[ror onri l-lotrtr6g Ynopovodeg * 0r nupqveg rtov porqprorov RNA notrugepoorirv e(vor notrupeprl oupntroKo -400 kD pe yevxl Oopn 02pB'o. e H rordtruol enuetrelrot on6 rtg unopovodeg p xor p'. To Otr05v(upo r4g RNA l-lotrupepooqg Anorele(ror ono rov lluprjvo rou Ev(rlgou ror tov llopoyovro lypo * H poxrrlptorq RNA notrupepriorl uno6rorpeiror orov 0z0p'o nupnvo rou ev(upou nou roro),uet Tn UeToypo- qr1 ror tqv unopovodo o nou onoue(ror privo yro r4v evop[r;. ffi ffi @*83 Meroypoefi oTouq flpoKqpudlreq Etooytrlyrt * H Ueroypoqn npoyporonore[ror Ue Qopo 5'npog 3' o' dvo ergoye(o nou 6Xet qopo 3'npog 5'. H Meroypotprl llpoyporonore(rot ge Zeuyopropo Booetov oe gro u0uootr[60, A(euyoporou DNA * H RNA notrupepoor; OroXopr(er toug drlo rl,drvoug rou DNA oe pLo eqlgeprl <quootr[do> rot Xprloqionolei rov 6vo rtrdrvo og erpoye[o yro vo roreuOuver q ouvOeo4 luog oupdrlpolUolrnq otrtr1trouXrog RNA. * To Uflroq rqg EuootrOog ervor 12-14 bp evtir rou uppt- d(ou RNA-DNA p6oo oq rpuootrOo 8-9 bp. H Avr(dpoon qq Meroyporp4g'EXerTpio lro6ro " H RNA notrupepdorl npoo06veror oe pto 06011 unorr- vr1rr1 oro DNA yro vo oXrlpor[oer 6vo rtrerord oupntro- K0. * H RNA notrupepooq [envo q peroyporprl oqou ovor[er ro d(xtrtovo rou DNA yro vo oX1por(oer pro peroyporprrq rpuootri6o. 510 KEOAAAI0 19 Meroyporpq otoug l1poroputirreg ffi 19 r.t I rss Ew tp6)'eta Keqt€vou : Richard Gourse
tl 0 nopdyovToq o Tpofiororel lg rdulqreg npoodeolq oro DNA r4q RNA notrupepoorlg, 6ror drore n ouyy6- vero qg yto ro yevrro DNA vo ger6verot ror, ovr[Oero, 1 ouyy6vero Trlq yro Touq unouvrlrdg v' ou[over. l16q Bp(oxer H RNA l-iotrugepdorl rLg Atrtr1trouX(eg rov Ynoruv116v; * H np6odeon rrlg RNA notrupepoolq otoug unoxrvlreg e(vor rooo ypnyopl tlore potrtrov dev nporeuor yr' ontrr1 6roXuor1. * H RNA notrupepoorl npoodeveror oe ruXo(eg 06oerg oro DNA ror lg ovro),trooer pe dtrtreg otrtrntrouxieq 6otq drou ppe0ei evog unorrv4qg. To 0tr06v(u;ro Tpononoe[ror Koro rr1 Arepyooio Avoyvdr- pronq Kr Anetreu0epoorlg on6 roug Ynorrvrlr6g s OTqv Il RNA notrupepoorl npoo6everor o' rlvov unoxr- vt1it1, dLoXropt(er roug rtr6voug rou DNA yto vo DrlpL- oupyrloet Uro UsToypoQrKn Quootr(60 xr evotopordrver vourtreoridro oro RNA. * H rvop[n ono rrlv RNA notrupepoo4 dev elvor novrore enuuxqg. * 0 nopoyovrog o(ygo ouvrlOoq onetreu0ep6veror ono qv RNA notrupepoor; orov 1 veoouvre0eqlevrl otruoOo RNA onorrrloet pflrog -1 0 pooeolv. 0 fiopoyowoq Irypo Etr6yXer rrlv l-lpdodeol oro DNA Avoyvop((ovrog IuyxerpLp6veg Atrtrqtrou1ieq rov Ynoxr- vrlrdv s 'EvoS unortvrlrrlg npoodropl(erot onri qv nopouo[o ppoXeuilv ouvorvenxrirv oltrrltrouXuirv oe ouyxerprp6- veg 06oerg. e 0t ouvorvtTrreq otrtrqtrouXieq Tou urroKrvrlrrl ouvrl0oq onore),ouvror on6 gro noupivq oqv oqeqpro, evo e[0Uep5q Ue otrtrr]troux(o nopepqepl Ue rnv TATAAT orq 05or; - -1 0 xr 6vo otrtro e[ogepdq ge gro otrtrr1trou- XLo nopegqeprl pe r1v TTGACA oq 0eor1 --35. * 0t Eexttptorol unoxrvlr6g ouvrlOog drorp6pouv ond rr1 ouvorvelrrl otrtrr;trouX(o oe Uto n neproorirepeg 0e- 0etq. e H onoretreogortx6rrlro rou unortvqq pnope( eniolg vo enlpeo(eror xor on6 enrnprioOero orotXe(0. H Anore),eopotxdn1ro rov Ynorrvqrtlv Mnope( v' Au[o- ver 11 vo EtrLordveror ono Merotrtro[4 e 0r uopvlnr6qD UeToMoqerq nou pertirvouv qv orroTt- leogortxoqro rou unorrvlrrl ouvrlOtrrg pertilvouv tr1 oupg6prprooq npoq nq ouvorverx6g otrtr1trouX(eg, evtrr or uoeflrdq, perotrtro[erq 6Xouv ro ovrf0ero onor6tre- 0u0. * Merotrtro[erq oqv -35 otrtrntroux(o pnope( vo enrqped- oouv rr1v opXrxl npdodeol i1g RNA notrupepoolg. * Merotrtrd[etq onlv -1 0 otrtrr;trouX(o pnopel vo enrlpeo- oouv Trlv np6o6eol 11 n1v ovridpoorl onodroro{qg nou perorp6ner 6vo x),erord ofgntroro oe ovorXrti. l-lotrtreg lleptoXeg qg RNA l-lotrupepoorlg 'EpXovror Ape- oo o' Enogrl pe rov DNA Ynorrvrlq q H do[rl rou o70 ol],0(er orov ouv66er0r Ue Tov nupflvo tou ev(upou, enupenowog ortg neproXtlg yto npoode- o1 oro DNA v' otrtrqtrenr6pooouv Ue rov unorrvlrfl. * l-lotrtr6S neptoXdg rou 070 otrtrrltrenrdpouv Ue rov urroKr- vrlTn. * H unouovodo o enfor1g oul.rpotrler on]v 0rc, , -- -- rou unortvrlq" H Anorunoorl e(vor Mrl0odog YmInS Avotruoqg yra :: Xoporrrlprop6 rtov Atrtrrltrenrdpooetrlv RNA l-lotru pepoorl c - Ynouvrltrl ror, [evxdrepo, D NA- l-l poreivtirv * 0r 0uv0rvtTrK6q otrtrltrouxreq -35 ror -10 nop6Xouv ro neprooorepo onpeio enoqnq yto rrlv RNA notrupepoorl orov unortvrlrfl. 6 T0 qgelo enoqrlg pp(orowor rupro:q or4 pro ntreupo rou DNA. 0r Atrtr4trenrdpooerq Mero[0 rou I-lopoyovro Iiypo rot rou fluprlvo ir;q RNA llotrupepooqq Atrtro(ouv Koro 11 Atotpuyrl rou Ynoxrvlr1 " Mro neproXrl rou otylro rorotroppdver ro 0toutro e[ridou rou RNA rot npener vo perouvr;0ei yto vo dreurotrtivet rr1 ouv0eorl rou RNA. * Avenuuxe(q evdp[erq ouv1Oog ou;rpofvouv nporoU ro 6v(u;ro oXqpor(oet 6vo npoyporrr6 oupntroro entgrl- KUVorlq. & 0 flopoyowoq orypo ouvrl0oq onetreu0epoverot ond qv RNA notrugepooq g6trrg 1 veoouvre0ergevil otruol- 6o RNA vo qrdoer oe Urlxoq -l 0 nt. H Kpuorotrtrrrl Aoprl Ynoderrvuer'Evo Movr6tro yto 11 Meror[vr1o1 rou Ev(ugou e To DNA petouvelrot 6rop6oou evdg droutrou oqv RNA notrupepoorl Kor npoypoTorrote( pto on6roprl orpoqn oro evepyo r6vrpo. * Atrtroyeq orrl 6roprlpgtrloq ouyrerprp6vov eu6trrrrov e[oprnporov p6oo oro dv(upo etreyXouv qv e(oo6o rov vourLeond(orv oro evepyo x6vrpo. Mto Iropoqpdvl RNA llotrupepoon Mnope(vo iovopXioer # Mro oTopoTrlpeq RNA notrupepoorl pnopei vo [ovopXr- oer rn UeToypoErl xripovrog ro RNA peroyporpo yro vo Dqproupy1oer evo rorvoupyro 3' oxpo. H Borrqprorf RNA l-lotrugepool Teppor((er oe Atorpudg 0eoerg € Ynopxouv 6uo ro[erq otrtrr1lou1uirv reppolopou. Erei veq nou ovoyvtop[(ovrot prlvo ono rrlv idro rrlv RNA notrupepoorl Xop(q rr1 oupperoXq rovev6g rurroprrou nopdyovro ouv40rrlg ovorp6povror a "evdoyaveC reppouoreq" (intrinsic terminators). Atrtreq onououv 1-rLo xurroptrrl nporelvrl nou ovopo(eror po (p) xor ovo- tp6povtot oq (TepuoTtoTtig e[opr6gevot onri qv p,. * 0 evdoyevnq reppouopog npoUno06rer ovoyvtirpro4 ptog otrtrqtrouX(og reppouopou oro DNA nou rordrxo- norei pro 6opn Qoupr6rog oro RNA npoi6v. 6 T0 orlpoTo yro repgolop6 pploxovroL rupltrlg gSoo oe otrtrltrouXreg nou 6Xouv n0n UeroypoQe( on6 qv RNA notrupepdoq xr 6rol o repporLoprig ppforeror un6 rov 6treyXo rou erpoye(ou ro/r1 rou RNA npoirivrog nou peroypoqer n notruUepoon. llrirg AeLroupye( o l-lopdyovrog p; € 0 n0p0y0vT0g p etvot pto nptrlrefvq nou npooddveror oro veoouvre0eq:6vo RNA rot perorrve[ror xoro pqrog tou RNA yro v' otrtrrltrentdpooer pe qv RNA notrupepo- orl Kor v0 qv onetreuOepdloer on6 ro o0gntroro enrpl- KUV0rlq. #wa :,!E*i ffi g*ee 1;ia;: .dg€!r14 qGL&&; .-ffi+. Ei,E*" Fg@. 4E&*; l I l 4t."7i4., ouvey[(erut ou1v enopevr1 oelido 511
ffi H Ynepetriro:oq Ervot lqpovrx6 XoporqptorLxo fllq Me- roYporllq " H opvlnrr; unepetr(roo1 ou[dver qv onoretreoporuo- qro ronourlv unortvrlr6v eneroq poqSd qv ovr(6poor1 onodroro{1q. a H peroypotprl dqproupye( gnpooro ono ro 6v(upo Oel- rdg unep6trtreg ror n[otrr rou opvrllr6g unep6],rxeg, or onoleg np6neL v' otpoLpe0oUv onrl rr1 yupootl xot qv ronoioopepool. qlE}i H RNA l-lotrugepool rou 0oyou T7 Elvor 6vo Xplorlio l-lporuno IuoTlUo & H orK0y6vsr0 ro:v RNA notrupepootirv tou T7 e(vor g.rovo notrunenr(Oro trovo v' ovoyvrop[(ouv roug unorrvrlr6g rou qoyou ror vo enuel,ouv notrtreg ond ng dp0orrl- proqreg tov notrupeptirv RNA notrugepootirv. * 0r rpuorotrtrrr6q dog6q rov RNA notrupepoorlv qg oL- roy6verog rou T7 pe ro DNA rouronororjv qv neptoXrl npoodeolq rou DNA, oro evepy6 r6vrpo xor uno0et- rvuouv ;lovre),o yro rri 0rorpuyrl rou unorrvqq. ffi O Avtoytr-rvro;rdg yro rouq l-lopoyoweg liypo Mnoper vo Pu0p(oeL qv'Evop[1 * Io E. coli eXer enrd nopdyovreg o(ypo, xo06vog ono roug onoloug o0ei qv RNA notrugepoorl vo (errvfloer oe pro opodo unoxrvrltrirv nou npoo6top((ovror ono ouyrerpLp6veg otrtrntrouX(eq -35 xot -1 0. * 0t evepyrirnroq ToJV 0rorpopertr6v nopoydvrtov otylto pu0pi(ovror ontl Dtorpopelxou g Lr4Xovropoug. ffi Or flopdyovreg l(ypo Mnope( vo Elvor Opyovrop6vor oe Axotrou0ieg * Mro oKotroue[o nopoyovrov oiypo dnproupye(ror orov dvog nopoyovrog oiygo onoueiror yro rq peroyporprl rou yovr6[ou nou rodrronorel rov enoplevo nopoyov- tu, * To npdrruo yov[dto rou qdyou SP01 peroyporpovtor ono rnv RNA notrupepoon rou [evrorr1. e 'Evo 0rt0 ro nptirLgo yov(6ro roDrronotel 6vov nopoyov- ro o(ypo nou trlOei r;v RNA notrupepoorl vo peroypo- ger ro evdtdgeoo yov(dto. 19.1 Erooyoyfi * H Ueroypoqrl npoyltotonoteirot pe qopo 5'npog 3' o' 5vo er- poye(o nou eXet qopo 3'npog 5'. H pretoypo<pr1 nopdyer pto oluo(6o RNA ge raut6o4- pry aMqAouy[a Ue rov 5vo x].6vo tou DNA, nou Uept- x6g gopSg role(ror x<o6rxonor4rrxrig xArivog (coding strand). Aut6g o r<Ltovog SXer <popd 5'-+3' xor e(vor o u pnlr1papaux6q (co m ple mentary) (6nLodn (euyopcbver) ge ro eKpoye(o (template), nou SXeL <po- pd 3'-+5'. I-t' out6 o nop6protog pe ro RNA x).rilvog * Auo ono ro ev0Lopeoo yov(6ro roduonorouv unopovo- 6eq evoq nopoyovro o(y;lo nou o0e( qv RNA notrupe- poorl vo ueToYpouer ro oQrlro yov(6r0. H Inoproyeveon Etreyxeror ono l-lopoyovreq lypo * H onoproy6veoq 6rope( evo porrrlpro o' evo prlrptro rurropo nou urpiororor truor1 rr dvo onopro ttou oTit- treu0ep6verot. I Koee [eXopror6 6ro;rSprogo npoXoper oro enopevo oro0ro ovdnru(qg ouv06rowog 6vo rorvoupro n0p0- yovro olypo nou erronl(er rov npolyorlpevo, * H rnrKorvovlo Uero[u rorv Duo dropepropdrtov ouvrovi- (er qv eniloyri rou rorotrtrr1trou Xpovou yro Trq ovTrKo- tootooetg ttrlv nopoyovrtov oty|to. 0 Avtrepponopoq Mnopei vo Elvor'Evo Pu0prolxo Iup- tsov * 'Evo o6untroKo owrrep[oTropou enup6ner orrlv RNA notruUepdol vo 6ropooer 6topdoou reppouordtv. * 0 Qoyoq trou0Do xpnotponorc[ ouorrlporo ovTtTtpuoTr- olrou yro vo puOpioer rooo ro npdrgo ooo Kot T0 otIiUC perdyporpo, otrtrd ro duo ouorrlporo treuoupyouv p' ev- retr6g drorpopenro0g plXovrol.roug. * H rpooDeq nopoy6vrotv oro veoouvteOetg6vo RNA ouv66sr rtq nporelveg ovTrTrpgoTroUo0 oq 0do1 iou reppolor1 6rop6oou proq 0qtrroq RNA. * AvnrepUortopdq rnq peroypoqrlg oulrpo[veL en[or1g roL oro oneprivro rou rRNA. 0 Kuxtrog rou Borrqplorou Ayyetrrorprjpou RNA * Iro porrrlpto, n UeroypoQn ror 11 ;leroqpoon npoyU0- ronorouvtor rouroXpovo (ou(euyU6vn Ueroyporpl/Uerc- Qpoon), ro069 ro p$ootoporLo [errvouv q perogpoor evoq mRNA npLv otrortrr1pol0e[ 1 ouv0eofl rou. * To poKTrlproKo mRNA e(vor ooro069 Ue Xpovo rlpr(orqc trQov trenrtiv qq topog. s 'Evo BoKTnproro mRNA pnope( vo e[vot notruotorpovtro ge notrtrrlg rordrreg neproXdg nou ovTrororxouv oe 6ro- qoperLro yovi6ro. nepitrnUn ffi ag.i*,#,* 5! L PJ't ,fiFts{ t:t &:!( l xolefuor xtrrilvog p4-expoyeio (non-template strand), evri: ow6g nou l,enoupye( og er<proye(o yro rrl o[v0e- on rou RNA rq}'ekor rLctrvog exproyeio, 6no:g tpofue- ror ornv EIKONA 19.1. H oUv0eoq tou RNA rotoLUeror on6 ro 6v(upto R- NA notruyep6o4 (RNA polymerase). H peroypocprl [e- rtvd 6rov q RNA noluprepdor; npoo6e0e( orov unoxrvqr{ (promoter), puo'er6rrcl neproXrl, orrlv opXtd rou yovr6(ou. O unorrvqr4g nepil.oprBdver to npd:ro (erlyog Bdoecov nou preroypdgeror oe RNA (ogerqpio - startpoint), roOrig Kor rrq neprBdl.trouoeq Bdoerg. An6 ourj q 06oq, r1 RNA noluprepdo4 perorrve(tor xo- rd pqrog rou erpoye(ou, ouv05rowog RNA prdXpr vo qrdoer oe Uro otr).4},ou1(o repporror{ (terminator), 6nou retrerc0ver ro perdyporpo. 'Erot ;uo peroypogrxri pov66o (transcription unit) erre(verqr qn6 rov unort- 512 KEOA,A|O 19 Meroyporprl oroug l-lpor<oputirreg Boorxeg cworeg
K),6voq un-eKuoys[o 5' TAC * CG ET,qC G GTCAATG CAICTAC CT 3' .l';':;,:j !:.: i1:t-i:ii:l i.; lt & I'jp.'i-l G jrt +,,$*i * A vrlrri 6oq rov repUorrmrl. To Ko(pro Xoporrqptortx6 rnq Ueroypo<pu<qq provddoq, 6noq <po(verot orqv ElK0- NA 1S.2, e[vor 6rt onotel,e[ 6vo rprrlpro DNA nou Xpnorpo- noe(tor og expoye(o yro rnv nopoycoyrl ev6g provo6trof prop(ou RNA. Mro peroypogrxrl provd6o pnope( vo rto6t- ronoe( neptoo5rcpo on6 5vq yov(6ro. Or oMrll,oufeq nou Bp(orowor npw on6 rnv oee- rqp(o ovogdpowor ovo6rx6g (upstream), evdr outdg nou Bp(or<owor perd qv ogeqp(o (g5oo orq Ueroypo- q6pevq ol,r1LouXio) rog xo0o6rx5g (downstream), Iuyyd 4 oMrlloul(o tou DNA ypdeeror 5rot c0ore vo <po(verot pr6vo o rd.cbvog prrl-erproyeio, o ono(og (6no:g npoovo<p6p0r1re) 51et rrlv (6to oMrlloul(o pe ro RNA. Or O5oerg rov Bdoerrlv optOpo0wot Kot npog rtq 6fo roteu0Uvoerg on6 qv ogerrlp[o, nou ovopd(etot + 1, or de oprOpro( ou[dvouv npoq rnv r<o0o6u<q <popd. H Bdoq Trprv rrlv oqeqp(o oprOprefuot -1 Kor ot opvnrt- r<o[ oprOgo[ oufdvouv nrlyo(vowog ovo6trd. (Kop(o Bd- oq 6ev 61et rov opr0;r6 0). To opXrr6 npoi6v rnq Ueroypoqrlg nepil.o1-tBdvet to ou0ewrr6 5' drpo ror ovoprd(etqt rporoyev6g per6- ypqeo (primary transcript). To npcoroyev4 pretdypogo rou rRNA Kqr rou IRNA utp(orowqr pto 6epyoo(o top(- Uovonq rord rqv ono(o otrLr1trouX(eg'rd:v drpov toug r6Bowor on6 ev6ovourd.edoeg. To cirptpro npoi6wo trrrv -35-10-1 +1 +10 --.+ Avo0rro KoOoDLr<o IVleroyporprrq povodo elvor 1uo otrtr1trouXio DNA nou peroypoqerol o' evo evrolo RNA, [extvtlvrog ono rov unort- vrlq r<or retreulvovrog orov repportor1. wNw{s=Ktrtirvoq erpoyaio METATPAOH RNA getoypotpo ElK0liA 19.1 AeLroupyro qg RNA notrupepoorlg e[vor v' ovlypoqet 6vov r],6v0 rou d(r],orvou DNA oe RNA. H otrtrr1trou1ro rou RNA ervot I oupnlqpupattr4 ge rov xtrtrlvo tergoyeLo xat raur 604 p4 ye rov <otdtronotrlrtxo rrtrovo = 5'UACGCGGUACGGUCAAUGCAUCUACCU onepov(rov rRNA r<or IRNA e(vot moOepd, trre 1p6vo (or1q nop6poro Ue ro Xp6vo yevtdq tolv Borcrqp(ov. Av- r(Oero, ro npcoroyevrl getdypogo rou mRNA 66Xowot oXe66v dpreo4 en(Oeor1 on6 ev6ovouxledoeg r<t e[u:- voux.).edoeg. 'Eror, o prSoog Xp6voq (onq tou Borcrqpto- r<of mRNA e(vor pr6vo 1-3 Lemd. Iroug europutiteg, to pretdyporpo rRNA ror IRNA utp(orowot Korepyoo(o t<ot ro npoi6wo nou npor[mouv e(vot oroOepd, 6nog mo Borrqpro, oLl.d to mRNA e[vqt noLti nto oroOepd on' 6,r oro Borcrrlpro. Ot rpononoqoetq Kot 11 ono66ptqoq rov mRNA 0' ovoLu0oriv ctrlv Ev6trya 19.23. H preroypogq e(vor ro np6to ord6to orq Srgpooq rcov yovrd(rov xor ro Brlpo sro ono(o oorefuot nto ou;yd p(Opuoq. Or puO;umtxo( nopdyoweg ouprd r<o0op((ouv ov 6vo ouyxer<prpr6vo yov(6ro 0o pretoypo<pe( pSoo qg RNA noLuprepdonq. En(oqg elSyXouv rot petoyevSore- po mddro rnq Ueroypogqq r<ot dMo mddto qq 6r<- qpoonq ro:v yovr6(olv. Ario orlprowu<6 epor(proro oletrd Ue rn Ueroypo- qq efuor: o t-lcirS Bp[or<er q RNA no].uprepdorl unortvrlt6g mo DNA; Aw5 eivor pr6poq ev6g yevu<6tepou epotrlpo- rog: l-lig (eXcop((ouv or nporefveg ng ouyrerptpr6- veg 06oerg np6odeoqg mo DNA on6 ttg dl,l,eg oMqLouX(eq; o flcirS oLLrllentdporiv or puOprorrr5g npoteiVeg trle rqv RNA noluprepdo4 (r<or pero(U toug) rbme vo evepyonorrloouv 11 v' qvome(trouv ouyrerptpt5vo Brigoro rotd rrlv Svop(r1, r4v entprlruvorl ri tov rcpprotropr5 qq uerqypoeriq ; Ie our6 ro reqdtroro 0o neprypdtloupe rtq otr4Le- nr6pdoerg qg Borcrqpror{g RNA no}.ugepdoqq Ue ro DNA, on6 qv oplu<ri enogrl rrlq pe tov unoxtwltti, r1 Ueroypoen Kor rnv oneleu06ptoon qS on6 to DNA p6- l,rg oloxl4pto0e( ro pretdypogo. Ito KeEdl.oto 20, 0' ovol0oouge tg ow(motleg owr6pdoetg prero[U rov europutoru<c0v RNA notruprepootirv Kor rov ergoye(<ov roug. To Kerpdl,oro 26 nopouotd(et dtdtpopouq rp5- nouq Ue toug ono(oug or pu0ptmtrSg npolteiVeq Kot ot nopdyoweg prnope( vo po4Orloouv ti vo nopepnod! oouv rn Bonrqpror<q RNA notruprepdorl vo getoypdtlet 6vo ouyrerprpr6vo yov(6ro. Iro Ke<pdloro 27 e[erd(ou- 9.1 Erooytoyq 513
19.2 H Meroypogri Zeuy6polpo Bdoeorv A(euydporou DNA llpoylroronoreiror pe o€ Uro (Ouoo^i6o,) pe nrilq or [elotpror6g pu0;uorrr5g owrdpdoerg gnope( vo ouvdud(owor oe nol,unLor6repo 6(rcruo. To Kegd- Loro 30 ovoL0er np6o0eroug rp6noug prlOpuoqg, pero- [ti dMtov n Xpnorporofiorl gu<pc0v RNA, r<or qvo].Uer nrilg ourdg or oMqlenr6pdoerg prnope[ vo ouvddowor oe peyolUrepo 6(rcuo. roypogrx{ quoo}.[6o", mrlv ono(o ro DNA nopo6rxd 6toXrr:p(eror orouq 6Uo provo0g drc0voug rou Kor o xi.cbvog extrroye[o Xpnorpono€(rot yro vo xqreuO0ver q o0vOeorlrou xl.drvou rou RNA. H oLuoGo rou RNA ouw(0erot on6 to 5, npog ro 3, dxpo pre npoo0qr<q v6ov voux.l.eon6(ov oro 3' drpo rqg ou(ov5;revrlg oi.uo6og. H 3'-OH opd6o tou reLeuro(ou vourlsort6(ou nou npoor60rlre or4v ol,uo.(6o owrbpd pr' 6vo eroepl6;revo 5'rpupoorpopx6 vourLeoo(6ro. To etoepX6prevo voux.l,eoo[6ro Xdver ng 6Uo re].u<6g gco- o<popu<6q oprd6eg (y r<or B), evr6 4 opd6o o Xpnoqonor- efuot oro goorpo6eoreprr6 6eopr6 nou ro ouvdder arnv qluo(6o. H roXUr4ro qg ouvoLx4g ow(6pooqg yro rr1 Borcrqpror<f RNA noLuprepdorl prnope( e(vor dog -40-50 vouxLeor6ro/6eurep6l,emo orouq gToC yro ro nepro- o6repo prerdypogo, nep(nou (oog pe qv to1tirqro pe- rdgpooqg (15 opwo[do/sec), oMd no].ri ;ur<p6repq on6 rrlv rolrir4to owrypoqqg rou DNA (-800 bp/sec). H RNA noLugepdorl 6qpuoupye( rr1 ;reroypogrxrl <puooL6o pr6).rg npoo6e0e( o' dvov unorrwlrn. H ElK0l{A 19.4 nopouord(er qv RNA noLu;repdoq vo perorrvefuor r<ord prqrog rou DNA, pre q guool(6o vo peroxrve(ror po(( qg Kor rnv o).uo6o rou RNA v' ou[dva oe pulrog. H 6repyoo(o rou (euyopcilUoroq rov Bdoerov Kor rrlq npoo0{r<r1g Bdoecov ;r5oo orq <puooA(6o roro.)Jeror xt enr4pefuor on6 r4v (6ro ulv RNA noi,uprepdo4. H 6opr1rrlg quooL(doq 96oo oro oritrrnLoro rnq Ue- royporp4g go(veror ar4 Aemopepn dnorln qg E|K0NAI 19.5. Ko0rig q RNA noLuprepdor; perorrvefuor r<ord prj- Koq rou DNA erpoye(ou, ro 6(r),ovo (erultyetor oro pnpoorrv6 pSpoq qg guool,(6og (onUe(o [erutriyprorog e H RNA notrupepoorl droXropr(er roug duo rtrdrvoug rou DNA oe pro eqrlgepl ,,rpu00L[60" Kor xprlouon0re( rov 6vo r],tirvo org ergoye(o yro vo roreu0uver rr1 ouvOeo4 prog oupntr4prrt- porxrlg otrtr1trouXLoq RNA. * To Unroq qg quootrrdoq e(vor'12-14 bp evdr rou uppr6(ou R- NA-DNA p6oo orq quootrOo 8-9 bp. H peroypocprl Xprlor;ronore( q ouprnL4prollolx6rqro ro-lv Bdoecov, 6ncog xor or dMeg owr6pdoetg noLuge- prol.rori, dqLodq 11 owrypogq xoL 4 ;rerdgpoorl. H .,',, ': ,', . ono6(6er q yevrKr1 opxn rnq ;retoypog4g. H o0v0eo4 tou RNA npoyporonore(ror pr6oo oe Uro (Ue- Kodtrdg x.tirvog Ktr6voq eKuoye(o 0r rtrtirvor rou DNA dtoXrop((ovror yro vo oX4porLoouv Uro UeToypoQrKrl rpuootrido. To RNA ouvrlOeror pe oupntrrlpotporxo (euyoputlro rtov pooeov pe rov evo ono roug r),(rvoug rou DNA. 514 KEOA Al0 1g MeToypoQ1 oroug l"lporoputirreg w;.@,TWrx"= ffifl.M-ffix ''r*er'ffi"x-ffi EIK0NA 19.4 H geroyporpq npoyporonorefror gdoo oe Uro quoq- tri60, orqv onoio RNA ouw(0eror pe (euydptopo rov pdoetov 1r, 6vuv on6 roug rtr6voug rou DNA oqv nprioropo [erutrryp6vrl neproXrl. Ko0tig perouve(ror q quootrbo, ro D(xtrrr:vo DNA [ovo- oXrlp0ri(eror n(oo u1g, exron[(ovrog ro RNA un6 popqrl povrlg nol,uvouxl,eo16xrl g otruo6og. To DNA onodroTdoor.o, I H oUvgeo4 Tou RNA [euvd mr1 quoo],6q Boorxdg evvoreg J + 0uootri6o peroypoqftg +,
Merorw4oq rou ev(Upou '+ 05oq npdo6eolg rou RNA EIK0NA 19.5 Kord q Ueroyporprl, I rpuootri0o Drorrlpeiror p6oo oq porqproxrl RNA no)upepdol, rou [erutrfyer xor [ovoru)ryer ro DNA rot ouv06rer RNA. - unwinding point) ror ro DNA enqvefurd)veror oro n(- oro rprlpro (o4pe(o (ovorullyprmoq - rewinding point), To pqrog rrlg <puool,[6og e(vor -12-14 bp, evcir ro ;uixog tou uBpt6(ou RNA-DNA pr6oo on1 rpuooLbo pr6llq 8-9 bp. Ko0rig ro 6v(u;ro petorwe(ror r<ord prrlxog rou eK- poye(ou, [ovoq4qpor((eror ro 6[r],rovo DNA ror ro RNA erron((eror o g eLeri 0ep4 nol,uvo u x.leorr6rnj ol,u o(6o. Ie rdOe 6e6op5vq onypf , ro rel,euro(o 14 prBovouxi.e- or(6to rou ou[ov6pevou RNA Bp(orowor oe oUpntror<o pe to DNA xo/r1 ro 6v(upro. 19.3 H Avri6poon Tnq McToypoefiC'Ex€l Tplo lrd6to " H RNA notrupepoorl npoo06veror oe pro 0eorl unoKrvrlTrl oro DNA yro vo oXqporloer 6vo r],eroro oupntroro. " H RNA notrupepoorl [exrvo q Ueroyporp1 orpou ovo[[et ro di rtrrr:vo rou DNA yro vo oXqporloer Uro UeToypoqrKrl rpuootrido. * Koro r1v enrpfruvor1, I Ueroypoqtrl rpuootrrdo perortvelror roro pr;roq rou DNA ror 1 otruo[do rou RNA entplruveror oe roreuOuvorl 5'+3' pe npoo0rlrrl vourtreortdlrrrv oro 3' orpo. * H Ueroypoql oroporo rot ro dlrtrtovo rou DNA [ovooXlporr- (erot gotrrg 1 RNA notrupepoorl onoouvdeOel ono ro DNA er- poyelo oe Uro eeon repportolrou. H ow(dpoon rnq Ueroypognq drorp(veror oe rp(o orddro nou cpo(vowor ornv i', ' : (1) 6vop[n (initiation), orrlv ono(o ovoyvrop((eror o unoxrvrltrlg, 6q;rroupyekor Uro guoo).[6o r<or (erLvd 4 oJv0eorl rou RNA, (2) enrp{xuvon (elongation), orrlv ono[o 11 guool(6o ;."tetoxrve(ror r<otd pr1<og rou DNA, r<or (3) repponop6g (termination), xord tov ono(o to RNA perdypo<po oneleu0epcbverou<or r<).e(ver q rpu- oo).(6o. H (6to q 5vop[r1 prnope( vo 6Lor<pr0e[oe noMd Bqpro- to. H ovayvdtpon rou expoye[ou lexwd pe u1v npoo6e- ENAP:H Avoyvtirptorl rou expoye(ou: 11 RNA noLugepdol npoo0everoi oro 6[x],rovo DNA To DNA {erutrryerot orov unortvrlq llotrrj ppoXe(q qtruo6eq ouw[0ewqt Kr oflelsueep(tvovror EflIMHKYNIH: 1 H notrupepdorl ouv06ter RNA TEPMATIIMOI: H RNA notrugepdon Kqr ro RNA onetreu0epdrvovror ElKOllA 19.6 H peroyporprj 6xer rp(o md0rq: To 6v(upo npoo66- veTor oTov unouvrlr4, onodroroooer To DNA xor nopop6ver or(vr1- ro xord qv 5vop(1: Merouve(ror rord plrog Tou eKpoye(ou Kqrd r4v enrp4r<uvq rqr 6r(ororor orov reppouogd. oq rnq RNA nohupepdo1g oto Sklavo DNA oe pn aA- l4louy[a DNA nou xaleftcl. unoxlrpr1g @romote!. To 5v(upro oXrlprot((er np6ro 5vq xtreror6 o0prtroKo (closed complex) oro ono(o ro DNA nopopSvet 6(r<Lro- vo. 'Enero, (eruLfuer ronrd ro tprrl;ro DNA'rou unorr- vqrrl nou neptloprBdver rn eSon Svop(r1g r4g peroypogqg yro vo q4qpot(oer ro ovol;6r6 oripnAoxo (open complex). O droXroptog6g rou 6u<Lcirvou DNA ro- Ormd rov r<i.c0vo-er<proye(o 6ro0Sorpro yro (euydpcopo Bdoecov pre eroepX6trrevo prBovoux).eor(6ro xor oriv0eoq -rcov npcbrrov vour.treon6rrc0v 6eoprdlv oro RNA. H qdon ouq prnope( vo nopo'ro0e( qn6 qvenrule[g on6nerpeg Svop[4q, orrg ono(eg ro 6v(upro qndprer BpoX6o pret6- ypoqo, runxd rdro rulv -10 vouxleond(cov (nt), evcir e(- vot or<6;rq npoo6e6epr6vo orov unorrvrlri. To 5v(upo ou;yd r<dver 6ro6oXxorig 66xtroug ovenrulcirv Uerqypo- grilv r<or lovopX((er q o[v0eoq rou RNA. H <pdo4 rqg Svop[r1g re^.er6vet 6rov ro 6v(upro retrrd r<oropOcOoer v' enercrefuer qv ol,uo[6o Kor vo onopor<puv0e( on6 tov unorlqrrl. Krivoq erpoye(o DNA lii .: H AvTiDpooq rrlg Meroyporplq'EyerTpLo Iro0ro 515 Boorxeg eworeg
H enLp-rrp<uvoq ouv(ororor oe npoo6eurrrrl preror<fvq- or'l rou ev(0pou trre 6tordpo(r1 tou (euyopri-rprorog rov Bdoeov oro d(x.tro.rvo DNA, ogrlvowog erreOerpr5vo rov r<Lr.rrvo-expoyeio yro npooOrl<q voux).eorL6(rov, Kor Ue- tot6nroq rnq peroyporpLxqg guooLf6og nro ro0o6rxd. Ko0rrrg perorrve[tor ro 6v(upro, o x],covog-exproye[o qg npoorrrpLvd (etuALyprdvlg neproXrlg (euyoprrtver pre ro veoouwe0eLpdvo RNA crro o.r;pe(o qg o0(r1oqg. Nou- r).eori6ro npom[Oewor ol.rorono].Lr<d oro 3' dr<po rrlg oulovdpevrlg oLuo(6og RNA, o'Xqpor((owog 6vo uBpG 5Lo RNA-DNA ;r5o'o mqv lerulryprdvrl neptoXrl. l-1(oot on6 rrlv [erufuypr6vr1 nepropl, o r<].ovog-er<poye(o rou DNA (euyoprrrveL pe -rov ouOeyrrrd r<),rrivo "ro[pr" rou, yto vo lovooXripor(oer rq 6rn).r1 dLrr<o, evctt o ou(ovdpe- vog N).rrlvog tou RNA [enpoBdtr].eL ond ro dv(upo. H nopo6ooLorr; on6r,l.rr1 6l r1 enr;rrp<uvor1 e(vor ;rovo- rovq 6tepyoo(o, o-rqv ono(o -ro 6v(up_ro npocu0e(ror r<otd pqrog rou DNA pe oroOeprl rolirqro nou owrmotle( ornv rrpooenKn rcuv voux).eorr6(olv, dXeL oMdler ro re- Leuro[o XpdvLo. H RNA no].uprepdorl npoyporonore( nor-ioq 11 ordpq ror oropord oe ouyKeKprpdveg oMrl- trouX(eg. H em6nrorl rou 3' dxpou rou RNA ond ro evepy6 r<6,wpo trrnopei vo nporol6oer "ofiroeox6prlor)" rrlg noLupepdoqg r<oL ogo(peoq ;_reprr<rr.lv vour<LeorL- 6(ov on6 trlv cu(ov6prevq o).uo[6o rou RNA fiprv rr]v enoverr[v4oq. O rep ;.rottopr6g nepiloprBdvet ovoyvcirptor; oMrlLou - Xrorv nou 6(vouv mo 5v(upio ro orlpro vo oropotrloeL r4v nepondpot npooOqr<r1 vour<).eol6iov ornv o).uo(6o rou RNA. H guo'o}.(6o xoroppdel<oOriq ro uBp(6Lo R- NA-DNA onodroroooeror Kor ro DNA lovobq;rtoupye( 6vo E(xLrr:vo, 11 611 puoupy(o goo<podLeoreprotv 6eo pur-rv dLoxdmeror rar ro orjpn).oKo Ueroypogqg 6tolrrip((etor oro ouorolKd on6 ro ono(o onopr[(eror: RNA no].upe- pdoq, DNA r<oi DNA ;reroypo<po. H aAA7Aouyfu rou D- NA nou xareu9lva r ArjEn rnq psraypaErlq ovolto(erat reppau.orq. 'l 9.4 H Borrnplorf RNA lloAuUepdon AnoreAeirot on6 noAAdq Ynopovd6eq ". 0r nuprlveq rov Borqproxtrrv RNA notrupepoorirv oivor notru- pepfl oupntroxo'400 kD pe yevLrrl 00prl 0208'0r. ., H rorotruorl enustrefrot ono rLg unopovodeq p ror B'. OL r<o).rjtepo XopqKrnprotl6veg yevettrd xor BtoXqpru<d RNA no).upepdo'eq npo6pXowor on6 Bor<rr1pro, t6(<i.rg on6 ro Escherichia coli. Oo16oo, oL p6veq BorcrrlpLo- r<59 RNA nolupepdoeq yro rtq ono(eg 51ouv npoo6ro- pLore( uUnlrlS ovd).uo11g rpuoro).Lrr<6g 6op6g, npodplowoL on6 60o Oeppr6gil.o Borcrqptor<d e[d4, ro Thermus aquaticus xat Thermus thermophilus. Ie 6o ro Borrrlpro, p6vo 6vog r0nog RNA noAugepdorlg ei- vor uneU0uvoq yrq rq orivOeoq rrov rRNA, mRNA xor IRNA, qw[0ero on6 toug europucirreg, srouq ono(oug ro rRNA, mRNA rqt IRNA runxd preroypdtpowor on6 6rogoperu<59 RNA noluprepdoeg, ng Pol l, ll xot lll. l-lepfuou 13,000 g6pro RNA noluprepdorlg undplouv oro rUrropo rou E. col/, nop6rr o orprB4g opr0;rdg norrf],er ovd],oyo Ue trq ouv0rlr<eg oU[r1oqg. Irqv npoyllorrr6rnro, oe rd0e 6e6opr5v4 orryprrl 6ev ep- nA6Kowqr 6).eg or RNA noLuprepdoeg orq psroypo- <prl, 6l,eq 6prog e(vor npoodedepdveq oro DNA, e(re et6rrd e[re ;rr1 er6rrd. To n)r1peg 6v(u;ro 11 otro6v(upo (holoenzyme) rou E. coli 51er propror<6 Fdpoq -460 kD. To o).odv(upro (qzpp'oo) pnope( vo droXroptme( oe 6Uo oumotrxd, rov nupr;vo rou ev(riprou (ozpp'to) Kor rov nopdyowo o(ypo (notrunenr(dro o), nou epnL6reror er6rxd orqv ovoyvcilprorl rou unoKrvrlrrl. H oUorooq rou oe unoUo- vd6eg ouvor!((erot orrlv EIK0NA 19.7. Ot uno;rovddeg B ror B' go(( rpoyUoronoroUv r4v r<ordl,uoq rou RNA ror qnore^,o0v to preyol,rirepo rpulpo r4g pd(og rou ev(0- ;rou. Or oprLvo[rrdg o].].r1].ouX(eg rot ot rpro6tdmoreg 6op6q rouq e(vot ouwqpnp6veq, x6tt nou rqxuer yro nq peydleg unopov66eg rtov RNA no).upepoociv r<or on6 rtq rperq enrxpdreeg rqg (or1g: Bor,rrlpto, oplo(o xr eu- t<opuc0reg (B).6ne Ev6rrya 20.2). Auto uno64l,cbver 6rr or Boorr5g t6r6rrpeg rnq Ueroypogrlg e(vor roweg prero- [ti rtov no].upeprirv RNA no].ugepood:v 6].cov rrov opyo- vrogd;v. Or B xor 9' Uo(( oXq;ror((ouv ro evepy6 r5wpo tou ev(6prou, rov r<ripro 6(oulo 6ropr5oou rou ono(ou 6rSpXeror ro DNA xo-rd tov r<0xi.o rqg preroypogrlg, ro 6ewepe6owo 6(ou).o 6ro;r6oou rou ono[ou ro prBo- vouxi.eo{6ro rou unoorpriporog ero6plowor oro 6v(u- [ov[6to llpoiriv Aenoupy(eq rpoB B unopovddo (r51 kD) rpoA 2 o unogovd6eq ftA Iuvopgotrdyqorl Tou ev(ugou ;; ^; i'r ql7 ovoyvopron rou unoKrvrlnl xoOeg(o) npooddver rdnotouq 1|i'-onoqrdq l- Kurotrurtxd x6wpo glrpoC p' unopovddo (1 55 kD) o unopovo6o (1 8-70 kD) Etdtxorrpo rou unorrvrlrrl or unopovd6o (r 0 kD) 'Ev(ugo rou E.coli =460 kD 0t RNA notrupepooeg rov euporqpiov eXouv ne.- re tunoug unopovo0ov: oL o, p, p' ror o eXouv potrtrov oroOepc U6ye0oq oro dlorpopetro erDrl porrrlp(ov, evtir 11 o norrltrer ne - prooorepo. rpoD rpoZ 516 KEOAAAI0 19 Meroyporprl oroug llpoxopurireq Booxdq 6woreg
uo orrlv nope(o rouq npoq ro evepY6 KSwpo Kot ro 6[- ou^o e[66ou 6ropr6oou tou ono(ou to veoouweOetp6vo RNA eyrorotre(net ro 5v(upro. Ie oup<pcov(o pre ro p6Lo ourrilv ttov unopovddcov oe 6l.eq ow5q ng Lettoupy(eg, prerotrtr6(erq oro yov(5to rpoB xot rpoC nou xtodronot- oriv lg unoprovd6eg B ror B' en4ped(ouv 6tro ro ord6to rqg pretoypogqq. To 6tprepdg nou oXrlproil(etot on6 ng 6Uo q unopo- v66eq Xp4orpre0er toq u<p(cr:pro Yto rrl ouvopprotr6yrlo4 rou KewprKo6 ev(0pou. H C-retur{ neplolli (C'termi- nal domain - CTD) rov o unopovd6tov en(oqq 6pXetot dpreoo o' enog( ;re to DNA rou unoKtvrlrrl rt Stot oup- BdMer orrlv ovoyvcilptorl rou unortvqr{ (B}.5ne nopo- rdrco). EntnL6ov, ot unoprovd6eg o Kot o e(vot ot rUpteg enrgdveteg qq RNA nol.uprepdorlg yt' oAtrrlLenrdpdoetq pe nopdyoweq nou pu0pr((ouv qv 6vop[q rrlq Uero- ypoqnq. H unoprovddo to en(or1g no(et p6lo orrl ou- voppol6yrlorl rou ev(6pou, 6nog en(or1q Kot oe ouyr<erpq.rdveg pu0ptorrxSg ettoupy(eg. H unopovdEo o e(vot rotd r0pto tr6yo uneU0uvrl yo rrlv ovoyvdrpton rou unortvrltrl. H xpuorotrtrtr<{ 6opf rou nup4vo rou Bor<rqptororj ev(rlprou 6eiXveL 6tt 51et 5vo oXrlgo oo 6oyr<dvo xoBouptorl, Ue rn Uto 6oyrdvo vo oXq;roil(etor tcup(cog on6 rq B uno;rovd6o Kot rnv dtrl.q rup(rog on5 q B' unoprovd6o, 6nog go(verot ot4v Ell(0NA'1S.8. O npoteUorv Dfoutroq yto to DNA Bp(oretot mqv enrgdveto eno<pqg rov unogovd6tov B t<ot B', ot ono(eg oroOeponoto0v tou g 6toltoptoprdvou g, provoti g r<).cirvouq orrl Ueroypo<prxrl quool(6o, 6nog rpo(vetot orrlv EIK0NA 19.9. H 05or1 rotd).uor1g Bp(oxerot orq Bdorlrqq q4totrrqg nou oXrlptot((etot on6 rq (ooY6vto" roJV 0 rot B'. 'Evo H ovodrrl onogl rou nuprlvo qg RNA notrupepo- o1g, onou qo[verot ro oX11.to ,,doYrovdtv ro$ouptou, Tou ev(u- Uou. 0r 0 (ruovo) rot p' (poQ unopovo0eq i1q RNA notrupepoonq eXouv evov 6[ou],0 yto ro DNA erpoyeto. H o I tpolverot npootvl, q o ll ruprvrl rol 1 o xornvl. tlpooopltoopdvo ono. K. M. Geszvain ror R. Landick (ed. N P Higgins). The Bacterial Chromosome, American Society for Microbiology, 2004 , t, H 6opr1 rou nuprlvo rrlg RNA no),upepoolg ono ro porrlpro Thermus aquaticus, Ue U g unopovoDo pntre rot q $' npootvq. Aopq ono: Protein Data Bank 1HOM L. Minakhin. et ai,, Proc. Natl. Acad. Sci. US/ 98 (2001):892-897. on6 to 6rjo roroluttrd t6wo Mg2* nou e(voL crnopofurl- ro yrcl ro prllovLopr6 rrlg rord).uoqg e(vot ogrXro npoo- 6edepr6vo oto evepy6 Kevrpo rou ev([pou (B].6ne Evoulta 19.18). To dMo L6v Mg2- tptovet crro evepyd x6wpo oe oUpnLot<o pe ro etoepx6prevo tpt<prooqopLxd vour.treori6to (NTP). Onoq npoovotp6pOqr<e, o nupqvog rou ev(rjpou rov tuKopuorov dlet qv (6to pootr<q 6o- prrl pe ro Borcr4ptor<6 dv(upo (Erx6vo 20.2). En(o1g ne- pr51er r<onoteg entnp6o0eteg unoprovo6eg KClt nepLLoprBdver oMrl).ou1(eg nou 6ev undpXouv oro Bo- rcrrlpLor<6 6v(upo. Or r<ripteg 6Lotpop6g ovdi.-ieoo mo Bo- r<rrlproxd Kor ro euKopuortKo dv(upo Bpior<ovtoL oXe66v onor).etorLro mqv nept<pdpeto rou ev(0;rou, uoKpro ono ro evepyo Ktvrpo. 'l 9.5 To OAo6vCuUo AnoreAeiror on6 Tov Tov nopdyovTo o Tnq RNA l-loAuuepdonq Iluprivo Tou Ev(tlpou Kot a H Borrlptorl RNA nolupepooq unodtotpe[toL orov 02BB'o nuprqvo rou ev(upou nou roro),0et rl1 UeroYpotp1 KoL Tl'lv uno- povodo o nou onouoltot ltovo yto rr1v 6vop[q. * 0 nopoyovro-Q o rpononote[ rtq tbtorlreg npoodeorlq oro D- NA qg RNA notrupepoorlg, ersL 6ore q ouyy6veto Tlq rpoq ro yevlr6 DNA vo ptet6verot xot, ovr[0ero, 1 ouyyeveto qg npoq Touq unouvrlrdg v' ou[ovel. O nuprlvog rou ev(tlptou 61et yevtr<q ouyy6veto Yto ro D NA, xu p(r.og ).6ytr-l rllercrpoorotLxrrtv oMqenL6pdoe- oiv 1.rero[[ tqg nporelvng, nou e(voL Bootr<r1, Kot rou DNA nou e(vot 6(tvo. Otov npooddverot mo DNA ror' our6 rov rp6no, to DNA nopopevet oe 6(r<trovq lfr.li To Otrodv(uUo qq RNA tlotruUepoonq Anoretreiror ono rov l-luprlvo rou Ev(upou roirov I-lopoyovro o 5'17 Boorxeg eworeg
/ i r::,1 , .T -"' I llopoyovrog o 0 nupqvog rou ev(ugou npoodeveTot odtorplrog oe roOe DNA. 0 nopoVovrog o perdlvet rr1 ouyy6veto yto npoode- oq ove[opqrq ono rqv otrtr1trouXio ror npoo6[6et eldtroqro yto roug unorrvlreg, Uopqn. O nup1vaq tou ev(l1tou 6yeLqv xav6trya va ouv06teL RNA an6 6va DNA expayeb, oMd 5e pnopel v' av ayva p [(er uno xwryt 6q. H Uopqri rou ev(ripou nou e(vqr uneri0uvq yro rnv 6vop[q rnq Ueroyporprjg on6 rouq unoKrvqrdq ovoprd- (eror ol.odv(upo (o2BB'roo) (EIK0llA 19.10). Arorp6per on6 rov nup{vo rou sv(rlprou enetdri nepdXer dvov nopd- yowo o. O napayowag o$pa 6yt 1t6vo efaoEah((et 6tt q Baxtqpnxi RNA nolupepdorl evepyonoe[ rn pera- ypaqi an6 optopdveg 0ioerq, aMa en[or1q peuitvet xat qv np1odeory oe pr1 edxdq a&rltouy[eq. Xop(g nopd- yowo o, 11 oro0epd ouyydverog yro rrlv np6o6eor1 tou nupfvo rou ev(Uprou oro DNA per6verot rord -104 rpo- p5q r<or o 1p6voq npr(onq rou ougnl,6rou e(vot <1 6ewep6Lenro. Aw(0ero, ro otro6v(upro npoo66vetot fiouq unoKrvrlrSg noLrj nro orptXrd, pre oroOepd o'uyy6- verog xotd pr6oo 6po -1000 gop6q UeyoUropn Kor Xp6vo ngr(onq 6o9 r<or oprer6q cirpeg. 'Etor o nopd- yowoq o ouorcrorrKd onomo0eponore( q pq etStrrl np6odeoq rou unorwrltj. H roX0rrlro pe rnv ono(o ro oLo6v(u;ro npoo66ve- ror oe 6rorpoperu<59 oMrlLouX(eg unorrvqrc0v nopou- ord(er peydl,eg 6ror<updvoetg ror yr'our6 onore)re[ orlprowrr<4 nopd;rerpo arov ro0optop6 rnq "oy0oq rcu unoxwt1r4", drllodTl rqg onotetreoponx6rqrog ev6g (e- Xoptoro0 unorwrlq oq npoq qv 6vop[4 rnq Usroypo- qnq. H ouXy6rrlro rqg Svop(qg nox(trer on6 -1lsec yo ro yov6ro rou rRNA un6 rSovrx6g ouv0{reg 6o9 < 1/30 min yro rdnoroug dLLoug unoxrqt6g. O nopdyowog o ouv40cog onetreuOepc0verot 6rov 11 otruo(6o rou RNA gtdoet oe ;rr1<og -10 nt, oEtivowog"rov nupfvo tou ev(Uprou une0Ouvo yro rrlv entpr{ruvot1. 0 o onomo0eponore( q gr1 el0rKn npdo000n I 9.6 I'ltilg Bpiorer n RNA lloAuuepdon rtq AAAnAouxiaq Tov Ynomvnrdrv; a H npoodeo! rrlq RNA notrupepoolg oTOUS ulloKtvrlTeg eivc. rooo ypnyopn tiore gotrlov 6ev nporeuot yt ontrl OtoXuo1 ," H RNA notrupepool npooDevetot oe ruXo(eg Odoetg oro DN" xor trq ovrotrtrooer pe otrtreg otrtrr1trouXteg 6oq orou Bpe0s. evog unorrvlrrlg. H RNA nol.uprepdoq np6ner vo Bp(oret unortvqt6g ;.re- oo oro nLo(oto tou yovt6uol-rorog. I-1cog [eXorp((ouv c. unorrvrlrdg on6 trg 4 x 106 bp nou onopt((ouv rc un6Lorno pSpoq tou yovtdrcbporoq rou E. coli', N nopouotd(er ontrd prow6Lo yro to ntrrg q RNA nouprepdorl 0o pnoporioe vo Bp(oret rtq oMqlor-- 1(eg rorv unorrvqrcov prSoo oe 6eq rrg oMq).ou1[e: trg ono(eg pnope( vo npoonel.doer. To otrodv(upo r,': RNA noLupepdonq ewon[(et ro Xptopr6ooLio Ue rL- 1o(o 6rdXuoq ror npoo66veror pr1 erdtrd sro opvllr(c cpoprtop6vo DNA. Ie our6-ro pow6lo, to oo5v(ut-: 6r(ororor noti yprlyopo. H 6tdXuorl 05tet 6vo ovrr-rrc- ro 6pto yro rr1 oroOepd qq roXUtrltog otiv6eorlq r-: rd(r1g rou <108 M'1 sec'l yro or6Xo 75 bp. l-ldwoc - npoyprortxr; moOepd npotrl0rlttrrlq toXUtqroq 1fo'- ward rate constant) yro rdnoroug unortvlt5c, in vtt': rpo(vetot vo e(voL >108 M'1 sec1, oro 6pro dLdxuo1c * xor ndvo qn6 our6, H 6nuroupy(o rot rotompocc- f-tog oerpdg oupnL6rcov 5og 6rou (r<ord tUXrl) rl RN: no).u;repdorl ouvowqoet 6vov unortvqrq xot 6rlptouc- y4oer 5vo ovotXr6 orjpnlor<o u<ov6 vq rpttd[et RNA E: 4rov oXerrrd opVn 6ro6Lroo(q. 'Etot, o Xp6vog no, onoLre[roL yto tuXo(ouq r<rix].oug dto6olrr<qq oJvdi- onq Kor 6rdmooqg oe Xolop6g 06oerq np6o6eoqg e .- vot noAU peydoq ror 6e prnope[ vo epprrlveUoet nc. Bp(or<er r; RNA nouprepdoq rov unoKrvnrq tr1g. Iuvo noq, n RNA no).u;repdoq npSner vo XprlotUonotei ci.- o prSoo yLo v' ovo(qrrloer lg O5oeLg np6o6eo4q rqc H Eu<6vo 19.11 6e(yyer 6rt 11 5to6u<oo(o nt0ov6v en.- rolUverot enerdrl o opltr6g m6Xog qg RNA notru;,a- pdonq e(vor oL6xLnpo ro yovt6(<r-r1.to, 6Xt pt6vo g.: ouyrerptpdvq o).r1Lou1[o unor<tvqq. Au[dvowoq r: pdye0oq rou or6Xou, 11 moOepd rqg toX6tqrog 6toX-- orlq oro DNA ou[dver ow(orotlo xoL 6ev e[vot ntrec, neproprmu<1. l-169 pretor<tve(rot ro 6v(upro on6 gLo t-- Xo(o 06o4 np6o6eorlq mo DNA o' 5vov unoKtvfr- I[prgorvo pe opretd 6e6o;r6vo, toutrdXtorov tpe(g 5Lc- <popertrSg dtepyoo(eg 6topoptptivouv l'lv toltitqr: ovo(qtrloqg unortvqtq on6 tqv RNA noLuprepdor I-1purrov, ro 5v(upo prnope( vo petortvrl0e( oe pto 1.tovc- 6tdoroq tulo(o 6to6poptrl r<otd prlxog rou DNA (,.oI,- o0r1or1 - sliding"). AeUrepov, enet6rl ro Xpropi6otr.rl.tc 6Xet 6or6oLridrl muXcor4 6otrrr1 oro Borrqptox6 nupr- voer66g, ;16l.19 ouv6e0e[ oe pro oMq].ouX[o rou Xpo[o- I ffiy r-"*W L"Ah!** '%ffi92 I To revrpLxri dv(uUo npooddveror oe onoLodrlnore DNA To olo6v(uUo npooDdverot mov unoKtvlrn 518 KEOAAAI0 19 MeToypoQrl orouq flporopudrreg Boorxdg eworeg
EIK0NA 19.11 llporew6pevor Urlxovrouoi yro ro ntirg n RNA notruuepoon Bpioret 6vov unortvqnl. (o) otr(o0non (p) dLorplportxrl perotpo- pd(y)ev6oneproXrxrlouvdeoqxordrdotoolnovonnOnon.llpooopUoog6voon6:C,Bustamante,etal.,J. Biol.Chem.274(1999):166665- 1 66668. orilgorog, to 6v(upro Bp(orerot nMov nto xowd oe dtr- trsg O5oetg, yeyov6g nou petcilvet to 1p6vo nou onqtte[- ror yrq 6r6mooq rt enovonp6o6eorl oe pto 6IIn 06on ("dtotUrluortrrj getocpopd - intersegment transfer" i (ovonndnon - hopping"). Tp(rov, evril e[vot ouv6ede;r6- vo prq erdrd oe p[o 06o11, to 6v(upro gnope( v' otr].d[et O5oetq qro DNA pr6Xpt vo Bpe0ei o unoxtvqtfg (.dpreor1 prerogopd - direct transfer"). I 9.7 To OAodv(upo TpononoteiTot Kord Tn Arepyoofo Avoyvtipronq Kt An€Aeu8dptrlonq on6 Touq Ynortvnrdq * '0T0v n RNA notrugepool npoo66verot o' 6vov unortvrlrrl, dto- Xop((er iouq rtrtirvouq rou DNA yto vo dlptoupyrloet gto pero- vpoqrrrt rpuootrido rt evootgor6vet vourtreorldto oto RNA. & H evopfi ono qv RNA notruuepdoq 6ev e(vot novrore enttu- xts. * 0 nopoYovroq 0tYU0 ouvflOoq oneLeu0eptlveroL ono rrlv R- NA notrupepoorl otov 1 veoouvre0etp6vrl otruotdo RNA ono- rqoer pflrog -10 pooetrlv. Mnoporipe rdlpo vo neptypdtpoupe ro orddto rnq Ue- roypogrlg oriprgovo pe ttg olLrllent6pdoerg preto[ti ttov 6rorpoperrrrov ptoptpcov tqg RNA nolu;repdoqg Kor rou DNA er<proye(ou. H ow[6pooq 6vop[qg prnope( vo neprypoge( pe Bdoq ttg nopopttitpouq nou ouvoQ(- (owor orrlv : o H ow(6pooq oloev([prou-unoKtvrlrrl [ertvd pe to oXrlprolopr6 ev6q xretoro0 6Lpepoiq oupnA6xou, 6nog <po[vetot offlv Etr6vo 19.12(o). "K].etor6" orl- po(ver dtt to DNA nopop5vet 6ixtrtovo. O oXqgotL- opr6g rou xletoro0 6tprepoUg oupntr5xou e(vot qwt- orperrr6g. 'Etot, ouv4Oroq neprypd<petot on6 pto oro0epd tooppon[og (Ke). YndpXet preydLo e0pog orrg nprdq qg mo0epdg tooppono(og Yto ro cDCq- prottopr6 tou xl,etoroU oupntr6rou. o To xtreror6 orjpnloro petorp6netot oe ovotyr6 o01t- rrloxo pe "oflodtdroqn, ;rtog ptr<priq neptopiq DNA pr6oo orrlv oMqLou(o 5nou npoo6dvetot ro 6v(u- pro, 6nog qo(vetot orqv Etr6vo 19.12(9). l-to roug neproo6repouq unoKtvrlr6g, 11 perorponrl on6 to ovorXr6 oro xtretcn6 oriprnloro e(vor ptq owtorpe- nrj ror owri q ow(6poor1 prnope[ vo neprypoqe( on6 11 oro0epd npool0rlrxrlg toX0r4roq (forward rate constant) (kr). Oo16oo, pepxo( unoxwqrdg (n'1' ol unortqt6g tou rRNA), 6e drlprtoupyo6v oroOepd ovorXrd o0ptntroro rot our6 e(vqt to xLet6( yto 11 priOprori roug. O nopdyowoq o no(et orlprowtx6 p6Lo orqv owidpooq ri€nq GMne Ev6r4ra 19.19). Or preroBdoetg nou ou;tBo(vouv on6 tqv 6vop[4 ;t5- Xpr rnv entprlruvorl en(o4g ouvodeUowot on6 orl- ;rowrr6g oMoy6g orq 6ogrl Kot rn oOmoorl tou ougnl6rou, Ot oMoy6g oro oXrl;ro qq RNA notru;repdorlq ouvo- 6efouv ug t<tv4ttx5g ;retoBdoetg nou neptypdgnKov nporlyouprdvtog, ro0rilg r<ot q ;rerdBoon oro o6pntroro enrpulruvoqg (nopouotd(erot firlv EIK0NA 19.13). Ito xtretor6 oUpnloro, to ol,o6v(upro qg RNA noLupepd- oqg roLrimer nep(nou 55 bp- DNA, errew6pevo on6 -- 55 5og -+ 1. To 6[x].tovo DNA npoo65vetot rup(tog xot6 priKoq rqq puog ntreupdg tou oloev(ripou, 6nou SpXetot o' enoqrl ;re nq C-refur6q neptoX6g rov o uflopovd6tov roOdrq Kor Ue rtq neptoldg 2 xat 4 rnq o unoUovd6og (BL6ne nopor6to). Kord q getdBooq oro ovotXr6 o6pr- nlor<o, q drop6prpoorl t6oo qg RNA noluprepdoqg 6o'o Kqr rou DNA oMd(et. Ot nto dpoonxSq oMoy6g orrl 6o- Uri rou oupnl"6rou qo(vowot crqv Ex6vo 19.12: (o) ptto xdprUn rou DNA -90 potp6v, nou enrpdnet orov xtrtivo "i1l ? To 0tr06v(upo Tpononore[ror Koro rr1 Arepyooro AvoyvtirpLolq rr Anetreu06ptlorig ond rouQ YnoxLvnte c 519 Boorxfg eworeg
eKpoye(o vq npooeyy[oer ro evepy6 x6wpo rou ev(6- Uou' (B) dvorypro tov x.trcirvrov rou DNA rou unoxwrltl pero[6 --11 rqt +3 ovoEopxd pre qv ogerrlp(o rnq Ue- roypocprlg r<or (y) r<tre(orpo rov (ooyovrcbv> tou ev(0prou ciore vo neprruxl,cioer to rpui;ro rou unoKrwlr4 nou Bp(- oKeror r<o0odxd on6 rrlv orperlp(o preroypocprlg. Iuve- ncilg, ro oqge(o enorpriq rou ulroKrvrluj mo ovorXr6 oUpnl,oro erre(vowor on5 --55 oe -+20. To en6prevo Brigo e(vot vo evooUorco0oUv rq 5[o npdrro voux.treot(drq Kor vo oXqpronore( rivog <ptrlorpo- 6eorepx6g 6eopr6q prero[ri roug. Aur6 drlproupye( 5vo rproroy6q o0pnAoxo (ternary complex) nou nepr6ler RNA, DNA xor ro rlv(upro. Iroug neproo6repouq unoKr- vqr6g, qXqpror(eror puo oLuo(6q RNA nou SXer pr4r<og opr<er6g Bdoerg, Xrr:p(g getor<(vqoq rou ev(rjprou xotd prrlrog rou erpoye(ou. M6Ltg npoore0e( puo Bdoq, un6pXer rdnoro nt0ov6- rrlro ro 6v(upo v' onel,euOeprioer trlv oluo[6o rou RNA, rorolqyowog oe npoi6wq overtruxo6q (errptonx{g) 520 KEOMA|019 Metoyporprl mouq flpoxoputirreg EIK0NA 19.12 H RNA notrupepoorl nepvoer on6 op- rerd orddLo fiprv rnv enqrlruvorl. 'Evo r],erotri 6q:e- prig orjpntroro gerorp6neror oe pto ovoLXrrl poptp4 u 6neno o' 6vo rproroy6g oupntroro. l-lpoooplroop6vo on6 S. P. Haugen, W. Ross ror R. L. Gourse, Nat. Rev. Microbiol. 6 (2008): 507-51 I dvop{4q (abortive initiation). Merd rrlv one- Leu05prooq rou ovenrruXorig npoi6woq, ro 6v(upro [ovopf(et rrl orlvOeor1 RNA oq 06orl * 1. EnqvoloprBov6prevor r0xi,or ovenruXd:v ev6p[eov 611 prro u pyo rlv o].ryovo u r].eor(6 ro nou ouvrlOtog 51ouv pqxog tr-r6vo Liyeg Bdoetg (nop6n prnope( vo rptdoouv oe grir<og ro 20 nt), npw ro 6v(upro npoyprorxd roro<pdper vo 6rog0yer on6 rov unorrqrl. Or otrtr4l.enrdpdoerg Ue rrlv RNA noAupre- pdoq te).u<d e[otre(rpowor r<ord q 6rorpuyq rou unoKrvrlq. M6Xprg 6rou 4 oluo[6o rou RNA ensKro0e( oe 15-20 nt, ro 5v(upro yevxd 61et nepdoer on' 6treq lg pretoBdoerq nou Xo- porcrqp(ouv 6vo orjprnLor<o enrprlruvoqg. Or d0o nro govepSg pretoBdoerg e(vor 4 qneLeu- 05poor1 rou nop6yowo o [ono6(6eror mrlv Ex6vo 19.12(y)l Kr o oxqporropr6g ev6g ou;r- nL6t<ou nou rorol,o;rBdver pr6vo -35 bp DNA, ow( ralv -70 bp nou e[vor Xoporrrlptmu<6q uov oupnl5rov rov unortvqrrriv. MoLov6rr 4 oneLeuO6pcoorl rou o ouvqOoq ougBo(ver xo- rd q 6roquyrl rou unorwrlrl, our6 6ev e(vot npoononoripevo yro tq prerdBooq oqv en[- prrlruvoq. Ie optopr6,veg neprmdloerg o o SXer rouronor4Oe( oe oU prnLor<o enq-rr1ruvor1g, oi.- )d q oUvdeo{ rou Ue ro 5v(upro (ocog owrro- tomp((er enovonp6o6eorl orov nup{vo rou ev(0;rou r<otd 14 gdo4 rrlg enqlrp<uvor1g. I 9.8 O l-lopdyovroq o EA6yx€r rnv np6o6€on oTo DNA Avoyvrrrpi(ovToq luyrerprpdveq AAAnAouxieq Tov Ynorrvnr6v * 'EvoS unorLvrprlg npoo6ropl(eror ono rrlv nopouo[o ppoxerotv ouvorvelrtlv otrtrrltrouXrtilv oe ouyrerprpeveg 0eoerg. * 0r ouvorvelx5g otrtr1trouXreq rou unorrvlrrl ouvlOtog onore- Lo0vror ond ;-uo noup[v1 orrlv oqerrlp(0, 6vo e(opepeq pe otr- trritrouXro nopepqepfl pe qv TATAAT oq 0601 --10 rL 6vo otrtro e(opepeg pe gto otrtr1trouxio nopepQept Ue qv TTGA- CA oq 0eo1 --35 * 0t qexoproTo( unouvlr6g ouvrl0rog dtotpepouv ono q ouvol- velrq otrtr1trouX(o oe pro fi neprooorepeq 0eoerg. * H onoretreopolr6rrlro rou unorrvqrrl gnoper enloqg v0 cnl- peo(eror rot ono enrnpooOero orotXe[0. Avodtrd DNA I-lepro1r1 3 tou o Tlsproxn 2 Tou o lleptoXrl 1.1 rou o E(opepdg -1 0 Ktrtivog 91-expoye[o Ko0o0rrd DNAfiepLoXr1 3,2 rou o Ktrrirvog ergoye[o Boorx6g eworeg
.aciexet 10v 0 Kot -5: --3: -30 -20 -10 1 +10 +20 +30 -- ,., ...:;iJ:.i'ttti:,: oxluoT((tTot oTtq 10 Bdoetq, .' -- :-:- .,cKorxov€rrtq0T10Q6q0116ro-356(l)qr0-55 -5: -40 -30 -20 -10 1 +10 +20 +30 . .:.:. :..':. r:.,::t, oxnuoll(tT0t ortq 15-20 -5a -40 -30 -20 -10 '1 +10 +20 +30 H RNA notruUepool opxtKo epXetot o' enoqrl pe -,- ,- :ro -55 eolq +20.'Orov dtfororot o o, o nuplvog , .--,, oloretrtrerot oro -30. MotrLq ro ev(upo perortvloel - -, . - --: ','rvLro o0pntroro enLglruvolg. --. - -:,).qAouXio DNA nou l.ettoupyio qg e(vot v' ovo- -:.leror an6 npare'tvtq, o unoKtvnrnq 6tog6pet --: .: cMq).ouX(eg tcov ono(ov o p6og e(vot vo trle- -: . : loowoL. Ot nLrlpo<pop(eg yro tr1 Lettoupy(o tou ,*:...rlrrl nopdXowor oneu0e(oq on6 rlv oMqLou- -- ::.l DNA: n 6oUn tou eivot to or1trro. Aut6 e(voL . : ., .oorxd nopddetypro ptog cis-evepyoU 05o4q, - --: op[m1Ks ortq Eu<6veg 2.16 rot 2,17. Aw(Oeto, - .,:co(dpreveg neptoXdg ononorjv to v64ptd toug ,-,: lroorou q nLrlpo<pop(o prerorpep0e( orrl uopan . r-l.ou dMou vour<Leivtxoti oE6oS n npottelvqq. -,oq rpdnog vo oledtdoouge Svov unor<tvr1t1 0o --:. Jro ouyr<erpttrt6vq oMrlLoul(o DNA v' ovoyvop((e- -: :lro r1v RNA noLuprepdoq. Kd0e unortvqtrlg 0' --::iorjwov on6 tr1 ouyxercptp6vq otrLr1i.ou1(o, r1 --,:.c1rorov, 0o rrlv nepildprBove. Iro Borcrqptor<6 yo- i.-;-io, ro etrolroro prrl<og nou 0o prnopotioe vo no- : rr(01 dvo enopxdg orlpro e(vor 12 bp. (OnoLo6rlnote :::4r.rrepq oMqtrouX(o e(vot nt0ov6 vo oXqprottore[ op- -..:,q rpop6g -onLcog xord t[p1- n 6rot vo npor<o].5oet -.-5q orlporo. To e).d1Loro prrl<oq nou onottekot yto -: . cbrxrl ovoyvcoptorl ou(dvet pre to pr5yeOoq rou Yovt- - ,.roroq - 6vo np6B).npo oro europu<ortrd yovtdttrt- ..::c), H oMrltrouX(o tov 12 bp de Xpetd(erot vo e(vot ouveX6prevrl. Av 6voq ouyrerptp6vog opt0p6q Bdoeolv 6toXrop(er 6rio oro0ep6q BpoXftepeq otrLfltouX[eq, to ouvotrrr6 pulr<oq touq 0o prnoporioe vq e(vot )try6tepo oTro 12 bp, enetdrj q (6to 11 an6orao4 StoXtoptogori onoretre( 6vo tprrlgo rou otiporoq (6orol rot ov q evdt6- peor'1 oMrlLouX(o e(vot doXerrl). It4v npoyprortr6vlto, 11 RNA noLuprepdoq ovoyvop[(u tLg DNA oMq].ouX(eg rov unoKtvntriv oe preyd).o Fq0U6 ptSoo "dpeoqg ovd- Yvoonq" ouyrerptpSvtov Bdoeov oro DNA on6 ouyre- r<pr;r5vo opuvo(6o tou oloev(tiptou. Ot ewunootordg 6torpop6 g tcr4ioq trr:v 6totp6ptlv Bor,crl ptoxtirv uno Ktvn- rdlv ono6(6owot rqrd noLf or4 6totpopenxrl onotetr'e- o1-rottr6rqro ovdyvoo4g tcrlv otrtrrllouXtrirv toug on6 ng optvo(tx6g oLl.qlou(eq rov unopovddtov o Kot o. Or on6netpeq yto rnv ovoyvtirptorl tov tdtorltov rou DNA nou e(vot onopofurlreg Yto rrlv np6odeoq qg RNA noluprepdonq (er<(vqoov ouyt<pivowoq ttg oMt'1- ),ouX(eg 6torpopettxrilv unortvrltc0v. Kd0e onopofurlrl voux.treot6rrrl oltrr1Lou1(o 0o 6npene vo undplet oe 5oug roug unorwqr6g. Mto t6toto oMrll'oufo Xopo- rcqp((eror ouw4p4p6vq (conserved). Mto ouwrlprlpS- vr1 oM4l,ouX(o 6ev efvot ovoyromxd ouwqpqpSvq oe rd0e (eXoprorrl 05o'r1qq, 6r1}'odri enmpSnerot xdnoto norril.opropg(o. l-kiq ovoltioupe ;rto oMrl).ouX(o DNA yro vo npoodtop(ooupe qv 6Xet ouwqprlOe( opr<etd rilore v' owrnpooonefet 5vo ovoyvtr:pfotpro onpo; Auvqnr6g Odoetg ovoyvrirptorlg tou DNA ;rnopef vo npoo6roprorotiv ovotpopu<d pe pto eft6ovu<eup5vr1 oL- Lql,ou{o 11 ono(o owrnpoooTle0et rq Bdoq nou Bp(oxe- ror nro ouy3r6 oe xdOe 0dor1. Mto ouvolvenx{ oAtrqtrou- ;1fo (consensus sequence) xo0opi(etot on6 t4v eu0u- ypdpptorl 6trtov ttov yvomc0v nopo6etyprdtov 6tot rilore vo preytoronotrl0e( q oproloy(o rouq. l-to vo y[vet ono6erc4 puo oM4trou{o oq ouvotvertr4, rdOe ouYKe- rp4l6vr; Fdon qq np6net vo entxpote( or4v ow(orotXq 06o11 r<ot to neptoo6repo npoYpqrtxd nopode(yproro np6ner vo 6ro<p5pouv pr6vo rotd pto 4 6Uo owtroro- mdoetq on6 rq ouvowenrrl oM4trou1(o. 'Evo ewunootox6 Xoporrqptonr6 trrlv unorwrlrcbv oro E. colie(vot 11 iMeulr1 et<terupivrlq ouwrTp1oqq aA- lqAouyuhv oe 6).11 rrlv 6rcrooq ttov 75 bp nou oler(etor Ue rqv RNA noluprep6oq. 0o16oo, rdnoto BpoXSo tprl- proto pr6oo orov unoKtvrlrl 6Xouv ouwrlpqOe( rot e(vot (onr<r1g oqproo(og yto t4 l,ettoupy(o tou. H ouw1p1ory p6vo noAA Fpaxeuiv ouva:euxtisv aM7ouyuitv e(vatru- nx6 yapaxr4ptwtx6 tuv pu1lttottxtitv 06'oeuv (6nuq ot unoxlrySg) t6oo ota npoKapu@rtKd 6oo xot ora euKa- puanxd yovtdubpara. Apr<er6g oMrll.o u1(eq tov Bor,mq p rorcilv uno rtvltriv ouverogdpouv ornv ovoyvdrprori toug on6 to oLodv(u- Uo rnq RNA nol,u;repdonq. Ario oLl'r1i.ou1(eg 6 bp, yvoord og ororleio -10 G10 element) rot orotlefo'35 (-35 element) (6norg rot to prlxog rnq "6toqrnUotxtlg" oMrltrouX(og nou nopeprBdMetqt ;reto[rj rouq - (spac- er>> sequence) e(vor ouvqOtog oqpowu<6tepeg on6 ou- 169 ro0out6g ttq oll4l,ou1(eg ovoyvrirptoqg. Ot oll,rll,ouX(eg rov unoKtvrlrciv nou Bp(orovtot mqv o<peqp(o r<or oxp$rig 5[nl.o t1g, ot otrtr4]'ou{eq Ydpo) on6 ro ororXe(o -10 [yvotmd oq "6teupup6vo orotXefo - :i.* Ol-lopoyovroqItyUoEtreyXeLUvtlpoobeorloroDNAAvoyvopi(ovrogluyrexprpeveqAtrtrltrouXieqrovYno<1,'- - 521
'l0D (extended -10 element) mqv ovodKn nl,eupd ror "6rqxoprorig" (d iscriminator) mrlv xo0o6u<r1 nLeupdl r<or or 10-20 bp og6ocog qvodrKd rou morXe(ou -35 [yvoor6g oq uororxe(o UP, (UP element)], en(oqg o],- Lqlenrdpotiv rord erdu<6 rp6no Ue rrlv RNA noLu;repd- orl Kor ouveroqSpouv orrlv onorel,eoporrr6rlro rou unorlrprl. . :rouq neptoo6repouq unoKrvqr6g ovoyvrop((eror pro neproXrl 6 bp nep(nou 10 bp qvo6trd rnq oqe- qp(oq (r1 npoygoru<r1 on6orooq on5 r1v oqeqp(o 6rog6per eLogpd on6 unoxwqrrl oe unorwqrrl). Au- r1 q e[opepqg o].].q].ou1(o ouvq0rog ovoprd(eror ororXe[o -10, r<ow( Pribnow (Pribnow box) ri oMuirg xourf TATA (TATA box). H ouvowenrrl oMqLouX(o TATAAT ouvor.l;((eror orn UopQn Tao Ags T+s Aoo A.5o Tso or 6e(rcreg 64Lrirvouv rnv eKorofiro(o ovo),oyfo ulg oulv6tep4g Bdorlg, nou norr0.et on6 45"/" 6tog 96%. (Mro 05or;Xcop(g ep<povr1 npo{gqoq yro xdnoro Bdorl 0o 6rlLcov6rov ge ro oriprBoAo N). H ouXv6qto epr<pdvroqg owrmorle( mr1 o4proo(o our<ilv r<ov (euyc0v Bdoeov yto rrlv np6o6eor1 qg RNA noluprepdonS. 'Etor, ro opXrx6 noLri ourrrn- pqpSvo TA ror ro rellr6 oXe66v on5l,uro ouwn- pqp5vo T orqv oLL4LouX[o -10 oulvd e(vor ro(ptog orlproo(og yro rnv ovoyvriprorl rou unoKr- vqrrl. I{pepo yvrop((oupe 5rr ro ororXe[o -10 flpoyUoronore( er6u<69 yro rrlv oLL4Loul(o enoE6g mrg neprolSq 2.3 Kor 2.4 rov nopdyowo o (BMne nopordro). 0o16oo, ouq q neprofi rou unoKrvn- ri e(vot 6ir<).tovq oro x.l,eriyr6 oriprnLoxo ror ;rov6- r<Lovr1 oro ovorXr6: ouvenc(lg, or oM4Lenr6p6oerq UeroEU tou morXe(ou -10 rcot qg RNA notruUepd- o4g e(vor noLUnl.oxeg ror oMd(ouv oe drogoper- xd o166ro rnq 6repyoo(og 6vop(r1g rnq peroypogrlg. . To ouwnprlp6vo e[opepdg -35 bp ovodrrd rrlg oqerlpfog ovoprd(eror orotleio -35 C35 element). H ouvolerrrrl oMqLouX(o e(vor TTGACA. Ie nro tre- nrotrrep{ Uopen, n ouwnpnprivrl o},r1}.ou{o 61er os eqnq Ta+ Gze Aos Cs+ Aqs Ot Bdoerg oe our6 ro morle(o ollrl).enrdporjv oneuOeiog Ue rnv neproX4 4.2 tou nopdyowo o (BL6ne nopordtro) ;re nop6proto rp6no 16oo oro ovorXr6 6oo ror oro xLeror6 oriprnLoro. r H on6moon ;rero[ri rtov O5oeov -35 ror -10 e(vor 16-18 bp oro -90% rov unoKrvrlrciv. Ing un6lor- neg neprmdroerg e(vot '15 bp ro lly6repo xor 20 bp ro neproo6repo. Molov6tt 4 (6n 4 aMrlLouy[a mo peya)thepo pipog u1q napepBaM6pevrlq nepoyfig e[vot oyenxd ao4pawq, 4 on6otao4 elvat xa[pn enetdr1, t6yu r7q elLxoet6o0q E1oryq rcu DNA, xa1o- pGet 6XL 1t6vo tov xardM4io 6nyapop6 rav 6Ao neprcydtv r4q BNA ndtupepdo1q nou a)rLr1ient- 6poiv, aMa en[oqq Katro ye@petpm6 npooavarcit- o1t6 tuv 50o 06,oeav. o H oqetqp(o ouvrlOog (>90%) e(vor truo noup(vr1, ou- 522 KE0MAI019 Meroyporprtmouq llpoxoputirreg yyd 4 r<ewprrq Pdon qq oMr;),ouX(oq CAT. Oordc: q ouwrlpqol ourrlg rrlg rprn).6rog 6ev e(vor tdoo lr- yd).r1 urore vo Oecope[tot oq unoxpeorxd oqpro. o Opropdvo (e[yq Bdoeov ornv neproX( pero(U r: o<peqp(oq Kor rou morXe(ou -10 ovoyvrrrp((or,.:: on6 qv nepropl 1.2 rou nopdyowo o (B).dne .:. por<drco). l-to nopddetypo, Uro oMr;).en(6poorl o : . r<r1 yro t4v o).).4LouX(o prero[U ev6g roro].oi:r:- youov[vr1g orov x.,rrtvo prrl-expoye(o 6tjo O5oetq .:. 0o6u<d rou orotle(ou -10 e(vor e(orpelr<d of lrc,l . rq yro tov r<o0optopr6 rrlg oro0ep6rqroq r3- ovotXro0 oupnl6r<ou. 'Etor, oL 6ro<pop6g orqv o).1.-- Loul(o rou unoKrvrlrrl oe O6oetg nou 6ev e(vor n:, - ouwrlpqpdveg prnope( vo eu0Uvowor yro rnv ere::- yevq roXU rov 6Lo<p6prr-lv unorrvqrov. o Or pdoerq oro 6eupupr6vo orotXe(o -'10 npooeyyil: ,. ror on6 qv neptoXll 3.0 rou nopdyowo o (B).dne :: poxdtrrr). H oM.r;LouX(o TGN oro ovo6rx6 dr<pc .:- ororXe(ou -10 ouprpret6Xet oe oMq).ent6pdoerg L-.:. repo orlUowu<6,9 yro qv 6vop[q rnq peroypcc*: 6rov 11 oMr;),ou1[o rou ororle[ou -35 orov unoKt,,-:* 6e pord(er noLU pe 14 ouvorvelrcl. Aur6 ovo6err,,,: norg ororleroOerotjwor or oMrlLoul[eq rou uno(.,-. 14: prrpr; oprot6rlto Us rrl ouvorvetLrrl oMqAc-,.: oro 6vo ororXe(o ;inope( v' owroroOpuore( ond ue,:. ).r1 oprot6qro o'6vo dMo orotle(o. o H neptoxn -20 bp ovo6u<d rou ororle(ou -35 cr-:.- Lenr6pd pre ro CTD rotv 6Uo o unopovddcov, OL i . nrcuoerg ourcbv rov ol.Lq).en16pdoerr-rv c:-. evepy6tqro rou unoKrvrlrrl prnope( vo e(vor op(,:: oqprowrr<69, ou(dvowog rn Ueroypoerl rou).:, . orov rord p-uo rd(r1 prey60oug yro no).ri er<gpoi:*i. vouq unoKrvnt6g, n.X. urlv yovr6(rov tou rRNA 'O :. or ouyKeKprpdveg oMrlLouX(eg protd(ouv noAu p. ouvorverrrrl, 11 neptoXrl ourl ovo<p6peror c,Dq c.: . 1e(o UP (UP element). H 6o9( ev6g unoxtvrlrq, pre ro enrpen6pevo eupcl norril.otrropg(oq (on6xlrorlq) on6 ro B6trrroro, nop.-- oro(eror mqv I 9.9 H AnoreAeopoTrK6TnTo Tov YnoKlvnTd,v Mnopel v'Au{dvel 6 vo EAAordveror on6 MerdAAoIn + 0r uopvrllx6qD UsToMoqerq nou per6vouv qv onoretreopc. - rriqro rou unorrvrlrl ouv1Oog pertirvouv q ouppopqrr:c- npoq rtq ouvorvenr6g otrtrqtrouXieg, ev6 or ,,0ertr6g,. perc;. tro(erq 6Xouv ro ovri0ero onor6treopo. * MerotrtroEeLq oqv -35 otrtr1trouXro pnope[ vo enrlpeooo-, rrlv opXrrl npoodeoq qg RNA notrugepooqg. * Merotrtrolerq orqv -10 otrtrr1trouX(o pnope( vo enqpeooc-, qv np6odeorl 11 rrlv ovldpool onodroro[lq rou UrToTpsr: evo rLeroro ougntroro oe ovorXro. Boorxeg evvoreg
l-leproxn 3.0 tleproXrl 2,3-2.4 fleptoXq 1 .2 TOU O TOU O IOU O IroLXeio UP -35 Ext -10 Uls +l -t n0txs[0 Tou DNA Kor T0 ouoToTrKo qq 6oUnq Trlq RNA notru;repoolg nou ouppereXouv oTnv ovoyvtirptorl rou unortvlrrl ----,-, 1: tlpoooppool.t6voono:S.P.Haugen,W.RossrotR.L.Gourse, Nat.Rev.Microbiol.6(2008):507-519. - ,- --::.c getoMd[eotv pnope( vo npooq6pouv - -: - -: :: ryerlKo |re q trettoupy(o rou unoKtvrl- - -. .:'-i.cEelq orouq unoxtvrltdg enqpe6(ouv to - -.: : :-: eKepooqg tou(tov) yovtd(ou(tr.rv) nou -, : -, ,*c[g v' oMd(ouv outd xo0outd ro yovt- ' -,: -::.l.ro. OL nepLoo6tepeg prerotrd[etg ovo- -: -:, :. ;e Borcrqpto tog oloydg tqg oMq).ou- - : - - - : l-','ouv oe onotreto rl Uey6).q ;re(tooq qq . --: ::j-: :tDV yettovtrrirv yovt6(ov. E(vot yvoor5g - , :: , nrxeq peroM6[erg (down mutations)' AMo- - - - : :r-'.-iou1(og euO0vowot Kot Yl'o 0erlx6g pe- -:--: iarc (up mutations) ottg ono(eg undpXet -. -: : ". -3-ioYpoQ( on6 tov unor<tvqrq. -- l- :. ,,8ettrd,g,, doo xot ot "opvr1ttr6g" pretoMd- .. - :: --. .:L oe oXeoq pre q ouvry1to1tiv4 onotetre- - -- .- -:: evdg ouyrer<ptp5vou unoKtvt'1tq nou - - - - I : .:.rL.tret notrU. 'Etot, prto oMoyr; nou ovoYvo- . -.-- -: ::i.1rtr<q petdLl.o[r1 o' 6vov unor<tvr;tr| pno- . : - -- , lrcUovrr-loe( not6 o' 6vov 6Mo unor<tvqtrl (o - - - : r. :. cto rorooroorl 0o pnopo[oe vo e(vot o16- :-r:: cnortleoprottr6g on6 tq petol'LoyprSvrl -,: -- -:- :cLJrou unor<tvr1q). Ae6o;r6vo on6 in vivo - . :-: - : .r.;q ovoyvcop((ouv 11 ouvoLtr<r1 rote[Ouvot1 - - r . :.-: -rou npor<ole[ 11 pretdMo[q. - -. ::,,.oEeiq rou ou(dvouv ttlv opot6tqro rov - . - " " - <ot -35 Ue rtq ouvotvettx6g oMrlLoulieg 11 . :- - : - , :- pretolti toug on6oroorl rdto on6 17 bp - - : - : :- lc. ouv rlv evepy6rrlto tou unortvrltrl. l-lo- . -. -: : *iroMo[etg nou getovouv ttlv ogor6tqro - - -:l-::.i nLeupog Ue rrl ouvotvettr<r1 oMrltrouX(o - - :::::::icuv rr1 preto(0 toug on6orooq (on6 to . - -: :-, '7 bp) o6rlyoriv oe petcol-ttivrl evepy6qto -' ,-.n, Or opvqttr6q getotrtrd(etq ouvq0og - .: -:*,c',/Tot ortq TIto ouwrlprlpr5veg O5oetg tou . - ," -:- :riBeBottrrvowoq rnv t6to(tepr1 oqpoo(o ou- - - . ::ceolv Yto rnv onorel'eopottr6qto rou uno- l;coo, undplouv onopodtrdg elotp6oetg oe - --: -: :ouq rov6veq. -: r,opddetYpo, 6vog unor<tvqtrlq ge ouvotvettxdg - , ..'.cr1[eg oe 6].o to oroLXe[o nou neptypdqrlKov nq- :: -:.:(erot mqv Etr6vo 19.14. Aev undpXouv 6ptolq -:-: r. rpuotxo[ unortvqt6g mo yovt6(trl1-to tou E. coli . : -:, ,rro( unoxLvqt6g nou totptd(ouv on6Luto 5le 11 - - r .::.<rl oMlLoux(o oe 6).eg ttg 06oetg srnv npoY- -:- . ::-:l e(vot oo0evdorepot on6 unortvqtdg pe tou- - l.:, evo ).d0og to(ptoopro oro ouvotvettrd e(opepri -10 r<ot -35. Aw6 ou;rBo(ver enet6rl npoo56- vowor 16oo orptXr6 orqv RNA noLuprepdo4 rilore ornv npoyl.tonx6tnro nopeuno6((ouv tr1 6ro<puyf rou uTloKl'- VNTN, l-ro vo ro0optoroUv ot on6l'uteg entmriloetg rov Ue- roMd[e<ov rov uTloKtvnrcilv, pretpdtot in vitro q ouyy6- verq rnq RNA nouprepdorlg Yto rov dypto rot ro pretotrl.oyprdvo t6no rtov uTloKtvntriv' H etepoy6veto tr1q roxfunroq Ue rnv ono(o q RNA nol'uprepdorl npoo- 66verot in vitro oroug 6totpopetxorig unorwrlrdg o1e{- (etor r<oLd ;re q ouly6rrlro rnq pretoyporprlg 6tov r' ow(orotlo yov(6to errppd(owot in vivo. Enercrefuowog ourl qv ovdl.uorl, ;rnopei vo npoodtoptore[ to orddto oro ono(o prto pret6Mo[r1 enrlped(er r1v ono6onx6tqto ev6g unoxrvntn. ALI'd(et q ouyy6veto rou unorwqtrl yro np6o6eoq rqg RNA nol'uprepdorlg; Arpqvet pev ro 6v(u;ro trqv5 vo npoode0e( oM6 ov(rqvo vo (erwrloet rrl Uerqypoeri; Atogoponoe( trlv enrppoq ev6q enxou- pu<ori nopdyowo; Metpdrwoq ttg t<tvtlttt<69 oro0epdq Yto ro ox!1gort- opr6 ev5g x.tretoto0 oupntr6rou Kot rnq gerotponrig rou oe ovotXr5 orjptnl.ot<o, prnopo6ge vo dtorp(voupe to 6rio ord6to rrlg ow(dpooqg 6vop(r1g: . Ot opvnrtK5g geroMd(etq orrlv ol.i.r1}'ouX(o -35 ou- vr'1Ooq get6vouv r1v toX0rlto oxnUortopoti tou xtretorori ouptntr6rou, oLl'd 6ev ovoqr6Mouv rn Ue- rotponf rou oe ovotXr6 oUPnl'oxo . Ot opvnrtKdg pretoL),d(elq orrlv o).trr1l,ouX(o -10 prnopei vo petc0oouv e(re tov op1tr6 oXqpottopr6 ev6g xtretmori ou;rntr6r<ou e[te r1 petotpon{ tou orrlv ovotXrrl Uopqi efue xot ro 5Uo. H ouvowenrrl oMr1tr'ouX(o orq 06o11 -10 qnotelefuqt onorletonrd on6 (e0yq Bdoeu:v A-T, Stopr6prpoon TIou 9on06 orov opXtr<6 6roXoptopr6 tou DNA oe provo0g xLrivoug. H Lty6tepq ev6pyeto nou onorefuor q 6totd- po[r1 tov (euydrv A-T oe ox6on Ue ro (eUyq G-C orlpo(- ver 611 dvo ttrrripo pe (e0yq A-T onotte( prtrp6teprl noo6rqto evSpyetoq yo to 6toXcoptqr6 tov xi.rirv<ov' Ot otrr1Lou{eq op5oo4 y0po rot r<o0odtrd on6 tqv o<pe- tqp(o en(oqg enqped(ouv qv 5vop[r1' Ax6p4, n np6rn neproX{ nou preroypdqerot (on6 -+1 Udxpt -+20) enq- pedEfl rrlv toXriqro Ue rnv ono(o q RNA noLuprep6oq onogot<p0verot on6 Tov unoKtvnrrl r<ot yt'our6 enr6pd orqv roXti rou unoKtvnr4. 'Etot, 4 ouvotrrr4 toXtig tou unorwqtq 6e ;rnope(ndwore vo npoBtre<p0ei on6 ttg ou- IroLXe[o UP 'i:':.t g 4noat^eoporLrorrlro rolv Ynoxrvlrdlv Mnopei v'Au[overot 1 vo EtrtroroveTot 0[o l,ra :):::- 523
vorvenKdq oMqAouXieg rou, ox6pr4 r<r ov tr4gOoriv un6_ qrlro enrnp5o0ero ororXe(o nou ovoyvrop((er 4 RNA no_ Aupepdo4 elc6g on6 trg oMqi.ouX(eg -10 ror -35. . Molov6rr 4 opor6rqto Ue rrl ouvorveru<r1 oMrlAou_ {o e(vor lprlorpro epyo},e(o yro rrlv ovoyvcilprorl unoxr- vrlrrirv pr6vo Ue Bdon qv oMql,ouX(o rou DNA, 6nog en[o4g xoL nop6rr ot (runrKo(> unoxtvqrdg nepdlouv eudtdr<pneg oMrltrouX(eg -35 r<or -10, noMoi unoKrvrl- tdg 6e 6ro05rouv eurprvrl morlefo -10 ror/ri -35. Ie notrilig on6 ow6g rtg neprmc0oerq, o unoruvrlrlg 6ev ovoyvrop((etor on6 g6v4 rrlv RNA no).uprepdor; xor qnorre( pto enrKouprKri npcoreivrl (,evepyonor4riq, - activator, BL6ne Kegdioro 26) nou unepBo(ver rrlv ove_ ndprero tov evdoyevcirv oMql,enrbpdoeov pero[ti RNA notrupepdo4q Kor unoKrvrlrl. Eivot oqpowrr6 en(or1g vo_rov[oou1,re 6rr "g6trrrm4 evepy6qroD 6e orlpo,v . ";r5yrorr1 evepy6rqro,,. llol,Lof unorrvqt6g dlouv e(e_ ).t10e[ pre oMqtrouXfeg nou on6louv no],rj on6 to vo e[- vor ouvqrverrr6g oxptBrig enerdrl 6ev efvor ro Bdtrnoro yto ro rUmopo vo grrd;yer 16oo nol,[ npoi6v nou KoJ_ 6monorekot on6 ro RNA pietdypoeo rouq. l9.lO l-loAA6q t-teproxdq Tnq RNA l-loAupepdonq'EpxovTor Apeoo oe Enogfi U€ Tov DNA ynorrvnrfi * H doHl rou 070 otrtro(et orov ouvdeeror ge rov nuplvo rou ev_ (u;rou, enup6n0vr0q oTrq neptoXeg yto npoodeoq oro DNA v, otrtrltrenrdpooouv Ur Tov unorrvr;r1 er llotrtreq neproXeg rou 070 oitrrltrenrdpouv pe rov unorLvrlrl. s H unopovodo o en(orig ouppotrtret orr;v ovoyvtirprol Tou uro_ rrvrlq. Onog npoovorpdp0rlKe orqv Evoulra 79.g, opr<erdg neproX6g orrlv unopovddo tou nopdyowo o Kot rrl CTD r;g unogovddog o Splowor o, enogrl pe ro DNA rou unoKrvrlr4. H rowono(qor1 ptog oerpdg drorpoperr_ t<c6v ouvorverLxov otrl.rlLoulLrrtv or ono(eg ctvoyvop(- (owot on6 o).o6v(upo trie dro<popelxoUg nopdyoweg o (dntog qo(veror o-Trlv ' , ., ,.) uno6r;i.rr;vrr o-i. : [6ro9 o nopdyowog o npdner vo dpleror o, enoef *. ro DNA. En[or1g unodrl].rrrvet 6l ot drogoperLxo( nop:- yoweq o npdneL vo npoo6dvowor Ue nop6pioLo rpo:: mov nuprlvo rou ev(rjprou, dror c6me oL enLgoveLs: ovoyvirtprorlg yro ro DNA ftov 6rorpoperrxrr:v nopoyo,,- roJV vo rono0erotiwoL ire nop6;roro rp6no otore ,,,n npoyporonorotiv ng xpforpreg enogdg Ue rov unoKrvr_ q r<owd ortg oMrlLoul(eg -35 xor -10. Enrnp6o0ereg ev6e([erg drL o nopdyowog o(ypro ec Xeror oe dpreorl eno<p4 Ue rov unor<rvr;r4 16oo orrl _35 6oo roL o-cr1 -10 ouvorvelrq oMrlLouX(o nporlLOov on: unoro-roordoerq orrov nopdyowo o nou KoroordMo", peroALd[erg orrq ouvorvelr<6g oMq].ou1(eg" Otov 1.ric perd).).o[r1 oe Liro ouyxercprpr6,vrl 0dorl rou unoKrvr.lr- nopepno6((er r;v ovoyv<rrpro4 on6 rqv RNA noLuprepc- on Kor puo owroro0prorLrq ;rerdMo{4 orov nopdyowc o enrpdner orrlv noi.upepdorl vo lprloLponorqoer ro f.lr- toMoypr6vo unoxrvqq, q nro nr0ovrl eEnynol eivor or. to olerrxd (e[yog Bdoeorv rou DNA dpleror o, enoer pe ro optvolti nou sxer owrKqtooro0e(, I uyr<p(oerg tlv oMq).ouyurtv opxertrtv Boxrq pto r<u_r,, nopoydwov o un6.6er(ov ouwqpqpr6veg neproXdg o-rc o70 Tou E.coli .,,, , ,, ,.', nou oMqenr6poriv dpreoc pe unorrvqtdg" Aur6g or ev6e((eLg rexprqprot0qxov onc rqv avoyvriprol Uroq xpuoroMLr<qq dopnq rou oLor,,- ([pou qg RNA no].upepdonq oe oUpnl.or<o pr, dvo r<.r.c- ol-lo rou unor<rvqtr1. To Borcrqpro Thermus aquaticus xc. Thermus thermophilus derlvouv notg or neprol6q npoc_ 6eo4g oro DNA rou nopdyowo o mulrr.lvowor or ovr_ {dprqreg neproX6g orrg nptrrreivLr<dg neproleq 1 2 2,3-2.4, 3,0 r<oL 4.1 -4.2. H Eu<dvo 19.'16 nopouord(er ro rprrlproro rou nopc- yowo o nou exouv dgeoo pd),o mrlv ovoyvcilprqn roL unorrvqrrl. Auq q eLrdvo de(pet q 6op4 -rou xrlpLcu nopdyowo o oro n).o(oro rou oLoev(0prou. Atio Bpolec rp4proro qg neproXqg 2 rr evo rprrlpo qg neproplq .1 (2.3, 2.4 KoL 4.2) eplowor o' enog4 ;re Bdoerg oro oror- Xe(o -10 r<or -35, owroro(1org. H neproX4 1.2 rou nopo- yovro o Spleror o' enorpr'1 pe rqv nsproX4 rol unoxrv4rq r<o0odLnd on6 ro ororXe[o -10, evot rl neproxn 3.0 dpXerot Ue qv nepropl rou unoKrvrir4 prd).rg ovo6L- rd rou oroLle(ou -10. Ko0etrr(o ond outdg trg neproldq oX4porf(er Bpoldo rp4poro o-ti].u<og orqv npc,-rteivn Ynogovd6o/ yov0ro M6ye0og (# aa) Kord npoo6yyrorl # tov unorw4rdrv Aliflou1(o rou urtoKvlrn nou ovoyvropi(eror ILypo 70 (rpoD) 613 1000 ILypo 54 (rpoN) 477 5 IryiLo S (rpoS) 330 100 Irypo 32 (rpoH) 284 30 ILypo F (rpoF) 239 4A I(ygo E (rpoE) 202 20 TTGACA-16 bp 6o9 1B bp-TATAAT CTGGNA-6 bp-TTGCA TTGACA-I6 bp eog 1B bp-TATAAT CCCTTGAA-13 erog 15 bp- CCCGATNT CTAAA-1 5 bp-GCCGATAA GAA-1 6 bp-YCTGA Iiypo Fecl (fecl) 173 1-Z ? Elx0NA 19.15 0t nopdyoweg o rou E coli ovoyvcop((ouv unorrvrlr6g pe drorpopelrrlg ouvorvelreg otrtrr1trouxreq. 524 KE0AAAI0 19 Meroyporprl oroug ilpoxopudrreg Boorreg eworeg
H 6opq rou nopoyovro o oro yevrrorepo ntroioro rou o),oev(upou: otr).qtrenrdpooetq -1 0 rot -35 0 nopoYovrog o enerrel- vrT6r Kgr or neptoXrlg rou ouvfleovror p' euetrrrroug ouvdereg. To oXrlpo 6XeL npoooplioorel ono: D. G. Vassylev, elal., Nature 417 (2002)'. 712-719. AoUrt on6 Protein Data Bank 1lW7 D, G Vassylev, elal.,Nature 417 2002).712-719 Mro r<puotoMu<q 6o;11 tou oLoev(0prou oe oripnlot<o 1.t' 5vo xldopro tou unoxtvrlni, oe ouv6uoopr6 pre nsrpdpto- ro Ue unoKrvqt6g oro ono[o ot r<].ri:vot tou DNA roro- oreudmqxov ri:me vo nept5Xouv oto(ptomo (eUyr1 (,,etepo6(xLrovo)), 66erqov 6rr o o70 6pXetot Kotd r<rjpro L6yo o' enogq ;re Bdoerg orov xtrcbvo-prr1 expoyeio tou ororXe(ou -10, oro 6reupup6vo orotXe(o -10 t<ot orrlv ile- ptoXrl tou 6tolcoptorrl xot ouve{(et vo 6rotqpe( ow5g rg eno<p6g t<ot getd to (etritrtypro rou DNA outtlg qg neproXqq. Aur6 entBeBouilvet 6tt o nopdyowog o efuot or1trrowx6g orrlv ow(6pooq onodtdtoEns FnEnS) H Xprio'n o-ehroetdriv pro{Brov e(vot t<owr1 oe npo- retveg nou ovoyvop((ouv oMql.ou{eg 6(r}'orvou DNA (BI6ne Ev5qto 28.6). Apwo[5o nou on5louv pero{U roug 3-4 06oetg Bpiot<owot ornv (6to nLeupd qg o-51't- Koq Kor yt'out6 e(vot oe 05or1 vo 6p0ouv o' enoq4 pe yenovrrd (e0yq Bdoeov. H ElKOl'lA't3'17 de(Xvet 6n r' oprwo[5o rord prlxoq ptog n].eupdg tqg neptoplg 2.4 trlq o-6lrro9 5pXowot o' enogrl pe rg Bdoerg ortg 06- oerg -12 5og -10 rnq -10 otrtrr1).ou{og tou unoxtvrprl. H neptoXrl 2.3 potd(et pre npo:teiVeg nou npoo66- vouv 1.rov6rtrrovo vouxleivtxd o(5o rt epnl,5retoL or4v ow(6pooq rn[nq. Ot neptoXSg 2.1 ru 2.2 (ro nto ouv- flpnudvo rpqgo tou nopdyowo o) egnl'6rowot oqv otrLqtren(6poon Ue rov nuprlvo tou ev(ripou. Eu<d(erot 611 6tror or nopdyoweg o npoo66vowot onq (6eg ne- ptoX6g rou nuprlvo trlg RNA no).uprepdoqg, on6re "ov- royrr:v((owotD yto neptoptopr6vo Udpoq tou nup{vo rnq. I-1op6l o nopdyowog o 6XeL neptoldg nou ovoYvo- pi(ouv ouyr<enpt;tdveg Bdoetg oro DNA tou unortv4trl, q N-teLrr<q neptoXrl tou eerJOepou nopdyowo o (ne- proXrl 1.1), evepyolwoq Log ouroovoorotrttrrl neptoXtl, onorpUnret qv noptoXrl np6odeoqg oro DNA. M6vo 6rov q 6Lopr6p<ptr:or1 rou rpononotq0e( on6 11 orjvdeorl rou ue rov nuprlvo tou ev([p-rou, prnope( o nopdyowog o vo npoo6e0e( erdLr<d ortg oMqLouX(eg tou unortvqtrl H oduvopr(o tou etrerjOepou nopdyowo o v' ovoyvolp[oer oMqtroul(eg tou unortvqt4 dleL orlpo- o(o: ov npoodevdrov oe unoKtvrlr6g u.tg e).e00epq uno- provo6o 0o pnopoUoe v' ovome[].et rqv 6vop[q tqg 06on -13-12-11*10-9-8-7 ElK0$iA 19.17 T' oprvo[60 ot2.4 o-5trtro tou o70 6pXovrot o' enoqn Ue ouyrsrprpdveg pooetq orov rodxonoulrtrd rtrtivo qg -1 0 otrtrqtrouXiog rou unortvrlrfl. 19.tr0 [lo),treg l-leptoX6q qg RNA llotruUepoonq'EpXovrot Apeoo oe Enoqrl pe rov DNA YnoxLulnl 525
neproxeg flp6o0eonq oro DNA tlH N-rdwr1 neptoXrl npoo6Sver nepLoX6g npdobeorlg DNA mov ele00epo o To DNA erronl(et qv N-re)x1 neproXrl rirov oXqgor[(eror ro ouurroKo pre ro DNA EIK0NA 19.18 To N-retrxd orpo rou o epno6(er Trq neprox6q rp6o6s0nq DNA vo ouvoeeofv Ue To oTdxo Touq. Mdtrrq oxrlUoTr- ore( 6v0 ovorXro otjuntroKo, ro N-rs,xd orpo ourlpeuor 20 A po- rpud ror or 6Uo neproX6q rpdodeonq Tou DNA 0r6xouv 11 1uo ond qv dtrr1 roro -15 A. Ueroypoqnq on6 ro ol,o6v(ugo. H EK6vq 19,18 ono6(- der g4qporrd qv oMoyri dropr6pgooqg rou nopdyov- ro o rotd ro oxnporropr6 rou ovorXroU ouprnL6rou. 'Orov o nopdyowog o npoo66verqr orov nupr]vo qg noLuprepdonq, n N-rellrq neproXrl ourlpe(ror -20 A por<pud on6 rtg neproX5g np6o6eorlg rou DNA ror or neproX6g np6o6eo1g rou DNA on5louv preto(ti roug -15 A, nr0ovr6g rilore v' ononrloouv nro enlgrp<uoprdvrl 6topr6pgcoorl, rotd).Lr1l,q yro rnv enog4 pre ro DNA. MeroLLd(erq e(re oqv oLL4LouX(o -10 efue mrlv -35 ovoor6tr.ouv rrlv np6o6eor1 oro DNA puog xotroBro;r5- vrlg erdoXqq rou o70 Xcop(g rqv N-rel,u<ri neproXrj, yeyo- v6g nou uno64l,6ver 6tr o o70 6pXeror rour6lpovo o' eno<p4 Kor Ue trq 6rio d.)l.r1lou1(eg. Aur6 rorprd(er pre rg nlqpotpop(eg on6 rqv rpuoroLllxrl 6oprl rou olo- ev([pou (Etr6vo 19.16), orrlv ono(o e[vor oorpdg 6n o nopdyowog o 61er gdMov enr;rrlxuoprdvrl Uopen, eKrer- v5prevog oe -68 A rov 6Uo orpoec0v rou DNA. MoLov5t 11 neproXq 1.1 rou nopdyowo o 6ev SXet ovoLuOe[ pre r<puoro].troyporp(o, Brorpuorx6g prerp4oerg qq ew6nrorlq rnq oro oLo5v(upro Kor oro ovotXr6 oUpr- nloro unodql.<irvouv 6r oro eAe00epo olo6v(upo 11 N- retur<{ neproX( (neproXri 1.1) Bp(oretor orov rewpr6 6(ou).o DNA rou ev(0prou, puproriprevq ouoroorrrd q 06- orl nou 0o xoroldBer o unoKrvnr{q 6rov q4qprorrme( ro oUpnloro rnq Ueroyporp{q (ElX0ilA 19.19}. Orov ro oLo6v(upro oXqproiloer 6vo ovorXr6 oripntroxo oro DNA, q N-reLtrrl nepropi rou nopdyowo o eKroTr((eror on6 rov r6pro 6(oulo. Iuvendlg, q ew6nroq qg oe q46or1 Ue rnv un6trotnr1 nporelvr1 e(vor notr0 euprerdB],4rr1. A].- trd(er 6rov o nopdyowog o npoodeOe( oro rewptr6 6v- (upo ror (ovd 6rov to oLoSv(upro npoo6e0e( oro DNA. H 5fur<o rou DNA np6na vo trretorwq0e( nepfnou xo'rd 16 A on6 t4v opXxrl qg 05or1 nporeql5vou vu ero6),0er mov xripro 6[ou].o tou DNA rr 6nemo vo perorrvrl0e( (ovd 6ore vo enmpdr.fer oT o DNA vo ero6l0er oro 6(ou- l,o t<otd ro oxrlporropr6 rou ovorXro6 ou;rnL6rou. H El- (01{A 19.20 nopouord(er owrl q perorfur1or1, B}.5nowog 526 KE0A Al019 Meroypogrt mouq l-lporoputlreq EIKONA 19.19 0 nopdyovtog o 6Xet gro enqrlxuop6vrl 0opr1 nou exre(veror rord p1rog qg enrqoverog rtrlv unogov66rov Tou ttu- prlvo 6rov oXrl;iouore( ro otro6v(ugo. oe eyxdporo roprrl rotd pr4rog rou dlovo t4g 6).u<og rou DNA. l-loLot6repo enu<pcnoiloe 11 dnogq 6rr o nopdyowog o efuor q povo6rrq unogovd6o rrlq RNA nolupepdorlg nou npoo66veror orrlv nepropi rou unorwrlrq, I(pepo yvrop(oupe 5rt or C-refurdq neptoyiq rov 6Uo unopov6- 6rov o pmope( entorlq vo no((ouv orlprowx6 p6lo orqv enogf pre ro DNA rou unorwrlrq ;r5oco rqg np6o6eor;g rouq oro ororle(o UP (BL6ne Ev6rqra 79.8). Enerdq or oCTd ouv66owor XoLopd Ue rnv un6l,orn4 RNA nolupre- pdon (FMne Eu<6vo 19.14), ro 5v(upro prnope( vo rprdoer oe opxerd onopoKpuol,l5veg neproX6g evril e(vor ox6pr1 npoo6e6epr6vo qro ororye(q -10 Kot -35. 'Etor or oCTDs nop6Xouv rwqr5q neprol6g yr' oM$.en(6ppooq pe psro- ypogu<orig nopdyoweq npoodedepr5vouq oe 6r6gopeq onoordoerg ovodxd rqq oger4pioq rnq Ueroyporp{q 616- gopov unorwrpcilv (BMne lGgdloro 26,7o Onep6vto). EIK0NA 19.20 To DNA opXtrd 6pXeror o' enorprl pe rov nopoyov- ro o (po0 ror rov nuprlvo rou ev(Upou (yxpr). Merouve[rqr nro po0rd p6oo orov nuprlvo rou ev(Upou yro v0 npoyporonouloer enoq6g pe rrlv otrtr4tr0uX(o -10. M6)rg onetreuOeptoOe[ o o, ro etj- poq Tou nepdoporog nou nept6Xer DNA ou(dver. Avor0ntooq pe r4v d6eLo rov Macmillan Publishers Ltd: Nature, D. G. Vassylev, et al., uol 417, pp. 712-719, copyright 2002. 0toroyporpfo pe qv odero rou Shigeyuki Yokoyama, The University of Tokyo. N-TetrrKn neproxn U-
19.1 1 H Anorrjntoon Eivor Mi6o6oq YrlnAfq AvdAuonq Vro To Xoporrnprop6 rurv AAAnAenr6pdoeov RNA lloAupepdonq- Y no xw nr i't xaq I €,v rx6repo, DNA-I'I p an ei' v thv * 0r ouvorvenreq otrtrntrouXreq -35 rot -1 0 nopSXouv T0 nepro- oorepo orlge[o enoqrlq yro qv RNA notrugepoorl 0T0v uIIoKt- vnTrl. = To orluelo enoqnq 9p(orovror rupfog oq pto ntreupo rou DNA. H txov6r4ro qq RNA nouprepdo4q (ornv npoyport- xdrqro, r<d0e nporeivqg) v' ovoyvorp((er to DNA pno- ce[ vo XqpoKrnprore( pe onorUnooq (footprinting). Vro oMqloul(o DNA npoo6e6epr5vrl oe Uto nptotelVrl r6meror peputitq on5 pro evdovoux.tredoq nou enrG 0erqt oe (eltrrptoroIg grrrogodreoteptroUg 6eopotig ldoo mo voux.treivrr6 o[6. Yn6 r<otdtr].r1].eg ouvOr1- <eg, rdoe (e1oprm6g grj:ogodremeptr6g 6eopr6q .rnope( vo ondoer oe rdnoro prdpto DNA. Ot O5oetg :ou ndnrowor ovoyvtr;p((owot Xpqotpronoui:wog D- A o4pooprSvo orov 5vo x).rivo, p6vo oro :vo dr<po. [lp6retror yro rrlv (6to oppl nou iraoppd(eror ornv oMqLoriXqoq rou DNA: - peprr<r1 ndr,Uq ev6g telrrd oqpoopr6vou *op(ou oe Uro eunoOrl 06o11 dqpuoupye( 5vo doopro povo6u<oU prqxouq. H 6e(Xvet 6tt petd rrlv Korep- .'ooio pre voux.tredoq to onoop6vo r<i.doproro )NA pnope( vo 6toltoproroUv ovdloyo pe to *r'lKoq rouq pie rlLer<rpog6pnon oe nr1rr1, (o0e x).oopro nou 6Lotr;pe( 6vo orll.rooprdvo :r<po nopdyet pto po6revepVn (rr:vn. H 06ot1 :1q (rrrvqg owtororle( mov opt0pr6 torv Bd- trocrJv oro xi.dopo. To Bpoxurepo r^doporo *erorrvo0wot roXUrepo, 6ror q on6moorl :16 ro orlproop5vo drpo unoloy[(etor on6 :c xdrro pr5pog qg nnKqq, Iro e).eU0epo DNA, r<d0e eunoOnq 6e- :-rog ondet roul.dltorov oe rdnoto p6pto. H =L<dvo 19.20 6e(Xver 61 6rov to DNA Bpi l(rror or oripntror<o Ue Uro npore[vq, ot 05- leiq nou roLUnrovror on6 tqv npotelvrl .cccpuldooowor ond qv n6Qr1, 'Etot, 6tov 5.ro owLdpdoerg rp6(ouv nopdl,).q).o -dvo H onorunool ovoyvtrtpi(eL rtg 0e- -.r-q onou npood6vovrot npoTelvsq oro DNA ono -,' npooroo(o nou nop6Xouv rlvovrL qg 6qgtoup- , .rc evropcirv. DNA prdprupog 1op(g ropr(o np<oreTwl ru 6vo nerpogqn- 16 prfupro pre pr6pro DNA npoo6e6e;rdvo mqv nporreivr; 0o 64;uoupyrl0e( dvo XopoKrnprorrr6 npdruno. 'Oruv gro npoodedepdvrl nptoretvq ovoore(Aer rqv np6oBoo4 qg vouri,edonq oro DNA, or 6eoiro( orryv npoo6e6e;r6- vr1 oMqAoulfu 6e ondve oro rretpapaut<6 1t$pa Ket ctu- t6 to t1t4pa rqq ftnKrrlq 6ev nepdya o4paopiva xid- oparu DNA. Iro prdptupo, onder r<dOe 6eog6g dqpnoupytitwog pro rl,(poro (ovrirv, orrlv ono[o x60e Bdoq owtnpooo- neJerot on6 gro (c0vq. Irqv ex6vo, undpXouv 31 Bd- oerq. Iro npoororeuUdvo xtrdopro, 6eopro( 6e ondve orrlv neproXri 6nou e[vot npoo6e6etrr6w1 11 npcoteivq, on6re 6e dr1;uoupyoriwor (c0veg nou owtnpoooneUouv rl6oproro tcov ow[morlrov preye0c0v. H onouo(o turv (ovrrlv 9-18 orrlv eu<6vo rou'ronoe( ptto 06oq np6o'6e- o4g q ono(o roltjmet rrlv neproXtj nou Bp[oretot 9-18 Bdoerg on6 ro or1;roopdvo drpo tou DNA. Iuyrpfuov- roq rrl Lop(6o tou ;rdprupo Kor rnv netpogotu<r1 op(- 6o pe pro ovt[6poor1 o].].q].orixnong nou rpdlet nopdtrtrr1tro, prnope( vo "6tqpoore6, dpeoo q owforotXq oMrlloul[o, ovoyvrop((owog 6ror tr1 vou t<].eottdu<rj o],- Lql,oufo qg 06o119 np6o6eo4g. 'Oncog npoovo<p6p0r1r<e (BL6ne Ex6vo 19.13), q R- NA nol.uprepdo4 npoo6Sverot srrlv neptoXti rou unoKt- .AJffinffihnffih I*stu@htr@ + /V/Va/1. + /-,/,/VV. }a b + .ra/./vv1. *ry+ /L / HAEKTPOOOPHIH DNA olgoogdvo oro / Ilto gorPtd on6 dxpo rou evrjg rtrtivou m ffi- ro olpoopdvo ry ffi drpo 1ETpAMATTKH ffi ffi 1HKTH NHKTH ffi ffi MAPWPA: ffi To DNAnou 6ev To orlpntroro RNA rduUepdonq- unoKrvnrn ddXerot peprxq en(Oeoq on6 qv DNAdoq I 0r (tiveg nou onouord(ouv ovoyvrop(ouv 06oerg npdoDeonq Anopovrrlorl rou DNA xor onoDrorolrl oe lrovoug rtr6voug ffi eivoL npoodedepdvo ffi sqynougepoorl ffi eXer (tiveq oe Oeoerq ffi fiou owrororxouv oro :::: onoorpo Kooe deol]ou llro rovro oro orll-roouevo oxpo ' ; :: H Anorunorq e[vor M50o6og Ytpllrlg Avoluqg yro Xoporqpto;.ro Atrtrqtrentdpooeov RNA llotruuepoolq-Ynoxtvrlq xct Dl',]i-': *..., * , 527 Boorxeg evvorcg
v4q on6 -55 Scog +20. To oqpre(o oro ono[o q RNA no- Lupepdorl.6pXeror oe dpeorl enoqrj Ue rov unoxl4rrl rquronororiwor Ue rponono(qor1 rrlg reyyu<rjg rnq ono- t0n<oor1 g, 1p4 orprono uirwog owr6 poonl pro no u rpono- notoUv ouyrerptpr6veg Bdoerg oro orjprnl,or<o RNA noLuprepdo4g-unor<rvqri. Me ouq qv npooriyyron, ro ne(po;ro ;rnope( vo ercrel,eore( ge 6tio rp6noug: o To DNA pnope( vo rpononotrlOef nprv npoodeOe( 4 RNA nol,upepdorl. Ie ourrl qv nep(nrooq, ov 11 rponono(qo4 egnod(oer qv np6odeoq r4g RNA no- ).upepdoqg, rouronomfuor trlo Bdo4 6nou q enoErj efuor onopofurlq. o To o0unLor<o RNA notru;repdoqg-DNA pnope( vo rpononor4Oe(, on6te ro np6runo rov npoororeu- pr6vrov (alvcbv ouyr<p(verot ge ro ovr(morxo rou eLerlOepou DNA r<ot rou 1-trl rpononorqpr6vou oup- nl6xou. Kdnoreg (6veg e(ogov((owor, rouronor6v- rog 6tor 06cerg ottg ono(eg ro 6v(upro rlXer npoororeUoer rov unoKtvrlfi on6 tponono(r1or1. iL- Leq (6veg 6Xouv ou[r1pr6vq 6woor1, towonorciwog O5oerg orrg ono(eg to DNA np6na vo 6roqpr10e( oe 1.rro orepeo6op4 mqv ono(o e(vor neproo6repo erre0erp6vo orov nopdyowo n5Qqg. Aurdg ot otri,oy5q or4v euoroOqo[o onord].utfov r1 yeolprerp(o rou oupnL6rou, 6ncog ouvotp((eror orqv El- K0l,lA 19.22 yro 5vo runx6 unorrv4rri. Or neprol5g oro - 35 ror oro -10 nepr6Xouv to neproo6tepo on5 -ro o4pe(o enorpfg yro ro 6v(upo. MSoo oe our6g trg ne- proXdg, or (6teg oprd6eg O5oeov re[vouv'16oo v' ono- rp5nouv rrlv np5o6eon qv 61ouv nporlyoug6vrog tpononot4Oe( 6oo xor ov eprrpov((ouv ou[qpdvrl ri prer- cr:pdvq euoro04o[o orrlv rpoTronoirlorl trrerd rqv np6o- 6eoq. To orlpefo enogqg 6e oupn(nrouv n],ripog pre O5oerg prero,trtrd(eov: ouprBo(vouv ndwcog orrlv (6ro ne- proproprdv4 neproX4. A[r(er vo orlpreuoOe( 6rt or (6reg 06oeLq oe 6rorpopen- roUg unoxlqr5g nopdXouv noMd on6 ro orlpre(o eno- rpr'19, 6oro Kr qv rl Bdoq nou Bpioreror oe Uro ouyKe- xprpr6vr1 06oq 6rog5per. Aur6 unodrlAdrvet 6n undpler rivog r<otv6g ;rr1Xovtotrr6g yro rnv np6o6eo4 qg RNA noLuprepdoqg, nop6rr 11 ow(6pooq 6ev e(optdror on6 r1v nopouo(o ouyrerprprSvrov Bdoeov oe optopr6vo on5 ro orlpre[o enoqrjg. Aw6 ro ;row5].o e[4ye( yror( xd- noro on6 ro o4pe(o eno<pr1g 6ev e(vor 06oerg peroMd- [errlv. Enrn].6ov, 6e Bp(or<erot r<dOe prerd].],o(r1 oe oqpe(o enorp{g: or preroMd(erg ;rnope( vo enrlped(ouv q yettovu<q neproXrl 1o:p(g mqv npoygolr6rnro vq Splowor o' eno<pri pre ro 5v(upro. E[vor r6ro[repo orlpowx6 6n ro nepdproro nou oKo- LouOo0v q orporrlyrxrl rng npo-rpononoqorlg rouro- nororiv Odoetg or4v (6to nepropl Trou npoorore[eror on6 ro 5v(upro Svowt enqK6Lou0qg rponono(qor1g. Au- rd to 60o nepdgoro perporiv 6ro<poperu<d npdyproro. H npo-ponono(r1oq ovoyvop((er dleg ttg O5oerg nou npdner v' ovoyvcop(oer ro 6v(upro yro vo npoodeOe( oro DNA. To netpdgoro npooroo(og ovoyvrop((ouv 6Leg rg O5oerq nou npdygort Splowor o' eno<pq oro 6rprep5g otipnloro. Ot npooroteu;r5veg 06oerg nepil.oprBdvouv 6leg ug 06oerg ovoyvc0proqq r<or peprrdg enrnpdoOereg 05oerg, yeyov6g nou uno6rlLtiver 6rr ro 5v(upro npcbto ovoyvcop[(et puo oprd6o Bdoecov onopo(r4tov yro rnv "npooye(oon) Tou Kr Snemo enercrefuet ro oqpe(o eno- grlg rou o' enrnp6oOereg Bdoetg. H neproXrlrou DNA nou e(vor (eruLrypdvq oro 6rpe- p6g oUpn).oxo prnope( vo rouronorlOe( d;reoo pe Bdon rrq Xqpu<5q oMoy6g orq 6ro0eo4-r6tr1rd qg. 'Orov 6ro- lcoprororiv or x.trri:vor rou DNA, or o(euydpcoreq Bdoerg yfuowor eud.troreg oe owtdpoorrlpro nou 6e prnopotiv vq rrq npooeyy(oouv or4 6rntr4 5Lu<o. T6roto neLpd;roro nepiloprBdvouv 06oerg pero[ri -9 xoL +3 orrlv opXu<r] ov- ildpooq rri[nq. I-r'qur6 11 neproXrl nou [eru]lyeror rord rqv 6vop[4 neprAopBdver ro 6elr6 dxpo rrlg oMq].ou- X(oq -10 ru ercre(veror pr6Xpt pr6).rg prerd r4v oEerlp(o. Av r<orrd[oupre rpro6tdmoro ro orlpre(o enocprlq qvodrKd rqg oLLr'1louX(oq -10 dronrmrirvoupe 6tr Bpi- oKowor 6Lo orq puo nLeupd rou DNA. Aur6 <pofveror oro xdrcrr Srdypogpo qg Eu<6vog 19.22, oto ono(o ro EIKSNA 19.U2 Mto ntreupo rou unoxtvlrrl nepL6Xer to orlpe(o enoqlg yro ro RNA. Atrtrr1trouX(o -35 t ALtrnlouxio -1 0 J+ t,I, ,1. l. I lt^,t+Iitl, I K06rKoIornrtK{iq t Ktrtivoq lxxxxllu.IffiKtrdlvog erpoyeio To neprooorepo orlge(o enoqlq pploxovror oq gto ntreupo rou DNA Tpononouloerq nou egno0[(ouv r1v npdodeorl qg RNA notrugepoorlg 06oerg rinou 1 RNA notrugepoorl npoororeuet ond qv rpononoiqorl l4ero),tro(erg nou xoropyouv 11 perdlvouv qv evepyriqro rou un0Krvlrr1 J + 0 ti ililil1 528 KEOAAAI0 19 Meroyporprl oroug l-lpoxoputilreg
: ^Joro enoglq onpercovowor oe pro drnArq dtrrro nou j 1.,,'rror ond q prLo nLeupd. To neproodtepo Bp(or<ov- :trr orov r<ovo prq-er<pioye(o. Aur6g oL Bdoetg (or,rg ovo- .,ropi(owor r<otd tov opXrr6 olqpotropr6 evdg xLer- :.or-i 6Lpepo[g oupnLdr<ou. Aw6 0o enSrpene orqv R- ', - .rouprepooq vo n).rloLdoer to DNA on6 14 pro ntreu- - r (or v' ovcryviop(oer ourr; rqv dQrl tou DNA. (o0rig [errvd ro (erritrLypro tou DNA, ;rnopef v' ovo- .*pLoro[v r<or dtr].eg O5oetg nou opXrr<d Bp(orowot --',, oMrl nl.eupd tou DNA. | 9.1 2 Or AAAnAenr6pdoerq Mero(U rou llopdyovro o Kor rou llupfivo Tnq RNA IloAupepdonq AA deouv KoTd Tn Aroguyfi TOU YnOKIVNTfi n ','.0 n00t0Xl rou o rorotroppovet ro dtoutro e(o6ou rou RNA .r. lrprnrr vo perouvl0el ylo vo dteurotruver q o[v0eo1 rou t.i . - ,.nTUXe[q evop[erg ouvq0tog ouppo[vouv nporou ro ev(upo ,,i-porlorr evo npoygortro o0gntroro enLprlruvolg. " I .opoyovroq o ouvnOoq onetreu0epdrveror ono qv RNA :,,tpepoo1 potrtq q veoouvre0elpdvq otruo(60 RNA tptooet :: plxog -'10 nt. - rA noLupepdoq np6neL vo ouprBrBdoer rrg ovdy- ..: !'ro dvop[q pe rtg ovdyxeq yro enrprnruvon. l-1pco- -: , o 6(ouLoq Sroguyqg rou RNA ornv --:iuolx6rqto e(vot r<oreArlprpr6vog on6 rprlpo tou -r:ryowo o, ro ouv66rq (linker) nou ouv6Set rtq ne- - r ,.3Q 3 r<or 4. 'Eror, 11 6to<puyr1 rou unoKtvrlrl np5net : .oorloprBdver ovodLo-rdro(q rou nopdyowo o, .- cri(owdq rov on6 ro d(outro e[66ou tou RNA - -t vo prnope( vo ouvexrore( 4 otlv0eoq tou RNA. ' : - .3oov, q 6vop[r1 onone[ orevq np6o6eor1 p6vo oe - -. <a<prpdveg oMq).ouX(eg (unor<tvr1r5g), evcb q enr- --,-;ol onotre( np6o6eor; oe dreg Ttq oMrlLouXieg - - - :o ouvowrloer to 5v(upo Kord tq preroypogtl. H 6e(Xvet nurg enil.Uetot out6 to 6il.qprpro -.r* rlq owtorpemrlq np6o6eor1 tou nopdyowo o - :, rupqvo rou ev(Upou. Oncog npoovogSp0lKe, ro . --ic ouvrl0og u<p(ororor r<rirLoug oventtuXtov - ::ieov xord q 6Lepyoo[o r4g 6to<puy4g on6 tov --:..'rlll. Ev6oto oupBo(vouv outo( ot oventruXe(g . -- ' :., ro dv(upro 6e perorrvekor ndv<o oro expoye(o, : -' : -oMov rpoBd ro npdlto ).tyo vour<i,eor(6ro tou . :::5Lr<oU DNA, npoBdl).owog ouroUg roug provo6g . -- ,o,Jq novco orqv enrgdverd rou, oe pto dtepyoo(o -:rrl orq <paoqoq tou DNA - DNA scrunching". Ztn r- ,. '.aro. p' 6vov prrlXovtopr6 nou 6ev SXer onooorp4vr- , . .r.r1pcog, to 6v(upo dro<peUyer on6 ouq q <pdor1 --, r','enrruxov r<rjr<.trrrlv r<ot nepvd orrl rpdorl Tnq ent- _ -'- , rplyopo I f novo ww@w ",:1lk!'> / --*ru I i U rpnvopo * : ^m.:@E i ,*oypnyopo I i ^d!%' :*-Ef*eB ! n*rypnyopo I i#@r....... I To revrprxri 6v(ugo repgorl(er rr oneheu0ep6veror EIK0NA 19.23 0 nopdyowoq o ror o nuprlvoq rou ev(ilgou ovo- ruxtrtirvovror oe dtorpopenro oqpeio qg Ueruypoqnq. Molov6rt q oneLeu0Spoorl rou nopdyowo o on6 to o0pmtroro 6ev e(vor onopokrlrrl yto q 6roguyi on6 tov unorwqr], 11 drdoroorl tou nopdyowo o on6 tov nuprlvo rou ev(titrrou ouvrlOoq ouprBo(ver rout5lpovo 11 Lryo perd rq 6roquyrl. O nopdyowog o e(vor oe nep(o- oero or o16o4 pre qv RNA nolupepdoq, ouvencig 11 oneLeuO6pcoon rou on6 ro oLo6v(u;ro 6e y(verot onL6g yro vo ro xdver 6ro06otpro yrq Xp{on o' enrnp6- o0ero owlypoqo rou otroev(riprou. Irqv npoy;rorr6q- ro, ot nopdyoweq o owoyrov((owol yro neproproptdvo owfuporpo rou nupfvo qg RNA noLuprepdo4q, yeyo- v6g nou 6(ver q 6uvot6rqro v' oMd[er ro npSruno t4g peroyporp4g (BI6ne Ev1trTra 1 9.1 9). O nuprlvoq tou ev(6;rou (<,xewpx6 tiv(upto", core enzyme) cro rprprep6g oriprntroxo (nepd.o;tBdvet DNA, veoouweOery6vo RNA r<or RNA noLuprepdor;) ouorool- rd e(vor "x.l.et6ofr5vog" pr6Xpr vo o).oxlr1po0e( 11 enrprl- KUVon. 'On<oq 0o 6oripre, outl 11 e[6].t(q npor0met ev ;rdper on6 rov rp6no Ue rov ono(o ro 6v(upro neprKu- xl6ver to DNA ru ev p6per on6'rqv of[non qg ouyyd- verdg rou yro ro orl;rntroro Ue ro veoouwe0ettrr6vo RNA. KevTprKo 6v(uUo ono0lreupevo oro DNA 0 nopdyovroq o ouv06eroL pe ro revrprxd 6v(upo To otrodv(upo perorrve[ror npog Touq unorLvlr6g To revrptxrl ev(u;ro ouv06ret RNA 3.rtr,rltrenrdpooergMero[0rou[lopoyovrooKorToutluprlvorrlqRNAtlotruUepoonqAtrtro(ouvKoroqAroquyl:0i-'.].,.,-.- 529 Booxfg eworeg a
To gdppor<o prgopnu<(v4 (UdLoq rrlg orroy6verog rou owrBrorrroU prgol.rur<(vq) epno6((er q preroypo<pq on6 q Bor,m4pror<r1 RNA noluprepdoq. E(vor ro rUpto owrBtoru<6 evow(ov qq gupror(coonq. H rpucroMrr{ 6opr1 qg RNA noluprepdonq oe o0gnl,or<o Ue rn pt- <poprnx[vr1 e[4ye( q 6pdor1 rou cpopprdrou: n prqognr- r[v4 npoo6Sveror o' 5vo Ori],or<o rqg B unopovddog, >12 A prorpud on6 ro evepy6 r6wpo, oe ;uo 05o4 6nou nopoxrr:trrJer q 6to6popuj rou enrgnKuv6gevou RNA. Epno6((owoq r1v endrrooq qg oLuo(6og tou RNA ndpo on6 2-3 vouxl,eor(6to, 6tor5nter rrl Uero- Ypoen. 19..I3 H KpuoroAArrrl Aopri Yno6errv0er 'Evo MovrdAo yro Tn Merorivnon Tou Ev(0pou e To DNA perorrve[rol dropeoou evog droutrou orrlv RNA notru- gepooq ror npoygoronore( pto onoropq orpotprl oro evepy6 r6vrpo. x Atrtroydq oq dropoptptrlorl ouyxerprpevov eudtrrrrtov e[oprr1- porov p6oo oro ev(upo etreyXouv qv e[oodo rrov vourtrrol- d[ov oro evepyo xevrpo. Me q droLeUrovoq r1g xpuoro).).rx4g 6ogrjg rov ev- (tipuov on6 Borcrr;pro r<or (tipre oe oUpnloro pre NTP ror/n to DNA, onorcr4oope noMdg nnpogop(eq oxe- nrd ;.re rq 6o;14 r<or 14 l.enoupy(o rrlg RNA nolupe- pdonq rord rrlv enrprrlruvoq. H Boxrqpror<r1 RNA noLugepdorl 6Xer 6romdoerg -90 x 95 x 160 A, evo-l or s0A RNA noLupepdoeg rov opXo(rov Kot rov europu<orrirv e(vor eloEpcbq preyol0repeq, rup(rog ).6yo np6o0e- rov rgqgdrcov o1.uvo(6cov r<o/r1 unopovd6ov rou ev- ton((owor orqv neptg6pero rou ev(ri;rou. MoLorowo, ot nuprlveg rov ev(U;uov prorpd(owor 61r pr6vo r<owr1 6opr{, mrlv ono(o undpler 6voq "6[outroq" eripoug -25 A nou meyd(er ro DNA, oMd r<or r<ow6 pr;Xovt- qr6 yto npooOrjrq voux.l.eou6[ov. 'Evo gowriLo owo( rou 6roUl.ou orq Bor,irrlprorcg RNA nolupepdo4 ono6(6eror orrlv EIK0HA 19.24. H oG Loro ouyr<pore( -17 bp DNA. Ie ouv6uoopr6 pre to -13 nt DNA nou Bp(orowor orrlv neproXq rou evepyof x6v- rpou rou ev(0pou, owtorotloriv ornv npoororeup6v4 neproXrl tru'p<oug -30-35 nt rou noporqpe(ror oe on(> tunc0oerg tou oupnL6r<ou enrprlruvoqg. H o0Aoro enevdUeror pre Oeru<d gopr(o, Xdprl oro ono(o oM4Ae- nrdpd pe rtg n^erupopvrlnr6g qrr:orpoprxdg opd6eg tou DNA. To rorolurrx6 r6wpo (catalytic site) oX4pro- r((etor on6 pro o1rotrrr1 ovdgeoo mrg 6rio ;reydleg unopovd6eg rt otXl.roLor((er oro (oqy6vro" rou to D NA t<oOcbq ero6pleror mqv RNA noi,uprepdo4. H RNA noLupepdo4 neprBdLLer ro DNA Kor oro evepy6 rdv- rpo rnq nepdXer 5vo 6vo rorolurx6 r6v Mg2+. To D|.lA ouyrporefuor orq 05oq tou on6 rov r<o0odrrd ouvde- rrjpo (clamp), evoMorcrtrq ovol.roo[o yro 6vo on6 to ooy6vro. H EIK0HA 19.25 onerrov((er q orporp{ 90o nou ovoyr<d(eror vo npqyporonorfoer ro DNA orrlv e(oo6o rou evepyo0 r<5wpou elon(og ev6g 6rnlovoU npcoreivr- ro0 re[Xoug. To pr{r<og tou uBprdu<o[ RNA neprop[G- ror on5 6vo 6).Lo npo:reivu<5 eprn66ro, yvom6 og rdlu;r;ro (lid). To vourctreor(dro ntoreUerot 6u ero6p Xowor oro evepy6 r<5wpo on6 xdtco, prSoo rou 6eure- poyevorig 6torlLou (mqv RNA nol,uprepdoq rnq E0pnS ovoprd(eror "n6poS", pore). H rpuool(6o nepiloprB6va 8-9 bp uBpr6(ou preroyporptxrl DNA-RNA. To xdLuppro Movr6tro nou 6e[Xver q Dogrl qg RNA no- truUepoolq 6top6oou rou rup(ou 6roul,ou. 0r uno;-rovodec rordrronorouvror Xpopolro coq e[lq: B po(, 0, ruovo, o npootvo, oll, r[rprvo, o, 16rrrvo. Ipoooplroogrlvo onc K.M. Geszvain rorR. Landick (ed. N. P. Higgins). The bacte- rial Chromosome. American Society for Microbiology. 2004 FcTp,,l 530 KEOAAAIO 19 Meroyporprl oroug llpoxopudrreg Boorxeg ewoleg
Avo6rxrl DNA 'Evo nporeivtro re(Xoq e[ovoyro(et To DNA v0 :,poqr1 oro evtpYo K6vTpo. Nouxtreor[dto pnopel vo oro evepy6 rSvrpo 0toireoou evog nopou oqv npto- :.-ir rLq BdoeLg DNA rot RNA oro 5vo drpo tou : :- tpl6ne Etr6vo '19,25) r<ot n Fdon tou DNA l:.cxeroL mo dMo dt<po rou uBpt6(ou ttvd(etot llpoor(0erot rowo0pu Bdoq xot ondve 0eogo( 'R11i-: lrt;ii,l lltr;111. To 6v(uuo gerortvefiot rot ovodrlptoupye( touq 6eoU00q il ;, r. ll ll:t,i, , -:- - "-..... --, ;:.r.i;il+1li.it:; ', Noudeot(6tq KtuIon tou ev(Upou ir I !1,_! 2To DNA Ero6pXoTot oro ooydvto : .: e[o yto vo enttpSqet to (euydptol-to Ue ro et- : -o'vo NTP. ^N r A . ,-^-^i -'(E* h -^avia rr,lr Ftwr,lnt- :1..q ro DNA unoore(qEn, n rpox(q tcov [elcopt- , - . ,.i.!vcDv U6oo crro 5v(upo dev neptop((erot nlSov :-: :- 5uoKoUqr(o 'rnq 6tnI(q 5trxog, enrp6nowog oto - ,: . : rpoYuoronotrloet rq orpoqn 90o oro evepy6 r6v- -: : I:.c1q, ouprBo(vet ptto peyoLr1 oMoyrl dtopt6ptpoorlq -- l : ro dv(upto nou nepAoptBdvet to "ouvderrlpo" o EIK0NA 19.26 H Ueroxivnon rrlq notrupepdorlg evdg vourhei- vrKorj oE6oq rpoUn0e6Tet Oporion Kot 0v06rlUtoupY(0 6eop6v pe vourtreotldto oe oT00ep6q e6oetq oXettrd Ue Tn Oogn rou ev(rlgou. To vourleoridto oe our6g rtg 06oetg d0d(ouv Kdee Qopd rou To 6v(uUo UeToKtvs(Tot KoTd Uto pdoq ndvto oT0 eK' uoYe(0. vouKleivtKov o[6ov nou Bp(or<owot prto 06oq nto rdrco Kord ilnKoq tou t<.trtilvou. To ;reoo[o n].o(oto 6e(1vet 6tt ow6 np6net vo onpo(vfl 6tt pr6tr'tg npooreOef n pdon Kot nprv ro 6v(uUo petorwqoe[ oe oxSon pe to vour]'ei- vrx6 o(ti, ot enogdq nou 61ouv 6r1prtoupy40e( pe er6x6g 06oetq np6net vo ondoouv 6tot cilore vo Unope( vo Eo- vodrlpuoupyl0oriv pe rtq Bdoetg nou Kqto^,ogBdvouv ng O5oetq our6g prerd q ;retor<[vqor1' Optop6veg rpuoroLLtt<69 6opdq rrlg RNA nolupe' p6onq npooq6pouv oqpowtx6g ev6e([erq yto to nc0q ro 6v(upro 6rotqpe( rtq enog5g pe ro un6orpopd tou ro0drg ondet Kot [ovo6qproupye( 6eoprofg qrn 6tepYo- ofq rou r<[x].ou npoo0rixqq vour],eott6(ov rt evcir uq(- orsrqt petot6ntorl. 'Evo eudNrro t;r(trro nou rol'efuqt 0qtud nupo66qonq (trigger loop) qo(verot 6tt 6ev efuot mulo;t6vo Tlptv rl'lv npoo0(xr1 tov vourl,eort6(tov, dr- d muXrivetot pr6l,tq to NTP etodLOet oro evepy6 x6v- rpo. 'Orov olorlqpo0e( o oXrl;ronopr6g tou deoproU rot r1 pretor6ntoq tou ev(rigou orrlv en6prevq 06o11, 11 Or1Ltd nupo66tr1or1g (erutr'iyerot rot n6Lt, 6roq-r1 yto tov en6- prevo r0xl.o, 'Etot, 1.tto Sopu<r1 oM.oyrl orrl OqLtd nupo- 66rr1oqg ouwov((et Tr1v orotrouo(o tcrrv oupB6wov rqq rot6).uor1q. :: cXqprot((et 6vo on6 to ooy6vto tou ev(fpou nou -ouv ro xo0odtr6 DNA orn 05or1 tou. " : :nd to 6tl.qpr;roto roOe notruprepdonq voux.e'i- :ieoq e[vot 6tt to 6v(upo np6net gev vo 6qpt- -:r,, oerxr5q eno<pdg pe to un6otpopo Kqt Ue ro : . lMo en(or1g xot vo 6tor6r.l.tet out6g ttg eno<p5g , : :.Q [ovooltlpot(oet oe rdOe r<0ro npooO4r<qg -'r::16(ou. Ag ovoloytmo[pre trlv rotdorooq nou . s3Qerot oTnv Mro notrupepdorl *::onote( pto oerpd et6trrov enorpov pre ttq Bd- : :,.l Bp(orowot oe ouyxerptp6veq O5oetq. l-tq no- :...*c, q enogrl "1" dtlutouPYe(tot pe q Bdorl mo :: :ou ou(ov6pevou x.l.<ovou, evco 11 enocptl "2" pt - ::]r orov rLdlvo er<proye[o nou e(vot ougn].qptl1'to- - . :: -. qv en6pevq B6oq nou 0o npomeOe(' Entoq-- . :: -. qv en6pevrl B6oq nou 0o npomeOe(' Entoq- -: ..:lr, ocn6oo, 6rt ot pdoetq nou rotol'oprBdvouvl. el, wv .: :rq 06oetg arouq r<Ltovoug ttov vout<LeivLxriv - i: --, oMd(ouv r<dOe <popd nou npoor[0etot 6vo vou- -: rovo.r ror to xdttrl nl,o(oto r1g etr<6vog 6e(lyouv -. l.c rordoroorl: prto Bdo'r1 np6rettot vo npomeOe( , :. :r(ov6;tevo xtrtrlvo. H dtotpopd e(vot 6tt mo rdttr; - r l.c o oufov6pevog x}'tovog SXet ener<roOe( rord pto : --- -l yeo-lpretp[o Kot rov 6rio ouprnL6rolv e(vot oxpt- :-: - i6Lo, qMd oro xdto nLo(oto ot enorp6g "1" Kot : :ooyUoronoro6wor pe Bdoetq tolv oLuo(6<ov tcov l* il H Kpuorotrtrrrl Aoprl Ynooltrdrver'Evo Movr6tro yto q Meroxtvlon lou Et'[-*:- 531
19.1 4 Mlo lropornp6vn RNA l-loAupepdon Mnopei vo lovopxioel r Mr0 or0U0Tlp6vq RNA notrugepool pnopei vo {ovopXioer ng pet0ypogrl xdpovrog ro RNA peroyporpo yLo vo Drlgroupyloer 6vo roLvorjpro 3' oxpo. H RNA noAutrrepdoq npdner vo e[vor oe 05or1 vo Xerpr- ore( rotomdosrq orrq ono(eg n Ueroypogq ovoor6).- Leror. I-ro nopdderypro, qur6 prnope( vo ouprBe( 6rov ro DNA SXer unoorei BIdBn. 'Evo np6runo oriorqpro yro r6roeg neprordoerg nporUnrer ge 6ror<onri qg enrpr{ruvoqg in vitro e(opc0wog 6vq on6 ro onopo(- rqro np66popro vouxleor(6ro, lepropr6g nou enmpd- ner rn gOopd rou dxpou rou RNA. 0o16oo, to [6ro onorSLeopo 61er xou<dOe ouprBdv nou nporoLef ].d- 0og euOuypdppton rou 3' dr<pou rou RNA Fre ro evep- y6 r<6wpo: r<dr lperd(eror yro vo lovoronoOerfoet ro 3'-OH rou veoouweOer;rdvou RNA oro evepy6 r<6wpo 6ror c6ore vo prnope( vo 5e10e[ en[Oeor1 on6 ro en6pre- vo NTP Kor vo oxrlpor(oer rpologo6eoreprx6 6eog6. H enoveu0uypdprgtorl ennuyldvetor pe ronq rou RNA to dxpo d:qre vo rono0erqOe( orq ocoarrl 06oq yro npoo0qr<r1 enrnp6oOerrr;v Bdoeolv. Molov6rr 11 11 xonrl e(vot Spyo rqg (6ro9 rrlg RNA noLupepdorlg, 6eye(peror nolri on6 enrroupxo0g no- pdyoweg, noL0 6to6e6opr5voug Kor oro rp(o BroLoyrxd Boofi.ero. Iro E coll undpXouv 6Uo'r6rotor nopdyov- req, or GreA xor GreB. H RNA nol,upepdoq ll rov euKo- purorrriv Xpno$onote( yro rov [6ro or<on5 rov TFllS. O TFIIS 51er pu<pri opor6rrlro oq npoq rnv oLL4l,ou1(o ror rq 6opr4 Ue rouq nopdyoweq Gre, otrld npoo66ve- ror oro 6ro rprl;ro rou ev(rjprou, ro 6eurepoyevrl 6(ou- ).o (n6po) qg RNA no).uprepdo4g. Or nopdyoweg Gre/TFllS enrp6nouv or4v noLupe- pdoq vo x6rler prepu<d prBovouxLeot(6ro on6 ro 3'rcLt- rd drpo rou RNA npoi6wog, ennp6nowog ritor mo roro.l.urtr6 rdwpo qg RNA nol,upepdoqq vo eiloveu- Ouypogrore( pre ro 3'-OH. Ko0Svog on6 roug nopdyov- reg erodyer gto orev4 nporeiVrrrl neproXrl [orov TFIIS e[vor gto r<op66],o geu6opyripou (zinc ribbon), oro Bo- rrqproxd riv(upro 6vo onerpoer66g one(popro (coiled- coll)l Bo0rd prdoo or4v RNA noi,u;repdor1, qrdvowog noL6 rowd oro rorolurrr6 r<6wpo. A(o 6(wo oUrvo- [5o or4v ropugrlrou nopdyowo nl,qord(ouv fio npo- -reUov Korqluttx6 r6v ;royv4o(ou rou evepyori xdwpou ennp6nowog rrlv e[oo6o ev6g 6ew6pou r6woq Uoyvq- o(ou ror gerorp6nowog ftor q 06o4 rordluoqg oe pr- Bovourtredoq. Iuvomxd, 4 enrpqxuv6prev4 RNA noLuprepdoq riXer rqv xov6trlto vo [eru],(yer Kor vo [ovoru].(yer ro DNA, vo ouyKpors( roug 6toXroproprdvoug r<).c0voug tou DNA ro0oig ror ro RNA (npoi6v), vo rorol,fer rqv npoo0rlxrl prBovouxi.eon6(rov orqv ou[ov6gevq otruo[6o RNA, vo enmrlpe( r4v np6o6o out4g qg ow(6poo4g Kor -Ue rrl 532 KE0A Al0 19 Metoyporprl oroug l-lpoxoputirreg ouv6popr4 ev6q r1 60o enrxouprxcbv nopoy6wov- vo enilUer npoBLriproro nou dqproupyo0wot on6 qv rorq Ltycov nt on6 ro npoiSv RNA rot u1v enovdvop[n rnq otivOeorlq. 19..l5 H BorrnploKn RNA t-toAupepdon Teppori(er o€ AtoKprrdq @6oerq * YnopXouv d[o ro[eLq otrtrr1trouXuirv reppologori. Exelveq nou ovoyvopf(ovror povo ono qv [6ro rrlv RNA notrupepoorl Xo- p[g rr1 oupperoXq rovevoq rurroprrorj nopoyovro, ouvrl0otc ovotptlpovroL oq "evdoyevelg rep;louoreg,, Atrtreq 0n0rT0u, pro rurroplrrl nporelvr1 nou ovopo(eror po (p) xor ovorpepov- Tot trq <<TepUoTroreg e[oprc0pevor ono rrlv p". * 0 evdoyevrlg repponopdg npouno06reL ovoyvtrlprorl ;rrog otr- trqtrou1rog repportopou oro DNA nou roduonorer pto dogl qouprerog oro RNA npoiov. * To o1poro yro reppolopo pplorovror rupirrrg p6oo oe aAAq- Aouyieg nou eyouv 1764 peroypagei ono qv RNA notruUepo- o1 rr dror o repporropog pp[oreror un6 rov 5treyyo iou erpoye(ou ro/r1 rou RNA npoidvtog nou geroypotper I notru- uepoolt. M6Lrg 11 RNA no).uprepdorl [er<rvrloer rrl pretoypoqq t<ord pujr<og rou erpoye(ou, ouv06rer RNA, pr51pL vo ouvowrloet prto o).).qLouX(o repporrorq (terminator) . Ie out6 to orlpe(o, ro 5v(upo oro;lord vo npoo0drer vour<).eor(6to orov ou{ov6pevo r}.rrlvo RNA, one}.eu- Oepcbver ro oloxr1pcopr5,vo npoi6v xor 6rfororor ond to erproye(o tou DNA. O reppouopr6g npouno0dreL 6rL SLor or 6eogo( udpoy6vou Trou ouyKporoJv ro uBp(6ro RNA-DNA np5ner vo ondoouv ror getd vo [o- vooxrlUorrore( ro 6(x).rrlvo DNA. Meprrdg eop6q e[vor 6Uor<o].o vo npoo6rop(ooupe q 05o4 tepprorLoprorj yro 6vo RNA nou dlet ouwe0e( oe (orwov6 rlirropo, ener6q ro 3' dr<po rou prop(ou Unoper vo 6toonoore( on6 pro 3' efovouxi.edorl r1 vo rone[ ond pLo ev6ovouxedor1, lcop(g v' ogrloer rov6vo (Xvoq qq 05or1q dnou npoyprolr<d reppdttoe 11 RNA no).u;iepdorl: out6 e(vor rdro(tepo rorvd oroug eur<opurrtreg. l-r'ourd ro ).6yo, ot O5oeLg -reppolop.roU oulvd loporcrrlp((owoL xo).0repo in vitro. C)016oo, 11 rrov6rr;ro rou ev(Uprou vo reppot((er in vitro enrlped(eroL noLU ond nopog5rpoug 6norg q rowrxrl roltig xot q 0eppror<poo(o mqv ono(o npoyporonorefuor r; ow(6poor1. 'Eror, o tepporLop6g oe prro opLoprdvq 06,o1 in vitro 6ev onoreLe( on66er(q drr 11 o6v0eoq 0o reppolore( orrlv (6ro 06o11 Kor oro x0ropo (in vivo). 0o16oo, 6rov ro (6ro 3' dr<po ovrXveUeror 16oo in vivo ooo xor pe roOopd ouororLrd in vitro, our6 yevr- rd Oeorpeftor utg Lcrluprl Svdetlq 6tr npdrenor yro rnv ouOewrxrl 05or; tepprorLoproU, Ot i.li,,..::. ., " Ko[ r'jir-:,:... i'. 'rrr oUVoqJ((oUvro 60o r<rjpro lopor<rrlptmrr<d rcov evdoyevdrv repporror6v Boomeq eworeg Boorxeg eworeq
Ixe6ov otreq ot onopo(rrlreq lrliltrouXieq yro tov repgotlopo Ppiorovror otr1v fleptoxn nou usroYpdQnKe H rpoupr5ro oTo RNA Unopei vo onotTtiTot 0t otrtrtltrouxieg DNA nou onotTolJvTot Yto Tov -.:-lrLolto $plorovrot ovootro Uq otrtrntrouxtog rou repportorrl, , ,,-lorrouoq uroq QoupKer0q oro RNA unope( v0 elvot 0n0p0i --tt ntri nsic terminators). l-l ptrttov, ot ev6oYeve(q repUort- : .q, or ono(ot e(vot ove[dprlrot on6 rov entrouptr6 nop6yovro po (rho factor), Yto vo oXqpot(oouv trl 1.-repotoyq doprrl rou RNA nou preroypd<petot onot- :JV llo goupx6ro (hairpin) n).oUoto oe G+C. 'Ero4 11 -.,aypagn eoptatat an6 rc RNA npoi6v xat 5ev xa)o- .*#.w E*G ffiiG b#& L&th U}4F $r$4 "6RqeAg. h* wtrg6lE h&r& ilfl-E ffi ffi neproxrl iloriotq oe G-C oro gtoxo ffiq [Jels il&T TMovoxtrovn oend U EIKONA 19.28 0r evdoyeve(g repponor6g nepiloppdvouv notr[v- dpogeq neptox6g nou oXl;roti(ouv goupx6req Kot notKfiouv 0s g4rog on6 7 etrrq 20 bp. H 6opr1 Utoxou-Ontrtdq nepiloUFdvet Uto neproxn ntroUoto oe G-C nou oxo)rou0eltot on6 pto ostpd K0T0- tro(nov U. p(etu anLriq pe lleyyo n1q a&rlLouy{ag tou DNA xa- td q peraypacpfl. 1o 6e0repo XopoKrnptsrr6 efuot pro oetpd xotdtrotnov U (T oro DNA) nou orol'ou0o0v to pr(o1o qg qoupr<6tog oMd nporlyo0wot qg npqYpm- rrlq 05oqg repUortopoti. Ito yovtd(topto 'cou E. coli undpXouv -1100 otrtrr1LouX(eg nou totptd(ouv pre owd ro r<pttripto, yeyov6q nou unodqcilvet 6tt ndvo on6 to gto6 pretdypqqo rou xurrdpou tepprot(owot o' ev6o- yeve[g reppottm6g. Ot repportor6g nou efopr6wol qn6 rov p (rho-dependent terminators) xoOop(owot on6 t4v ovdyr<r1 npooOrixqg tou nop6yowo p in vitro. En(oqg, pretoMd(etq 6e[lyouv 6rt o nopdyowog epnl'd- Kerol fiov reppronop6 in vivo. Ot tepgoror6g nox[trouv no]li og npoq rnv onotetre- opcntr6rrpo. Og per6ypogo nopopiooqg (readthrough transcripts) ovog6powot to pretdypogo nou 6e oropo- roUv orovtepgonon(. (O 6poq nopoB(ooq lpqoqlonoef rot ornv Evot7ta 25.14ytovo neptypdtpet rrlv xotooroNi rodu<ov(cov tepporropori on6 dvo ptBooropdtto). En(oqq, o rcpponop6g prnope( v' onotpone( on6 et6u<orig entKou- pu<oUq nopdyoweg nou oMrltrenrdpo0v pre to RNA ro/r1 rqv RNA noluprepdorl, rotdoroorl Yvoorn oq qwlrep' ponopr5g (antitermination). 'Eror, 6nog rot orqv nep(- rrroorl qq 6vop[r1q r1 qg entptir<uvonq, o tepporrop6g prnope( vo pu0ptore( oq Unxovtotl6g yro tov 6treyXo rqg 6rgpoorlg rrov yovt6(tov, 6noq 0o 6ori;re oro Ke<pdtroto 27, Zrpaqyxiq tav @dyuv. YndpXouv Kqt dMot nopoM4Lrqro( preto[U 6vop[qg Kor repportopro0. Kot or 6Uo 6epyoo(eg qnomo0v 0rt6- orpro deoprdlv u6poy6vou (opXu<r1 ui[r1 tou DNA r<otd qv 6vop[r1 rot 6tdorooq rou uBpt6(ou RNA-DNA orov tepgottotrl6) r<ot pnope( vq XprlotUonoto6v entnp6o0e- reg nproreiveq (nopdyoweq o, evepYonotqt6g, roro- orol.e(g rot nopdyoweq p) nou otrq}'ent6po(v pe rov nuprjvo tou ev(0;tou. Qor6oo, ev<ir 11 6vop(q Boo(etot pr6vo orrlv oMr[ren(6poor1 prero[U qq RNA notru;repd- onq Kor tou dtxtrtivou DNA, o reppottopr6g nepilopBd- ver Kot ovoyvtilptorl onpdtov oro ;retdypotpo on6 qv RNA nol.ugep6or1. Ot orlpretor<6g pretoMd(etg nou trretcbvouv rllv onore- treopour6qro rou tepgottogoti ouvrlOoq ouprBofuouv p6oo oro p(oxo rqq rpoupr5rog ri ornv nLo0otq oe U oMrllouX(o, 6v6et(4 Yto rrl onUoo(o outriv ttr:v oMq- trouXtc0v oro prlxovtopt6 tou reppottopro0' To uBp(6to RNA-DNA 6Xet preydtrrl ouvetogopd ortg duvdpretq nou ouyrpototiv to otigntroro entpfruvoqg' Euvendrg, to ondotpto tou uBpr6(ou 0o prnopoUoe v' qnooro0eponot- qoet to o(prnLot<o en4.uiruvorlg, o6rlydrwog oe repUo- rrotrt6. Atrtrntrenrdpdoetq rrlq <pouprdtog pe rrlv RNA noluprepdoq ri 6uv6pretq nou oot<o0wot on6 ro crxnpo- nopr6 qq goupr6tog ro0tirq ro RNA (enpoBdMer on6 ro 6(qutro e(6dou tou RNA, prnoporjv np6oxotpo vo eu- Ouypoprpr(oouv pre trd0og rp6no to 3' dt<po tou RNA pe ro evepy6 x5wpo tou ev(Uprou. Aurl q I'ov0oopr5u1 eu- Ouypdprpuoq, oe ouv6uoo1.t6 pe to oouvti0toro oo0ev6g uBp(6to RNA-DNA to ono(o oXrlpro{(etot on6 to (eUyr1 Fdoecov rU-dA RNA-DNA nou npox0mouv on6 ro 4tt1- Uo Ue ro rqtdlotnq U, onosroOeponote[ to otipntroxo enq.rlruvorlg. 19.15 H Bonrlptorrl RNA l-lotrupepdon Tepuqr(et oe Aroxptt6q 06oerq 5:B lGUu UA GG AA I H RNA notrupepdoq ror ro RNA onetreu0ep6vovrot ..'}..I. , [ }'^
H onoteLeoporrr6qro rou rspUorrol,-tou in vitro not- x0,er nol.ri, n.X. on6 2% 6toq 90%. H onoreLeoponr5q- ro rou repUorropori 6ev e[oprdror pr6vo on6 trg oMqLoul(eg on1 goupx6ro KoL rov oprOpr5 ror ng 06- oerq rov roroLo(nov U xo0o6rxd qg goupr<5rog, oMd ror on6 otr).r1LouX(eg 16oo ovodrxd 6oo ror ro0o6u<d rr;g 06oqg repponopori. Aw( vo reppor(oer, ro 5v(u;ro gnope( onl6q vo rdver puo no6or1 npororj (ovop1(oer qv enr;u1r<uvo4. Aut6q or 06oerg no0orlg prnope( on6 p6vegroug vo ennetroUv puOpuoru<o0g oronoUg (Bl,6ne Ev6trya 26.13). To ov 11 RNA noLu;repdorl 0o oroprorl- oer Kor 0' one).euOepciloer rov r<Lrrlvo rou RNA r1 ov on).c09 0o oroporrloer Trprv [ovop1(oer rq ;reroypogri (5rll,odn q brdpr<ero qg noUorlg ror 11 onoreLeogour6- qro drotpuyrjg on6 rrlv noriol) r<o0op(owor on6 6vo no),[n].or<o orivo],o r<tvrpxoiv r<or Oep gobuvotrru<r6v no- poy6wcov nou drlproupyorlwot on6 ro lopor,rqprorrxd qg goupr6roq Kor rou nLo0otou oe U t;rq;rotoq oro RNA, 6no4 enioqg xot on6 rrg ovodrrdq rot ro0o6trdq oMqLoul[eg rou DNA. l-ro nopd6erypo, no0or1 prnope( vo oupBe[ oe 0doerq nou ;rord(ouv Uev Ue repprorrordg, lopor<rqpi(ovror 6puog on6 on6oroorl prero[ri rqg rpoupr6rog Kor rnq oerpdg U preyoLUreprl on6 q Bdtrrr- orrl yro rov reppotropr6. l-loLri Lry5repo e(vor yvroord oXerrxd pre ro orlproro Kor rouq enrroupu<orjg nopdyoweg nou egn),6Kowor srov repportopr6 rcov europurorrxc0v RNA noLupepo- ocilv. Kd0e rd[r1 no].uprepdonq go(veru vo Xpnoqonor- e[ (e1opror6 prlXovropr6 (BMne KeEd],oto 21). 19.16 l-l6q Aelroupyei o llopdyovroq p; * 0 nopoyovrog p e(vor pro npotreivrl nou npoo66ver0r oT0 vr- oouwe0eLp6vo RNA roL perouvelror xoro gqrog rou RNA yro v' otrtr4trenrdpooer U0 Tlv RNA notru;repool Kor v0 Tlv ors- LeuOeptiroer ond ro oupn),oro enrplruvorlg. O nopdyowog p e[vot onopo(rr1r; nporelvq oro E. coli nou npor<ol.e[ tepironopr6 rnq Lleroypognq. H ouyrdwpotoq rou prnope( vo grdver 6otg xor -10% qg ouyr<5wp<oonq rnq RNA no).upepdonq. Tepprorrop6g ove(dpqrog on6 rov nopdyowo p unooy((eror 6rr ouprBo(ver oXe66v oroug l.rrooriq repponordq rou E. coli. H - nopouord(er 5vo prow6Lo yro rq Ler- roupy(o rou p. l-lptito npoo6Sveror oe pro oM4louXio prdoo mo prerdyporpo ovodrrd rrlq 05or1g tepporLollori. Aurq 4 oLLq).ouX(o ovoprd(eror 06oq rut (or<provrj;uo yro: rho utilizalion - XpnorUono(non p). Iq ouv61eto, o nopdyowog p peror<rve(ror rord piqr<og rou RNA drog 6rou npo).dBet -rr1v RNA nolu;repdoq. Orov 4 RNA no- Lupepdoq qrdoer orl 05oq teppotrollori, o p e[ovoy- rd(er ro 6v(upo v' one),eu0epc6oeL to RNA. H noUoq 534 KEOAAAI0 19 Meroyporprl oroug t-lporopudreg {p ra EIK0NA 19.29 0 nopoyovrog p npoo66veror oro RNA oe pro 06- q rut Kor peroron[(eror xord prlrog rou RNA gdXpr vo rprdoeL ro uppiDro RNA-DNA mlv RNA notrupepdorl, 6nou one),eu0eptlver ro RNA on6 ro DNA. on6 r4v nol,uprepdorl orrl 06oq repporropro0 6(ver 1p6- vo orov nopdyowo p vo Uerorontore( oro uBp(6ro xor e(vor o4 1.rowx6 Xopor,r4 prorrr6 ro u tep porrollo 0. E6ci BL6noupe Uro oqpowu<rj yevrxri opXr1. 'Orov yvrop((oupre q 06oq mo DNA mqv ono(o enevepye( pto npore(vq, 6e pnoporipre vo ouUflepdvoupre 6rr owr] oupn(mer Ue rnv oL.4Lou{o rou DNA nou oplrrd ovo- yvop((er. Or 6rio O5oerg prnope( vo drorpSpouv xor 6e Xperd(eror vo undpXer pro ro0opro1.r6vn oX6on prerolri roug. llpdyporr, or 06oerg ruf oe 6rogoperrr6g prero- ypogrx6g ;rovd6eg Bp(or<owot oe 6rdqopeg onomd- oerq nprv on6 lg O5oerg repprorrol.ro6. Mro nopdproro 6tdrproq y(veror on6 roug nopdyoweg owrrepUorr- o1-rori (BL5ne Ev6qta 19.22). E(vor dyvcrlmo rr ouvrord orrlv npoypqrrx6rqro puo 06,o1 rut. Korv6 Xoporrrlproru<5 tov 06oeov rut e(vor H RNA noLupepdon Ueroypdqsr DNA 0 p peroroni(eror rord prlrog rou RNA H RNA notrugepdorl rdver gro norjorl orrl rpouprdro roropqvnpotropo(ver 0 p [etutr(yer to uppGro DNA-RNA tTepgortog6q: dtro ro oumortxd one)eu0epdrvowot Boorxeg eworeq
Bdoetg C 41'/o <- H onotrotQr1 onorp6net rov TepuortoUo '+ A 25lo u 2oo/, $ 14Yo H e60n ru1 6Xer pro gtrtrltroux(q iltrguoro 0e C Kor QToXn 0e G nou npolyetTot TIq rpoYUoTtKnq(ov) 0eo1q(eu:v) TepUoTt- ouou. H otrtrl^ouxio ovTtoTotxel olo 3' oKpo Tou RNA. 6n q oMrlloux(q rouq e(vot nl.o0oto oe Kordtrotno C Kor Qr@xn oe Kord^otno G Kor 6ev 5xet 6ewepotoytl Eopui. 'Evo flopddetyUo 6(vetot ornv ElK0l'lA 13'30. H C e(vor Ue dtogopd rlnto Kotvrl Bdorl (41%), evcb n G n It- y6repo Kotvn Bdon (14%). Ot 06oetg rut nox(trouv oe pnKoq. Qg yevu<6g rov6vog, rl onoretreogottx6tqto Uroq e6onq rut ouqdva pe to prirog rnq neptoxiq nou e[vor ntro0oro oe C/<ptoxn oe G. O nopdyowog p e(vot pr6l,og ttlg otKoY6vetoq rcuv e[qreptirv eturoocilv nou e[oprc0wot on6 to ATP. Kd0e unoprovd6o 61et pto neproXrl np6odeoqg RNA rot ptto neproXrl u6p6Luo4g ATP. To e[oprep6g treroupye( nep- vdrwog vour.treivtr6 o[ri 6ropr6oou qg onrlg oro p6oo qg 6o;ujq nou oxnUor(erot on6 trg neproX6g np6o6e- oqg RNA ttrlv unogovd6tr:v {EIK0NA 1$.31}. H 6o;tt1 tou p 6(vet xdnoeg evde((erg yto ro n6q 0o prnoporlos vo tret- toupye(. Oq[vetot 6rt ru],(yet RNA on6 ro 3' dr<po yUprrl on6 ro e[coteptr5 tov N-retrtt<citv neptoXcbv rot onpril- )yer ro 5' drpo rqg npoo'6e6epr5vr1g neproXqq oro eoo- rep116, npoq Uto 6euteperiouoo neptol{ nou npooS6vet rtg C-rerr6q neptol6g rou RNA. H op1trci popei rou p eivot 6vog dtoreroprprdvoq SorcrrjLtog, oI'- Ld r1 np6o6eon rou RNA tov petorp6net oe xtretor6 6o- rrr0trro. Merd rqv np6o6eoq orq 06o11 rut, o p Xprlotponoe( qv evepy6qro rng elrrdorlg nou 6to06tet Yto vo pero- ronrme( xotd prrlxog tou RNA, to0o6prevog on6 tqv u6p5l,uor1 rou ATP, ;r5Xpt vo Etdou t4v RNA nol'upe- pdoq. Iq ouv6Xeto, Xdprl crrrlv evepy6t4to efuxdoqg, [eru],iyer q 6(r].ovr1 6oUn xo/ri oMrl]'entdpd Ue rnv RNA noLuprepdorl oupBdMowoq qrnv one).euO5pocn1 rou RNA. O nop6yowog p npSner vo Uerorontsre[ r<otd gti- Koq rou RNA on6 14 06o11 rut oro npoyl.tottt<6 o4pe(o repportoporl. Aw6 npounoOSret 6n 0o petortvefuot ro- XUrepo on6 rqv RNA notrupepdorl. To 5v(upo xdvet pto norioq 6rov <ptdoet o' Svov tepgononl Kot, ov rrlv npo- grdoet o nopdyowoq p, eq ouprBe( tepprottopt6g. Iuve- ncirq, rl 4 norioq 6Xet oqproo(o Yto rov teppottog6 nou e[optdtot on6 tov p, 6nrrlg Kot orov ev6oyev4 repUort- opr6, ener6{ 6(vet to Xp6vo yt' dltro onopo(tr1to ou1.t- Bdwo. H o6(eu[r1 1.rero[U ;retoypoqriq r<ot pretdgpooqq 61er orlprowu<5g npoercrdoetq Yto rn 6p6on tou p' O p np6net npc0to v' onorcrnoet np6oBoorl sro RNA ovodt- rd on6 ro otipilr,oxo preroypogriq rt d'nemo vo UeroKt- vq0e( r<ot6 prrp<og tou RNA yro vo npogtdoet qv RNA no)ruprepdoq. 'Erot, n Spdon tou eprno6((etot 6rov to prBooroprdno preto<ppd(ouv 6vo mRNA. To ouyrexptptd- vo pow6Lo el4ye( 6vo rpotv6gevo nou npoBtrrlpfutoe roug npriroug Bor,mrlptoro0g yevenor5g. Ie opropr6veg neprm6oerg, 1.uo oouv0ettr4 petdMo[r1 (nonsense mu- tation) o' 5vo yov(6to ploq UeroYpogu<r1q provddoq Bp6- 0r1xe 6rr onotpdnet trlv dr<rppoorl rov endpevtov yovrS(rov orrl provddo o16;ro Kt ov Kq06vo on6 to 60o yov(6ro e(Xe tn 6u<rl roug t6to(tepq 06o11 np6o6eoqg mo prBoocol.tdtto, qorv6ptevo yvtrlor6 og notrlx6rqro (polarity). M6oo oe Uto Ueroypogrrci povd6o, ot 06oetg reppo- rropro6 nou e[optc0wot on6 tov nopdyowo p ouvrlOog rotrUmowot on6 ptBootoprdno nou Ueto<ppd(ouv (ElK0' NA 19.32), on6re o p 6e prnope( vo enevepy{oet oe ro0o' 6rrdg RNA notrupepdoeg. Oor6oo, oouv0ertr6g pretotrl.d(erq oneLeu Oep rirvo uv p tBooco;rdno prSoo oro RNA ev6g notruyovt6tot<of onepov(ou, enttp6nowog orov p vo repgoiloet rrl UeroYpogrl np6olpo rt eprno6(- (ouv tr1v 5r<ppoorl onpror<puopr6vrov yovt6(rov rnq pero- neprogdq RNA 0 eloUepqq 0om[trtoq rposddver RNA 3', 0 p exet pto N-relLrq neptoXrl npooDeorlg RNA ror gro C-re),rrn noptoxrl ATPoonq 'Evo e[oUepeq uno Uopqn evog dtoreroppevou borrutr[ou npood6vet RNA roro Urlroq rr1q eloreprrnq enupovetog ttrtv N-retrtr6v neptoXtilv. To 5' orpo rou RNA ervor npoodede;rdvo oe pto deurepoyewl 06on oro tooTept- 16 rou e[opepo0g. I I f i"i i n6q Aeuoupyel o llopoyovroc p 535 AU CGEUACCUCAUAU COSCA*{U CEU C}AqA C{i CIJA C'UCGACiAGAMS$CffUOU CUU
il: i il:,l i-,ri:i:,1: i,5ill[:Hl wrpd(l)p0 ^@ H dpoott rou nopoyovro p pnopei vo ouvbeoer q peroyporpri ror rr1 peroqpool orov evoq p-e(oprtlpevoq TsppqTroTll( pp[oreror rovro oe pro oouv0elrq perotrtroi1. ypogrrlg provd6og nopdrt r' ovorXrd nLq(oro ovdyvro- oqg roug nepeiXov oM4louX(eg dyprou r0nou. l-ror( ro oroOepd RNA (rRNA r<or IRNA) 6ev unoBdL- l,owor oe notrrx5r4to; To IRNA e(vot Bpox6o xr dXouv ercretop6veg deurepoyeve(g 6op6g nou ntOovdv ono- tp6nouv qv np6odeoq rou p. Ttrrrjproro rtov rRNs en(- o4g 61ouv errerop6veg 6o;159, ol,Ld ro rRNA e(vor nol,0 prorprjrepo on6 rq IRNA, ogrlvowog oprer6g eu- rop[eg yro rn Spdon rou p. To r<6rropo 6prcog 61ouv ovom[[er 6vov dMo prrlXovropr6 yro v' onorpd,nouv rov np5opo reppotro;r6 rov rRNA geroypdqov: 6ro0rirouv npolteiveg nou npood5vowor ortg ovol.ro(6preveg 0d- oeq nut (nut sites) mrq neproX6g-o6rlyorig 16oo rcov 165 6oo Kot rov 23S rRNA preroypdqov, oXrlpori(ov- rog oripntroxo owrepponopori (antitermination com- plexes) nou qvoor6l ouvrrl Spdoq rou p. Ot peroMd(er.q rou p nopouord(ouv preydl,rl notrr- Lo;ropg(o oq npoq rtg enrmcioerq rouq orov reppol.- opr6, To Boou<6 onot6l,eopo ourc6v trov pretoMd{ecov e[vor onorul(o rou repprorropori. Om6oo, o Bo0p6q rqg onorul(og, 6nog errrpdror on6 ro nooosr6 ralv nopoBrdoeov in vivo, e[oprdtor on6 ro ouyxexpqdvo r6no nou onorel,e( or6Xo yro repporrqr6. l-1opo;ro(og, 4 e(dpqoq on6 tov nopdyowo p in vitro nou<il.er. Me- prxo( (p-e[opr6prevoQ repporror6g qnonorjv olenrd ut|lnMq ouyrewprioerq rou p, ev6 dMor treroupyo0v e((oou xoLd oe 1op4l6repo en(ne6o. Aur6 unodrll,c0vet 6rt 6rogopenro( repporrm6g ononotiv drogoperu<d en(ne6q rou nopdyowo p yro rep;ronog6 ror yr'our6 owonorp(vowor drogopenrd qro unol,erptrronrd en(- ne6o rou nopdyowo p nou onop6vouv orct preroMoy- 536 KEOA Al0 19 MeToypoQn orouq tlporoputilreg :;- :a: 4..:t.:)!.,, .. To prpootopdrro ouoreud(ouv mRNA n(oo on6 qv RNA no)tupepdoq To prpoorrrpolo egnod((ouv qv npooxritrqoq rou p ro/rl rt1 UeToK[vnon 0 p npoo- rotrtroroL otrtrd to prpootrtpdlo egno6[(ouv 11 petox(vr1oq rou IuveX((eror 1 peroypogrl To ptpootrlpdrro drimowor oq 06011 qq [erdMoqr]q -0 p onoxrd npdopoorl orrlv RNA notrugep6orl pr6vo oretrSXq (ouvr1Oog 1oporcr4p((owor. oq preroMdy- proro droppoqg, leaky mutants). Kdnoog preroMd(etg rou p Urops( vo xorooroLo0v on5 prerotrld[erg oe dMo yov(6ro. Aurl 11 npoo6yyroq npoo<p6per 6vov e(orpenr<6 tp6no yro rrlv ovoyvcbproq npcoreivc0v nou ol.tr,4Lenr6poriv Ue rov p. H B unoprovd- 6o r4g RNA nolu;repdonq epnLdxeror oe 66o r0nouq peroMd[eo:v. llprirov, or preroMdlerg oro yov(6ro rpo B pnope( vo perdtoouv rov reppolopr6 oe puo p-e[op- rriprevq 0dor'1. AeUrepov, or peroMd(erg oro yov(6ro r- po B pnope[ v' onororoorrjoouv rqv rrov6rqro teppottopoti rnq perqypoqTiq orrq p-e[opt6peveg 06- oerq oe Borcr4pro trre ;reroMd[erq rou p. Ae yvotp((oupre oor6oo noro Lemoupyrx6qro 61er q oM4).en6poor1. 19.1 7 H YnepeAiroon Eivot Inpovrrr6 XoporrnproTtK6 Tnq Meroypogfiq e H opvnlrn unepetrlrtrloq ou{over rqv onoretreopoTrKoTlTo ronoulv unouvrptiv enerDl poq0o rr;v ovr[6poo1 onodtdro- Ers. 6 H Ueroypoq1 dqproupye( pnpooro on6 ro ev(u;ro Oertxec unepetrrxeg xor n(oo rou opvqrrx6g unep6trrreg, or onoleq np6net v' orporpe0ouv ono q yupooq xor rlv ronoiooUepoort T6oo ot npoKopuonr<5q 5oo Kor or euKopuco-rLr<6g R- Boorxeg evuoleg
',: roluUspooeg Qo(vetot 6rt ln vifro evepyoilotouv -- JoroypoQq onoteLeoprottx6tepo 6tov ro expoye(o . ,:i unepetrolp5vo. Meptr<6q Qop6q, q onotetreopo- - ^,irnro rou unoKtvnrrl evtol0etot ewuntootoKd on6 -- . opvrlttr<rq unepe[rtrlotl' I-tor[ optol-ttivot unoKtvn- -:l enrlped(owot neptoo6tepo on6 to Bo0p6 tqg - .aoel(xtooqq on6 dMoug; H nto ntOovrl e[r1yr1oq e(- ,l. ort 11 e[dpqorl ev6g unortvqtrl on6 tr1v unepe]'[- --oq r<o0op((e-rot on6 qv etreUOepq ev6pyeLo nou : .cLre(rot Yto rqv q(r1 tou DNA cn'o otipnor<o tivop- :-l Me q oetpd rr1g, 11 eleU0eprl evSpyero e[optdtot r 3 rllv oMrlLoul(o tou DNA rou unoKtvnrrl: 6oo nto -'.c..loro oe G+i eivot 11 oLLqoux[o tou unortvqtrl -: - owtorotXe[ orrl 05or1 qg pretoypoqtr<r1q quoo]'G l:l rooo nto orlpowtKrl yto tov unortvrlrl 0o e(vot 11 - ,ce).[root1, q ono(o 0o tov ouvSpdpret crrqv r1(r1 -:- DNA. ',1ooto[ro, ot ent6pdoerq rnq unepe]'(xtr-ro4g orrlv . . ':vorlro ev6g optopr6vou unoKtvntrl e(vot nol0 nto -:'.-n,or<o (ntnpo. H e[dpqorl dto<popettxt'tlv unoKt- - ;v on6 ro Bo0pt6 qg unepeL(Koonq enqped(etot ,-.rrq on6 oMrlLouXieq DNA 6[tl on6 rr1 rpuooL(6o, .-..5q q unepel(xcr-ror1 otrI.d(et q yeopretp(o rou oup- - :(ou enrlped(owoq ttg yc^lv(eg Kot rlq onomdoetq -.::[Lt rr.ov Bdoeolv mo 1c,-lpo. 'EroL, 6to<pop6q mo :::,o rqg unepel'lrolorlg prnope( v' oMd(ouv ttg o]'trr1- . 5pooetq preto[ti ttrlv Bdoetov rou unoKtvnrrl roL rcov --.. .c(er-,.rv tqq RNA nolutrtepdorlg. Entn}'5ov, enet6rl - ::opertxd pr5pq tou Xpclprootrlprotog 6Xouv 6to<pope- - - l-q Bo0proIg unepetr(r<.rlonq, ro onot6treogo trlq - .:eit<rr.lot1q onlv evepy6tqto ev6q unortvqtrl prnope( . : rrqped(eroL on6 rq 06o11 rou unoKtvllrq oro Xpirl- - - l!f-lo. (oOoq q RNA no)'uprepdorl ouveXtrlq (etutrtyet rot ::,:ru,Lyet to DNA evul npoXtr.lpel mo ex;royelo (no- ::-:lc(etor orqv ELt<ovo 19.4), e(te o)'6x}r1po to otipr- - :{o rlq pretoypoqqg npdnet vo neprorpo<pe( yUptrl r-: :c DNA, eite ro (6to to DNA vo neptorpoqe( yfpro r-: rcv o(ovo trlg 6trtr<og. l'ltme0etot 6tt nLo xowd - - , itpoYUortr6tqto e[vot to deritepo: -ro DNA nepvd- : ..oco on6 to riv(upo dniog 4 B(6o pr6oo on6 6vo --: -tovt, ',lLo ouv5neto rnq rleptorpo<pqq tou DNA nopouotd- -. :i orlv ,:r,l:l:lrii' it: :':. >To prow6Lo ttov 6[Supcov ne- : - ':i'/ (Nvin domain nodel) Yto rn peroYpotp(, xoOr'trg -. :rrverot q RNA no).upepooq oe crtr5or1 pre q 6tnLq ' ..rc. dqptoupye( 0ettr<6q unepSltreg (nto orptXro tu- . *avo DNA) prnpoord rnq Kot orpr-1vet opvqttr6g une- :':..req (pepir<d [eru].typi6vo DNA) nioo qg l-to r<dOe :-:ccpq iqq d).tr<og nou 6totp6XeL r1 RNA notrupiepdoq' :--.cupye(tot +1 orpotpq pinpoord r<ot -1 orpoqq nG - - l-t out6 11 petoypo<pr1 61r prdvo enrlped(etor p6vo r o r'lv rontr<r1 6oprl tou DNA, oMd en(or1q enrlped(et . r. :rlv npoyl.lottr<r1 6opr1 tou DNA' To 6v(upro DNA yu- : r:I. nou etodyet opvqttr<6g unep6tr'tr<eq oro DNA' t<oL l',,A tonotoopepdorl l, nou otpotpe( opvrlttr<69 unep6trt- . .: ond to DNA, e(vot onopo[trlro Yto v' onotp5qouv .-. ouooopeuoq tonotroytt<cilv nr5oecov roto r1v nope(o - e ileroYpo<pr1g r<or rnq owtYpoQrlg 'Etot, ov ovooro- (Apvnw6g unep6trtreq) MeroyporpdUevo YnePetri*u:- DNA 1Oet.^ec.,e;e;.':- H yupdon etodyet 0etu6q unep6trxeg 19.18 H RNA l-loAupepdon Tou Odyou T7 Eivor Evo Xprioruo l'lp6runo l0ornpo EIK0NA 1g.33 H getoypotpf drluroupYei nto orptlro ru]rtyp6vo (Oerxd unepetrxop6vo) DNA Unpootd on6 qv RNA nohugepdorl' ivdr io DNA n(otr: on6 ro 6v(uUo y(verot trtyotepo orptxtd rultyp6- vo (opvnrtro unePelrrtrlP6vo). Le( 11 evepy6rnro rnq Yupdorlq Kot rrlq tonoioopepd- ons bt npoxtrqOo0v 1-reydLeq oMoy6g orqv unepeL(xto- on'rou DNA, enrlped(owoq rrl UeroYpo<pr1 r<ot tqv owtypog4. Aut6 ovoL[OnKe nponYoupr6vrog mo ntro(- oro rnq owtyporprlg (pMne Ev6rrya 14'2)' * H otroY6veto rotv RNA notrupepootirv rou T7 e(vot povo noLu- nenr[Oto rrovo v' ovoyvtrtpi(ouv roug unortvrlr6q Tou QoYou ror vo entreLo0v notrtreq ono rtg Opoorrlptoqieg rov notru- peptirv RNA notrugePootitv. * bL rpuorotrhtrSq 6ou6q rov RNA nolupepootilv Tnq otKoYe- veroq rou T7 ge ro DNA routonoto0v qv neptoXl npoobeolq rou DNA oro evepy6 r6vrpo xot unoDrltrtfvouv povretro yto 11 DtotpuYll rou unortvrlq. Optopr6vot Borcrqpto<pdyot (n.X. f3,T4, N4) gttdXvouv rtq 6ireq touq RNA nou;repdoeq, nou onoretroUwot on6 provdg nolunenttdtxsq oLuo(6eq' Aut6q ot RNA nou ;.repdoeq ovoyvtr:p((ouv p6vo Ltyouq unortvrltSg oro DNA tou <pdyou, oMd entte}'ofv noM6q on6 ttg evepy6tqteg tolv notru;rep<i:v RNA notru;repoocov' 'Etot,'onotetro[v np6tuno ouorrlproro Yto rrl ;re6tq ouy re r<p tpSvtr:v e tto u py rc6v tq g pietoy p o<pr1 q' 'l-to nopddetypro, 11 RNA noLuprepdorl rou T7 e(vot puo ;rovrl notrunen-rt6tr<r1 oLuo(6o <100- kD Iuv06ret hNn Ut tol0tqto -300 vour<treot(6to/Seutep6Lento orouq 37"i,'notrrj u(.rrlLdtepq on6 t1v toX0rqto t6oo rr1g nolupreporiq RNA noluprepdoqg tou Borcrrlptoror-i [evtorrl 6oo xot ttrlv ptBoocol.tor(ov nou petotppd(ouv ro mRNA rou. 'Etot, n UeroYpoqq nou roteu0rjvetot on6 tov T7 0o unoBoM6tov oe UeroYpoptr<r1 notrtxorn- to, e[5].t[q nou onotp6netot on6 to yeyovdg ort Il us- H ronoioopePciorl I Xotroptivetttg.. J opvnTtKeq unepe^tKeq I A(rtrtovo DNA (10,4 bp/orpoan) .10..!8 H RNA flotrupepoorl rou odyou T7 Etvot Evo Xprlotlio tlpotuno ldorqFo 537 Boorx6g 6woleq
roypogn on6 rrlv RNA noluprepdorl rou T7 ouprBo(vet oe peroyev6mepo ord6to rqq Lo(;rco[4q, 6rov 11 Srrppo- orlrou p efuor nepropropr6v4. H RNA nolupepdo4 rou T7 e(vor opr5Loyr'1 pe rrg DNA r<or RNA no).upep6oeg enerdri or roroLwu<o( nupriveg rot ttov rprcOv ev(Upuov 6Xouv nop6proteg 6opr5g. To DNA Bpi oKerot oe Uto unotrdun (palm)" nou nepgdMerqr qn6 "Sdrcrul.o (fingers), rt Svov (ow(Xerpo (thumb), (FMne Er5vo 14.7) ru ro 5v(upro Xprloq-ronoe( novo;.ror6runo xotol,ulr6 gr11ovrog6. Irlgepo 6loupre oprer6g rpu- oroMr5g 6o;169 rcov RNA noLuprepooc0v rrov T7 ror N4. H RNA noLupepdoq rou T7 ovoyvcop(et ng oMq- LouX(eg-or61ouq rnq sro DNA pre np6o6eo4 orrg Bd- otrq rnq pe[(ovog oriLoxoq, 5nog qo(veror orrlv Ell(0NA 1 9.34, 1pr1or1.rono rciwog pro grTir<i etdm6rr7t aq (s p e c ifi c i - ty loop)nou q4q;ror(eror on6 puo B rop66Lo. Aur6 ro XopoKrnprorrr6 efuor provodrr6 yro rrq povopepe(g RNA noLuprepdoeg (5e ouvowdror orrg DNA no).uprepdoeq). 'Onog ong noLuprepe[g RNA no].uprepdoeg, o unoKr.vrl- rfg onoreLe(tor on6 ouyxerprpr6veg Bdoerg oro DNA ovo6rrd rqg 06oqg peroypoqrlg, ;rol,ov6n or unoKrwl- 169 rou T7 onore).orjwor on6 Lry6tepeg Bdoerq ouyKpr- trrd pe unortvqrrig nou runrxd ovoyvrop((owot on6 noLu pepe(g RNA no).u prepdoeg. H perdBooq on6 ro oritrmLoro rou unoKrvl1r1 oro oUpntroro r4g enrp{ruvoqg enmuyldverqr on6 6rio oq- gowxSg oMoySq dropr6pgoorlg oro 6v(upro. llptinov, 6noq xor pe rrg notruprepe(g RNA notrugepdoeq, oup- Bo[ver "pdoqol, tou erpoye[ou oro evepyd x6wpo xor ro 6v(u;ro nopoprdvet npoo6edeprdvo orov unoxrvqri r<oOrig 4 notruprepdoq npoyUoronols;( avenruyetq, ouv- 06oeq (abortive synfhesrs), drlproupyriwog BpoX6o pre- rdyporpo pr{roug 2-12 nl. H nepropl np6o6eorlg rou unortvqt{ 0' onoretrotioe eprn66ro mo olqpronopr6 ovenmulrriv npoi6wrov, coor6oo onoporpUveror Ue ne- ptorpogti nep(nou 45 gorp<irv, enmp6nowoq orrlv no- Lupepdo4 vo dtorqprioer enog6g pe rov unorwqrj evcb ouw(0eror ro op1rx5 RNA perdyporpo. Aur6 e(vor ovd- Loyo pre rrlv erc6nroq rou ouv66rq rqq neproplg 3-ne- ptolnq 4 rou nopdyowo o on6 ro 6(oul,o e[66ou rou RNA r<ord r' opXrxd srddro oriv0eorlg rou RNA mqv noLugepq Borcrrlprorrl RNA noLuprepdorl. To RNA ovo- 6Ueror orqv enrqdveto rou ev(riprou 6rov 61ouv ouwe- Merox[vr1or1 rou ev(U gou..-...] H RNA notrupepdol rou T7 eXer pro Oqtrro erdtr6- TIToq rou npood6ver rtg 06oerg -7 6rrlg -11 rou unouvrlrrl, evdr or 06oerq -1 6tog -4 eLodpXovror oro evepyo r6vrpo. 538 (EOAAA|0 19 Meroyporp1 oroug flporopuc0reg 0e( 12-14 vouxleor(6ro. 'Eneno oupBo(ver puo or6prrl peyoL6tepq oMoyrl 6top6pgooqg, xord r4v ono(o puo unopovd6o yvoorrl og neproXq H geroxrve(ror ndvo on6 70 A on6 q 06o11qg oro o6pntroxo 6vop[r1g. Au- r{ 11 tepdorro 6ogu<rj ovo6ropydvoorl rou oupn}.6r<ou enrpr{ruvo4g 6r1;uoupye[ puo oripoyyo p6oro rqg ono[og pnope( vo e(5l0er to RNA prerdyporpo, r<oOrilg Kor pro 05oq np6o6eonq yro ro 1.rov6r<Lrovo Url-eKpoye(o DNA rnq peroypogtr{q quootr(6o9. I9.19 O Avroyulvrop6q yto Touq Ilopdyovreq o Mnop€i vo Pu0pio€t Tnv 'Evop{n * fo E. coli SXer enro nopoyovreq o, ro06vog ono roug onoi oug o0ei rrlv RNA notrupepool vo [errvqoet oe gro ol.rodo unoxrvrlrdv nou npoodrop((ovror on6 ouyrerprpdveg 0trtr1- trouXieg -35 ror -1 0. * 0r evepyoqreg rrrlv drorpopeurdrv nopoy6vrorv o puO;rl(ovror onr! 6rorpopelxouq UIXovroUoLtq. Irrg en6peveg ev6rqteg 0o nopo06ooupe pepu<d no- podety;roro yro rn pU0proq rrlg Svop[qg, rqg enr;rq- KUVonq Kcu rou tep;,rologoU. 'AMo nopodery;roro 0o 6o0oriv oro Kegdloto 26 r<ot 27. O roropepropr6g opl.ro6rotrltcov prero(ti tou nupr;vo tou ev(0ptou, nou e(vor uneU0uvog yro rnv enrpul<uvor1 rrlg o).uo(6oq, Kor rou nopdyowo o, nou e(vor une00u- voq yro rqv entLoyq rou unoKrvl'lrr1, unorfvr1oe to epot- rrlpo ov 0o unrlpXe ndvo on6 6vog tUnog nopdyowo o, o r<oO5vog er6rr6g yro 6rogopelr<q o;rd5o unoKrvn- t6v. H i r :, 6e(1ver rqv opXrl ev6g ouorrlprotog To o,lo6v(upo Ue To 070 ovoyvtop(er gro opddo unorwltriv H owrxordmool rou nopdyovro o nporole( qv 0v0yv6pron 6torpopetrrlg ogridog unoxlqrtiv ond ro 6v(ugo 0 nopoyovroq o nou ouv66er0r Ue rov nuprlvo rou ev(upou ro0opi(er rrlv o;rodo rov unouvlr(rv or0uq ono[oug opXi(er 1 UeroypoQn. To p tptitr)ro erdrrdqrog npooDdveror orn
oro ono(o Uto owrKqrdsroorl rou nopdYowo o oLI'6Eet rnv enil.oY{ rou unoKtvnri. To E coll oulyd lptlotponoe( evoLtr'otcttrofq nopd- yoweq o Yto v' owonorptOe( oe nep$oMowr6q 11 6to- tpoqir<6q oLloy6q. Ot evotrtrot<ttro( nopdyovrsq nopouotd(owot offlv IlK0t'lA 19.36 (ot nopdyoweg o ovo- prdiowot on6 to proptor6 gdpoq tou npoi6wog { on6 q iettoupy(o ttov yovtS(tov TIou UeroYpdEouv)' O nto 6q0ovognop6yowoq o, un6 xovovu<6g ouvOrlreg une[- 0uvog yto rn UeroYpoqq tov neptooot6pt':v yovt6[tov' eivqr o oTo flou rtodtronoefuqt on6 to yov(6to rpoD' O evoMorrtr6g nopdyowoq os (ot8) Xpqotponoe(tot yto rrlv nopqYoY( noMrirv npoi6wtov oXen(6pevov pe ro stress. Or oH (o32) xor oE (o2a) onoffoUwqt Yto rnv Tlopo- yoyrl npoi6wtov onopo(tqtov yf ondwnon oe ouvOr1- itg nou (e6tnLcivouv npcotetveg oro 1urr996ntro4to *oi *o nep(ntroopto, owtoro(1coq' O oN (os') ouv06tet npoi6wo rup(tog onopofu4to Yto rrlv evotrrg6rroorl rou oiir-or. 6 qi*r (ors; nopdyet Aiyo npoi6wo-onopokqto yto rn peroqop& -o, ot6{pou, evci o oF (o'z8) er<qp6(et npoi6wo onopokqto yto tt1 otivOeoq trov-poorty(tov' H ondwrlon rov ononrultopr6vtov ((edrnl'opr6vov) nproreivrilv efuot pto on6 ug TIto ouwnprlpr6veg pu0pt- orrr<6q onowrioerg oe ol'6xl'r1pr1 q Bto)roy(o' ApXtr6 *o^oL,l.peqre u:g ondwnon crrrlv ou[4oq qg Oeppo- xpoo(oq ir.or yt'our6 ovopdorrlre ondwrlo4 Oepylroti s'hock, heat shock response). Xoporcr4p((erot on6 r1 oriv0eoq ev6g ouv6Lou npoteivrilv, nop6potrov Kol' oro rpio Broioytt<d Booileto, nou npoorore0ouv to r0rro- po ot O neprBoMowu<5 stress. l-1oM6g on6 out6g nq npcotetveg Oepprtxoti shock e(vot oanep6veq (chaper' ons) nou'1r",,irrou, to en(ne6o rov oTloTlruX<opr6vu:v npoteivdrv nporoLtilwoq rnv EllovomUxoon 11 onodt6- told toug. /ro E. coti,4 enoytoyrltov npoteivriv Oep- prxori shock npoYgorollote(tot mo en(ne6o rnq Ue- rcrypoQnq. To yovbto rpoH e(vot 6vog puOprtorqg nou cnoretroi Yto rrlv evepyono(qor1 qg ondrrrrloqq 0eppt- xo6 shock. l-1poi6v tou e(vot o d2, e(vot 6vog evotr"tr'o- rnx6q nopdyowoq o nou ovoYvop((et touq unortult6g trrlv Yovtd(tov oePPltr<o6 shock, H ondrrrqorl Oepgxoti shock (r<up(trtg oonep6veg rot npolrcdoeq) eL6yXetot Ue ndr(v6poUn pU0ptorl' To rtret- 6( yto tov 6)reyXo tou d2 efuot 6tt 11 6to0eoqr6rr6o cr rrilv ttov rurropontrooptottt<cbv nporcoodrv Kor firEPG vri:v eqopt6rot on6 ro ov rtthonototiwot on6 onontuXtop6veg npotetveg' 'Etot, ov etrqrro)Oofv ro en(ne6o rov onortuxop6vov npoteivriv (efue enet6ri ot npotetveg Oepprroi shock ttq enovontulti:vouv eke ener6q rtg 6toono0v, e(te enet6rl 1ogr1tr'drvet 11 Oeppo- rpoo(o), 6e 0' onopoxp6vouv nepott6pto ttq npote6oeg nou 6tqono0v rov o32, o ono(og 6tot 0o enqv6tr0et oe quorotroytrd en(ne6o, Enet6rl ot o7o rot cr32 owoyov(ov- io, y,o to 6to06otpro nupdvo tou ev(tipou, rl.pretoypo<pq on6 touq unoxwqtdg tu:v yovtS(tov Oepprtxo0 shock ent- orpdqet oe Bootrd en(ne6o roOcilq ro en(ne6o rov o24 ^oi .p' "nor6p1o*ot oro cpuotoLoytt<6"Etot, ro otlvotro ttov yovt6torrirv npoi6wrov nou nopdX0rp<ov t<otd tq 6rdpxeto tou 0eppxo6 shock e(oprotot on6 rqv toop- pon(o preto(0 o7o rot d2. Iu;rBot6 Ue rn onpoo(o rou o*oyru,o1.,oti ttov nopoy6wtov o e(vot t<ot to YeYov6q 6rt 11'ouyr6wpoorl rou o7o e(vot ;reyotrtkepq on6 tqv ow(orotXrltou nupqvo qg RNA notrupepdorlg un6 ouv- Orir<eq oris ono(eg 6e ouprBo(vet enoyu:y1tou d2' O o32 6ev e(vot o povodtr6g nopdyowoq o nou eMyXer qv on6wrlorl rov onornuXo:pr6vtov npolrcivdlv' o oi'endvetor on6 q ouoodrpeuon oTlofiruxop6vtov npoteivdlv otov neptnl'oopnr6 ldlpo Kot rnv e(oteprrtl pepppdvn. 'Onog r<or pe rov o32, q nptote6Luorl e(vot to nLiof y,o qv enoyoyri rnq ueroYpoqqq ro:v oE-e[op- rrirpevov unorw4tcilv. To nep(nLot<o r0rl'ogo nou eu- 0Uvetot Yto rn pU0prorl tqq evepy6rnroq tou oE ouvorf.r((etot mrlv Ell(ONA 19'37. O oE npoo6Svetot oe uro nbcoretvrl (RseA) nou Bp(oretot ornv eooteptr<ri Uepppdvn. H RseA e(vot nopdSetypro evSg nopdyowo ili-; (anti'sigma factor). 'Otqv e(vot npoo6e6epr6vrl NEPINAATMA c Krirlrpo ond qv OegS + K6rlruo onri n1v YaeL ( 0o onetreu0ePtivetot oE I v i:il'l 2V a.l r:.':i : iilll r :;l iii:II r i:i.]ill';.., H RseA ouvt(Oerot orrlv eoolreptrrl UeUppovl H rurropontrooportxfl neptoXfl rrlq npoo66vet rov nopoyovro oE' H RseA ro$erot 6tu6o1wo, np6ro orov neptnloogtro ltilpo rot ptero oro ruuopontroopto, To ruuopontrooporLro ro$tpo oneeu0epdlvei tov oE. RseA /-(ot i 6ro no06Yowoq Xpnoit '::-- 070 0t nepLoodrepeg onopotrnTeq ettoupYteq .: --S ss Iionrn rpdon/Ueptr6q onovqoetg stress . - 032 OePutr<o shock '--:-: sE l-leptntrooutr6q/eEorunopteg flporelveq '::', 054 Evor:-rgdtoorl rou o(titrou ._:= sF luvoeorl poorryitov/Xnuetoroqio .:: srecl MeroBotrroUdq/Uerorpopd rou otorlpou Enoq ono Tov 070, to E. co/i 6Xet oprero[q nopo- , Meq 0 nou endyovrot ono ouyxexptp6veg neptpolJrovrtr6q ,,,r01reg. (0 optOpoq oro ovo;to rou nopoyovro 6r1trdrvet U Uo(o :"), NTIAPONAA'MA it.l* 0 Avtoytrlvtol-tog Yto Touq tlopdYovreq o Mnopei vo Pu0liiE :'.' 539
oro oE, n RseA eUno6[(er rrlv np6odeorl rou oE orov rupnvo rnq RNA nolupepdorlq Kor rrlv evepyonofrlon Tov uTroKrvrlrd)v rou oE. Auro[ ot unoruvr1"r6q preroypd- rpouv npoi5wo onopofurpq yro rrlv enovomdlcrlorl rov onodroroypr6vov neptnloopu<tilv nputreivcbv ri yro 11 6rdoftoon roug. Iuvendrg, 11 neprn].ooptr<ri ondwrlorl 0epprro0 shock e[vor Uro npoooprvrl ondwrloq noA(v- 6poUnq prj0pto4q nou eL6yXerot on6 ng ouyrewpdl- oerq rov yovrdtorc0v npoi6worv rnq. H pri0puoq rou oE owonorpfueror oro enfnedo rov ononruxcotrlSvrov xot ono6 roroypSvov neptnAoopru<riv n pcoreivc0v nopd tr:v owiororXrov xurroponl.ooportrrilv npcoreivcirv. l-1c69 yvcop((er 11 RseA n6re vo oneLeu0epciroer rov oE; O prllovrqr6g neptLogBdvet pu0;u(6pevr1, 6ro6o1r- rrl npolre6luon rnq RseA. H ouoo6peuorl ononruxo- pr6vov npr.ureiv6v evepyonoe( pro nportedoa (DegS) orov neprnloopx6 Xcipo nou r<6Ber ro C-reLu<6 dxpo qg nporcivqg rseA. Aur6 to r6qtpro evepyonoe( pto dMq npo:redoq, q RseP, ourlq gopd or4v Kurropo- nloollotrrrl n).eupd rnq eooreprx4g pepBpdvnq. H RseP r6Ber qv N-relu<q neproXrl qg RseA, oneLeu0e- pc6vowog relrrd rov oE, o ono(og prnope( nL6ov vo npoode0ef orov nup4vo rqg RNA no].uprepdorlq Kqr vo evepyonor4oer rn Llerqypoqrl. 'Eror, 11 ouoocipeuorl onomulcopr6vov npoteivdlv mrlv neprg6pero tou Bo- mqp(ou evepyonore[ rqv oprd6o trov yovr6[tov nou e].6y- Xowor on6 rov nopdyowo o. | 9.2O Or l-lopdyovTsq o Mnopei vo Elvol Opyovtrrpdvot o€ AroAou0ieg s Mro oro),0u0[o nopoyovrov o 6qgroupyeuor orov 6vog nopo- yovToq 0 onorre(ror yro Tn UeToyporprl iou yovrDrou nou rotdr- ronoreI rov enoUevo nopoyovTo. e To npdruo yovfdro rou tpoyou SP01 peroyporpovror ono qv RNA notrupepoorl rou [evror1. * 'Evo ono ro np6rpo yov[6ro rodrronore( evov nopoyovro o nou o0e( qv RNA notrupepoori vo peroypo(.rer ro evdtopeoo yovrDro. c Auo ono ro ev6ro;.ieoo yovidro rodrronorouv unogovo6eg evog nopoyovro o nou o0e( rrlv RNA notrupepoorl v0 UETo- ypoQeL ro oQrl.ro yovr0ro. Ono.lg oro E. coli, nopdyoweg o Xprlorponoto0wot eu- p6og yro rr1 p60pror1 qq 6vop(r1g rnq Ueroypoenq crro Borcrrlpto Bacillus subfilrs. To yovLd(rrrpo rou B. subt/r,s ro-l6Lronore( rouldXrorov 1 8 6ro<popeur<oUg nopdyov- req o, ouyKpw6gevo pe rouq emd rou E. coli. Aev e(vor oouvrlOtoro vo undpXouv neptoo6repor nopdyoweg o on' 6,1 orc E. coli.Irrlv npoypro-rLr6rqro, ro yovtd(topro tou Strepfomycetes coelicolor xo6rronoe( >60! Iro B. subtilis, rdnoror on6 roug nopdyoweg o e[- vor nop6weg oro B).omu<d x[rropo, evut dMor nopd- yowoL pr6vo ong et6rr6g ouvOrp<eg rrlg ).oipro(r1g on6 540 KE@MA|0 19 Meroyporprl oroug l-lporopuoreg qdVo n rord r1v oMoy4 on6 Bl,omrrrl ovdmu(r1 oe oroptoy{iveo4. H rtipro RNA no},uprepdoq nou eprnM- Keror orn rpuorotroyx4 Bl,ooru<tj ovdmu(4 nepdXer tq 6reg unopovd6eg ru 51er r4v [6to ouvo],rr1 6o;ui pre rqv oeBB'crlo rou E coli, oMd enrnL6ov 6rq06rer rot dtrr1 pro unopovddo nou ovopd(eror 6. O r0prog nopdyov- roq o (oA) rou Borcrqp(ou ovoyvop((er unorrvrltdg pre rg 6reg ouvorvenr6g oL,r1).ouX(eq nou Xpnorponororlv- rot on6 ro 5v(upro tou E. coli un5 rg o6r1y(eg rou o70. Or evoMorcrtrdg RNA notrugepdoeg nou neptSlouv 6ro- goperroUg nopdyoweq o Bp(or<owor oe no).ri pu<p6rc- peg noo6rqreq Kor ovoyvrr:p((ouv unorrvrlr6g pe drogopertrdg ouvorveru<rig oMrl).ou1(eg ong neprolSg - 35 ror -10. Meromriloerg on6 qv 6r<Epooq ;uoq opddog yovr- d(rov oe gro dMrl oprddo e(vot 5vo Xopohrnproru<6 t4g ).oipuo[qg on6 Borcrrlprogdyo. Aur6 ouprBo(ver r<ord rrl Lo(Uo:[n rou B. sutilrs on6 ro qdyo SPO1 r<or rou E coli on6 <pdyoug 6nrrrg or T7, N4 11 o L. Ie 6leg rrg neprmri- oetg er,m6g on6 rrg onLoriorepeg, q ovdmu[r1 rou 96- you neprloprBdvet evoLl,oydg oro np6runo Uero- ypogns rord tr1 drdprero rou l,or1.roy6vou rUrLou, or ono(eg prnope( vo en6pXowor on6 q oriv0eorl Uroq no- Lugepdorlg nou rto6rxonorefuor on6 ro qdVo n on6 rq 6pdor1 enxoupxriv nopoy6wrov nou xtr:6rronotorjwor on6 ro Edyo rc eA6yXouv t4 Borrcrlproxri RNA noLupe- pdoq. Kord rr1 Lo(grrr[4 rou B. subtl/ls on6 ro gdyo SPO1, rq 6torpoperu<d orddro qg ),o(pro[r1g etr5y1owor ;t6oo qg nopoyoyrig vSov nopoy6w0rv o. O Lorlloy6vog r<Ur<Log rou SPO1 51er tpfo ord6ro 6rgpooqg rrov yovr6(rov. Apreoo rord 14 Lo(purl[r1, prero- ypdrpowor to nptilpo yovidlo (early genes) rou cpd- you. Metd on6 4-5 Lemd, ro npcirtpro yov(5ro nqUouv vo peroypdrpowot, on6te ntr6ov preroypdgowot ro ev- drdpeoo yovidro (middle genes). Iro 8-12 Lemd on6 q ).o(pro[q, 4 petoyporprl rcov evdrdpeoov yovr6(<ov ov- rtKo0(ororqt ond q geroyporp4 trov 6lpl;trov yovrdiov (late genes). To nprilq-ro yovbro preroypdrpowor on6 ro o).o6v(u- Uo rou Borcrqpiou (evrorrl. Ouoroonrd 6e 6torp(vowor on6 rq yovbro tou (evrorrl, rcov onol<ov or unoruvqr6g 6Xouv r4v ev6oyevrl rrov6rqro v' ovoyvcop((owqr on6 rqv RNA noi.upepdoq ozBB'tooA. H 5r<qpoorl rov yovt6(rrlv rov rpdyrov onore(tor yrq rrq evotrLoy6q rnq Ueroypog{q on6 ro ev6tdpeoo mo 6{.rtpro yov(6ro. Tp(o puOprmrrd yov(6to, ro 28, 33 ror 34, eL6ylouv t4v nope(o rnq perqypo<p4q. Or Ler- toupy(eg rouq ouvorl((owor or4v EIK0NA 19.38. To np6runo qg pU0ptorlg gord(er UE Uro ol,uorbcorq ov- r(6pooq rl xoropp6xrq (cascade) mnv ono(o ro riv- (upro rou (evrmq preroypdqer dvo npcilrpro yov(6ro rou ono(ou ro npoi6v e(vor onopo('rrlro yro q geroyporprl rcov evdrdgeoorv yovr6(cov. Merd on6 ouq rrl pero- ypoqn, 6Uo on6 to evbtdpeoo yov(6ro xrodrronoro0v npoi6wo onopo(rqro yrq rrl pretoyporpri rov 5r.!r1.uov yovr6(tov. Odyor ;re preroMd(erg oro npd:rpro yov[6ro 28 odu- voroUv vo ;reroypdqrouv ro evdrdpreoo yov[6ro. To npoi6v rou yovr6(ou 28 (9p28) e[vor 1.uo npotetvrl 26 Boorrfg ewoeq
i):.:,.?t;,tt;;,. unoKrvrlT6q rou gdyou ovoyvrrlp[(ovrot ono To [JoKrnptoKo otro6v(ugo To np6rpo yov(6to 28 r<odrronore( dvo veo nopoyovro o nou onoUoKp0vet ro Boxrrlproro o 0 nuprlvoq tou evordueoou ev(tuou gp2B geroypoqer ro peoo(o yov(6to rou qoyou To evbrdgeoo yov(0to 33 xot 34 xtodtronoto0v yro npoltelveq nou ovnroOtorouv u1v gP28 0 nupnvoq Tou ijiilii:l ev([pou gp33-9p34 uer0YpoQer r0 oqIu0 yov(0ro rou goyou 19.21 H InoploYdveon EAdYxeTol on6 Touq llopdyovreq o * H oroptoy6veol btotpet evo porrrlpto 0' 6v0 UrllptKo rurropo nou utplororot truor1 rL 6vo onopto nou 0netreu0ep6v6T0t, * Koee qsxoptoTo 5t0p6pt0u0 rpoxopet oro enopevo orodto ovonTuqnq ouv06rovrog evo rotvoupto nopoYOVTo 0 fi0u erron[(et Tov ltp0!Y0uUev0 nopoYovTo o. & H rnrKorvrovlo UsToq[ Tov 600 dtopeptoporolv ouvrov[(et qv enLtroyq TOU KoTotrtrntrou Xpovou Yto Ttq ovTtKoTooToostq Tov nopoyovrorv o. 'Evo r<o16 nopddetypro Yto rrl Xp4otpono(non rnq evoMoyrlq tov oLoev(ripov npoq dLeyXo rov oI o- ycrrv o'rr1v 6r<tppooq ttov yovLd(cov d(verot on6 r1 ono- proy6veoq (sporulation), 6vov evotrXot<ttx6 rp6no (tor1g oe noMd Bor<tqptor<d e(6q. Av oro5lorloet 11 Lo- yoprOprrxrl o[[4o1 enetdrl petdlvourot to Opemtxd oro ;.16oo ovdmu[r1q, 11 Bloorrxq 96oq (vegative phase) oU[r1o4g outtrtv ttov Borcrrlpitlv 0o 6toxone(. Autd nu- podote( tq onopLoySveorl, 5vo ovonru(tor5 orddto rord to ono(o to xUrropo e(vot ov0ercrtr6 oe noMd e(6r1 neptBoMowtt<<.6v t<ot 6to-rpo<ptt<ov ntdoecov (no- pouotd(owot orqv ::i"' .l::l ;rl r:r). Kotd q onoproy6ve- on oro B. subtitis, dvo on6 to Ouyotptxd yovtdttol-toto nou npox[ntouv on6 rlv owl.Ypoqrl tou DNA onopo- vriverot orn Uto dr<pq rou rurrdpou, npooxoMrlpr6vo orov Kurroptr<6 n6Lo. IXrlpor((etot 6vo 6tdcppoyl-to nou 6r'lprtoupye( 6rjo oveldprrlto dtopep(opioro: to prrlrprr<6 r<frropo KoL ro npoon6pto (forespore) . To ou[,ov6prevo dLdrppoyl-io noyrdeUet rpnpo ev6g Xptr-r- proocol-iotog oro npoon6pto Kot pretd prto tpovoAoxd- oq (translocase - SpollE) owle[ to un6]'otno Udpoq rou Xpopooto;rorog ;.16oo oro npoon6pto. TetrLxd, to npoondpto ;re to eyxol.no;r6vo 1p<opr6oopd rou nept- BdLetot on6 6vo or<Lqp6 nep(BLqpro xot our6 to on6pto e(vot oro0ep6 o1e66v en' o6ptorov. H onoptoydveoq 6topxe( nep(nou orcro <rlpeq. Mno- pe[ vo 0eopr10e( tlg nprot6yovn UopQn 6totpopono(- qoqg, r<otd trlv ono(o 6vo prqtptr6 r<urropo (ro B).o- onr<6 Borcrrlpto) 6qptoupye( 6Uo dtorpopettr<d 0uyorpt- xo r<rjrropo pre [eltlptor6 nenpo:ptdvo: to prqtptr<6 16r- ropo nou teLtr<d trfetoL xot to on6pto nou one]'eu0e- purvetot, to ono(o 6Xet ewetrolg 6torpopetrr<q 6oprl on6 to ou0ewtr6 BonrrlPto. H onopLoy6veol nepil.ol.tBdvet pto 6poorrr<r1 oLl'oyq orrg Btoouv0ettr<6q evepy6qteq tou Borcrqp(ou, crrnv ono(o epn).5Kowot noMo yov(6ro. Ot oMoydg orrlv 5r<- epoon trov yovt6(cov nou odrlyotiv retrtrd oro oxrlport- opr6 tou onop[ou nporo].o0wot r<up(tog on6 oMoy5q orrlv dvop[q rlq UeroYpo<pr1g, Kdnoto on6 to yov(6to nou tretroupyo0v orrl Btroortr<r1 gdoq odpovonoto[wot r<oro tr1 onoptoyeveorl, oMd to neptoo6tepo ouveX(- (ouv vo er<qpd(owot. Qm6oo, nol').d yov(6to etdtrd yro rrl onoptoydveorl er<<ppd(owot ;t6vo r<otd tq 6tdp- i,.I: H Inoptoy6veorl Etrdyxerot ono roug llopoyovrec o 541 Ell(OilA 19.38 H UeroYpoQn tolv yovtd[rrlv Tou QdYou SP01 etrey- XeroL onrl d6o 6to6oxtx6q ovruoromdoetg tou nopoyovro 0 nou 0trtr6(ouv qv etdLrdrrlto rnq 6vopfiq. kD nou owtt<o0tord tov nopdyowo o rou (evtorri crrov nup4vo rou ev(riprou. Awfi r7 awuardotaorl e[vat rc pova6x6 oultBdv nou ana:reftat Yn rn peruBao4 an6 t7v 1xEpaory ruv nptitryav mryv 1xEpaorl rav evddpe' oav yovt6fuv. Arlprtoupye( 6vo oloriv(upro nou o6uvqte( vo peroypdUet to yov[5to tou [evtorrl rot ow(yt'owd preroypdqer etdtrd to ev6td;reoo yov(5to. Aev yvtop(- (oupre ndlg to gp28 onoporp6vet ro cr43 o6te n ouprBo(- va oro o noLunem(dto tou (evtorri. A0o on6 to evdtdpeoo yov(6to epntr6rowot mrlv en5prevrl evotr).oyq. MetoMd(erq mo yov(6to 33 n 34 nopeprnod((ouv rrl UerqYporpri tolv 6tltprtov yovtS(<ov. To npoi6wo outtilv ttrlv yovt6(rov oXrlprot((ouv 6vo 6t- prepSg nou owtro0tord q gp28 orov nuprivo rrlq R- NA noLuprepdorlq. En(or1g 6ev yvcr:p((oupre nc0g ot gp33 xor gp34 erron((ouv tr1 gp28 (f onoto6rlnote un6L).etppro oA tou levrmri), aMa p6ttq npoode)olv orcv nuprlva rcu ev(i1tou, e[vu Lxavd va evepYonou fioouv rq peraypaEn p6vo orou;, unoKtvqriq tuv 6tltt- pav yovt6[av. Ot 6to6oXrr5q qwtKqroordoetg tou nopdyowo o SXouv 66o entmc0oetg. Kd0e <pop6 nou oLd(et n uno- provd6o, q RNA notruprepdoq ;rnope( v' ovoyvop(oet gto v6o t6(q yovr6[tov evdr tout6Xpovo no0et v' ovoyvtop(- oer rrlv nporlyoripevrl td[4. Euvendrq, owo( ot 6tot<6- meg entgripouv 6pomtr6q oMoydg mrlv evepy6qto t4g RNA nol.uUepdonq. I Boorr6g evvoleg
Ippoti(erot 6Ldrppoygo rUnopo trtjeror To onopLo onetreu0eptiverot ElK0ilA 19.39 H onoproydveon repDoppovsr Trl Oroqopono(norl evdq p)oorKou poxqpfou o' 6v0 UnTpKd xunopo rou trUeTor Kr 6vo on6pro rou qfletreueep6veror. Kerq ourrlq rqq nept66ou. Iro r6loq rnq onoproy6ve- or1g, -40olo rou BoKrrlproKoU mRNA e(vor erdrx5 yr' ourn rn Qdon. N6eg pop<pdq RNA nolupepdonq evepyonoro6wor oe rUnopo nou npoyporonoro0v ofloproySveon. ne- pr6xouv rov i6ro nup{vo rou ev(ripou 6ncoq ror ro B}.o- mtxd x0rropo, oLl,d 61ouv dtogoperu<69 npcoretveg orn 06on rou Bl.oonroU nopdyowo o (oA), Or oMoy5g orn peroypogxf er6x6r4ro ouvo(r((owor qrrlv Ell(BllA 19.40. H Boou<r1 op14 e(vor 6n o nopdyowog o r60e 6roprep(oprorog 6ro6oXu<d erron[(etor on5 Svov vdo no- pdyowo o nou nporole( q pretoypogn Uroq 6rogope- ttxrlg optddog yovr6(ov, Mero[U rrov 6ropepropdrov undpXer enxowrov(o ciore vo ouyxpovroro6v or oMq- ydg oro npoon6pro Kqr oro ;r4rptx6 xrirropo. H or<oLou0(o rrlq oroproydveorlg opx((er 6rov or ne- prBotrtrowx6g ouvOr1reg nupodororiv 6vo r<0xl,ropro gto- ogo-ovoFer66oo4g (phosphor-relay), orov ono(o puo gororpropu<ri oprd6o prerorp6petor rord prirog proq oer- pdq npolrcivrirv prSXpr vo grdoer aro Usroypo<px6 puO- 542 KEOA Al0 19 Metoyporpn mouq llpoxopurilreg gror{ SpoOA. Ie ouq q 6epyoo(o epn},6rowor notr- Ld yovr6rord npoi6wo, rov ono(rov r'1 nolunLor6rrp6 clwrKoronrp((er q Xpfon o4pre(alv el6yXou, 6r1no611 orryptor0nov Kord rq ono(o ro Borcr4pro enrBeBordloei 6rr entOupe( vo ouvel(oer orrlv o66 qq 6togopono[- r1oqg. Autrl 11 puOgronr{ nope(o 6ev np6ner vo },oprp& vetor odu<oroL6y4to, roOrig q rel,rr<q on6qoo4 eivor oprerdrlqq. H evepyono(qorlrou SpoOA Ue qooqopul(oor1 orp ;roro6ore( qv 5vop(q rnq onoproy6veorlg. Ee qoorpo puLuopSvq Uopen, o SpoOA evepyonoe( rn peroypo- gr1 6rio onepov(rov, r<oO5vq on6 to ono(o getoypdrperor on6 pto dro<popenrri Uopen r4g RNA nolupepdorlg tou [evrorrl. To 5v(upo rou fevrorq, Xprlorponord:wog ro yevrr6 nopdyowo oA, pretoypdqer ro yov(6to nou ro6tronop[ rov oF rqr, un6 qv ro0o6r1yr1or1 ev6g 6.]t- l,ou nopdyowcr o, oH, getoypdrper en(or1g ro yov(6ro nou ro6rronote( 6vqv np66popro tou nopdyowo oEs nou ovoE6peror rr:g npo-oE. T6oo o oE6oo ror o npo-d nopdyowor nprv ro olqpolo;r6 tou 6rogpdyUqroq, otrl.d evepyonoro(wor opy6repo. H peroypo<pr1 nou roteuO0veror on5 rov oF ovo- qrdMeror ener64 6vog nopdyowog owroryl.ro (SpollAB) npoo6dveror oe qur6v, eprno6((owdq rov vo g4rlpor(oer 6vo oLoriv(upro. Iro npoon6pro, com6oo, 6vog nop6- yowoq owr-owto(ypo (SpollM) r<oroor5Mer rov Kqro- oroMo. H o6povono(4oq tou owr-owro(ypo etr6yXeror on6 pro oetpd ouprBdwov googopul(coo4g/ono<pro- oEopuL(rooqq, ornv ono(o to npriro Brlpo e(vor n ono- goorpopuL(roor1 on6 Uro eoogcrrdoq nou ovoprd(eror SpollE. H SpollE e(vor 1.uo nproteiV4 qg eooreptx4g pepppdvnq nou ouooopeUerqr orov Kurroprr6 ndLo, pe onot5treopo q evepy6r4to groogordonq nou 6ro0era vo oupmurvc6veror neplso6repo oro npoon6pro. Iuvo mrrd, 11 onoeoogopul(roor1 evepyonoe( rov SpollAA" nou Ue rq oerpd rou ercron((er rov SpollAB on6 ro oF. H onel,eu0Sproorl rou oF rov evepyonoe(. H evepyono(qor1 rou oF orlproro6ore( qv dvop[r1 rnq KuTropoer6u<r1g 6r<Epoorlg rtov yovr6(<ov. Yn6 rrlv xo0o6tiyqoq rou oF, 11 RNA no).uprepdo4 pretoypdger qv npdrrq opd6o yovr6(orv rnq onoproy6veoqg, O Ed efuot unefOuvoq yro 6trr1 q preroypogrl oro npoon5- pro. O oA 6ev rotoorp6geror xord 14 onoproydveorl t<ot, ouvencig, to Bloorrr6 oAodv(upro, EoA, nopop6va oro rUropo nou npoyporonororlv onoptoy6veoq, H or<oLouO(o ouveX(erot xoOdrq oro npoon6pro no- pdyowor npoi6wo ro ono(o npo6pXowor on6 unorrul- 169 nou ovoyvop(owor on6 ro EoF (BI6ne ElK0l*A 13.41). l-ro nopdderypo, ro EoF etr6yXer q orlv0eo4 ev6q prero- ypd<pou nou rcodrxonoe( rov oc, o ono(og ;re q oerp6 tou oXqpro{(er to o},o6v(upro ilou peroypdger ro riore- po yovbro rnq onoproydveoqq. To EoF ovoyvrop((er eni oqg 6vov unonvrlrq nou eLdyler t4v 6rrppoorl ev6g npoi6wog une0Ouvou yrq rnv enrrowov(o pre to 6ropr6pr- opo rou prqrpu<oti rurdpou. To ouyrerptprdvo npoi6v, yvoor6 og SpollR, erxpfueror on6 ro npoon6pto prSoo orrl pepBpdvrl nou 6ro1op((er rq 6Uo 6roprep[opoto. In1 UepBpdvn, o SpollR evepyonote[ rqv nporeivq SpollGA 11 ono(o r<5Ber rov ovevepy6 np66popro npo-oE, pretorpS- Btroortrri poxqpro AvTrypoqo DNA Cq c(q ct^r-i ffi ffi' 1 .>
MHTPIKO RTIAP0 To oLodv(ugo nepr6Xer nP0:n0Pl0 tov gH n rov oao I /,---f,- "r' 0 oK evepyonoreftol rr exron[(et rov oE I Y 0 evepyonorq96vog oK unoorrlp[(er rn usToYpoQn TOV orlruov yovt6urrv H onoproy6veorl nepLtroppover 6ro6oXrr6g otrtroy6g oroug nopoyovTeq o nou etreyXouv qv erdrrorqro rrlq RNA notrupepo- --( yto TIV evop[1. 0t orotrouO(eg oro pqrprro rurropo (aporepa) ror ro npoonopo (6e(La) oXer((ovrot pe orlporo nou 0tonepvouv ro : orppoyuo (entorl;.ro[vovrot Ue opt(ovflo B6trl). I ,fuJI I 0 evepyonotlg6vog oG unooqp[(et q UsroypoQn rov ri{.rrprov yovt6[tov ffiw 0 oE evepyonore(ror n erron((er rov 043 ITo yovOto oK geroypdrperor on6 rov oE I I I #t nowdg rov finv evepy6 Uopgri, oE, oro prppu<6 r<(rrro- po. (O oE nou nopdyetor oro npoor6pro onodoprefuo$. H oxoLouO(q ouveX((eror 6rqv o oE fio ;rqrpu<6 rftropo owtxo0(ororor on6 rov oK. (H nopoyoyrj rou ox eivfl oprerd nep(nLox4, ener6{ ro yov[6r6 rou 6qpr- oupye(rot on6 6vo ouprBdv ronoerdrro0 ovoouv6uo- opori!). 'Onog ror o oE, o oK ouw(0eror og ovevepy6 np66popo pr6pto, ro npo-oK, 'Eror o oK np6ner vo evep- yonotqOe( pre r6rprpro qg np66poUnq UopAnq nporo0 pnop6oet v' owr.Koroorrloer rov oE Kor vo pretoypdget ro 6r.trrl,ro yov[6ro oro prrlrpu<6 r0rropo. O ouyXpovr- opr6q owcbv rov 60o ou;rBdwcov qro 60o 6toprep(opro- to ouwov(eror on6 brogopetxd or1;roto, Iuvomrxd, r1 evepy6qro rou oE oro truppx6 rUrropo e(vot onopo(- rnrn yro rnv evepyonofioq rou oG oro npoon6pro rot q evepy6qro rou oc ononefuor yro rq 6rlpuoupy(o ev6g o{porog nou peroS6eror 6top6oou rou drogpdyporog yro vo evepyono(oa rov oK. To rUrtrtogo n1q rptoorpo- ovogeroDdoqg evepyonore( q oUv0eoq tou sF 661 169 snPo-E 0 oF owrKoerord To 043 mov nupflvo rou ev(Ugou I V To rewpud odpntroxo rou oF geroypdqer ro np6tgo yov(Dto qg onopoy6veoqg 0 oG e[vor npor6v rtov np6qrov yovr6(rov n1 g onoproy6veorl g 0 oG evepyonorefuor ror erron((et rov oF nP0x0Pt0 &.ry flK0NA 1S.41 0 oF nupodorei U orjv0eon rou en6pevou nopo- yovTo o oro npoondpro (oG) rr evepyonoLe( ro SpollR, nou e[o0e[ qv SpollGA vo K6qrer rov npo-oE. 19.21 H Inoproy6veol EtrdyXeror ond roug flopoyorrec o 5tE +@SpollR l@p
Y [lporedluorl gK EIK0NA 't9.42 H dromoupou;rcvn pu0gron Tnq ofloproy6veorlq ouvTov((er To ouyxpovrouo rtov yeyovdrov oro urlrprKd KUTTopo KOr ro rrpoofidpro. Iuvenrilg, n onoproy5veon el6yxeror on6 Uro oKo- LouO(o orrlv ono(eq or nopdyoweq o Kdee 6toprep(opro- roq evepyonoro0wor 6ro6oXxd on6 toug ofupo F, E, G r<or K, r<o06vog on6 rouq ono[oug xmeuOUver rrl Uero- ypoAn ev6g ouyrerptprSvou oUddqg yovr6(rov. H oro- Lou0(q pnope( v' ovqfiopqoro0e[ on6 5vo oroupcor6 np6runo o4trrdrcov nou 6ronepvo0v ro 6rd<ppoy;ro, ouv- 66owoq qv 6rEpoo4 trov yovr6[rov oro dtogopelrd drogepioproro, 6ncoq eo(veror orrlv EIK0NA'l 9.42. Ko0drq evepyonororiwor vrior nopdyoweg o, or noLro( nopd- yovreq ercron((owor, evepyonordlwoq Kr onevepyonor- rbvrog drogopetrr6q opd6eg yovr6(cov oro 6Uo 6roprep[oproto. | 9.22 O AvrrreppoTrou6q Mnopei vo Eivor Pu0prorr16 EupBov * 'Evo ouUntroro ovTrT€pUoTrouotj enup6ner ornv RNA notrupe- poorl vo Dtopooer 6roprloou repportor6v. " 0 tpoyoq IoUB6o Xpr;orgonorel ouor1poro ovruepporLopo0 yro vo pu0pioer 16oo ro nptiulro ooo xor ro 6QLpo peroypotpo, otrtro ro 0uo ouorrlporo trenoupyo0v pe evietr6q Drorpopel- roug l.u1xovro1roug. e H npoodeon nopoyovrov oro veoouvre0erp6vo RNA ouvdtier lg nporelveg ovTrTeppoTropou oq 0eor1 rou repponorrl Dro- Ueoou Uroq 0r1trroq RNA. * Avnrep;loloUoq TIq peroypoqrlq ouppolver enlo4g ror oro onepovro rou rRNA. O owLrepprotto;r6g Xpnorponorr(tor yro rov SAeyXo rnq Ueroypoenq oro onep6vro 16oo tov gdy<ov 6oo Kor. rcov Borrqp(tov. 'Onog qo(veror ornv fifftii :i i.lj, o owrrepporropr6g ovogdparor oe rponono(4or1 rou ev(riprou, to ono[o prnope[ ntr6ov vo nopoBrdoer Svov repprororrl ror vo nepdoer oe yov(6ro nou Bp(or<owot roOodrxd. Ito nopdderygo rnq ex6vog, evcb 11 nporo- 0opro;r5vq o66q yro qv RNA no).upep6or1 e(vqt vo reppo{oer oro r5l.og qg neproXrig 1, o owrepport- opr6g rorolrlyer oe ouveXr(6prevn Ueroypoe4 6topt6- oou rnq neproplg 2. Iuorrlporo owrrepUorro;ro[ e(vot r<otvd orouq LoprB6oerSe(g Borrqprogdyouq (<pdyor nop6prorot pe ro qdyo l,dpB6o nou neprypdeowor mo KegdAoto 27). Aw(Oero on6 roug tpdyoug nou prord(ouv mov T7 tou E. coli Kor rouq rpdyoug SPOl rou B. subfl/is nou neprypdgqxov nporlyougdvog, o LdgBdo 6ev rcodrxo- nore( ofre 6trr1 rou RNA no).upepdo4 ofre roug 6r- roUg rou nopdyoweg o. MdLLov lpqorpronore[ qv nolupepq RNA noLuprepdorl rou [evrorrl yro 6].q rq peroypoqrlrou. Ie o0wopro 6rdorrlpo perd q ).o(pr<l- (r; on6 ro rpdyo, (er<rvd n Ueroypoen oe 6rio npcir- pouq unoKlvnr6q, Pn rot Pr-. 0o16oo, oe ro06vo on6 ourd to onep6vro undpXouv repprortm6g prerd rrg orpeqp(eg rnq Ueroyporprlg ror nprv ro neptoo6tepo yov(6ro nou ro6rronororiv rtg nprilrpreg Lenoupy(eg. O tepprono;r6g rnq peroypoenq oe our6g lg 06oerg po- rord:ver 14 ).o(puo[q. Av 6prcog 11 RNA no],upep6o4 no- poBtdoer rouq reppqrrm5g xor UeroypdQer to npcirrpo yov(6ro nou e(vor uneU0uvo yto rnv owrypo<pr1 rou yo- vrdrdrprotog rou gdyou, 4 ovdnru[q rou ].d;rB6o 0o ouveXrme(. H npcirtq on6qooq yro reppotopr6 el,Syleror on6 qv npcoreiV4 qwrrepuorrol.rori "N", qv nprilq npr.,:ret- ul nou nopdyeror un6 rov dLey1o tou Pr-. H N oXqpo- r((er o0trrnl,oKo Ue rrg npcoreiveq rou [evrmq nou xoloUwqr nopdyoweg Nus (N utilizing substances - ouo(eg lp4orprono(r1onq rnq N) rpononorcilwog qv R- NA noLuprepdoq rord r6roro rp5no riore vo Urlv owo- noxp(veror nL6ov orouq reppotm5q. Irqv npoyUo- rrx6r4ro, ro oriprnLoxo rou owrrepUotrotrroU oXqprorl- (etot ndvro oro veoouwe0erpSvo RNA oe pro oMqlr.ou- X[o nut (N utilization site - 06oq Xprlorprono(4onq rnq N). Or O5oerg nut onorel.o0wqr rup(cog on6 otrl,qlou- 1(eg RNA nou ovo;rd(owor boxA t<ot boxB,6nou ouo- otr:pe0owor or nopdyoweq rou [evrmrl NusA, NusB, NusE (prBoorotrrorrr4 npcoreivrl S10) ror NusG. Or nporefveg owrreppqtogoU nopopdvouv npoo6edepS- veq oe outriq ng 06oetq tou RNA oq dyo 1.t6vt1to oult- nltoxo avrlrepparLolJoti r<oodrg n RNA nolu;repdorl ouv05rer ro 6rjo perdypogo npoq ro 6e[rd xor r' opr- orepd. 'Eror, to veoouwe0erpr5vo RNA ouv66er rpuotrcd ng nporeiveq owrrepporroporj nou e(vor npoodedeprS- veq orn 06o1 nut pe rnv RNA noluprepdoq nou ni.q- ord(er rouq repUortor6g. Mol,ov6rr o or<prBqg 1.r4Xovropr6g Ue rov ono(o ro oripnLoro owrrepporr- opro( e;rno6[(et tov rep;rorro;r6 6ev e(vqr or<6pr4 yvro- m6g, ro 65or;ro ratv npo:teivdlv repporrol.roU mqv RNA no).upepdo4 6rop5oou rou veoouwe0erp6vou R- NA e[qye(rr1v u<ov6r4rd rou v' owreppol((er oe 6ro- 6o1u<orig repprotrmdg nou Bp(oxowor oe on6moo4 erorow66cov q ordpq r<or Xil.r66cov Bdoerov r<o0o6rxd. 0"ffi 544 KE0MAI0 19 Meroyporpn mouq tlporopudrreg nP0rn0Pr0 Boorxeq eworcq m
lleptoXq 1 Ynouqqg +{-l-leptoXfl2 W H RNA notru[epdon rep[or((et To RNA Bp(orerot grlvo oqv neptoXrl 1 ANTITEPMATI:MOT Ttloo I neproXrl 1 rioo xot 11 neptoXrl 2 getoypdtpovrot 0 owueppoloUoq Unoper v0 etrdyqer Tl UeToypoq1 K00op((0vT0q 0v I RNA notruUepool 0o repporloet r] 0o nopoptooet , -, :pLopevo repportorl rot 0o npoXoprlott orrlv enopevn neptoXrl. - ieuro(o npolrefvq nou nopdyetot on6 ro N-owt- -.:-crrop6vo petdypogo ond rov dtro np<otpo unor<t- -:'-l PR, ovopd(trot <Q,. Onotg n N, n Q e(vot -: *:srvl ovrrrepporrouof. Awtreppror((et rn uero- ::cr1 on6 rov drl.rtpro unoxtvqrq PR, o ono(og nopd- . ivo prerdypogo nou rordtrconote( ttg npotetveg qg .'::"qq Kor rnq oupdg tou tpdyou. 'EroL, 1 6t<<ppootl ---, yovr6(ov tou LdprB6o npoyporonote(rot oe 60o :-:51o, xoOdvo on6 to ono(o etr6y1erot Ue owtreppo- -:-c (BL5ne Ev6rrlta 27.7 Kot - . ). H Q enL- -:.:a o-rqv RNA notrugepdoq vo nopoBtdoet rouq -1:Jorroreq mqv 6q.rrpr1 pretoyporpLr<r1 provd6o, oMd pt', :. 3^r0g 6Lo<popetu<d prqXovtopr6 on6 qv N. Aw(Oetq - : rrlv N, n Q npoo66vet DNA (orr1 0doq qut, Q uti- :a: cn site - 06oq lpqotprono(nonq rnq Q), o).Ld : -q Kor 11 N ouprnope0etot Ut rrlv RNA no).upepoorl . :. le rdnoro tp6no nopeprBo(vet mq 6pdo1 rov rep- -::.;rov oe 6Lo to 6q.rtpro onep6vto. Oofverot 6rt q : ::r1 rrlg Q nepil.opBdvet enttdXuvorl qg RNA noLu- -.:corlq ovdpreoo oe 06oetg no0or1g. (H ouvoLtt<t1 :-3.ro1 trlg ovdnru(rlq rou ).d;rB5o 0o nopouotome( - : Kegd).oro 27). -o onep6vto rou rRNA 0' ovoprev6tov vo 6e(1vouv --'..i<orqto, enet6q e(vot ptoxptd oMd 6e preto<ppd(ov- -: Oordoo, r<o06vo on6 to onep6vto rRNA rou E. coli nepdXer otrLr;l.ouX(eq rinou boxA xqt boxB flou ouvclp- poloyoUv oriprnAoro owrepgortopoU ildvo oro petd- ypqeo, mo ono(q opropr6vot ilopdyoweq Nus e[vqt rowo( pe ro <pdyo LdpB6o. Awd to otiprntroro 6ev ne- pr5louv 6vov nopdyowq {nou N i O, TIou Ko6tKonot- oUwot pr6vo on6 ro yovt6uiporo roJV qdyov, oLLd enopxoriv yro v' oTlorp5{.rouv tov np6opo tepgonop6 ortg oM4trouX(eg qq qoupr6rog Kot orouq ooOeve(g p-e[optc0;revouq repUortor6g nou tuXo(vet vo undp- Xouv prSoo oro 6oprxd yov(6to tou rRNA. O owLreppo- tropr6g e(vqr ndvro onopo(tqtoq yto rnv enopr<r1 nopoyoyrl rRNA, 61r p6vo 6tov o LdptB6o ptotr0vet to rrirropo. Euvenrilg, t<ot6 rrlv eE6fuEn rov Borcrqp(rov, ot nopdyoweg Nus 6ev enrl.6X0qr<ov yto vo 6eurol,0vouv qv Srgpooq rrov yovt6(trlv-rou LdpBdo oLLd prdMov yro v' qnorp6rllouv qv nofur<6qro oro onep6vto tou r- RNA. Or neprol6g-odrlyo( ttov onepov(rov tou rRNA ne- pr6Xouv oMqLouX(eg boxA nou ouvopl-totroyorjv roug nopdyoweg Nus r<oOcirg ot otrtrqtrouXbq boxA (enpo- Bdtr.ouv on6 ro 6(qu^.o e[66ou tou RNA. Onog rot pe rov qwreppotro;r6 cno <pdyo LdgBdo, outrl 11 6epyo- o(o otr)td(er trg r6r6rqrcg qg RNA noLu;repdorlg rotd t6roro rp6no rime vo prnope( nL6ov vo nopoBtd(et rouq repLlorrorSg. Qor6oo, o Urlxqvtop6g 6ev 6Xet o16- prq onooorp4vrore(. To RNA ovTrnpoooneuer lqneproxdgl +2 iti.ii: 0 Awuepgortopog Mnope(vo E(vot Pu0pLorLrc I"-3 ' 545
iiifiiii -,ovrspus],olt.,.. .. &Ynortvrlqq t @r"liilrw*.wRNA q,rX-, ty*fyyf Meroypogrr4 povrido Ynomvrlqg l-lptotetuq owueppottolrori fL rR1 tR, Mto npolrelvrl ovTtTspu0Ttouou pnope( vo npoodtooet orqv RNA notrugepoorl rrlv rxovoqro vo nopoprooer 6vov erdrxo rs:- poiloq. PL Pnr D ,R) pN pN p0 Ageoo nptirpo Ageoo npdrrpo '0rlrt1ro 19.23 O KtirAoq rou BorrnptoKo0 Ayyetuoqdpou RNA * Iro porr1pro, n Ueroypoqn rot 11 perorppoorl npoypoTonor- ouvror rourriXpovo (ou(euyprlvr1 Ueroyporp1/perotppoo4), ro- 0drq ro prpooropolo (euvouv q perorppoorl evog mRNA nprv otrortrr1po0er 11 ouv0eo[ rou. * To Borrrlproro mRNA e[vor ooro069 pe Xprivo rlpt(au1g triyruv Lenr6v rqg opoq. e'Evo Borrnprord mRNA pnope[ vo e[vor no],uororpovrro pe notrtr6g rtrldrxdq neptoxeq nou ovTroTorxouv oe Ororpopenro yov[6r0. To oyyelrog6po RNA SXer n1v (6Lo erroupy(o oe 6Lo ro r<Urropo, undpXouv 6pro:g orlprowrxdg 6ro<po- pdq oq npoq rrq ).emoprdpereg qg oUv0eorlq Kor rrlq doUriq rtrlv npoKopurorrxd;v Kor roJV euxopuortrotv mRNA. Mro orlpowrxq 6rogopd onlv nopoyoyrl rou mRNA e(oprdrot on6 trg O5oerg 6nou ouprBo(ver 11 preroyporprl ror q perdgpoorl: o Iro BqKrriprq, ro mRNA preroypdgeror Kqr Uero- gpd(eror oro povo6rr6 rurropu<6 6roprdpropro. Or 6fo 6repyoo(eg e(vor 16oo mevd ouv6edepdveg nou ouprBo[vouv rour6Xpovo. To prBooroprdrro npe oroLlc0wor oro Bor,cqpror6 mRNA npw o16;r; ole rl,qpoOe( 11 peroypoqri rou, ouvendrg ro ndtuoapdrto (polysome) ;rnope( vo e[vqr oxdprl npooxoLlq;.uivo mo DNA. To Bomqprox6 mRNA ouv4Otog e(vot omq06g xor yr'ow6 prerorppd(etor oe noLunem[bro pr6vo en( ],(yo ].emd. Aurl q Drep yooio roAefuor ou(euypdvn Feroypqgrj/perdgpo- o4 (coupled transcription/translation). o I' 6vo euxopurotrx6 r<6rropo, q oUv0eorl r<or 4 rrlpr(- Uovorl rou m R NA npoyporonororiwor onoxl,ermx6 orov nup{vo. To mRNA e(dyeror oro rurrop6dt+ o1-ro, pr6vo ogori ol,or).4pro0oUv ourd ro ou;rBdwo. 'Evo tunx6 europurrlrrx6 mRNA e(vor oXeurd oro- 0ep6 r<or ouvel((er vo petorppd(eror en( opxedg cbpeg, poLov6l undpXer peydln preroBLqt6rqro oq npoq rn oro0ep6r4ro erdu<rirv mRNA. H ElK0l'lA 19.45 6e(Xver 6n oro Bor,r4pro n UeroypG grl ror q perdgpooq qXer(owor orevd. H petoypogri [er<rvd 6tov 11 RNA no].u1-repdo4 npoo6e0e( oro DNA ror op(oer vo nopeUeror qrtdXyowog 6vo owiypogo rou ev6g xtrrilvou. l1ol,[ o0wopro, prBooroprdrto npo- llporetvq ovlrepporrogorj Tepporoqg 546 KEOA Al019 MeToypqqq moug l-lpoxoputirreq Boorxeg eworeg H nporeivn ovnrepponoporj 6[ver rq duvordqro oqv RNA notrugepdorl vo nupoptdoet rov teppottoq
S mi* rri"r'rl,.i I :r lil'rlr ji,;'ir,.)l': j.J :rtitr li rllli*lrtririiijrj lll:l,lir.i..r iir'i1.'r lt: :ti rliitl To 5' oKpo elvor rprQooQoprKd ^urd2,0 nrin l-{ ENA noiupepdrq reppot[(er mc 3' axpo , _ ;'- 6rr' rff$1, - - 1:a::::_-3: / I Y:.+a-:--r/ M -:"t::11)A i.i1:-z 3 C min H ono0rlpqon ouveXi(eror. ru prpoor*uorto olor<trnotirvolrv rq [€,ToQpoun ,a 'n /zrr=-,"{-l- /,r'" I , :,. ; Iuvotprl: oT0 BoKT1pro To mRNA UeToypoQsTot, - : rorppo(eror rot ono6opelror rour6Xpovo. ;<oMrr;wor mo 5' dr<po tou mRNA t<ot (er<tvotiv tr1 pte- rl:!pooq, nprv or<6pq ouwe0e( ro un6lorno prdpog tou * - v u 1.roro g. l-l oMontrd prBooropdtLo rtvo tjwou<otd prrl - . :q rou mRNA ev6o<o or<6;ro ouw(0etot. To 3' dr<po ::r mRNA 6r;;uoupye(rot 6tov repprotrme( q Ueroypo- :- To mRNA 6oo (er ouveX((er vo pretogpd(eror on6 -: crBooco;rdlo, oltrd opr<erd ypiyopo ono6oge(tor :-:- ouvo).trq r<ore0Ouvoq 5'-+3'. To mRNA ouw(0etoL, ...::gpd(eroL on6 to prBoooprdrro r<or ono6ope(tot, :'.: no).r-i ypnyopo, 'Evo [e1opror6 pr6pto mRNA ent- : -'",er ro nolri yro prepu<d emd. Iro Borcrrlpro, n Ueroypoerl not pretd<ppoon npoy- -:.cnoLo0wot pre nop6proto rol0qro - oroug 37oC, - r, - 5 0 vo u x..eot(6ro/6eutep6).emo. Aut6 npooeyy((er -:r.r rrlv roXritrlto qg npurreivoo[vOeor1g, nou o6pd ..:, 15 oprvo[doi6eutep6].ento, Iuvencbg, -1 ],ent6 ,.:.ii(eror yro vo petoypocpei Kor vo Uerogpoore( 6vo -:A 2500 bp, nou owrororXe( o' 6vo noLunem(dro 90 . I Acpdrou [exrvr;oer q dr<gpootl ev6g yovrd(ou, ro m- RNA rou runrKd 0o epqovtote( crro Kt nopo $m oe -1.5l,em6. To ow(ororXo notrunem[dto 0o epgonore( nep(nou 0.5 Lem6 opy6tepo. H Borcrrlprorcl petdgpooq e(vqt no^.U ono6orx4 rot ro neproo6repo mRNA ;rerogpd(owot on6 peydlo opr0p6 ogrXrd ouoreuootrr6vov ptBooroprot(cov. I' 6vo ouyrexprpr6vo nopd6erypro (trp mRNA), xdOe Lent6 npoyporonoro6wor nep(nou 15 evdp(etg rnq Ueroypo- qrjg r<or xo0dvq on6 ro 15 mRNA prdMov pretogpd(etor on6 -30 prBooro;rdrro mo 6rdleq.r;ro prero(0 Uoroypo- gqq rot ono66prqo4g. H oord0ero rov Tleptooot6p<ov Borcrrlptorc0v mRNA e(vor ewunootor<r1. H ono66pr4orltou mRNA opXt(et l,i- yo prerd on6 rr1 prerdgpoorl rou Kot, nt0ov6v, ptSoo o' 6vo l,em6 on6 qv 6vop(4 rnq Ueroypoq(S, To 5' drpo tou mRNA op1((er v' onodope(tor npoto0 ouwe0e( f Ueroqpoore( to 3' 6xpo. H ono66pqo4 qo(verot 6rt oxoLou0e[ to reLewo(o prBootoprdtro rqg noprnrig r<otd prrlxoq rou mRNA. Qsr6oo, npolcope( Bpo6ritepo, nep[- nou Ue rn Urori tolUrrpo rnq Ueroypogrlg r<ot rnq Ue- rdgpooqg. H oro0ep6r4ro rou mRNA 61er peydlr; en[6poor1 mqv noo6rrlrcl rov notrunenl6(ov nou nopdyowot. Iuvq0cog erqpd(etor oe oX5o4 pre to Xp6vo npt(otiq. To mRNA rdOe ouyrerptpr6vou Borcrrlprorof yovt6(ou dXer Xoporcrqprortx6 Xp6vo r1;u(oriq, Ue pr6oo 6po -2 Lemd. Aorpolrig, ourl q oepd ougBdwcov e(vor equcrrl 95- vo ener6{ 4 petoypoqrl, 4 ;retd<ppoo4 ror q ono66pr1- on npoyporonororiwor npoq rnv (6ro r<ote0Ouvor1. H 6uvogu<r1 r4g dxrppoorlq rov yovt6(tov otrttKonote(rot orqv rllercrpovu<r1 gu<poyporp(o qg EIKONAI 19.46. Ittq ouyrerpqldveg (dyvcooreg) peroypogxdq govd6eg, op- retd mRNA ouw(0ewo row6Xpovo r<or ro r<o06vo rp6- per noLLd prBoocoprdrro Trou onooXotrotlvrot pe rrl prerdrppoorl. (Aur6 owrororle( oro orddto nou nopou- ord(etor oro 6e0repo nlq(oro r4g Ex6vog 19.45). 'Evo RNA rou ono(ou 1 oOv0eoq 6ev 6Xet or6pq otrortr4pto- 0e( ouyyd ovopd(eror yew6pevo RNA (nascent RNA). To Bor,cqprord mRNA noxfi,ouv notr[ cog npoq rov opr0g6 r<ov nproreivci;v nou ro6xonotoUv. Mepu<d mR- NA g6pouv ;r6vo 6vo prov6 ORF, 6qLo6r1 e(vot govoor- orpovrx6 (monocistronic). A).Lo (q ntrerov6tqto) rpSpouv otrqtrouX(eg nou ro:6rronotoUv notrtrd nolune- m(6ro, 6nl,o6n e(vor notruororpovrx6 (polycistronic), Ie ow6q rrg neprmcboerg, 6vo ptovo6tx6 mRNA preto- ypdgerot on6 pro ogd6o yenovu<tirv otorpov(olv. Mto rStoro oumorl(o ormpov(rov onopr((et 5vo onep6vto nou eLdylerot rrlg evro(o yevetx{ provd6o (B}.5ne Keqd- Loro 26). 'OLo ro mRNA nept6Xouv rperg neptoX6g. H ro6tro- notrlrtrri neproXrj (ovorXr6 nLo(oro ovdyvcoo4g, open reading frame, ORF) onoretrekor on6 puo oerpd xo6txo- v(cov nou owrTrpoooneUouv rrlv oprwo[u<{ oMrl}.oufo rou nol.unemr6(ou. ApX(er (ouvrlOrog) pe AUG Kor re- Lercbver pr' 5vo on6 ro rp(o ro6rr6vto tepgottol-toti. To mRNA oor6oo e(vor ndwore prorpUrepo on6 t1v t<to6t- xonorrlurrl neproX(, roOrilg nepdler enrnp6oOeteg ne- -lil 'il 0 KUrtrog rou Borqproxo0 Ayyetrrorpopou RNA 547
t-a To yevvdpevo mRNA np0e!6x0uv ono ro DNA xor eivor xotruppevo pe prpoorogono Mnopotlpe v0 flopornpnoouUe Ttq UeT0yp0qx6q Uovddeq oT0 PoKrnpro. EuyevLr4 nopoxd)pnon rou 0scar Miller. at proX6g Kcr. oro 6Uo dr<po. Mro enrnp6oOerrl oMrlLoul(o mo 5' drpo, ovo6u<d r4g ro6txonorrltrrrlg neproXrlg, ovogdpetor coq odqy6g (leader) n 5' UTR (5' untranslat ed region, 5' operd<ppom4 neproXri -). enrnp6o0erl oMqLoul(o r<o0o6rxd on6 ro o,j;ro repgotroproU, q4r1- pror((er ro 3' dr<po ror ovoprd(eror oupd (trailer) ri 3' UTR. llop6rt 6ev x@drronotoUv notrunem(6to, ourrlg or oMql,ouX(eg pnope( vo nepr6Xouv orlgowrx6g puOpu- oux6g o6qy(eg, rdicog oro europuronrd mRNA. 'Evo nol,uororpovrx6 mRNA en(o4q nepr5ler ev6o- ororpoux6g neprol6g (intercistronic regions), 6nog go(veror or4v Ell(ONA 19.47. Aur6g ot neproX6g nou<fi,ouv notr6 oe prdyeOog, on6 trr6l,tg 1-2 vouxLeor(6ro ovdpeoo oro rro6rr6vro repporroUoU ev6g no).unel6(ou rqt ro ro:6rr5vro 6vopEnq rou en6pevou nolunenn6(ou 6oq 30 voux.l,eorbro qro Borcrqprord mRNA (xor or<5prr1 ne- ptoo5repo oro RNA trov gdyrov). Ie pto or<po(o nepi rroon, 6Uo yov(6ro orrlv npoytlotrr5rrlro enrKo- trUmowor, rftor riore rlteAeuro(o Bdon qS prog ral6r- ronotrlru<qg neproXrlg vo e(vot xor 11 np6q Bdo4 tqg en6prevrlg. O opr0pr6g rrr:v prBoo<rt1.ror[ov nou eprn].6xowot orrl petdrppoorl ev6g ouyrerprpr6vou ororpov(ou e[oprdror on5 trlv ono6our6rrlro rnq 06o119 6vop(qg orrlv 5' UTR rou. H 05o4 6vop(49 yto ro nptirro ororp6vto y[ve- tor 6ro0Sorprl U6fuq ouwe0e( ro 5' drpo rou mRNA. E' | ..1r: -r-C rT1..: -+ ! il H 5' UTR nporlye(tot H 3' UTR oxotrouoei ro rou rotOtxovlou tlvop{1q xo.r6rx6vro repportopou I H dloototpovrxrl onooToon I I norx[trer ono -1 eulq -40 pooerq I l-lilg prerogpd(owot ro en6pevo ormp6vro; Apoye, or 6rd<popeg ro:6tronor4lr6g neprol6g ev6g nol,uotorpo- vtxori mRNA;rerogpd(owor ove[dpqro rl pujnog q 6r<- gpoon roug ouv65ero6 E(vor rorv6g o ;rqXovropr6g rrlg 6vop{qg yro 6},o ro ormp6vro, ri Unnoq dtorpdper yro ro npcilro ororp6vro Kcu yro ro eooteprxd ororp6vro; H perdgpooq ev6g Bor,rqprorori mRNA npoXope( 6ro6oXu<6 6ro;r6oou rov ororpov(<ov. Orov to prBoorr:- prdtro npoo6e0oriv orrlv npc0rrl ro6rronorqnxrl nepto- Xt1, or en6preveg ow(ororXeg neproldg 6ev 51ouv r<ov getoyporpef. M6]lq y(va 6ro05orprrl n 06on yto ro 6eri- repo ptBooropdno, q perdgpooq 6Xer r16q npolcop4oer yro ro roLd pr6oo oro np6to ororp6vro. Tunrrd to pr- Booropdrto reppror(ouv q prerdgpoorl qro drpo rou npcilrou ororpov(ou (rot 6roXrop((owor oe unoprovddeg) t<t tivo v6o prBooroprdrto ouvoppoLoyekor ove[dptrlro mqv op14 qg en5prevrlg ru:Stronorqttxriq neproX4g. (Or 6repyoo(eq rnq 6vop(49 Kcu rou repprorropoti 0o nqpouoroororiv de(o6txd oro Kerpd).oro 24). 19.24 I'lepiAn$n MLo peroypogrrq povddo onore}.e(ror on6 ro DNP peto[[ Tou unoKrvnrq xor rou repprolorrl, dnou oplL- J ,-.KiL'0ii0f l "iiKll + 'Evop[rl Tepgortopdg 'Evop(rl TspUqTroUdq To porqptoro mRNA nepiloltpovet operogpooTeq Kor geroqpo(opeveq neprox6q. Ko0e rodrronoullrrl neproXrl 6XeL 0 . : qq o1;to ev0pElq rot olpo repportopo0.'Evo tunrro mRNA pnopei vo eyer oprertlg rodrronorllreg neproXeq, 548 KEOMA|0 19 MeToypoqrl oroug l-lporoputrtreg
(er rot teletc0vet, owtmo[1tog, n UeroYporprl. 'Evoq x).rilvoq tou DNA ournq tr1q neptoXqg Lemoupye( o:g erproye(o ytq rrl o6v0eo4 ev6g ougnLrlp<oprotrr<o0 rl,ri:vou RNA. H uBprdtr<r1 neptoXrl RNA-DNA e(vot ppoxeio Kqt npooopw(, r<oOtig n (UeroYpoqtrrl <pu- ootr[6q" ruvekot rotd prjxog tou DNA. To o]iodv(u;to qg RNA nol,upep6o1g nou ouv06tet Bolmrlptor<6 R- NA drorp(verot oe 60o ouorottr6. O nuprivog rou ev- (ri;rou e(vot 6vo noLupep6g nou nepr6Xtt rtq unopovddeg ozgg'o ot ono(eg enoproUv Yto rnv eflt- prr6uvorl qg oluo(6og tou RNA. O nopdyowog o e(- vor Urq povtl unogovdEo onopoitry4 pr6vo oro qr66to rrlg Svop(qq yto rrlv ovoyvc0pr;or1 rou unoxtvqtf . O nuprlvoq rou ev(0pou SXet prto yevtni ouyydveto yro ro DNA. H npooOrir<r1 tou nopdyowo o petci:vet tr1 ouyy6veto tou ev(rjpou Yto pn etdtxrl np6o6eor1 oro D- NA evrir ou[dver rq ouyy6vetd rou Ylo touq unorwrlt6g' H roXrlqto pe rnv ono(o 11 RNA noLupepdorl Bp(oxet rouq unoKtwltsq qg e(vot t6oo nol'ri ;reydl'r1 dlore 6e gnope( v' ono6o0e( onLcirg oe tuXo(eg ouvowrloetg pe ro DNA ptSoo onLrlg btdXuorlg. Ot ;retoypo<ptro(nopd- yoweq nou npooetrrUouv tr1v RNA nol'uprepdorl oro DNA xot xoteu0Uvouv qv 6preoq owoMoyrl rou ev(ri- Uou UetqEU rnq Utoq n rnq 6Mnq oMrlLouX(og DNA nt- 0ov6v no((ouv p6Lo orrlv ovo(r1r4orl rou unor<wr1ti' I-1oLLo( Bor<tr1ptoro( unortvqtdq touronototivtot on6 6Uo otrLrll,ouX(eg 6 bp ortg O5oetg -35 t<ot -10 oe oxSorl Us r4v oqerrlp(o rnq perqYpoqriq. lqv ovoyvc0- prorl rou unorwqt4, coot6oo, ougBdMouv en(o4q rot 6Mq entKoupxd orotXe(o rou unoKtvnttl ovodxd on6 ro orotle(o -35 (ro orotle(o UP) r<or yUpo on6 ro orot- Xeio -10 (ro 6teupupr6vo -10 rot ot neptoX5g tcov 5to1to- protcirv) . H on6otqorl ;rero(0 rov ouvqtvettrriv oi.Lrll.ouXrc0v oXe66v ndwore e(vqr 16-18 bp. To 6v(u- pro prnope( vo rqtrUntet 5og -75 bp DNA. To opltr<6 *xletor6, 6tgep6g o0gnLoro petotpdneror oe (ovot- Xr6, 6rprep6q o6gnloro pe 6to6oXtr<rl ono6toro[q ptog oMqlouX(og -14 bp nou [er<tvd on6 qv neptoX( -10 rt eme[vetu 6ro9 nep(nou 3 bp r<o0odtrd on6 to oqpre(o Svop[qg. H ou(qgdvq nepterctx6tlro rnq oMrll'oufoq -10 oe (eUVn gdoeov A-T ouprBdLtret mrlv ow(6poor1 ono6tdro[qg. To dtpep6g o0prntroro petotp5nerot oe rpq.rep6g pe npooOri r<r1 n p66 poprov ptBovo u rLeort6(tov. Euvt10 oq oupBo(vouv noLl.oilro( t<0x).ot oventtulriv evdp(ecov, xord toug ono(oug q RNA notr'uprepdoq ouv06tet rt oneLeu0epdrvet notr0 Bpole(g xtrc0vouq RNA Xop(q vo drorpe6yet on6 tov unortvqnl. Ito t5Log outoU tou orod(ou, o o ouvriOog onetreuOepc0verot Kot o nuprlvoq rou ev(fprou nou onop6vet ro],rimet p6vo -35 bp DNA 6vowr rov -70 bp nou rol'6met oro o6ptnf,oro 6vop- (r1g. 'Enemo, o nuprivoq tou ev(0prou petorlefuot rotd prqroq rou erpoye(ou, [etuL(yowoq to DNA xqt ouv06- rowoq to RNA pretdyPogo. O nuprlvog tou ev(6ptou prnope( v' ovoyvop(oet unoruvrlt6g pe 6 totpo petu<5g ouvowetrr6 g oMrllo u1(eg un6 rqv r<o0o6riyqor1 evqtrtroKrtr6v nopoy6wtov o. Eto E. coli, ouro( ot nopdyoweq o evepYonoto0wot un6 ow((oeg ouv0{reg, n.1' 0epprx6 shock 11 6MEnl o(cinou. H yeopretp(o tou oupnh6rou RNA nofuppti' onq-unoKtvtlrri e(vot oXettrd nop6proto yto 5)to ro olo riv(u;ro. 'OLot ot nopdyoweq o er<t6g on6 tov d' ovoyvop((ouv ouvotvetu<d orotle(o nou Bp(orowot ne- p(nou 35 r<ot 10 ovodtrd on6 rr1 06oq 6vop(1q rnq Ue- toypoqrig, npqYporonotri:vtog dpreoeq enoq6q ue Bdoetq owc0v ttov orotle[tov. O nopdyowog o70 rou E' coti 6yet gro N-reLrr<ri ouroKoroorotrtu<q neptopl nou onotp6net qv ovoyvcbptorl rou DNA on6 nq neptoX6q np6o6eonq DNA. H ouroKoroorotrttx4 neptoX4 erron(- (etor on6 DNA 6rov to oLo6v(upro oXrlproiloet 6vq o- vorxr6 orjprntroxo. H *roxrls" ev6q unortvrlrl neptypd<per q ou;y6qro pe rrlv ono(o q RNA noLuprepdorl (er<wd 11 pretoypoqf . Elet((etot pre to Bo0pr6 opror6rrltog ("ouuu6p9oorl") tcov orotle(u:v -35 t<ot -10 rou unortvqt( rot 6l,Lcov ent- rouptrcbv orotle(rov pe lg t6ovtr6g ouvotvelx5g oMrl- l,ou1(eq. H opvrlttr<ri unepel,(rcooq ou[dvet tqv toX0 oprop5vtov unorwrlttitv. H pretoypotprl 6qptoupye( Oen- r<5q unep6trtreq prnpoord on6 rrlv RNA noLuprepdorl rot oqrivet opvrprr<5g unep6trtreg n(o<o qn6 to 6v(upto. To B. subtilrs nept6Xet 6vo govo6tr6 pre((ovo nopd- yowq o Ue rl'lv (6to etdtt<6tt'1rq Ue ro pre[(ovo nopdyov- ro o rou E. coli, ol'l,d en(or1g nepdXet rot notrilouq etrdoooveg nopdyoweg o, rdnotot on6 toug ono(oug evepyonotorjwot 6to6o1trd r<ord rq dtepyoo(o rrlq oro- proy6veorlq. H onoproy6veor; pu0p((mqt on6 Utq oKo- trou0(o nopoy6wtov o, ornv ono(o owtxotoordoetg rcov nopoy6wov o npoYuoronototiwot oro npoon6pto Kqr ro Unrptr6 r0rropo. Arotrou0(eg nou nepil'opBd- vouv 6to6oXtr<n Xpnon 6to<popettxtilv RNA noLupepo- ori:v prnope( en(oqg vo puO;t(oouv rn UeroYpoqrl rord rq Lo(go[q on6 BorcrrlPtorPdYo. H Borcrrlptoni RNA nol'u;repdorlteppror((et t1 peto- ypoei oe 6rio trjnouq 06oecov. Ot evdoyeve(q repport- m6q nepr6louv Uto nl.oUoto oe G-C rpoupr6to nou or<ol.ou0e(tot on6 gto ntro0oto oe U neptoXq. ln vitro ovoyvcop((owot p6vo on6 tov nuprivo tou ev(riprou. Ot p-e[optcirprevot repportor6g onottotiv tov nopdyowo p 16oo rh vitro 6oo rot in vivo. O p npoo66vetot ortg 05- oerq rut nou e[vqr n].o0oteg oe rotdtrotnq C xot <ptcoX6q or 6 oo, npolyorlwot rnq npqYuorrxrig 06or1q repuort- opoti. O p e[vot pto e[opreprlq e].rxdor1 e[optcOprevq on6 to ATP nou UeroKtvefuqt Kotd ;r1xoq rou RNA pr51pt vo grdoat orrlv RNA notru;repdor1, 6nou nporotre( 6t6mo- on rnq RNA notruprepdonq on6 to DNA. Kqt moug 60o t0noug tepponopoti, q no0oq qq RNA nol'uprepdoqq nr0ov6v ouprBdMet orov teppottop6' O owtteppotrop6g lp4otpronote(tot on6 toug tropt- Bdoerde(g qdyouq yto q pri0pton rnq npo66ou'on6 to 6vo srd6to 5r<rppooqg to.lv yovt6(tov oro dMo. Tq no- Lunpolteivtt<d orjprnLor<o nou nept6louv rrlv npotetvrl N 11 Q tou gdyou l'dpB6o 6nrr:g en(or1g rot touq nopd- yoweq Nus prnope( vo ouv6e0ofv Ue rnv RNA notruge- pdorl 6topr6oou RNA rot (oog DNA 0r1Ltdrv, qwtmo[- Xoq, Kot onorpdnouv rov repUortog6 rnq petoypogdg. To oriprnlor<o owtrepUortol.to( nou nepdlet N enttp6- reoLr-,- 549
ner orrlv RNA noLutrlepdo4 vo nopoBrdoer rouq reppo- rtm6g or ono(or Bpiorowor mo drpo rrov dtrleoov npciuprov yovr6(rov, evri rq oripnLoro nou nepr6louv Q Xperd(owot oe preroyev5orepo md6ro rqg l,o(;ro[r1g on6 gdyoug. BrBAroypogio 19.2 H Meroypoqrl flpoygoronore(ror Ue Zeuyopo1l0 Booeov oe plo u0uootrrdo, A(euydprrlTou DNA Avoordnq*q Losick, R, and Chamberlin, M. (1976), RNA polymerase, Cold Spring Harbor Monograph Series 6. 'Ipeuva Revyakin, A., Liu, C., Ebright, R. H,, and Strick T. R. (2006), Aborlive initiation and productive initiation by RNA polymerase volve DNA scrunching. Science, 31 4, 1 1 39-1 1 43. 19.3 H AvriDpooq Tr1q MeToypuQflq'EXer Tp(o Irddro 'Ipeuva Rice, G. A., Kane, C. M., and Chamberlin, M, (1991). Footprinting analysis of mammalian RNA polymerase ll along its transcript: an alternative view of transcription elongation. Proc. Natl. Acad. Scr. USI 88,4245-4281. Wang, D. et al. (1995). Discontinuous movements of DNA and R- NA in RNA polymerase accompany formation of a paused tran- scription complex. Cell 81 ,341-350. 19.4 H Borqprorrl RNA llotrupepool Anoretrelror ond l-loii6q Ynopovodeg !.i ::'.': t. {.:, t.:: :. : i ; Helmann, J. D and Chamberlin, IVl. (1988). Structure and function of bacterial sigma factors. Annu. Rev. Biochem. 57 ,839-872. Shilatifard A., Conway, R C., and Conway, J. W (2003). The RNA polymerase ll elongation complex. Annu. Rev. Biochem. 72, ovJ-/ I 3. Campbell, E, A., Korzheva, N., Mustaev, A., Murakami, K., Nair, S., Goldfarb A., and Darst, S. A. (2001). Structural mechanism for rifampicin inhibition of bacterial RNA polymerase. Cell 104,901- 912. Geszvain, K. and Landick, R. (2005). The structure of bacterial R- NA polymerase, ln The Bacterial Chromosome Higgins, N. P (ed.). Washington, DC American Society for Microbiology Press, pp. 283-296. Korzheva, N., Mustaev, A,, Kozlov, M., Malhotra, A. Nikiforov, V., Goldfarb, A., and Darst, S. A. (2000). A structural model of tran- scription elongation Science 289, 61 9-625. Vassylyev, D. G., Vassylyeva, M. N., Perederina, A., Tahirov, T. H , and Artsimovitch l. (2007). Structural basis for transcription e- longation by bacterial RNA polymerase. Nature 448,157-162. Zhang G., Campbell, E. A., Zhang, E. A., lVlinakhin, 1., Richter, C., Severinov, K,, and Darst, S. A (1999). Crystal structure of Ther- mus aquaticus core RNA polymerase at 3,3 E resolution. Cef 98,811-824. 19.5 To 0tr06v(ugo Tl1q RNA l-lotrugepoonq Anorele[ior 0n0 Tov I'luprlvo rou Ev(upou Kor rov n0p0y0vT0 Iry1to 'Ipeuvo Travers, A. A. and Burgess, R. R. (1969). Cyclic reuse ofthe RM polymerase sigma factor. Nature 222,537-540, 19.6 l-ltig Bpiorer H RNA llotrupepoon rtq Atrtrr1trouXieg rov Ynorrvrlrdrv; ,Avoordnqoq Bustamante, C,, Guthold, M., Zhu, X., and Yang G. (1999). Facili- tated target location on DNA by individual Escherichia coli RNA polymerase molecules observed with the scanning force mi- cr0scope operating in liquid. J. Bio. Chem. 274, 16665- 1 6669. 19.7 To Otroriv(upo Tpononorelrot Koro q Arepyoo[o Avoyvroprolg rr Anetreu06proonq 0rr0 Touq Ynorrvqr6q 'Ipeuva Bar-Nahum, G. and Nudler, E. (2001). lsolation and characteriza- tion of sigma(70)-retaining transcription elongation complexes trom E. coli. Cell 106, 443-451 . Kapanidis, A. N., Margeat, E., Ho, S. 0., Kolckhonjia, E., Weiss, S and Ebright, R. H. (2006). lnitial transcription by RNA poly- merase proceeds through a DNA-scrunching mechanism. Sc" ence 314,1144-1147 . Krummel, B. and Chamberlin, M. J. (1989). RNA chain initiation by f. coli RNA polymerase, Structural transitions of the enzyme ir early ternary complexes. B i o ch e m istry 28, 7 B2S-7 842. Mukhopadhyay, J., Kapanidis, A. N., Mekler, V., Kortkhonjia, E.. E- bright, Y. W., and Ebright, R. H. (2001). Translocation of sig- ma(70) with RNA polymerase during transcription. Fluorescence resonance energy transfer assay for movement relative to DNA. Cell 106,453-463. Wang, 0., Tullius, T. D., and Levin, J. R. (2007). Effects of discon- tinuities in the DNA template on abortive initiation and promoter escape by E. coll RNA polymerase. J. Biol. Chem 282,26917- 26927. 19 B 0 ilopoyovrog l(ypo EtrrlyXer qv ilpocdeol 0T0 DNA Avoyvop((ovroq IuyKeKprp6veq Atrtrr'1trouXieq rov Ynonvqrtlv Haugen, S. P., Ross, W., and Gourse R. L (2008). Advances rn bacterial pr0moter recognition and its control by factors that d0 not bind ONA. ffafure Rev. Micro.6, 507-520 McClure, W. R (f 985). Mechanism and control of transcription initiation in prokaryotes. Annu. Rev. Biochem. 54,171-204. Bar-Nahum, G. and Nudler, E. (2001). lsolation and characteriza- 550 KEOAAAI0 19 Meroyporprl oroug l-lpoxopudrreq
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complex containing the lambda N protein. Genes Dev.9, 2831- 2845 0 son, E. R., Flamm, E. 1., and Friedman, D. l. (1982). Anatysis of nutR: a region of phage lambda required for antitermination of transcription. Cell 31, 61-7 0. 19.23 0 KUrIoq Tou BoKTnploKori Ayyetrrogdpou RM Brenner, S., Jacob, F. and Meselson, M. (1961). An unstable nr::- mediate carrying information from genes to ribosomes for pro- tein synthesis. Nature 190, 576-581. BLptrroyporpLo 553
@ Carol & lVlike Werner/Visuals Un - :: - H Meroypogt1 oTouq EuKopudrreq ffi Erooyoyrl * lto vq npoode0e( orov unorrvlrrl 11 RNA notrupepool npenet v' ovo(er 1 XpoUouvrl. 0r Europuorxeg RNA l-lotrupepdoeg Ervor flotrupepr1'Ev(upo * H RNA notrupepool I ouvO5rer rRNA orov nuprlv(oro. e H RNA notrupepoorl ll ouv0erer mRNA oro nuprlv6ntro- ou0. s H RNA notrugepool lll ouv06rer ptrpo RNA 0To nupn- vtlntroopo. * 'OtreS or europurrltrreg RNA notru;repooeg 6Xouv -12 unopovddeg rol e(vor oupntroro rtov -500 kD. * Meptx6q unopovodeg eivor rorvrlg Kor onq rperg RNA notrupepooeg. s H Ueyotr[rep1 unoprovodo qg RNA notrugepoorlq ll 6XeL pro CTD nou onore),e(ror ono notrtrontr6g enovo- trrlgerq evoq enro;le pouq. H RNA l'lotrupepdor; 'EXer evo ALpepq Ynouvlr4 c 0 unoxrvlrrls iqg RNA notrupepooqg I onoretre(rot onrj 6vov xevrptrd unoxtvrlrr; rot 6vo ovodtro orotXe(o rou unouvlrrl (UPE - upstream promoter element). * 0 nopdyovrog UBF1 rutrryet DNA yupto onri pto npo- teivrrrl dopr; yro vo rp6pet rovro rov revrprro urroKrvrl- rr1 ror ro UPE. s 0 SL1 nepiloppovet rov nopoyovro TBP nou egntrexe- r0r oTrlv 6vop(q ror onrl lg rperg RNA notrupepdoeq * H RNA notrupepoorl I npoodeveror oro oupntro<: UBFl -SL1 orov revrpmrl unorrvrlrr;. ilFry5. *:, F,F ffi H RNA notrupepool lll Xprlorponorel T6oo Ko0o6rroLr '0oo ror Avodxoug Ynorrvrlr6g * H RNA notrupepoorl lll 6Xer 6uo runoug unouvlrolv. e 0r eou;reptro( unorrvrlr6g eXouv ppoXeieq ouvorvsT.- xeg otrtrqtrouX(eg nou pp[orowor luioo oTn peToypoq.- rq povodo ror nporoLouv rqv 6vop[q 0e Urc ouyrerpq6vrl ondoroorl ovodxd. e 0r ovoduo[ unorrvlr6g nept6Xouv rperg ppoye[eq ou- vorvertreg otrtrrltrouxLeq ovodrro rrlg orperrlpiog oric ono(eg npoo66v0vT0r UsToyp0qxoI nopoyovreg. * 0t TFrrrA ror TFrrrC npoodevovror ong ouvorverx6g otr- trqtrouXLeg rt enupenouv qv npdodeorl rou TFlllB o-qr oqeqp(0. * 0 TFrrrB rp5per cog unopovo0o rrlv TBP ru enupdnel qr npoodeol rrlq RNA nolupepoolg. H Aqeqp(o yro rqv RNA l-lotrugepoorl ll * H RNA notrupepoorl ll yro v' opX[oer Tn UeroypoQr onoue( yevlrouq UeToypoqrKoUg nopoyovreq (ovolro- (ovrotTFlfi). ryE*3 554 KEOAAA|0 16 Iuoqporo EnLdtop0uloqg 20 ji ,$- .:$.'j t ,.1 {.!.: 5i .* .*: ..!' l-
ffi H TBP ervor0txoupevxdg l-lopriyovrog ffi ffi H Bootrrl luoxeurl luvoppotroye(ror orov ynorrvqrrl * 0r unoxrvrlrdg qg RNA notru;repooqg Il oqv oqeqp(o ouXvo 6Xouv pro ppoXeio ouvrrlpq;rrivrl otrtrr1trouX(o PyzCAPys. e To xour[ TATA e(vor rotvo ouorolrd rov unorrvrlrov rqg RNA notrupepooqq ll rur onoretre(ror ond evo ntrouoro oe A-T orropep6q nou pptoxeror -25 bp ovo- 6xo r4g orpeqp(og. * To DPE elvor rorv6 ouorortrd rov unouvqrov r4g RNA notrupepoqg ll nou dev nepr6Xouv rour(TATA. * 'EvoS revrpxog unouvqrrlg yro qv RNA notrupepoo4 ll nepilolrpover ro lnr rol, ouvr1Oog, efue 6vo rour( TATA e(re 6vo DPE. Mnope( enro1g vo nepteyer roL dtrtro eLdooovo ororXe[0. * To orroUepn ru.rv torovdrv rpononoro0vroL np6oxorpo rord q perorivlon Tnq RNA notrupepdoriq. * H CTD ouvrov[(er r4v enetepyoo(o rou RNA Ue q Ue- r0yporpq. a Av ouUpouv ptroBeq oro DNA, 0o enr6top0rrl0orjv rord nporiprlol ro peroyporpopevo yov6to. * 0 TFIIH nop6Xer ro ouvdeopo yro evo oupntroro ev(u- Uov enrDt6p0oonq. ffi O,EvroXur6g llepreXouv Aggi6popo lrorle(o nou ynopoq- Ootjv r4v'Evop[1 * 'EvoQ evLoXuqg evepyonorei rov unorrv4rq nou pp(_ oKeror rr0 xovro rou rot pnopef vo ppfoxerot 0e olror_ od(nore onoorool e(re ovodrro e(re roOo0uo ono rov unoxrvrlrrl. * Mro AUS (ovodurl evepyonorlnx4 otrtrritrouXro) rou (u- popUrrlro oupnepup6peror oov evtoXurrlq, otrtro trer- roupyel p6vo ovodrrd rou unoxrv4r1. o 0r evroxureq oXrl;ror((ouv oupntroro on6 evepyonot- qr6g nou otrtrqtrenLdpor1v opeoo 11 6ppeoo Ue Tov uno- xrvr1rr.1. ffi O, EvroXurdg AeLroupyorlv Au[dvovrog 14 luyr6vrpororl rov Evepyonorrlrov Kovrd orov ynoxrvqrrl * 0r evroxur6q ouv4Otrlg treuoupyouv prlvo oe cis 6Lo- UdpQoo1 oe oX6oq Ue Tov unoKrvt1rr1-oroXo. * Ko0e evtoXurqg unoXpetirveror vo treuoupye( pdvo 6rov ppe0ei xovrd orov unouvrlrrl. ffil H 'Erqpoor'1 rov lovrd(tov lXer((eror pe qv Anope8utrito- on o H onope0utr[too1 oro 5' dxpo rou yovr6(ou e(vot qno- po(qrr1 yro t1 peroyporprl. ffi O, Nrlo6eg CpG E(vor Pu0ptorrr<o[Ir6Xor * 0r vrloOeq CpG nou ppforovroL oroug unortvrlrdg rov r6rootjororo ertppo(dpevov yovrd[tov e[vor onope0u- truop5veg. * Nqo(6eq CpG pp(orovror enfo4g Kor oTouq unouvrlreg op toluivtov yovtd frrrv pe roroe16 rrrl rixrppoori. * To ovOpdlnrvo yovt0(rrllro nepr6Xer -29,000 vrlo(6eq cpG. o H Ue0utrioon prog vrlo0og CpG onorpener Trlv evspyo- no(qo1 rou oXert(dpevou unoxrvrlq, a Koroorotrr1 nporotre(ror ond nporeiveg nou npood6- vovTor or pe0utrrop6vo (euyn CpG" ffi nepiinrpn e H TBP e(vol ou0rolx6 rou nopdyovro rono06qoqg nou onoue(ror yro qv npdodeoq roOe RNA notrupepd- orlq oTov unoxrvnrq qg. o 0 nopoyovroq yro Tnv RNA notrugepoon lll e(voL o TF1D, nou onoretre(ror ond TBP ror -14 TAFs, ;re ouvotrurl Uo(o -800 kD. * H TBP npoo66veror oro rour( TATA oqv etroooovo ou- troro rou DNA. * H TBP oX4pol(er pro o6tro y6po ond ro DNA ror ro ropnrer roro -800. * To ovo6trd ororXe(o xor ot nopdyovTeq nou npoode- vovror 0r ouro oulovouv 14 ouXv6qro r4g evop(r1g. * H np6odeon rou TFrrD 0ro rour( TATA 11 oro lnr e(vot ro npdro p4;ro rrlg evop[r1q. a Atrtrot peroyporprro[ nopoyovreg npoodtivovror oro oupntroro ge xoOoproprlvn oerpd, enexre[vovrog rqv npoororeupdvq neproXrl oro DNA. * H RNA notrupepoorl ll opX((et rq peroyporp4 pdtrrg npoode0e( oto oupntroro. H'Evop[1 Arotrou0efrot on6 Ko0oporl rou ynortvqrrl rt Enq1xuvorl * 0r TFrrE rot TFrrH onououvror yro rrlv r4ir1 rou DNA ror rr1 peroxfvqol qq notrupepdonq. * [t0 vo [errvrloeL 4 ro0oporl rou unorrvrlrrl ror 11 enqrrl- ruvorl npener vo rpootpoputrtro0e(11 CTD. * Ie pepuoUg unorrvrlr6g, yro vo repporiorr rl ovenrTu- Xrlg 6vop[1 lpero(erot enrnpooOer4 gootpoputrioorl qq CTD. 2O,l Erooyrrlyri e lro v0 npoode0er orov unoxrvqq q RNA notrupepool np6ner v' ovol[er 1 Xpoporivrl. H 6vop[q rnq UeroypoAnq o' dvo er<proye[o Xprr;prorG vrlq nou e(voL 11611 ovotXr6 onotre( np6o6eoq rrlg RNA no).upepdoqq orov unoxrvrlll Kor Ueroyporpxu_lv no- poy6wotv orouq evroxurdg. /n vitro, preroypo<pq o, 5vo er<proye(o DNA onore( pro 6roqoperu<4 unooprd6o geroyporprx<ilv nopoy6wov qn6 exe(voug Trou oTror- roUwor yro rn peroypogri rou erpoye[ou rnq Xpopo- rfur1g (oro KeqdLoro 28 0o e[erdoou;re nc6g ovoryer q Xpropror(v4). KdOe nprrlteivq onopo[rr1r4 yto qv 6vop- En qq petoypogrlg, q ono[o 5prog 6ev e(vor 11 (6ro rpnpo tqg RNA noLuprepdoqg, op((eror oq Ueroypo- gtr6g nopdyowog (transcription factor). l'1oMoi prero- ypogrxo( nopdyovteg 6pouv ovoyvrop[(ovrog cis-evepyorlg 06oerg oro DNA. H np6o6eoq qro DNA, to016o,0, 6ev e(vor o provodu<6g rp6nog 6pdorlg ev6g UerqypogtKo0 nopdyowo, o ono(og prnope( v, ovo- yvopr(er 6vov dMo nopdyowo ryrqv RNA noluprepd- l:i.1 Erooyoyri 555 Boorxeg eworeg
on, 6noq en(onq Kor vo evoopor6verqr USoo o'6vo oUpnLor<o 6vop[r1q p6vo nopouo(o oprercitv 6Mov npcoreivcbv. H r<p(or;rq 6orrproo(o yro rn ouprpretopl qrn Ueroypogrrrl ouoreurl e(vor Lemoupyrrrl: yro vo Bp(oreror puo npore(w1 o' 5vo ouyr<erprprSvo unoKrvrl- fi 11 o;rd6o unorwqrriv np5ner vo e(vor onopo(rqt4 yroq peroypogrj. Mro o4gowu<{ 6ro<popd peto[ri tqg pretoypoqrlg Tov euKopuoru<rilv ror rov rrpoKopuorxrilv RNA e(vot 6tt oto prev Bor<rrlpro n Uerqypoerl npoyproronore(rot o' 6vo er<proye(o DNA, evrir orouq euKopurilreg o' 5vo erpoye(o Xpro;ro{vr1g. H Xptoprodvq oMd(er ro ndwo ror np6net vo LoprBdveror un6rlq oe r<d0e FriUo. H Xpro- porrvn npdner vo Bp(oretor oe ovoffi 6opu] xor, or6prrl xot t6re, to or,nopreprl rolv voux.Leoocopoilcov npdner vo 51ouv onopor<puvOe( on6 rov unorwrlrl 6ore vo gno- p6oet vo npoo6e0e[ 11 RNA no].uprepdo4. Aur5 preprrdg gop6q npouno06ter preroypoqri on6 6vo ouon4pd r'1 rpuq6 unorwrlrrl, efue orov (6ro e(te srov qwrouv0etr- 15 x).rirvo. Mro 6e0repq o4prowrrf 5ro<popd eivor 6rr n BoKrn- proxrl RNA nolupepdo4, Xdpn orov nopdyowo o nou 6ro06rer, pnope( vo dtoBdoet qv oMrlLouX(o rou DNA ytcr vo Bpe( rov unor<tvrpf Kqr. vo npoo6e0e(, evc0 pro europuamr{ RNA no},uprepdorl 6e ;rnope[ vo droBdoer to DNA. l-r'out6 11 6vop(q o' euKopuoru<or3g unoruvrl- 169 nepil,oprBdver peyd).o oprOpr6 nopoy6wov nou np6ner vo 61ouv npoo6e0e( oe pro nou<tl(o cis-evepy6v ororle(ov npoeroq.rd(owoq ro 66o9o9 yro rnv np6ode- on rnq RNA nolupepdonq. Awo( or nopdyoweg ro- LoUwor poorxoi UeroypoglKo[ nop6yovreq (basal transcription factors). M6).rg oupBe( ow6, q RNA no- ).ugepdo4 npoo66vetor oro oUpnLor<o Boorr6v peto- yporprrcbv nopoy6wov/DNA. H ouyrer<pr;r5vq neproXrl np6o6eoqg op((etor og xewprx6g unoruv4r{q (ri nu- pnvoq rou unoKrvrlrrl, core promoter) xor nept6ler 6leg trg onopofuqteg 06oerg yrq rrlv np6o6eor1 ror rq Lemoupy(o r1g RNA nolu;repdoqg. H (6ro q RNA nol,u- pepdorl npoo6Sveror y6pco on6 qv orper4pio (start- point) rqg peroypogrlg, qI d 6ev 6plerot oe dpreorl eno<pr1 Ue rnv er<retoprdw1 ovo6xrl neproXrl rou uTroKt- vrprl. Aw(O*o, or Borcrlproro( unoxwqrdg nou ovoLrl- 04rov oro KerpdLoro 19 ro0op[(owor r<up(rog oe oXSoq pe q 05o4 np6odeoqq qg RNA noluprepdorlq ornv neptoXq rowd mrlv o<peqp(o. Evcil to Borcnjpro 6Xouv pr6vo puo RNA noLuprepdorl nou preroypdger. Kor lq rperq r<ripreg td[erq yovr6(rov, oro euKqpuorxd rUrropo q preroyporp4 unodropekor oe rperq xoulyop(eg. Kd0e r<oqyop(o peroypdgetor on6 pro drorpopeur<q RNA nol,uprepdoq: . H RNA nolupepdoq I geroypd<per ro 18S/28S r- RNA. . H RNA noLuprepdoq ll preroypdqet mRNA ror Lryo prrpd RNA. . H RNA nolupepdoq lll peroypdger ro IRNA, to 55 ptBootopu<6 RNA r<or dMo ptxpd RNA. Aurr1 r4v err6vo SXoupe yro lq xripeg rd[etg rrrlv yovt6(orv. Onog 0o doripre oro KeqdLoro 3C :::::: reg ovoxoL0qlerq ond Bo0ptdcordq ouorolliec :' :. - pou rou yovr6rrlrporog (whole genome t I 'c ,-". . onor<d).ur.l.rov 6vov v6o rdopo owrouv0errxr.,.lv *:r r , - . tpcov (antisense transcripts), preooyovLdror<cov -:.r, : : rporv (intergenic transcripts) ror peroypdcpc, :-:: :- Xpropror(vqg (heterochromatin transcripts). Ae ,,' . - - -: - pe or6pq t(noro oXerLro Ur rouq unoKrvfro( l--- rorv rdl,eov 11 yro rr1 priOprorl roug, yvcop[(or*, ---: 6t notrl.d (Ud).).ov ro neproo6repo) ond ro c, , .:.: trr6vo petdypo<po nopdyowor on6 qv RNA t:c: -*.:: oq ll. Or Boorr<o( UrroypoqrKo( nopdyoweg e[vcL :-:: - rrlror yrq qv Svop[r1, oMd or neproo6repoL 5: ,.:: d(owor orrl ouv51ero. l-ro rLg rperq euKoputrtrL<.: :',,,- no).upepdoeq, or Ueroypoqtro( nopdyoweq. r:::- -: (6to to 6v(upro, e(vor unerjOuvor yro rrlv ovoyvog.l- -: oM1l.ou1(og DNA rou unoxrvrlrrl. I-ro 6).eg r.c : -. : purrrflr<5q RNA no).uprepdoeg, oL BooLro( pero.:- .: nopdyoweg 6qpuoupyoriv pro 6oprr; fiov unoKt,,-:- -: pdlowoq ro 0161o nou ovoyvop((etot ono :- . = " - no).upepdo4, l-ro rLg RNA no).u;repdoeg I roi :--: or nopdyoweg e[voL oletrr<d on].o(, evol yro :- , :', - no).upepdorl ll ouyxpotoriv 1,.uo opr<erd peyo.I.r :*::: Or Boou<o[ nopdyoweg evcovowot pe rnv RNA r:' - -. pdoq ll yro vo oX4por(oouv 5vo orjprnLor<o nc" -:: Bd).).eL rnv qeernp(o xor ro0op((ouv rq 0ec- -- 6vop(qq. Or Boorxo( nopdyoweg po(( ;ie qv R: -- Lupepdoq onoteLorjv q Boorxl ;reroypoqLr<1 ouci. -- Or unorrvqrdq yro rrq RNA noLuprepdoeg I xc. i: oKovror (r<up(orq) ovodrrd rnq oeernpiog. Ar,tL:. - noMo( unorrvrltSg qg RNA no).u;,repdorlg lll Bc.;. : roL Kq0odrxd tqg otpeqp(og (p6oo orr1 Ueroypc- . povd6o) . Kd0e unoxrv4rrlg neptdler XopoKrrlpr:...: oprd6eg BpoXeLo.lv ouvtrlplpr6vcr.lv oLLqLouxro . -: ovoyvrr;p((owoL on6 rrlv KotdMrl^n rd(n Boorr<i", -." toypogu<rov nopoy6wov. Ot RNA nol,upepdoeg . : ovoyvrr; p ((o uv puo crlettr<d neproproprdvrl o ;rd6 o r- : : . vq-rctrv r<or Boo((owot oe puKp6 opL0p6 enlKouptKr, -: poy6worv. Or unorrvqt5q ilou XpnortrlonoLorjwor ond rqv' :',: noLupepdoq ll eg<pov((ouv preyoLUtepq noLrt).opro:: - orqv oM4).ou1(o r<or opyovrr.rvowor on6 6rdgopc 3:::- -rrlproro (modular organization). 'O).oL or uiloKrvrlrr. :- - RNA noupiepooqq ll nepL).opBdvouv oMrl).ouxrec . : rd orqv ogetqp(o mrg ono(eg npoodSveror q pc, .- ouor<euq rot q noLuprepdorl r<o0op((ovrog rq ii-- 6vop[r19. A].].eg nro onogoKpuoprdveg otrtrq,o1,, : : ovodLrd (q ro0odxd) roo0wot evrolur6g (enhancer r<or r<oOop((ouv ov 0o er<<ppoore[ o unortv4qq Kor, /.-- p[rog, ov q 6x<ppooq rou 0o e(vor oLxoupevu<r1 q r<urr:- po-et6txrl (cell type-specific). O evLoXutrlg e(vot 1-l: L6rofrep4 05or; nou epnl.dxeror crq Ueroypogq xor rcr- ronorekot on6 qv nopouo(o otrtrr1trou1tiov or onorri dLeye(pouv rqv 6vop{r1 oMo ewon[(owor oe 6Ldgope I onoorooetg ond rov rewpu<6 unorrvqrrl" Irorle(o ra. evroXurcov oulvd e(vor mdloL yro roroerdrxn n XpovLxr pti0;uo4. Meprxo( evroXurdg npoo6Svouv peroypogt- 556 I(EOA Al0 20 H Meroypog4 oroug Europudlreg
EvLoXurnq Ynouvn"rnq -1 00 bp, nEPtdxet oPKtTeq notrri r<ovrw6q 06oetg npdo6eorlq Yto UeToYPoQtKoIq ono rov xevrptro unoKtvlrll Unopo( vo xuPoiveruL onri 1 006eq bP PdXPt kb H onootoorl rtrlv evloxUT{iv +-1 00 bP, nePtdXer dtdonoPreq ohtrr1)rouXieq nou nPoodevouv . rn Booun ueTqYpqQtKn ouoKturl 2O.2 Ot Europuortr6q RNA lloAupepdoeq Eivol floAuUEPfi 'Evquuo nopoyovreq , : . , .Evo runrxd yovioro nou peroypoqeror ono rrlv RNA nou;repooq ll exer 6vov unortvrlrrl r10u eK1e[veTot ovobtro rnq oeolc ln. onou (errvo 11 Ueroyporpn. 0 unorLvrlrflg nepulXet op*rii BpoXeo (-iO np) o,o,Xiiu otrtrntroux(oq.nou npooo6vouv preroypoqtro0g no- j0y0w0q, oroonopro oe novrrr ono -100 bp..Evoq evroxurflg nou nepr6xer p,o n,o'or1.,r,oyt1 ouorotx(o orotxe[ulv nou en[olq npoootlvouv -eroypoqrrouq nopoyovreq, pnopei vo ppiorerot o, onioioJn,.i*r,,i,u i*o,ouro6orv Lp eolq opxerd'v kb (To DNA unope( vo elvot etrtrul- Jsv. | 0v06r0Toyp6vo pe otrtro rpono eror 6ore or usT.yp.QrKoi nopoyoweq orov unorrvqq K.r T.v rvr'xuTn v otrtrrrtrenrbpo.v titore vo :xf Uorloouv 6vo peyotro npolreivtr6 o[pntroro)' <orjg nopdyoweq nou treLtoupyofv pre o]']'r1]'enL6pd- otrq'U,*ptlq'eprBetreLoq r<ot Bp(ot<owot xowd orov uno- .,ul.h ,u, dLLo, prnope( vo Bp[or<owot oe on6oroot'1 ,<lL,iolu (euyrr-rv B&oeolv. H r' ''r ",,' '" ' nopouotd(et ttg :iru,o6q rdt6trlteq rr,ov unoKtvrltotv xot trov evtoxuttilv' r"ll.o pugU,*tr<r1 Oeoq nou npoo65vet nepLoo6tepouq "o"ni*o,is nopd Oertxo0q pu0gtor'59 Ylo..to' dI'eYXo iq !,t oypoQnq r<qtrekot onootonrp4g (silencer)' 'OL unortvrlr6q nou er<ppd(owot t6tooUototo t<ot e[- '.'ot onopoittltot oe 6tro to t<trropo (ro yov(5to rouq Ue- .,^X ,p"pei ovocptlpowot tr;g 6lolerptcrtx6 yovi6to' ntrt'"r"irping genes) tixouv ovo6trd orotxe(o ol'trq- r.cuxloq nou ovoYvcDpiqo*ot on6 otroupevtrotiq evep- . onotrlt6g (u biq u itous activato rs)' Kovtlvoq ouv6uoopr6g -or'lriorTt opdyowo 6ev e[vot onopo(tqto ouorotLr6 1., uno*,rn-n, yeyov6q nou uno6qtrtovet 6rt q evop[r1 :no tqv nNl no).upepoorl ll prnope[ vo pu0prtore( pe .oMolq dtotpopettro0g tp6nouq Ot unortvrlt6g nou t.,ppa(o*o, un6 etdLr6g ouv0r1<eq dlouv orotle[o o]'- i."1ou1(og nou onotto0v evepYonotqr6q 6ro06otpouq -ouo or,q ouyr<er<ptp5veq ouv0rlreq' -- , "pVarroq ev6q evtolurl 11 onootonqrq protd(et -t -nu !*t*otXq ev6g unot<tv1tr1' 6qodq.onotetre(tot l:ro 1.uo notrA(o e(opqpotcov 6nou prnope( vo npoo6e- 3oiv 0ettxo( q opvqtir<o( pu0;rtot6q oe oqtXtd ou- ,otuooUgu.l ouorotl[o Ot evtcrlut6g 6e Xpero(etoL vo .o(orowot rowd orov unortvrltrl' Mnope[ vo Bp(ot<ov- .or ovo6tr6, p6oo o'5vo yov(6to 11 nepo on6 to 6pto avoq yovt6[ou Kot o npooovoto]'topr6q toug o'letLrd pe -o yov[6to 6ev 51et,o1proo(o Ot npulte[veq flou npoo- 5evowot oe orotxeto iou evtoXurl oMrlent6po0v pe .pr-"iv"s npoodiaepteveq oe orotxe[o tou unortvlrq' ro0 ouXvd ptSoto ev6to;reo0lv nou ovo;'to(owot ouve- vcpyonorrlr6g (coactivators)' ' H pretoypotpq tov europurrltcilv un6rettot TIto ouxvo ;e Oettxll p,igU,o.t. 'Evoq petoypoqtr6q nopdyowoq -opeXt-o, un6 toroerbtxd 5}'ey1o Yto vo evepyonotrloet ,uou unoo,untrl q prto ogd6o unorrvqtolv nou nept6Xouv .rto r<otvr1 otrq}'ou1[o or6Xo Aurl ri 6tepyoo(o nept- Lo;rBdvet opretd Briproro: (erw6 pre,dvotytrro rnq Xpo- ,.ii*., ro'ono(o orol'ou0ef 11 np6o6eor1 tov Booxriv ;;;l;;v nopov6wtov xor ^qr6nw qq nol'uprep6- [[;:'[Eeu,on r-,t et6txrl rorooro]'ri ev6g unor<rvr1t(- or6Xou e(vot trtY6tePo r<owt1' Mro euxopucottKn UeroYpoqrrci govd6o levtrd ne- or6xet 5vq uovo6tr6 yov(6ro Kot o repportoUoq flpoYUo- i;,i;;;,';epo qnb to t5trog rqg rtodtronotqtrr<r1g neptoXfq. O rep;rort4t6q 6ev 6Xer 16oo UeY6In puO;tt- *l^n'iihu*(o'6oo oro TlpoKoputotu<d ouorrl goto' ot nrun'r,oLugepdoeg I r<ot ll teppot((ouv oe 6tor<ptt6q otr- iniouxfts'oe xo0optop6veq owr6p6o.ets' op6 o tp6- ;;;;AuL-,opro6 rrlg'RNA notrugepdorlq ll 6ev e(vot oo.inq. lldwoq to <n1powtr6 ougBdv orrl dlptoupy(o ior-ilaopou ev6q mnruR 6ev e(vot ow6q ro0out6g o tepproro;r6q oM6 prto ow6poorl nS$nq rou rporoYe- uo,iq UetoVpdqou (BL6ne KeQdtroto 21)' " H RNA notrupepoorl I ouv0eret rRNA orov nuplvtoKo' * H RNA notrupepoorl ll ouv06tet mRNA oro nuprlvontroo;t0' s H RNA notruuepoon lll ouv06ret uLrpo RNA oro nuprlvontroolto' * 'OeS ot europuortr6q RNA notrupepooeg 6xouv -1 2 unopo- vodeq rot e(vot ouptntroro rcov -500 kD' - 1y1a,*ei unopovodeg e(vor rotvrlg Kot oTtq rpetg RNA notru- Uepdoeq. * it ueyor[repr1 unopovodo qg RNA notruuepoonq Il 61eL pto CTD nou onoreLelrot ono notrtroneg enovotrlQetg evoq enroUePoUg. Ot tpetg europutottxSq RNA noLu;repdoeg Bp[oxowoL ot oio,popt trSg 06oetg orov nuprlvo' ot ono[eq owt- 20.2 0r Europuortr6q RNA tlohuuepdoeg Eivot llotruuepn'Ev(uuq 55'',
ororxofv oro droeoporrKd yov(5ro nou Ueroypd- 90UV. H evepy6rrlto nou (eXop(et neproo6repo e(vor qg RNA nol,uprepdonq l, 4 ono(o ewon((eror orov nuprlv[- oro (nucleolus) r<or ovoLoprBdver yro rrl preroypogri rtov yovr6frov nou rto6rronoroUv ro 18S ror 28S rRNA. E(vqr uneU0uvn yro 14 orivOeol qg geyoUrepnq no- o6qroq RNA rou rurr6pou. To dMo rUpro dv(upro e(vor q RNA noLuprepdorl ll, nou Bp(oretor oro nupqv6ntroopro (nucleoplasm) (ro rpnpo rou nuprlvo ercr6q rou nuprlv(oxou). Awrnpo- oone6et ro geyotrUrepo prdpog tqg un6Lornrlq Kurro- pu<{g evepy6rnroq Kor e(vqr uneUOuvn yro 14 oUv0eo4 rov neptoo6reprov on6 ro erepoyev{ nuprlvrx6 RNA (heterogenous nuclear RNA - hnRNA), nou e(vor np66popro yto noMd mRNA ror 6Xr pr6vo. O rLoou<69 opropr5g rltov 5rr ro hnRNA nepil,oprBdver ro ndwo mov nup{vo ercr6q on6 ro rRNA ror ro IRNA (en(oqg, oritrrgovo Ue rov r.troorr6 opto;r6, mRNA Bp(orerot pt6- vo oro r<urtop6ntroopo). H e[5toor1 rou hnRNA pe oriylpovo ;ropror<d epyoLe(o onorol.0mer noLl,d RNA nou 51ouv prev purprl ouyx6wp<ooq oMd peydL4 oq- proo(o, 6nog en(oqg ror noltrd dLLo RNA rrov onoftov q l,enoupy(o pr6lrg op1(ooge vo rorovooUpe. To mR- NA e(vor to Lry5repo dgOovo on6 ro rp(o rUpro RNA Kor owrnpooone(er p5tg 2/"-5% rou rwtoponl,o- olloru<o0 RNA. H RNA notrupepdoq lll e(vor 6troooov 6v(upro on6 qv dnor,Lrq qg evepy6qroq, oMd nopdyer 5vo orlvoLo oro0epc0v, onopo(rqrov RNAs. Iuv06rer ro 55 rRNA, to IRNA ror dMo puxpd RNA nou onoretrofv ndvo on6 6vo r6topto rov Kurropontrooprorxcirv RNA. 'Otreg or europurotrr6g RNA noLupepdoeg efuor pre- ydtreg nprrlreiveq nou Letroupyoriv og o0prnlor<o -500 kD. Tunxd 51ouv -12 uno;rovd6eg. To r<o0op6 5v(u;to prnope( v' ovo).dBer e(oprtir;revq on6 ro er<proye(o geto- ypoqn rou RNA, 6e prnope[ 6puog vo npor<o].6oet 5vop- [r1 ent].ercttxd orouq unoKrvnrdg. H RNA no].upepdoq ll tou Saccharomyces cerevisiae e(vot lopoxtrlptmtr6 nopd6erypro yro rq yevx4 oUoroorl tov euxoputottxc0v ev((;rov Ue rrl ouyKexptpr6vq evepySrlro Kcu ilopou- ord(eror orqv E|(0NA 20.2. Or 6rio ;reyotrrirepeq unoUo- vd6eg e[vor o;r6],oyeg pre rtg B r<or B' unopovd6eg qg Borcrqprox{g RNA notruprepdonq. Tperg on6 trg un6trot- neg unoprovd6eg efuor rowdg oe 6leg lg RNA noLuge- pdoeg, 64).o611 e(vor ouotottrd rot ttov RNA noLuprepoorilv I r<or lll. Enrorlpro(verqr 611 6ev undpler rdnoto unopovd6o oXenr<r1 pre ro Borcr4pror6 nopdyov- to o. H Lemoupy(o rou nepdXetor orouq Boou<oUg ge- roypogrro[q nopdyoweg. H preyoLritep4 unoprovd6o rrlg RNA nol.upep6orlg ll SXer puo xoppofu-retrrx{ neprol{ (carbory-terminal domain - CTD) nou onorel,efuot on5 noMontr6g eno- vol,4rferg Uroq ouvorveru<4q oMqtroul(og emd optvo- (6ov. H oll.qLouX(o e(vor l.rovodrrrl orrlv RNA noluprepdoq ll. YndpXouv -26 enovol,rll.rerg oroug (u- gog6xrpeg rqr -50 mo 0r1tromu<6. O opr0pr6g ttov enovol4r.peov e(vor oqprowu<69 ener6{ or ono}.orgdg IXertxrl ge rn Borrnptorn unogovodo p l-lpood6ver DNA 'EXer CTD : (/SPTSPS)n [(uun n=26, novrlrr n=52] Ixenr<r1 ge rn Borrr1prorn unopovodo I tlpoo66veL vourtreor[0ro IxerLrrl pe rrl Boxrnprorn unogovddo o Kotvrl ror olg rpeiq notrupepooeq Korvrl ror oug rpe[q notrugepooeq Korvrl ror olg rpe[g notrugepooeq Konoreg unopovodeg elvor rorveg oe otreq rLc .-- [erg rorv europutolr6v RNA notrugepootirv. 0ptop6veq en,.-- . oXer[(ovror ge rr1 porqproul RNA notrupepool. Auro ro o'1--- e[vor ;-uo npooo;-ro[roor1 rou ltrerrpoqopllrou npor0nou Tl( ..:. Oopnq RNA notrugepoolg ll Uq (uUnq oe pro n1rrrl SDS tlmE - unogovoDeg vo droyroproro0v roro p6ye0og. rrou oTropoKp0vouv (runrxd) ndvo on6 ng ptodg eno- vo},r1r,Uerg e(vor 0ovorlq6peq. Or CTD ;rnope( vo e[vot nolri groogopu).urlprdveq oe rordLotno oepfurlg rl epe- ov(vr1g. E;rnL6r<ovror otrl pU0puon rnq ovr(6poo4g 6vop[qg (B].6ne Evotrlta 20.8), arrlv enrprfxuvoq qg preroypo<pr1g ror oe 6Leg ng mu15g qg ene[epyooiog tou mRNA, or6;rq Kor srrlv e[oyroyri rou mRNA oro rurrop6nl.ool-ro. Or RNf noLuprepdoeq rov Urroxovdp(cov Kor rs)v XLcoponl,oordlv e(vor pur<p6repeq ror prord(ouv Ue rn Po- rcrrlprorrl RNA nol,uprepdorl nopd pe rdnoro on6 to 5v- (upro rou nup4vo (enet6ti e[e].(10qrov on6 to euBor<trlpro). Aogdrcbg, ro yovr6uiproro roJV opyovr- 6[ov e(vor no].ti gxp6tepo, on6re q rontrrl no].uprepd- o4 np6ner vo pretoypdQet olerrxd L[yo yov(6ro ror o SLeyXoq rnq Ueroyporp4g pdllov e(vor ndpo noAti onLoUorepog. Aurd ro 6v(upro protd(ouv neptoo6tepo oro 6v(upro trov Borrqprorpdyorv nou 6e Xperd(etot t' onoxp(vowor o' 6vo noLunlor<6repo neprBdMov. Mro rUpro npornr4 6rdr<ptorl pero[U rov euKo- purorxriv ev(6prov ouvdyerot on6 t4v ondwqo{ toug oro 6u<ur<Lu<6 omonem(dro o-o1.rovn(vq (ro rofu<5 ou- mortr<6 mo e(6r1 rou UovrroptotS Amanita). Ouotoon- r<d oe 6Lq rq euKopuorrrd rrjrropo, q evepy5qro rqq RNA nolutrrepdonq ll ovoor5Merot yprlyopo on6 Xo- prrlLdg ouyxewprrloetg o-o1lovn(v4q. H RNA noLuprep6- oq I 6ev ovqor5trtreror. H ondvrqoq tqg RNA notrugepdorlg lll 6ev e(vor t6oo r<oLd ouwqpnp6vn: oro (rou<d rUrropo ovomSMeror on6 utp4ld en(ne6o, oLld otoug (ugoprixqreg ror ro 6wopro 6ev ovoor6l- Leror. 558 KEOAAAI0 20 H Meroypoqrl orouq Europu6reg
: C 3 H RNA l"loAupepdon 'Exet ivo Atueplt '-crrvnrfr . I - .orLvlrtlS qg RNA notrupepoorlg I onoretreftot ond 6vov .!, .31K0 unorrvlrrl rr 6vo ovo6rr6 ororxe(o TOu u[IoKtvrlTrl , r! - upstream promoter element). . I apoyovroq UBF1 rutriyet DNA ytlptrt ono gto nporeivtrtl : : -- yLo vo rp6per rovro rov revtptro unorLvrlrfl xoL to UPE. . - SLl nepttroppovet rov nopoyovto TBP nou egntr6rerot - -, evop[1 ror on6 ug rpetq RNA notrugepooeg. r - r,[ notrupepoorl I npoo66verot oto ouplntroro UBF1-SL1 . ., rtvTprKd unoKtvnTrl. - =iA no).uprepdorl I preroypd<pet p6vo to yov(dto : :c prBooco;rtr<6 RNA, on6 5vo pr6vo tUno unoKt- :. -'-i.cu1(eg t6oo rou ;reydLou 28S 6oo Kot rou pt- . ::, I8S rRNAs, nou opy6repo u<pforowot ene(ep- r l : Jr rotortrlrloerg Kor rpononotrloetg. YndpXouv -: ''.1 ovr(ypogo rnq Ueroypogrxrlg ptovd6og. =, :'-'.cooowor Ue pn peroypog6pevo 6toor4port- .: r;,lri poro (non-transcribed spacers) r<ot e(vor Avo0rx6 ororle(o unortvlq r60 -150 -140 -130 -120 -1 10 opyovtopr6vo oe uto oumotl(o, 6n<og neprypoqerol o-trlv Ev6t4ta 7.3. H opydvooq tou unoxwrfr( ror ro ou;rBdwo qg 6vop(nq nopouotd(owot ornv El- K0NA 20.3. H RNA notrugep6orl I u<p[motot tog o]to 5v(u;ro nou neptdXet entnp6o0erouq nopdyoweg onopo(trlrouq yro tqv 6vop[q Kt eluorporefetot on6 roug ueroypoqlKorig nopdyoweg oneu0e(og orov urroKrvqrrl un6 Uopgn ev6g ytyowto(ou ou1.t- ntr6rou. O unorrvqrrlg onoretre(ror on6 dUo [eXorptmSg ne- ptoX6g. O r<ewpx6g unor<rvqtrlg neptBdMer rrlv oge- qp(o, on6 -45 Srog +20, rt enopre( yto rrlv 6vop[r1 rrlq peroypogqq. l-evu<d e(vot nl.otiotog oe G-C (oou- vrlOroro yro Svov unorwrlui), er<r6g on6 ro p6vo ouv- rrlpqpr6vo otorle(o olilrlLoul[oq, pto BpoXe(o oLLql,oul(o nLo0oro oe A-T yripo on6 qv o<peqp(o. H onoretreopotrr6rlto rou KewptKo6 unortvrlti ou- [dvet no].6 on5 ro ovodtr6 motle(o tou unortvqtl (UPE, preprx6q gop6q ovopd(erot rot ovo6u<6 pu0pu- onr6 morXe(o, r1 UCE - upstream control element). To UPE e(vor dLl.q pro ntrotjotq oe G-C oMqlouX[o nou olet((eror pe rnv oM4trouX(o rou KewptKoU unortv4- tr1 rr ercre(veror on6 -180 6o:q -107. Aw6g o trinog op- ydvoor'1q e(vor r<otv6g orouq uTloKtvqr6g qg pol I oe noLld e(6r1, pol,ov6rt or (6reg or oLlql,oulieg 6tog5- pouv nol0. fltrorioro oe G-C n)oIoro oe A-T Kevrprx6q unortvlrrlg -40 -30 ,20 -10 +10 +20 0 UBF npooddverqr oro ovo6rr6 motXe(o rou unoKtvnrn To otro6v(ugo qg RNA no)tugepdorlq I nepfiouBdvet tov nopdyowo npdodeonq mov revrptr6 unorrqq (SL1) nou npoo66vetot mov xewptrd unorwlq 0r peroypotprrdq Uovo6eq rrlq RNA notrugepooqq I 6Xouv rivov revrprro unortvlrrl nou dtoXtopi(erot ono'70 bp ono ro , ., .: morXe[o Tou unoKrvr]Tr1. H npooDeorl rou UBF oro UPE npooyer qv npoodeoq tou nopoyovro CBF (core-binding factor) oTov Kov- '..- -.oxLvlrl.0nopoyovrogCBF(SLl)rono0ere[tr1vRNAnotrupepoollorrlvoqeqp[0. 28.3 H RNA l-lotrupepoorl 'EXer 6vo ALpepq Ynoxtqrfl 569 eworeq
H RNA nolupepdon I onore( 6Uo enrrouprro0g peroypoqrKoUq nopdyovreg. O nopdyovroq nou npoo66vetor orov rewprr6 unoxtvqr{ e(vor o SL1 (n TIF-1B rqr Ribl oe 6rorpoperrrd e(6r1), o ono(og ono- rele(rot on6 r5ooeprg npcoteivu<69 unogovddeq. 'Evo on6 ro ouorqrrxd rou SL'l e(vor q nptore'iv4 nou npoo66ver r4v otrtrqtrouyio TATA (TATA-binding protein - TBP), en(o4g onopo(rr1q yro rnv 6vop[r1 on5 rrg RNA no),upepdoeg ll roL lll (BL6ne Ev6rqra 20.6). H TBP 6ev npoo66veror oneuOe(og oe DNA nl,orioro oe G-C r<or 11 np6odeorlrou DNA e(vor oppro- 6r6rqro dlLtov oumorrxtirv rou SL1. flt0ov6v, r1 TBP otrl,qlenrdpd Ue rnv RNA noLuprep6o4, (oog Ue Uro r<orvf unopov66o 11 6vo Xoporcrqptmrx6 nou 61et ouv- rqprl0e( mrg noLupepdoeg. O SLI ennpSner or4v R- NA nol,upepdorl I vo [errvrioet on6 rov unorrvqtrl oe XoUnIn Boorrrl oulv6rqro. Boorr6 ro0qxovrou SL1 e(vqr vo e(oogol(oer 6rr q RNA nol,ugepdorl SXer ronoOeqOe( ooord ornv ogern- p(o. 'Ontog 0o 6oripre opy6repo, pro nqpepeep4g Let- roupy(o nop6Xeror orrg RNA noAuprepdoeg ll ror lll on6 5vov nopdyowo nou onotel,e(ror on6 tr1v TBP ror 6A- treg npolretveg. 'Erot, rivo rorv6 XopoKrrlprorrr6 qg Svop[qg ror on6 nq rpsrq no],upepdoeg e[vot 6tr orqpi- (owot o' 5vov "nopdyowo rono06r4oqq" (positioninQ factor) nou onorelefuqt qn6 TBP ouv6e6e;r6vq pre npo- retveg erdu<5q yro rov rd0e r0no unorwrlrrl. O or<prB4q rp6nog 6pdonq r<rlv e[oprc0prevov on6 qv TBP nopo- y6w<ov tono06rqo49 6roq5per r<ord nep(moorl: orov unorrqr4 yro rrlv RNA noluprepdorl I 6ev npoo6Sver DNA, ev6 fiov unoKrvrl{ yro qv RNA no},uprepdorl ll e(vor o r<riprog rp6nog yro rnv tono0Srrloq rou ev(tiprou oro DNA. H 6vop[r1 Ue uqrnln ouyy6rrlto onome(ro Ueroypq- 9116 nopdyowo UBF. llp5r<enot yro 6vo govodr6 no- Lunem(6ro nou npoo66veror o' 6vo morXe(o nLorioro oe G-C oro UPE. O UBF 61er 6Uo Leroupy(eg: 6eye(- per rrlv one).euO5poorl rou unorwqrq on6 r1v RNA no- Lupepdoq r<ot, 6nog 0o 6ori;re opy6repo, dreye(per ro SL 1. Mto 5v6er(r1 yro ro ndrg oLlrlLenr6pd o UBF pre tov SL1 6(verot on6 rr1 oqproo(o t4g on6oroorlq Uero- Ef UBF ror rewpu<ori unonvqt{. H ouyrerprpr5vq oMq- l,en(6poor1 prnope( vq rpoflonorq0e( pr6vo ov q on6moorl oLld[er rord 6vo dprro opr0p6 mporpc0v tou DNA 61t 6prrrg ov or nerpquoru<o( lerpropro( nepr- LoprB6vouv etooy<oyrl ror prori:v orpogriv. O UBF npooS6verot orqv eLdooovo o0l,or<o rou DNA xor rul(- yer ro DNA oe Uro 0qlld nou oXqpor(et orpo<p{ nep(,- nou 3600 orqv enrgdvero rnq nporetvrlg, Ue onor5treogo o xewpx6g unor<wrlrjg xor o UBE vo oup- nLqord(ouv, enmpdnowoq srov UBF vo 6eye(per qv np5o6eoq tou SL'l orov unor<rqq. H Etr6vo 20.3 6efiyer qv 5vop(q og oerpd 6ro6oXr- rcirv oMq).enr6pdoeov. H RNA noLuprepdo4 l, ocr5oo, ug(ororor tog oLoSv(upro ro ono[o nepdler rouq nepto- o6repoug r] 6).oug roug nopdyoweq nou onomofwor yrq rnv 5vopEn ror prdLl,ov enrorpore0eror oneuOe(og orov unoxw4trl. 2O.4 H RNA noAup€pdon lll Xpnorponorei T6oo Ko0o6rro0q'Ooo ror Avo6lro0q Ynorrvnrdq * H RNA notrupepoorl lll 6Xet 6[o runoug unorrvr[drv. * 0r eoolrepuol unorrvqr6g 6Xouv ppoXeteq ouvorverx6g otrtrn- trouXteg nou pplorovror p6oo oq peroypoqxrl ;rovo6o ror npc- ro),ouv n1v evop(q oe pro ouyrerptg6vl onomooq ovodtxo, a 0t ovodtro[ unonvrlr6g nepreXouv rperq ppoXereq ouvorvrTL- rdq otrtrltrouxieg ovodrrd qg oqerrlprog olg ono(eg npoobe- v0vTor UtToyp0rpuoI nopoyovreg. * 0r TFlllA ror TFlllC npooOevovror oTrq 0uv0rvrTrK6q otrtrltrou- X[eq rr enup6nouv Trlv npoo0eol rou TFlllB oqv ogerqp[0. q 0 TFlllB rpdper oq unoprovodo qv TBP Kr EnrTpener rrlv npoo- deon rrlq RNA notruUepooqq. H ovoyvrrlpror; rov unoKrvnrcllv on6 qv RNA no).upe- pdorl lll ovo6etrvUer ewunootord rouq oxetKoUq pc- Loug rtrrv peroyporptxrrtv nopoy6w0lv Kor r1g notrl- Uepdonq. Or unorrwlr6q rorordooowor oe 6[o x0prec roqyop(eg nou ovoyvop((owot pe 6rogopelrorjg rpo- noug on6 6rogoperu<59 opd6eg nopoy6w<ov. OL unox.- vqtdg yro to yov(6ro rou 55 RNA xor rou IRNA eivc. eourepLxo( (internal), 6r'1Lo6r1 Bp(or<owot ro0odLrd onc qv oqeqpio. Or unorrvqr6g yro ro yov[6ro tou snRNA (pur<p6 nupqvu<6 RNA, small nuclear RNA) Bp(r<owc. ovo6rxd rnq oeernp(og r<ord to ouprBoru<6 np6runc tcov dMov unoxrvrltulv. Kor ong 6Uo neprmo.loerq, rc [eXoprord ororle(o nou e(vor onopoirrlro yro r; ].e:- roupy(o rou unoKrvnr4 onoreLoriwor onor).erortxd onc oMrlLoul[eg or ono(eg ovoyvop((owor on6 Ueroypoei- roUg nopdyoweq, nou pre rr1 oerpd roug r<orcu0Uvou,. tqv np6o6eoq rqg RNA nol.u;repdorlg. Ot 6op5g rprciv r[nrllv unorrvqtutv yro rrlv RNA no Lupepdol lll ouvotU((owor ornv' : "ir --' ,. Yndplou, 6Uo t0nor eocoteprroU unor<rvr1rq. Ko06vog nepr6leL ;-rrc dtpeprl 6oprq, orqv ono(o 6Uo BpoXdo ororle(o oMr- A.ou1(og 6tolcop((owor ond pro preroBLqrrl oMrlLoulic Oct PSE TATA I 0L unouvrlreg yro qv RNA notrupepoorl lll pnopei , onoretro0wor ono 6rpepe(g otrqtrouXreq ro0odxo rr1q oQeqpioq T: boxA vo OtoXtrlpr(erot efre ono rc boxC elre ono rc boxB, eue o .: [eXorprmeq oiitroux(eq ovo6xo qq orpeqp(oq (Oct PSE, TATA) 560 KEOAM|0 20 H Meroypoqrl oroug Europudrreg Boorx6g ewomg
- - :<Lvnrqq tinou .1 Yto ro ptBoottlptr<6 yov(6to tou :: ::ctelenoL on6 prLo oL}qtrouX,rr boxA nou dtoXtlpG -. :. ond 6vo ev6tdpreoo orotle(o (indermiediate ele- --.=-l-lE) ond prto boxC otrtrr1ouX[o 0]'6r]'qpr1 q -.:.o1q boxA-lE-boxC ouXvd ovorptipsrot olq 5S eorrlre- : ,- cuOprortr<r1 neptold (5S internal control region - l- -: :rl (0pr1, prdvo ro motle(o boxC e(vot onopo(-rqto r rn peroYpoAn O r0nou 2 unor<tv4tqg yto to IRNA :-::e}.e[ror on6 pto oMq].ou1[o boxA nou 6toXtopi(e- -: :no pLo oMrltrouX(o boxB. Mto r<otvq op65o unoxt- .. *,'tUnou 3 yt' dMo;rtxpo RNA 6Xet tp(o orotle(o - .'-oux[og nou 6].o Bp(oxowoL ovo6txo r1]g oQtrn- : -.: To [6Lo outo orotle(o en(or1g Bp(or<owot Kot ot - -i:.-too unorwqrttrv qg RNA nouprepdoqq il' 3: emopepeiq oMqlenLdpooetq 6toq6pouv orouq , ,: rjnoug eooteptrcbv unoruvrltr.rlv, n opxrl dpoq e(- -- ,,cLvr'1. O TFrIC npoo6dvetot r<o0o6tr<d rlq opern- : r: eire ove[dpqto (unor<tvrp6q tlnou 2 yto IRNA) : -.- :. ouvduoop6 Ue rov TFrrrA (unoxtvqt6g tilnou 1 : 5S RNA). H nopouo(o tou TFrrrC enttp6net orov no- : r, 3',ro tono0Stqoqq TFrrrB vo npoo6e0e[ ornv oqerr]- , : Enerro enLorpore6etot 11 RNA noXutrrepooq lll' : i ouvo{,r[(eL to orobto tnq ow[dpoonq , : - -i tocDreptr<o[g unortvqtdq r0nou 2 nou Xpnotpo- - : .l -wot yLo to yov(6to tou IRNA. H on6mooq ;reto[ti - :,.1 <ot boxB ;rnope[ vo notrfter eneL6q noMd yov[6to ':'. - nepLdxouv 6vo prtr<p6 twp6vto' O TFrrrC npoo6eve- -r (lr oro boxA Kot oto boxB. Auto enitp6net orov -: 3 rro npoo6e0e( ornv oQernp(o. Ie ow6 to orlpe(o --::oi vo npoo6e0e( 11 RNA noLuprepdoq lll' - 5io<popo orouq eoorsptr<o6q unor<wr1t6g t0nou 1 : :o yov(6ro tou 55 RNA) e(voL 6tt o TFrrrA np6net vo -:::6e0eI mo boxA Yto vo enttp6!.reL mov TFrrrC vo 0r eocoreptrol unorLvrlrdq runou 2 rnq pol lll Xpnot- ponotouv roug nopoyovteg ouvoppotroynorlq TFrrrA rot TFrrC oro box,4 rot rc boxC Yto v0 ontoTpore0oouv rov nopoyovro rono06- qorlg TFrrrB, nou npooetrx[etrriv RNA notrupepoorl lll npoo6e0e( wo boxC. O TFrrrA e(vot Svog nopoyowog np6o6eorlg eL6tr<dq Yto rrlv oMq).ou1[o 55 nou npoo- S6verot orov unoKtvrlrrl r<ot oro 55 RNA olq ouvo66g npcltelvq (chaperone) roL yovt6Lor6q puOptmqq H deiper 6rt ;r6trLg npoo6e0e( o TFrrrC, n oKo- ).ou0(o rr.,.lv oupiBdwcov e(vot r<otvr1 pe roug unoxLvqt6q rrjnou 2: o TFrrrB (nou nepL6Xet rrlv otKoupevtr<r1 TBP) npood6veror ornv oqerrlp(o r<ot q RNA nolupepoorl lll ouv66etoL oro odpnl'or<o' Ynortvqt5q t[nou 1 Bp[ot<ov- ror p6vo oio yov(6to Yto to 5S rRNA. bt TFrirA t<ot TFrrrC e(vot nop6yoweq ouvoppoA6yq- oqg (assembly factors), pre provo6txd p6Lo vo Boq0o[v orqv np6o6eol rou nopdyowo rono06tr1or1g TFrrrB orrl oo.lorrl 0eor1. M6LLg npoodeOe( o TFirrB, ot TFrrrA xot TFrrrC lrnopti v' onopoKpuvOo0v ond rov unoKtvrlrrl lcop(g vo enqpeoor-e( q ow(6poor1 6vop[1q. O TFrtB napapivet npoiodedel.tevoq, Kowa o'tqv aEerqpb xat 4 napouo[a rcu enopxe[ 6orc 11 RNA noupepaorl lll v' avayvapbet Kat va npoo6e0e[ oil]v aqfirlp[o. 'Etot, o TFrrrB e(vot o 1'tovodt- r<6q npoy;rottr<6q nopdyowoq 6vop(qg nou onotre(toL on6 tqv nNR noLupepdoq lll. Aurn n oerpo oupBdwov e(qye( nog to ro0o6Lrd orotxe(o rou unoKtvnrq prnopo0v vo nporotr6oouv rnv npdo6eorl qq RNA noLupepooqg oqv oqernp(o, 611l'o611 nto ovo6trd. flop6lo nou Il tKo- v6qro UeroYpoQnq rov ouYKeKptpSvc'-lv yovt6(tov npoo- 6[6etot on6 rov eoortepLr6 unortvltrl, o]}'oy6q mqv neptoXrq opr6orrrg ovo6trd qq oqeqp(oq prnope( v' oMd- (ouv qv onoteleoporLr6rqto qg peroypo<prlg' O TFuC e(vot 6vo peydlo nporeivtx6 otipnlot<o 0r eooreptrol unortvrlr5q r0nou 2 rlq pol lll Xplot- -. :i0uv qv npoo6eorl rou TFrrrC ortg otrtrr1trou1leq rou box' rot -', - :oxB Yto v0 entoTpuTeu00uv Tov nopoyovro ionoOdqorlg TFr' ' -rl npoottrruet qv RNA notrupepooq lll. Aroeqp(o Kour( A Kouri B 2*.4 H RNA notrupepdorl lll Xplorgon0tsiT600 Kooodtro0q '0oo rot AvoDLxoug Ynoxrvltdq 56'l tr
(>500 kD), ouyr<p(ot;tou pey60oug Ue rnv 6to qv RNA noLuprepdoq, nou nepdlet 6(t unoprovd6eg. O TFurA ei vor pr5Loq puoq ev6ro<p5pouooq td(49 npoteivrriv nou nepr6louv 6vo prot[Bo np6o6eoqq voutd.eivxcbv o(dtov yvoor6 olg 6drcru),oq rfeu6opy6pou (zinc finger) (FM- ne Ev6trpa 28.6). O nopdyowog ronoO5tr1or1q TFrrrB onoretre(ror on6 tpetg unoprovddeg. l-lepil,oprBdvet rov [6ro nporeivrr6 nopdyowo TBP nou undplet t6oo orov KewprK6 nopdyowo np6o6eo4g SL1, rov ono(o Xcqoqonororiv or unorlqtdg rrlg pol 1, 5oo Kot orov ow(mor1o preroypo<pm6 nop6yowo TFrrrD, tov ono(o XprlorUonore( rl RNA noLu;repdo4 ll. lleptdlet en(or1g Brf, nou olet((etot Ue ro Ueroyporprr6 nopdyovto TFrrrB o ono(og Xprlouonoefuot on6 qv RNA notrupe- pdo4 ll. H rp(tq unopovddo, yvomri oq 8", e[vot ovo- Lc0oq.r1 ov 6Xer ;repu<dlq onodtorol0e( to 6(xLrovo DNA, yeyov6g nou uno6rlltivet 6n q trsnoupy(o trlg efuot vo evepyonor{oer rn Ueroyporpu<4 quooo}.(6o. O p6loq rnq B" (oog e(vor ouyrp(otprog pe rou nopdyowo o orrl Bor<rqpror<r1 RNA noLuprepdon (FLdne Ev6rryta 19.19). H ovodrni neptofi 6Xet ouptBottx6 p6l,o orouq uno- r<rvqr6g rpkrlg td[r1g qg RNA no],uprepdonq lll. Iro no- pd6erypro rqg Eu<5vog 20.4, undplouv tp(o ovo6trd ororle(o, ro ono(o enronq undpXouv Kot orouq unoKtvrl- 169 yro ro yov[6ro rou snRNA nou pretoypd<powot on6 rrlv RNA notruprepdoq ll. (l-o yov(6to yto pepxd snRNA ;reroyp6gowor on6 n1v RNA noLuprepdorl ll, evcir dMo geroypdrpowor on6 rqv RNA no),uprepdoq lll). To ovoSt- rd ororXe(o Leroupyo0v ;re nop6;toto rp6no orouq uno- r<wqt5g t6oo rqg RNA nol"u;rep6onq ll 6oo ror qg |ll. H Svop[q o' Svov ovo6tr6 unorrulti yto qv RNA no),uprep6oq lll prnope( vo ouprBe( oe pto BpoXe(o Tlepto- Xri nou Bp(oreror opr6otog nptv on6 rrlv orpeqp(o t<ot nepr6Xer p6vo ro orotle(o TATA. H onotetreoporu<5rr1- ro rrlq preroypogrlq ou(dver noLri on6 t4v nopouo(o rtov motle(ov PSE (eyyUg ororXe(o oLL4l,ou{og, proxi- mal sequence element) ror OCT (on6 q 1.6[q octamer = o]fiouep6q) tou evrqxutrl. Or nopdyoweq nou npoo- 66vowor oe owd to orotXe(o oLtr4l.ent6po0v ouvepyo- rrxd. To motle(o PSE nt0ov6v e(vot onopofurlto oe unoxwqr5g nou XcrlotUonotoUwqt on6 rqv RNA noLu- pepdon ll, evri oore( 6teyepnr{ en(6pooq orouq unoKt' v4t6g ng RNA notruprep6o4g lll. To ororle(o TATA npoo6(6et et6x6trlro yto tov tU- no rrlq noLuprepdorlq (ll n lll) nou ovoYvop(etot on6 6vov unorrvrlq snRNA. [lpoo66vetor on6 6vo nopd- yowo nou nepil.oprBdvet tlv TBP, q ono(o finv npoY- potrx6rqro ovoyvtop(et tqv oMrltrouX(o tou DNA. H TBP oMrll,enr6pd ge dMeg npotefveg, etdu<69 Yto rov tUno tou unortvqt4. H Lettoupy(o t4g TBP Kot rov oXetr(6prevov np<oretvc0v e(vot vo rono0etoUv ooqr6 qv RNA noluprepdoq or4v ocperqp(o. Or nop6yoweg Lettoupyo6v rotd tov [6to rp6no Kor yto rouq d6o t[noug unortvrltc0v qg RNA no].u- pepdo4g lll. Ot napayoweq npoo66vowu orov uftoKt- vr1t4 nporcA pnop6,oa va npoo6e0e[ 4 6n 4 RNA no/r.u pepdo1. Ip1;ror((ouv 6vo oripnAoxo npo6vo p- [qq (preinitiation complex) nou roteu0Uvet trlv np6o6eorlrqg RNA noLuprepdorlq. H RNA noLuprepd- oq lll 6ev ovoyv<op((et 1 (6ro qv oMrllouX(o rou uno- rrvr1r1, oLld npoo66vetot oe nopdyoweg ot ono[ot e(vor npoo6edepr5vot L(yo nro ovo6trd on6 r1v qqerrl- p(o. Fro rouq eooJreptxotig unortvqr6g ninou I xot t0- nou ll, or nopdyoweq ouvoppoL6yqoqg e[ootpo]t[' (ouv 6rt o TFuB (nou nepil.oprBdvet qv TBP) e(vot npoo6e6epr5voq l,(yo nto ovodtrd on6 rqv oEeqp(o, nop6Xowog 5rot ttg ntrrlpogop[eg tonoO5tr1or1q. l-to roug ovodtr<otig unoxtvqt6g, o TFrrrB npoo66verot oneuOe(og orrlv neptoXrl nou nepil.ollBdver to xowf TATA. Aur6 oqpo(vet 6rt ove[dpqto on6 q 06o4 rov oMqLouXuirv rou uTroKtvnrn, o(0 nopdyowog(eq) ei- vot npoo6ede;r6voq(or) rowd m4v ogeqp(o Ytq vo xoteu00ver(ouv) rrlv np6oSeoq qg RNA nol,u;rep6- orlg lll. Ie rd0e nepfntooq, 4 Xptoprot(vq npdnet vo 51er rpononorq0e( Kot vo Bp(or<etot oe ovotXrrl 6to- p6prpooq. 2O,5 H Agetnpio yro Tnv RNA l-loAuUepdon ll * H RNA notruuepdon ll yro v' opy(oer q Ueroyporpil onotrel ye- vLroug peroypoqrroug nopoyovreg (ovogo(ovrot TFrrX). o 0r unorrvnr6q rqg RNA notrugepoolq ll ouxvd 6xouv orlv ogerrlpio pto ppoXe(o ouvrrlprlpdvl otrtrntroux(o PyzCAPys. e To rourl TATA eivor rorvo ouorolr6 rov unoxtvlttiv qq R- NA notrugepoolg ll rot onoretre[rot ono 6vo ntro0oto oe A-T orropepeg nou ppiorerot -25 bp ovodtro rqg oqeqpioq. " To DPE elvor rorvo ouoronro rorv unortvrlr6v qg RNA notru- Uepoolq ll nou Dev nept6Xouv rouri TATA. e 'EvoS revrptrog unortvlrrlg yto rrlv RNA notrupepoorl ll nepL' tropBoveL ro lnr xot, ouvrlOoq, elre 6vo rourl TATA elre evo DPE. Mnopei en[o1g vo nept6Xet rot otrtro etroooovo orot- Xe(0. H Boorr<r1 opydvcrloq rnq UeroYpogtr<r1g ouoreuqq yro yov(6to nou ro6tt<onototjv npotelveg onoro].rj- gOqr<e on6 qv ovor<dtrugr1 6tt q r<o0opr1 RNA no).u- prepdor; ll rotq^.Uet Uev rl'l otiv0eorl mRNA oMd de gnope( vo evepyonot4oet rrl UeroYpqrprl Xopiq 6vo np6o0ero erXritopro. H rd0opoq owoU tou erluLl- oporoq o6rlyr;oe orov ro0optop6 ttov yevtro:v peto- ypoqrr<drv nopoy6vttr:v, 11 Baomav peraypaEmav nopoyow@v (basal transcription factors), prtoq opo- 6o9 nptoreivov onopo(rrlrov yto qv Svop(r1 on6 tlv RNA noupepdorl ll oe 6).ouq rouq unoKrvqt6q. H RNA no).uprepdoq ll oe ouvepyoo(o pre ourorig touq nopdyoweg onoretre( tI goorxn pretoypo<ptx( ou- or<eur1 nou e(voL onopokrlq yto rrl pretoyporprl r<dOe unorrv4rrl. OL yevtro( nopdyoweg neptypdgowot rrlg TFrrX, 6nou ro *X, npoodtop((et r<dOe [e1optor6 no- pdyowo. Or unopovddeg rqg RNA no).upepdorlg ll 562 KEOA Al0 20 H MetoYpoQn orouq Europutitteg Boorxeg eworeg
Kor ol yevrKo( pretoypo<ptxo( nopdyoweg efuot ouwrl- pnudvor geto[0 trrrv europutotcbv. Iqpe(o err(vqoqg Yto rrlv nopouo(oorlqg opydvo- onq rov unortwlrdlv e(vqt vo r<oOop(ooupe og nuprlvo rou unoKtvrlrq qv nro BpqXe(o oMrll,ou{o oqv ono(o prnope( vo evepyoTlorrloet q petoypo<pr1 4 RNA noLu- Uepdon ll. Oeropqtrxd, Svog r<ewptr6g unorwqtnq Uno- pef vo erqpome( oe rdOe r<rirtopo (pro],ov6tt, mrlv flpd[n, 5vog xewptr6g unoxwrltrlq on6 pr6voq rou Ko- rolriyet oe l.(yq 11 r<o06trou geroypotprl oro nAo(oto qg Xpopro{vqg in vivo). E[vot 11 etr6por4 oMrltrou{o nou ennp6net q ouyx6wpoorl rtov yevtxriv peroypoqxdlv nopoy6wrrrv srqv ogerqp(o. Auto( ot nopdyoweq eu- n).6rowot qrrl UnXovtK4 qg np6o6eorlq oro DNA rt ennpSnouv or4v RNA noluprepdoq ll vo evepyonotrloet rn Ueroypoqrl. 'Evog rewptr6g unoxtvqtrlq Lettoupye( pe 1oprr1Lr1 ono6otx6rqto. iortrtr'eg npoteiveg nou ovo- pr6(owor evepyonotnrSg (activators), ox6ptrl prto t6[r1 preroypogu<rilv nopoy6wtov, onoroUwot Ytq ro oroqr6 en(ne6o trenoupy[og (B].6ne Ev6trya 20.9). O' ovStrleve xove(g 6l.eq ot ol'trqtrouX(eq nou eprnL6- Kovror crr4v np6odeon rnq RNA nol,u;repdonq Kor rov yevrrc0v preroypo<prrdlv nopoy6wcov vo 51ouv ouwn- pqOe( orouq neptoo6tepouq r1 oe 6trouq toug unoKt- qr6g, 6ncog tqXriet yto rouq unoKtv4t5q rrlq pol I xot r4g pol lll. Kot' ovotroy(o npoq rouq Bor<tqptoxotig unorrvqt6g, 6tov ouyrp(vowot uTloKl.vqtdq qg RNA nol,uprepdoqq ll, 6rontarc0vetot 6tt ot o1.totroy(eg orrlv neproX4 rowd or4v oEeqp(o neptop((owot oe oxert- xd BpoXe(eg olLrll.ouX[eq nou epntr6rowot m4 tret- roupy(o rou unoKtvqtrl, 6ntog npor[ntet on6 ttg enrmdroetg pretolLd[ecov. H EIK0NA 20.7 defiyet tq 6o- ;r{ ev6q tuntrof rewptt<oti unortvrlrl qg pol ll pe rp(o on6 Tq nto rowd motle(q. l-evtrd, o euKopuort- 169 unorw4rnq yto qv pol ll nopouotd(et peyol'rirep4 6opu<r1 norril.[o on6 tov ow(orotXo Borcrqpror6. Ercr6g on6 ro rp(o rfpro motXe(o, noMd eldooovq orotle(o prnope( en(or1g vo oul.tpetdXouv ornv opydvtoorl tou unoxwrlr{. Ir4v oqeqp(o 11 oMrlloul(o 6ev e(vot ouw4p4pr6- TATM ......N20...... WCAYWYY ......N2+..'.. - - KoutiTATA lnr nuonvoc rou unoxrvnrn .a -r'r'"i +- nou Treprexet T0 KouTt I A I A nuonvoc rou unoxtvnm -loptgro<oulTHTH - evo rour[ TATA -25 bp ovoDtro qg Inr, To xourt TATA eXet rr1 ouvorvertxfi otrtrr1trouXro TATAA. H Inr dXet nuptl-ttdlveq (Y) nou neprpotrtrouv to dtvourtreor[DLo CA oqv otperrlpto. To DPE ppt- oreror ro0odrro qg otperrlprog. H otrtrtttrouX[o delXvet rov rodt- ronoulrtrd rtr6vo. w1. Oo16oo, q npd:tr1 Bdorl tou mRNA ouvn0oq eivaA ror nep$dtrLerot on6 nupqt6(veg. (Aw6 ro10er Kor Yrtr trlv evopr<trlpto ol,L4trouX(o CAT trr:v Borrqptoxtirv unorwqrri:v). H ouyrexptl.t6v4 neptoXr] rotrekot evop rrripro (initiator - lnr) r<ot prnope( vo neptYpqgef orn yevtnj Uopgli Py2CAPyE, 6nou Py owtototle( oe oTrot- odrlnote nuptpu6(vr1. H Inr nept5X*ot ovdpeoo ortg 05- oerg -3 Kot +5. I-lotrLo( unorwrltdg 51ouv ;rto ollqLouX[o nou ovo- ;rd(erot xouri TATA (TATA box). Itoug ovtinepoug eu- ropudlreq ouvrlOo:q Bp(oretot -25 bp ovo6txd qg orpetrlp(oq. E[vot ro povo6tr<6 qvo6tK6 motle[o tou unoruvrlr] nou 61et olenrd oroOepq 06o4 ovogoprxd Ue rnv oqerqp(o. H ouvowertr{ oMq}.ouX(o rou Kewpl'- ro6 orotle(ou e(vot TATAA rot ouv40tog qrol'ouOefuot on6 dtrl.o tp(o (e6yr1 Bdoe<ov A-T (BLdne Evdtrya 19.8). To rour( TATA ouXvd neptBdll.etor on6 oLtrr1l'ouX(eg ntroUoreg oe G-C, nou 0o pnopo0oe vo enqped(ouv tr1 l,enoupy(o rou. Efvot oXe66v novogot6tuno pe ro rour( TATA or4 06oq -10 torv Bor,crlprordlv unortwltriv. Ir4v npoyporu<6rnro, rl provo6u<r1 6togop6 5Xet vo xdvet pe qv ew6nroq tou (-25 6vowt -10, ercr6q on6 rn EUUn, 6nou to row( TATA Bp(or<etot ouvrl0coq oro -90). Awt- roromdoetg preprovopr6vrov vout<} eott6(clv mo rout[ TATA ;rnope( vo 51ouv Oettr4 11 opvqlr4 en(6pooq, ovdtroyo Ue rnv oproloy(o qg ouOewxrlg oMqLouX(og Ue rn ouvorvetxri oM4lou{o t<ot to Bo0;t6 on6xtrto4g qg preroMoyUdvnq oMnloul(og. Tunu<d, ot owtKoro- ordoerg nou etodyouv 6vo (eriyog Bdoeov G-C 51ouv nq Trro ooBopdg entmriroetg. Ot euxoputirteg 6to06touv en(oqg unoxtv4r6q ;1or- pig TATA (TATA-less promoters). Mel.6teg rov oM4- LouXrcilv rtrlv unoKtv4t6v 6qLc0vouv 6tt ro 50% ti neptoo6repo rov unoKtvrltdrv pnope( vo e(vot owoU tou r6nou. 'Otov 6vog unortv4tr1q 6ev nepdXet row( TATA, ouyyd qdpet to motXe[o DPE (downstream pro' moter element, xo0o6tx6 orollefo unoxlv4r{), nou Bp(oreror on6 +28 5coq +32. Ot tuntro( rewptxo( unorwqtdg onotelotiwot efue qn6 5vo rout( TATA ouv tnv neproXri lnr efue on6 trtto neptoXrl lnr ouv to ototle(o DP-E, proLov6tt undplouv ror dMor ouvduoopo( pe etrdooovo orotXefo. 2C..6 H TBP eivor Orroupevrr6q l'lopdyovroq * H TBP e(voL ouororrxo rou nopoyovro rono06rrlonq nou onoue[tot yto qv np6odeorl roOe RNA notrupepoonq oTov unouvrlq rr1q. * 0 nopoyovroq Yto Trlv RNA notrupepool lll elvot o TFllD, nou onoreLe[rot on6 TBP xoL -1 4 TAFs, pe ouvotrxrl go(o -800 kD. c H TBP npoo0Sverot oro rour( TATA oqv etroooovo 0utr0K0 rou DNA. q H TBP oXrlporL(et ;rto oetro yuptr: ono ro DNA rot ro ropnrel raro -800. AGAC +DPE ?S.S H TBP Etvot 0txoupevtrrlq flopoyovroc 553 Boorx6g ewoleq
Anopokqrrl npoun60eorl yro vo (extvrloet I UeroYpo- qq e(vor vo rpononotqOe( 11 Xpcoprot(vq rot v' ovo6top- yovoOe( orrlv ovoLarrq 6topr6pcpooq. En[or1g, oe 6treg rLg rd{eLg rov euKclpucortrcr:v unoxLvrlrcrlv, to 6noto oxroprepdg voureooull-toticov Bp(or<etot ndvo orov unoxrvrlrq npSner vo peroKrvrloe( 11 v' onotrlor<puvoei (BAdne Ke<pdtroro 28). Ie out6 to orlpre[o, 6vog nopd- yowoq rono0Stqoqq prnope( vo npoodeOei orov uno- xrvr'1rr1. Kd0e rd(q RNA no),upepdonq unopoqOefuot on6 6vov nopdyowo rono06tqoqg o ono[og neprdXet TBP nou oMqenrdpd pre dMo ouorortxd. H ot<poort- X(6o TBP owtorotXe( crro "TATA binding protein", 6q- trodrl "npore(vr1 nou npoo66vet tqv oLLqLouX[o TATA". H ouyxexprll5vq nporeivq opXrxd nrlpe ourd ro 6vopro ener6rl eiye Bpe0e( 6tL npoodev6tov mo r<our[TATA cno yov(ELo rrlq RNA no].uprepdorlg ll. Me- rdner-ro ovor<o).[<p0qre 6rL e[vot en(or1g ouorottrd rorv nopoy6wov-rono05rloqq SL1 Yto rrlv RNA no- ).u;repdorl I (BI5ne Ev6rqta 20.3) t<ot TFrrrB yto rrlv R- NA noAuprepdoq lll (BLdne Evotqra 20.4). Ze outdg rtq Srjo RNA no).uprepdoeg, q TBP 6ev ovoyvtrtp[(eL qv oLr;trouX(o rou KourtoU TATA (er<r6g on6 roug uno- xtvrlr6g rUnou 3 tqg pol lll): ouvenrig, to dvopd qg e(vot ncpontrovqttrd. Entntr5ov, nop6tt noMo( unoKt- vrlrdq qg RNA nolu;repdolq ll dev dXouv r<outLd TA- TA, en(or1q onot-ro0v rqv nopouo(o qq TBP. l-ro rqv RNA noLuprepdorl ll, nopoyowoq rono06rq- oqg e(vot o TFrrD, nou onoreLe(toL ond TBP ouvde6e- ;r5,vr1 oe 6tdgopeg d).Leq unopovd6eg (6tog 14), yvoror6g og TAF (TBP-associated factors, nopoyoweg nou oMrll.enL6porjv Ue rnv TBP). OpLolt6vot TAF e(voL ororxeroperprxo( pre qv TBP, dMot Bp(oxowot oe UL- r<pdrepeg noo6rqteg, eriprlpo nou oqpo(vet 6rt undp- louv nol.Lon).69 nopolLoydg TFrrD. Ot TFrrD nou neprdXouv 6rotpopetrr6g TAF 0o trlnoporioe v' ovoYvo- p((ouv unorrvqtdg pe 6totpopettroUg ouv6uooproUg ouwrlprl prdvov crrotle(ov, 6nog npoovop5pOqre (B1"6- ne Evorqra 20.5). MepLr<o( TAF dlouv toroet6tt<4 6x<ppo- oq. H ouvotrLr<r1 pd(o tou TFrrD tuntxd e(voL -800 kD. Ot TAF rou TFrrD opxLr<d ror6u<onotq01t<ov un6 popgn n.X. TAFIiOO, 6nou ot optO;ro( "00" dqtrorvouv 11 propror<r1 pd- (o rnq uno;iovd6og. l-lp6otpoto, oL TAF tqg RNA no).u- pepdorlg ll petovo;rdornKov oe TAF1, TAF2 t<.o.t<. Ir1 ouyrexpLprSvrl ovoporol.oy(o, TAFl e(vot o peyotr0re- pog TAF, TAF2 o en6pevog oe prdyeOoq r.o.t<. 'Erot, ot otrr6troyot TAF dto<popettru:v eL6o-rv 61ouv (6to 6vopto. H "::.. ' ' 6e(1ver d-rr ot nopdyoweg tonoO5-rq- onq ovoyvop((ouv rouq unoKtvlt6q pe 6totpopertr<6 tp6no rord nep(moor1. Itouq unortvqt6g yto qv RNA notruprepdoq lll, o TFlllB npoo6Svetot 6[ntro orov TFll- lC. Iroug unorrvrl-r6g yto rrlv RNA noLuprepdoq l, o SLI npoodSveror oe ouvduool-td pe rov UBF. O TFIID e(voL onox.,etsrrxd une0Ouvog yto rnv ovoyvtrtptol unoKtvrl- rov yto qv RNA notruprepdoq ll. I' dvov unor<tvr1tr1 ge r<our( TATA, 11 TBP npoodSvetoL et6Lt<d mo xout[ TATA, evc0 oe unortvqtdg ltop(g TATA, ot TAF ovoLopBdvouv ro Spyo rrlg ovoyvtoptorlg dMtov orotXe(rov rov unoKt- vrltcrrv, 6no:g 4 neproXrl lnr rot ro DPE. Ontog t<t ov et- Ynorrvrlr6q qg pol lll C/nruRnorupepooltrt Ynorrvnrdq rnq pol I Or^NAnorusepriqr ' :. Ie otrouq roug unoruvr1t6q, ot RNA notruUsp00€. rono0erouvror ono evov nopoyovro nou nepL6Xet TBP. o6pletoL crro oUpn).oKo Svop(r1q, enL6(olq rrlg TBP er- vor v' oMrl).enr6pooet Ue rrlv RNA no).upepdor1. H TBP 6Xer rrlv oouvrlOtorq L6t6qto vo npoodeve- ror orqv etrdooovo orjloxo tou DNA. (Itr1 ouwptmLxr nlerov6tqro, ot npcoreivsq nou ol.l.qlent6potiv pe rc DNA npood6vowot orr; pei(ovo o0).ot<o). H xpumoML- xr1 6oprl qg TBP unodrlltrtvet 6vo Lemopep5q powdAc yro rqv np6o6eor1 rnq oro DNA. Oncoq rpo(vetot orq'' . ,' . , q TBP neprrurLrrrvet rn [i[o ntreupd tou D- NA, olrlpor((owoq Uto "0610> yiptr; on6 5vo tprlprc rqg etroooovog oriLor<og, nou rdptmetot Yto vo Xopt' oet p6oo oe ourrl rr'1 o6tro. Irqv npd[1, ! eoorsptK- enrqdveto rrlq TBP npoo66vetot mo DNA Kot n UoYC- ).rjrepq e[oreprxrl enrqdveto e(voL dto0SotUrl Yto sno- edq ps dMeg nprotefveg. H 06oq np6o6eoqg tou DNA onoteLe(ror on6 pLo C-tel.tr<q neptoXq, ouwqprlpdv' perolrj rcov er6c,.lv, xot q peroBLrlrl N-teLtr<r1 oupd no" e(vor erreOerpdvrl yr' o).Lr1).enrdpdoetq Ue dl).eg npco- refveg. 'Evo pr6tpo rqq ouw(prlorlq rou prlxovtopot 6vop(qg rrlq Ueroypo<pr1q e(voL drt 11 oM4LouX(o npdc- 564 KE0A Al0 20 H MetoypoQn orouq Euxopu6req YnoKrvnTdq rnq pol ll
DNA ono rrlv ntreupo rrlg orevrlq o0troroq. H TBP onoretrelrot rno duo ouyyevrreg (40% rouroonpeg) ouvqplp6veq neptoX6g, rou errovl(ovrot oe ovotXro xot oroupo pntre. H N-tetrtrfl neptoxn .cLrfier notru rot etxov((erot npootvl. 0t D0o xtrtitvot qq 6Lntrttq :trLroq rou DNA Ue ovotXro rol orotrpo yrpt, H tptoroypotpto e[vot auyevrrn nopoxiipnol rou Stephen K. Burley. Seoqg oro DNA qq TBP e(vot r<otd 80% ouwqpqpr6vq uero[ti (0pqq r<ot ovOptonou. H np6o6eor1 qg TBP prnope[ vo pnv e(voL oui-rBorrl .ls llv nopouo(o orropep<r-rv vout<Leooul1.tor[rrlv. To '.,o u r<Leo ocr: 1-tdtto ouvrl 0 oq oXq por((owot rono Oerov- :og oMr1trou1(eg nLotioteg oe A-T roro t5roto tp6no oore or el,doooveg o0tror<eg vo t<oL-roUv npoq ro toore- cLrd (Bl.6ne Evorqra 70.9), on6te 0o prnoporiot vq flo- cepinod[oouv rrlv np6o6eon rnq TBP. Aut6 ntOovdv epprlveUer yrot( 1 nopouo(o ev6g vouxeooulpor[ou rrov uiloKtvlr( eprno6f(et qv 6vop(q trlg peroypogrlg. H TBP npooddvetot o"rrlv etrdooovo q0Loro xot <opmer ro DNA Kqro -80o, dnog go[verot ornv ':rr:'r " . To t<out( TATA r<o;rmeror npoq 14 pre((ovo o0Ao- <o, dreup[vowoq rr]v e].dooovo orj],ot<o. H nopopt6p- orr.rorl outn neptopl(etot mo 8 bp rou xourtof TATA. Ie r<dOe or<po rqq o),Lr1l.ou1(oq, 11 el"oootr.lv otjLor<o exer ro ouvrl0Lopevo e[pog tttv -5 A, o].].o oTo Kewpo rqq otrtrr1l.ou1iog 51et e0pog >9 A" H nopo;i6pgoroll rqq 6opr1g orqv npoypo-rtr6tqto 6e dtoXotp((et rouq x).rovoug rou DNA, r<oOtrtg 6totqpeirot to (euyoptol.to rcov Bdoeorv. H drroorl rnq Kdpqrnq pnope( vo 6totp6pet ovotroyo pe rrlv oxptBrl oMq).ouX(o tou r<ourLo6 TATA <or olet((erot Ue rnv onoreleoUorLr6tqto rou unoKt- VIT!. Auq q 6opu1 61er opr<etdg LettoupyLx6g npoer-rd- ceLq. H oMoyrl qg opyovoonq rou DNA crro Xurpo oe onoto6r;note ntreupo rou routto0 TATA enttpdneL orouq UeroypopLr<oig nopdyoweg Kot oqv RNA no}'u- gepdorl v' o).Lr1tren16pdoouv orevdtepo on' 6,tt 0o qrov 6uvor6 oro ypoUUtK6 DNA. H xopturl mo Nout( TATA owrorotle( evepyetord oro [erJ].ty;ro tou ev6g rpr-rou nepinou on6 pr(o ol'poel tou DNA KoL owLoroe- p((eror ond pto 0ertr<r1 etr(xotor1. H nopouo(o trlq TBP orrlv eldooovo orjlot<o, oe ,'.: ,,, " H rpuoro),trtrt1 6op1 rlg TBP oe oupntroro pe D- NA ono -40 p6XpL qv orpeqpio be(Xvet pto ropnl rou rourtou TA- TA nou 6reupuver rqv etroooovo ouLoro ere[ onou npoo6dverot 1 TBP H tporoypotpto elvot euyevtrl nopoXtirplol rou Stephen K. Burley ouvduoopr6 ;re dMeq npcotetveg nou npoo6dvowot orn pe[(ovo oU].or<o, 6rlprLoupye[ tLg npounoO6oeLg yto no- Lu oprO peg enorpdg D NA-npu-rreivc0v orrl ouyr<erpt;.rdvq neptoXrl. H np6o6eoq r<oOopqg TBP oro DNA in vitro npoorore0et -1 mpocpq trlg 6tnLrlg 6LLxog oro rour[ TATA, nou runtrd erre(vetot on6 -37 doq -25. H np6o- 6eo'r1 rou ouprntr6rou TFrrD orrlv ow(6poot'1 Svop(qg, oror6oo, rovovrrd npooroteUet rqv neptoxrl ond -45 6o9 -10. M5oo mov TFrrD tDS etrerj0epo np<ote'ivLx6 otipntro- r<o, o nopdyowoq TAFI npooS6vetot mrlv TBP, 6nou r<orotroprBdvet rrlv xo(trr1 entqdveto npdo6eorlq rou DNA, Irrlv npoyportx6tnro, rl 6oprrl qg 06oqq npdo- deorlq, nou Bp[or<erot orrlv N-te].Lr<r1 neptolrl tou TAF1, ;irpre(tor -rqv enLrpdveto rqq etrdooovog oij)'orog rou DNA. Aurq q popLor<r1 onopr(prr1or1 enttpdnet orov TAFl vo eL6yXeL trlv Lt<ovdrqro qg TBP vo npoo65ve- ror oto DNA: yto vo pnopeosL o TFrrD vo npoo6e0ei oro DNA, 11 N-te).tr<1 neprolrl rou TAFl npdnet vo erronrme( on6 tqv entrpdveto np6odeorlq o-ro DNA tnq TBP. E[vor o[toolpre(otro to yeyov6g 6rr no].]'oi TAF protd(ouv pe tor6veg: 9 on6 toug 14 TAF nept6louv prLo neptoXn nruXrlg tmovrr.lv (histone fold), ;roLovdrL mLg nepLoo6repeq neptTlrt.loetg oL TAF 6ev 61ouv ro rorol,otno outrlq tqg nepLoXqq nou e(vot uneU0uvo yro rrlv npdodeorl rou DNA. T6ooeptg TAF 61ouv xo noro ev6oyevrl Lxov6tqto np6o6eoqg DNA: ot TAF4b TAF12, TAFg r<ot TAF6 eivot (pror<ptvo[) o;-r6].oyor to''' 20.6 H TBP E(vor 0xougevx6g llopoyowog 565
rorovcbv H2A, H2B, H3 r<or H4, owrmo(1<og. (Or ro16- veg gXrlpor[(ouv ro Boou<6 oUpntroro nou npoo66ver DNA mrlv euKopuonKti lprrlpot(vq, BL6ne Ev6tqta 70.3). To etepodr;reprl TAF4b/TAF12 r<or TAFg/TAF6 oXqpor((owor on6 ol.l,qLen(dpooll rtov ow(morlrov unopovd6ov gSoo rcrlv pror(Balv rcov nruXri:v rov roro- v6r.lu).o.yrxo, ouvoppooloitv q Bool prog boprlg nou ;rord(er Ue ro oKrogep5g rov rorovcbv. Mtq rdroro 6op{ nr0ov6v eu06veror yro Un er6rrdg yro qv o},}.r1- LouX(o oLlrlLenLdpdoetg rou TF1lD pe ro DNA. Or rmovx6g muldq rov rorovcbv Xpnorponoro[wqt en(- orlq Kor oe drpepe(g o).LqLenr6pdoerg prero(ri dl,),rrrv nopoy6wcov TAF11. Mepxo( TAFrr Filope( vo Bp(or<owor oe dLlo ori1.r- nloxo ercrdg on6 to TAFrrD. Er6u<6repo, TAFrr "rUnou ror6vrlg undplouv en(or1g ror oe npcoreiVu<d oUprnLor<o TIou rpononororiv rq 6op{ rrlg popor(vqg npw on6 11 peroypogq (B).5ne Ev6tqto 28.9). 2O.7 H Boorri Euoreufi luvoppoAoy€iTot OTOV YnoKIVnTfi * To ovodlro ororXelo xor or nopoyovTeq rrou npoodrlvovror oe ouro ou{ovouv q ouXvorrlro qg 6vop[r1g. + H npoodeol rou TFrrD oro rour( TATA 11 oro lnr elvor ro np6- ro pqpo qq 6vop[qg. * AtrtroL peroypoqrroI nopoyovreg npoodevovrot oro oupntroro pe ro0opro;levq oerpo, enerre(vovrog ro prirog qq npooTo- reupdvlg neproXlg oro DNA. e H RNA notrupepoorl ll opX((er rr; peroyporprl ;rotrrg npoode0ei oro ougn),oro. Iro r<Urropo, unorrvrlr6g prnope( vo Bpe0oUv oe rperq BooLr<oUg rUnoug Xpropror(vr1g. O npoirog rUnog e(vor 6vo ovevepy6 yov(6Lo oe x,trerorr1 Xpco;ror(vr1. O 6eU- repog e(vor 6vo 6uvrltr<d evepy6 yov(6ro oe ovotXrq lprr;por(vq, nou ovogdperot yov(dro oe eypqyoporl (poised gene). Aw6g o rUnog prnope( Uev vo ouvop- pro).oyqoer 14 Boo'u<r1 ouoxeurl, o).).d o6uvote( vo npoXrop4oer crrrl Ueroypogrl rou yovr6(ou Xiop(g 6vo 6eUrepo orlpro yro tqv Svop(4 rrlg Ueroypoqrtq. To yov[6ro Oeppur<oU shock Bp(or<owor oe eyprlyopo4 droL oore vo prnope( vo evepyonoulOoriv op6oog pr6- Lrg ou(q0e( q Oeppror<poo(o. O rp(rog rrlnog e(vor dvo yov[6ro nou evepyonore(ror oe ovorXrrl Xpro;ror(vr1. M51pr np6oqoto 5ro06,ro;re eldlrmeg nL4pogo- p(eg olelr<d Ue rn oupperoxn pn rrodrxonor4tu<ov RNA peroypdrpurv (noncoding RNA, ncRNA) ornv evepyo- no(r1oq rclv yovr6(otv. H r<ordorooq dMo[e r<or orlprepo yvorp((oupre no).).d nopode(yproro oro ono(o q pero- ypoAn ncRNA puOpr((er rn Ueroypoqrl r<owLvrrtv 4 enr- ro).umd;ievcov yovr6(ov nou rro6rronoLoUv nporeiveg. 'Evo np6o<poro nopddeLypro neprypd<per q ou;rpreropl rrov ncRNA nou (errvoriv ovodu<d on6 rov unorrvrpq tou yovr6(ou fbpl tou S. pombe orrl oro6ror<11 ogo(pe- oq 6Uo orcroprepc0v voux,.eooogor(rov on6 rrl ouyKeKpr- p6vq neproXrl rou unoKrwlrrl. H nopoyroyrj ourcirv rrov Lenoupytxrirv ncRNA (nou ovog6powor en(or1g Kor oq rpumxd ooro0rl perdypocpo n CUT - cryptic unstabte transcr)pts) nr0ov6v elvor nro 6todedo;r6vq ouyr<pnu<o pe trg oplu<69 ercrrg4oerg. Ie preydlo opr0p6 evepyc0v unortvrlrciv nopdyowor prerdypogo nou npo6plowor ovodrrd on6 roug unor<rv4r5g (yvoord rog ovo6u<6 pre- rdyporpo rou unoKrvnrri 4 PROMT - promoter up- stream transcripts). To PROMpT peroypdgowor ror npoq lq 66o r<oreu06voerq oe ox6on Ue rov xq0o6rr6 unorrv4rrl xor [orog no((ouv pu0proru<6 pd],o or4 preto- ypoen. Or noMo[ p6],or rotv ncRNA orrl prl0prorl qg peroyporp(g 0o nopouoroorofv orqv Ev6trya 30.5. Anopofu4rrl npoun60eorl yro rn 6ro6txoo(o rqg 5vop(qg e(vor vo Aeroupyrjoouv ge ro0optoprdv4 oerpd or Boorro( pu0ptorxo( nopdyoweg c6ore vo ouvop[o- i,oyrloouv 5vo orign).oro oro ono(o 0q ouv6e0e[ 4 RNA noAupepdorl. H oroLouO[o rov ouprBdwov ouvorl(eror mqv EIK0NA 20.11. M5).rg ouvdeOe( 11 noLuprepdo4, 4 evepy6qtd qg e).5y1eror on6 UeroypqgrKorlq nopd- yoweq nou npoo66vowor orov evrq4ur1. 'Evog unorrvrlrlg yro l.1v RNA no),uprepdorl ll ouyyd onotel.efuor on6 Srio trinoug nepLoXrrlv. O r<ewptx6g unorlqrig nepdXer r4v (6ro rrlv ogerqp(o, nou runrxd routonorekor on6 qv neptoXrl lnr ror ouXyd yenoveUer g' 6vo r<our( TATA n 5vo ororXe[o DpE, ev6 prnope( en[- orlq vo neprdXer rr enrnp5o0ero, Lry6repo rorvd qror- 1e(o. H onoreleoportr6qro ror q et6rx6trlro Ue rnv ono(o ovoyvcop((eror 6vog unoxwqrqg, oo16oo, e!op- rcirwor on6 BpoXe(eq oLL4loul[eg nro ovo6rxd, nou ovoyvcop(owor on6 pto drogoperrrrl oprd6o Ueroypo- qu<riv nopoy6wov, yvcrlorc0v @g evepyofioLqtiq (activa- tors). l-evrxd, or oLLql,ouX(eg-m61or Bp(oxowor -100 bp ovo6u<d qg oqerqp(oq, ov ror preprx6g gop6g ond- Xouv neproo6repo. H np6o6eoq rov evepyonorqrcbv oe ourSg rrg 06oerg ;rnope( vo en4pedoer ro oX41rotropr6 rou oupn).6rou 5vop(r1g oe drdrpopo ord6ro. Or unoxr- vr1r6g e(vot opyovropr6vor pre Bdorl qv oppi rou ouv- 6uooproU ototXe(tov. Ardgopo ororle(o gnope[ vo ouverogS,pouv orq Aeroupy(o rou unoxrvqrj, oMd xo- v6vo 6ev e(vor onopofurlro yro 6l,oug roug unorwrlr6g. To npciro B4trro oqv evepyono(4oq ev6g unoxrvqrrl nou nept6let rout( TATA xot Bp(oreror oe ovorXr{ lpol- ;tot(vq evepyonoofuot pr6Lrg o TF11D npoo6e0e( oro row( TATA. Aut6 prnope( vo evroXferor on6 ovo6u<d ororle(o nou 6pouv pSoco ev6g ouvevepyonornrn. (O TFrrD ovo- yvop(Efl en(onq rrlv otrtrr1LouX(o tnr orrlv oEeqp(o, ro DPE ror (o<og r<or dMo ororXe(o rou uTroKrvnrry). 'Orov oro o0gntroxo npooreOe( rqr o TFrrA, o TFrrD gnope( nM- ov vo npoo6e0e( oe pro neproff nou errefveror or6prl nto ovo6rrd. O TFrrA prnope( vo evepyonoujoer rqv TBp o(powog rrlv r<otooroLri nou nporolekqt on6 rov TAF1. O TFrrB npooddveror xoOodrxd rou rouroU TATA, di- nAo orqv TBP, ener,re(vowog 6"ror rg enoqdg r<ord prrl- Koq rnq prog nl,eupdg rou DNA on6 -10 6<o9 +10. H 566 KE0AAAI0 20 H Meroyporp4 orouq Europurilreg Boorxfg eworeg
+ TFllE & .Evo 0uunl,oKo evopqnq yro TIV RNA notruuepoorl ll ouvoppotroye(rot oroug unorrvqreg ono uto repopxnu6vn oxotrou0(o :rrltrenr6pooeov ue ueTqyporprxoug nopoyovreg. 0 TFLrD onoreeirot ono rqv TBP ouv ro ouv6edepSvo TAF, onoq qo(verot oTo Tlovo ;xluo. l-ro troyouq ontrouorguorlq, oro unotrorno oxnuoro 11 TBP tpo(vetot uovn uq, xopig tov TFrrD. l-lpooopuooptlvo ono: M E Maxon' J' : Goodrich rot R. Ti1an, Genes Dev. I (1 994): 51 5-524' xpuoroMtr( 6ogri nou <po(vetot mqv E|(ONA 20'12 6e(- )yet 6n n TBP q(4got(et enoqdq orqv elr'dooovo oUI'o- io ro0oax6 on6 to row( TATA xt 6plerot oe eno<pr1 pe rr1 pe(ovo q0l.oro qvodKd on6 to xout( TATA, oe Uto neproXrd yvoorn oq BRE. Ito oplo[o, o o;t6troyoq no- pdyowoq tou TFIB npoYUoronoe( etdx6g yo trlv oMrl- iouXio enoq6g Ue rov unortvrlti mrlv neproXti BRE' eur6 to Bripro ntore0etot 6tt enrlped(er xo0oprorxd rrlv ro0r6proorl tr1g noLtr6trlroq rou unortvrlr{, 6nlo6ti npoq ro noti 0o e(vot orpoppr6vrl 11 RNA nol'upep6or1 rot nor6g 0o e(vor o rtrri:vog erqroye(o. O TFrrB nt0ov6v no- pSxeirnv ent<pdveto nou ue q oepd qg ovoyvcop((etot lnO .n, RNA noLuprepdoq, on5te prnope( vo op((et rrlv rmerj0uvorl np6o6eoqg qg notruprepdorlq. H rpuoroMtrci 6oUn tou TFrrB Ue rrlv RNA no}'upe- p6oq 6etXvet 6tt tpetq neptoX6g tou nopdyowo oMq- Lntopo6J pre ro 5v(upro' 'Ontog ono6(6etot oxqpotrr<d ornv EIKONA 20.13, prto N teLu<d xop65lo rpeu6opy0pou or,6 .o, TFrrB 6pxetot o' enoqri pe to 6v(upro row6 orrl 06oq e[66ou tou RNA. Aw6 nt0ov6v nopeprBofuet ornv 6(o6o tou RNA r<ot enqped(er q pretdBoorl on6 r1v oventtupj 6vop(4 mr1 Stoquyri rou unoKrvrlrrl' 'Evog entpr[<uopr6voq,,66*11oS' tou TF1IB eto6pX*ot 96oo oro evepy6 r<6wpo rrlq notrugepdonq. H C-teLtrd ne- proXq oM4Lent6pd pe qv RNA nolugepdoq.xot rov T- F1;D yto vo npooovorotr(oet ro DNA. Ko0op((et en(or1g tlv nope(o rou DNA ere( 6nou 0o 6tr0et o' enoqtl pe toug nopdyoweg TFrrE, TFrrF t<ot TFttH, toug ono(oug nr0ov6v'eubuypoprpr((et pre to otignLoro trov Bootxriv nopoy6wtov. O nopdyowoq TFrrF e(vot etepo-ot<togep6g nou onotel,efuot'on6 6rio t0noug unopov66u:v' H peyol'ti- reprl uTlopovddo (RAP74) 61et prto e[optrirprevrl qn6 to nie'evepy6tqto e)'trdorlg nou 0o pnopoUoe vo eprntr6- Keror orrlv onodt6ro(r1rou DNA r<otd qv 6vop[q' H pt- rp6tepq unopovddo (RAP38) 61et rdnoto oprotroy(o pe ng neptoX6g tou Borct4ptor<o0 nopdyowo o nou 6pXov- roi o''enocpri Ue rnv r<ewprr<r1 nol'uprepdorl xot npoo66- 20.? H Bootrrl luoreurl IuvoppotroYeirot mov Ynonwrn fil
Auo onorferg rou rpuoroyouq ougntr6xou TF;B_ TBP-DNA derXvouv ol o TFIB npooD6veror roro 1u1xog Tnq Ks_ roppevlg ntreupog lou DNA. 0r Duo rtrdrvot rou DNA Ue rpoorvo x0l r[rplvo, i1 TBP pntre ror o TFriB rrjrxrvo rot pop. H tporoyporpio e[vor euyevlrfl nopoXtirprlorl rou Stephen K Burley. vetor o<prXrd orrlv RNA noLuprepdoq ll. O TFrrF nrOovdv g6pveL qv RNA noLupepdorl ll oro ouvopl_roioyo,jprevo orjpnl.or<o rrlq Ueroyporpqg xor nopdler ro pdoo yLo rqv np6odeorl. To orjprn).oKo rrlq TBp r<or ro.rv TAF * : pe(v' oM4lenL6pd ;re rri CTD oupd rrlg RNA nor."*. poong xoL 4 o).).r;tren(6poor1 pe rov TFrrB iorr;g e..: en(ot|g o4powrx4 6tov ro 6rpep6g TFrrF/notruLrep::_ ouv6e0e( pe ro otiprnl.oxo. H npdodeorl rqg no).ugepdorlg enerre(ver rrc :.. oetg nou npoororerjowor xo0o6tr<d rou + 15 orov ().-'. vo expoye(o Kor rou +20 orov r<).rr.lvo ;rr1_enUoysro -_ 5v(upro enexre(ver ro oUpni.oxo oe 6).o ro gqroc :: - enerdfl mo ovo6Lx6 dpto undpXer trtpo enrnpooii:- npomoo[o. Tr y(vetor orouq uroKrvrlrdg 1op(g r<outf TATAr A::. po(r4ror e[vot or [6ror yevu<of UeroypoerKof nopo,, : ,_ teg, geto(U dMo-rv xor o TFrrD. H nepLoXrl lnr nope ,. ro orotle(o rono06rr1oqg, oro ono(o npoo6due-o, . -- FirD [.r5oo qg txov6r4tog ev6g 11 neprooordpcrrv TAF . ovoyviop[(ouv oneuOe(og r4v lnr. !],],or TAF rou -tr len(or1g ovoyvrop((ouv to morle(o DpE xo0o6tr<c :-:ogeqp(og. H TBp l,eLroupye( oe owoUg rouq uno(,,_. r5.g nep(nou dncDg oroug unorrvr;r6g qg RNA no).:*:, pdonq I rot roug eooreprrorig unorrvqrdg rrlg F.,,_ notrupepdorlg ilt. Orov undplet rour( TATA, xo0opi(eL rrlv ewor.:- rrlg o<perrlp(og. H ono,trorcprl rou nporotre( ovur!:.. : orq 05o4 Svop(qg, 1-ro).ovdrL 4 dnoro ouvo).rxq UaLa;* rnq Ueroypo<pnq e(vor olerrxo pur<p(. l-1pdypol, KCr:: _ or unorrvrlr5q Xop(q r<our( TATA 6ev dlouv 1-rovo6L<.: ogeqp(eg; 6ror, 11 dvop[4 npoyproronore[tor peoo ;.puo ouoroLX(o oqerrlpurrv. To xour[ TATA euOuypc_.. (er rqv RNA no).u;repdoq pdoro r;g oMrl).en(dpo;'.: Ue rov TFrrD r<or dLAoug nopdyoweg dtoL rr-lore vo <a, _ vder mr1 oroorrl 0do4. H npdo6eoq r4g TBp oro TA-_ e(vor ro r<[pto Xopor<rqpLorrr<d orqv ovoyvo,rptorl r: _ unorrvqtq, oMd 6rjo trreydi.or TAF (TAF1 r<or TAF2) a orlg dplowor o' enog4 pe ro DNA rowo orrlv oes:-. p(o xr en4ped(ouv qv onore).eoporLrdrqro rr.lQ o,1.. dpoorlg. . . H ouvopl.ro).6y4o4 rou oupnLdrou dvop[rig rlq r" NA no).upepdorlg ll epr<pov((er ;uo ev6rogdpouoo owit:. !n le q petoypo<pq rov npoKopurr;trrrv. H BorcrqpLc*- RNA noAupepoorl ouoroortxd e(vqr dvo ouvercrrx6 our:. 0poro1lo pr evdoyevrl trov6r4ro vo ovoyvop((er Kcl ,. lnpoodSvet ro DNA rou unoKrvnrrl. O nopdyowoq lnou e[vor onopo(r4rog yro rrlv dvop(4 oMd o1r yro :r . enrprlxuvo4, yiverot rpnUo rou ev(U;rou nporoI or:: npoo6e0e( oro DNA, nopdrL opydrepo pnopei vo 5.. ororot. H RNA no).uprepdoq ll npooddvetor orov unci.. vqrrl prdvo ogdrou nporlyou;rdvcrlg npoo6e0c_. [eXoprmo[ UeroypoerKo[ nopdyoweg. Or peroypo<p L,r : nopdyoweg 6to05rouv ovd).oyo pd).o ;re ro BoxrqpLc<: nopdyowo o, 6r-ilo6r1 enrpdnouv otr1 Boo.Lxrl noAuu. pdorl v' ovoyvrr;p(oer eL6rro ro DNA orrq oMrl).ouXLot rou uroKrvrlr4, oMd dlouv ovomri{er ;reyo}.0reprl o,,,r- {opr1o[o. l-lpdyporr, oL nopdyoweg e(vor xup[rr-lq unr_ 0uvot yro rr;v e16rxdrnro rnq ovoyvoprorlq rc- unoxtv4r4. Mdvo optoprdvor ou1lperdlouv o, efloe.. nproreivrlg-DNA (xor pdvo 11 TBp xor prepu<o( TAF npo. potonoloriv eno<pdg eL6Lr<dg yro rrlv oMqtroul(o). Iur,i Ko0oDrrd DNA 0 TFIIB npooDevetor oro DNA rot epleror o,eno_ Ql Ue qv RNA notrugepoorl rovro orrl 0eo1 e{oDol rou RNA r<or ro evepyo revrpo, ro onolo rot npooovorotrL(et novrrl oro DNA. Avnnopopotrtrere Us Tlv Etrovo 20.12, nou beLXvsr rr1 doprl rrlg notrupepoorlg roro q peroyporpq, 568 tGOA/lAt0 20 H Meroypog4 orouq Europutirreg I Merox[v1or1 Ev(rjpou
ndlq, ot oMrlhenr6pdoetg preto(ti npoteivdlv e(vot or1- Lrowrdq Yto rn ouvoptrro6yr1oq rou outrrnl6rou' ' tn viiro, ouvopprol'6yr1or1 prnope( vo TlpqYporonot- r10ei or<6po t<ot p6vo orov KewptK6 unorwqrq' oorooo in vivo ourl q ow(6poor1 dev enopre[ Yto rn peroYpo- on. rogtirc onono0wot rot ol'Lr1}'ent6p6oetg U' svepYo- ;"',n;A;, ovoyvtop((ouv ro nio ovo6trd Totxtt:ol evepyoilotnrdq oMqLent6poriv pre q-Bootrt1 ouoKeurl or 5,o,popo orddto rotd tq ouvopgo6ynorl tns (9I5- ne Evotqta 28.5). geto[ti rov UeroYpoqu<tilv nopoy6worv nou e[wtnpr 6e6e;r6vor orouq evtoxur6q rot tou unoKvrlrrl' ---ili"L"u-oioi nopdyoweq nou qtdvouv aro oti1.rtAc oo Jropqns e(vot or TFIE r<ot TFrrH' Apouv oro re)'ewoio *aO-itr1q 6vop[r1q. H np6odeoq tou TFIE nporolei il;;rn' ina rieiioxns nou npoor.oretetot ro0odu6 r<otd or<61.11 1r,o orpo,pf'-r1q 6tnl'riq d'Ltrog' doq to +30' o ri n tfrli o provo6tr6q yevx6q ;retoypoqrr6q nop6- ;;; ut .oLr.ii.Lgq ove(opqteq ev(upru<6q evepy6rrl- irs, trti"gO dl'tov ;rto ATP6orl' eLtraols-1ill no)rtx6trltog Kot Uto evepy6tlto r<wdorlq TIou Unopet vq ,pro,ilpr),,,iroet tr1 CTD rqq RNA nolu;repdorlq ll (orq oep(vn 5 rrlq emonemtdxqg enovdtrqrprlq)' O TFrrH e(vot 6voq oouvqOtorog nopdyowoq nou en(oqq.prnope( vo noiql, *anoio p6tro or4v entpt(xuvorl' ot oMrltrenr6pd- ;;G;", trre to DNA xq0odtKd tqq oqetrlp(og etuot ono- ootmteq vto rn 6tqquYrltrlq RNA noLuprepdorlg on6 tov ;;^,"n-,'i o TFrrH inions etrrntrSretot orrlv ent6t6p0oor1 tou DNA (Blene Ev6rrya 16'4)' ' - -:" gudi ypoprprtr6 erpoye(o, onopofurlteq. npouno0S- ot,iv* in'uetor<ffior1'rr1q nouprepdorlq e(vor q udp6- truot1'tou ATP, 11 nopouo(o TFrrE t<ot .n tu"qY-6-ll,o eiu<6or1g tou TFrH (nou nop6letot o19 rtq ufiouovo- ;;; Fb- rot xeo;. Aw6q oi npounoo5oerq nopor<dp- nrovrot o' 6vo u.,"p,ito'1'5vo exgoye(o' Aut6 ,r,oatlii,ut, 6tt or TFrrE r<qt TFrrH e(vot onopo(tnror Yl'o irlv onoorfuo(r1tou oUn O*t vo gnop6oet vo Eer<wn- oe, n pe.ooluqon rnq RNA no}'uUep-dor1g' H evepy6tq- ro eltrdorlg rnq uTloUovd6oq XPBB tou TFrrH e(vqt une0Ouvr1 Yto rrlv npoyprottr<ri onodtdtofq tou DNA' - ;-RNA'.olupepdorl rowoor6retot oe ;reptr<6 yov[- 6to 6tov op{(er rn UeroYpoQn' (To onot6eotrro 6e o,o,p6pr, noii'oto ttlv oventtuxq.eygpEn qq Borcrr1- p,ooni RNA nol,uprepdonq nou ovotr60r1Ke ornv Evot4- ii i s.'t z, ;toov6tt o prqlovtopr6g e(vot..5to<poperr6q)' Ie noMd yov(5to, n RNA noLuprep6orl ll teppoil(et pe- ; ;6 u*bn ot omoo4' To Bpqx6 RNA npoi6v qno6o- Ut"o, VpnVopo. l-to vo oupBe( entprqxuvor'1 U6oo orn 'ut-oVpoEion provd6o, onome(tot 5vo orSpnl'ot<o Ktvo- !O" *tu r<otreftor R-TEFb' To R-TEFb neptsXet r1v rwd- on COKS, q ono(o e[vot U6troq tqq ou<oy6vetoq rov o*oot.fx nou eL6yXouv rov Kurrctptr<6 x6xtro' O R-TEFb ,ftoorpopuLtOvet nepom6pto tq CTD (orr1 oeptvrl 2 qq emonemtdtrc'1q enovdMlqrlq)' M6Xpt on-gepo 6ev xoto- ,oo,iut o0te yiot( onot're(tqt outi 11 en6poorl oe uept- *o,ii' ,"o*,rr1t6q ror 6Xt oe dI'ouq o0te ncirq puOpti- (etot.--'H',pro,popuL(tloq tqq CTD e(vot onopo(tqq yto t1v onel'eu05pcoon qg RNA nolupepdorlq ll on5 rov unortvrlt( ooi -oug ueroYpoqtKo.Uq nopdyoweg 6tot dlme vo trrnope( vo xdvet rr1 petdBoorl ornv entprlKu- ,Ui"n lropq(, 6norq <po(verot^orrlv EIK0NA 20'14' To np6tuno rptootpopu)'(tlonq rnq CTD rotd t1v entp{ruv- ;n;fu;iorrol-,,*o' e[6yxetot rot Kq'rol0erot on6 notr- igs-nprt",utr5g rwdoeg Kqt Qooqordoeq' ot nepto- ,Zitpt, Boou<oi uerqYpoEtKoi nop6yoweq oneLeule- il";;"ion6 rov unor<wqtri oe our6 to.cnd6to' o Me- diatorol.l.rl}'ent6p6er6tr<dgeno}.upepdoqnourpdpet 2O.8 H'EvopEn AroAou0eiTor on6 K60opon Tou YnoKlvnT6 Kl EntPfiruvon * 0r TFIE ror TFrrH onoLtottvrot yto r1v rfl[r1 iou DNA Kot rn Ue- rorivqorl rrlq notruUePoonq. " rio ro [erwrloer rr ro0opol rou unortvqrrl rot 11 entpflruvorl nprlnet vo rpototpoputrttl0e( 1 CTD + Ie uepLro0q unoxtvlrdq, Yt0 v0 lepuoT(oet q 0ventruxlg evopEn Xpe to(eror eninpoo0errl tpo'rorpoputritloq rrlq CTD' * io o*ilir.pl rov toroviiv rpononorouvrot npooroLpo roro 11 perorivrlorl rrlq RNA nohuUepoonq' * H CTD ouvrov((et rrlv ene[epyoo(o rou RNA ue Tn ucToYpoQr]' =, Av ouUBotiv ptrdpeq oro DNA, 0o entdLop0trl0otiv roro nporf- Ulon To geroYPorP6Pevo Yovi6to' " 'O f i,,U nopdXet ro ouvdeopo yLo evo ouptnhoro ev(0ptt'tlv ent- 6toP0tlonq. An6 q ortYLi( nou 0o olqprottororiv o^tnptotoL vourl'e- oiio,noi at"uoi r<ot ;relpiirlv oneeu0epooq.tqg RNA nouprepooqg on6 tov unoxtvrlrl preootroBouv UeprKo .ri,o[ bnuo-o. Aw6 to ord6to ovo;rd(etot xaaaporl ,o, uno*rur1rr1 (promoter clearance) rot e(vot to rp(otpio preU,qOUeuo'$6po no, r<oOop((et ov 0o preroypo<pe( ;;;i;;(6t oe evplvopo'q 11 5vo.evep16 vov(6to' Aut6 ro Brlpto eL6yXetot on6 evLoXur6q' (Ouprq0elte 6rt to ^p iI,l,l- Bn u"'xLeta( q q pretoy pocpq q To Pool p^t:: uo, q'pre-oiponrl tou r<.etoro0 ouprntr6rou oe ovotxro' Bi.O"t rr,iinru 19.3). ot neptoo6tepot yevtr<o( peto- vpl,p*ot nopdyoweg onottotiwor pi6vo yto r1v np6o- Oton t,fq R'tlA noLuprepdonq fiov. unoKtvqtrl' o)'d geptr<oi 6pouv oe Uetoyevdorepo ord6to' Ot petoypo<ptt<ot nopoyoweq nou nOo96.evow11 1' evrcrlutdq ouv(Oorq eL6yXouv rov unoKtvnrn oxt lt-11? on ino,pll, ol).d pdlov pr6ool ev6q-ouvevepYonorntrl nou ,,poo66vero[ orov unoruvrltq 'Evog on6 rouq nto r<oLvouq ouvevepYonotqt6q e(vot o Mediator' 6vo rt9Ltl preyd).o nolu prep5g npoteivtr6 orj pntroro' ouwnpn Uevo onL .n (rjpq Stlq tov ov0polno, nou evonote( oqgoto on6 noMo0 g UeroYpoQtKo 0q nopoyoweq npoo6edepr6- ,./ouq o' evtcrlwdq. Tdoo to yov(6to oe eYpnYopon ooo . r, i" *tpvl yovidto xpetd(owot r1v oMrlLen(6poorq 20.8 H'EvupEn Arolou0eitot ond Ko0opon tou Ynoxrvnq xt EnrElxwol 5t I Boorx6c ewoteq I
Gr:G G v,i'Aip $ *ro Pol ll PP Y.iersg s RNA pol ll Ysprsgs RNA Pol ll Aeqolevf pol ll rou DNA Ynoxwqqg npooflKuoq r[q pol ll ond ro otiurloxo rpo6vopqnq H Qooqoputrioon qg Ser5 ond ro TFllH Por10o ornv npood)xuon ev(rjpov nou rotrUnrouv ro 5' 6rpo rou [erqypdqou Poly A site Anoqooqoputritoon ond q QtooQordon SerS (n.x. Scpl) Un qoogopu)rroprSvq CTD: <po[vetot 6tt 4 qoogopu- l.(roo4 6tor<6mer ourti qv oM4lenidpoorl, H CTD epntrSretot dpreoo { 6ppeoo orqv ene(epyo- o(q rou mRNA rord 11 orivOeorl rou Kot ogo0 onetreu- Oepo0e( on6 rrlv RNA noLuprepdon. Kd0e 06o11 tproorpopul,(roonq rqq CTD XprlorpeUet og o4pre(o ovo- yvdlpronq r1 oyr<upo66tnonq yto rn ofvdeon dtrl,tov npcoreivc0v Ue rrlv noluprepdoq. To 6v(upro oxrlport- oUoU rnq roL0mpog (tpovogepdorl rou Youovultt<oti, iexivllro rnq pol ll H rptorpopu)r(oor1 on1 Ser2 ond ro P-TEFb evepyonote( rqv enqgrlruvorl, q ouppoErl xor qv noluodevutr(trlq H Q(DoQopu)(oon orn SerT und Utq dyvrooq rwdorl Dev SXet Udxpt mryUnq rdnoro yvomd pdtro oqv drgpootl rtrtv yovt6lov nou rtoduonororiv nporetveq Enrp{xuvol on6 rqv pol ll Iuppotpti guanylyl transferase), nou npoo0Stet to rotdLotno G fio 5' drpo tou veoouweOetg6vou mRNA, npoo6Sve- ror or CTD nou 61er qoogopuLtco0e( arrl oepfuq 5 (r1 npdnrl <proo<popul,(trlon nou KotqLUetot on6 rov TFrrH). Aur6 (oog e(vot oqprowtx5 enet6ri enttp5rlet oro 6v(u- Uo vo rpononor{oer (rt 5tor vo npoqror6q;e0 ro 5' dxpo pr6l,rq ow6 ouwe0e(.'Eneno, q goorpopul,(oorl tqg oep(vr1q 2 on6 to P-TEFb odqye( oe npoo6trr<uor1 orrl CTD puog oprd6og npoteivcbv nou ovoptd(owot S- l-truxo(utrioon; 0toorloputritoorl on6 )dk8/Srb10, ERK %, )DCZCDKl) Anorpoorpoputrioorl onri q qoorpordol Ser2 (n.X. Fcpl ) noqooqoputr(ooq ond m QooQqrdqn SerT; Kon{, notruodevutr[toorl xor repporroprig l-lpdodeol rtov nopoydvttov roluodevutr[oonqmrgqooqoputruog6veg efiovqtrnuerq Ser2 ue nenttdu]tonpo]rtvtro0g 6eoUoUq oe trans 0roudpqoon EIK$NA 20.14 H rpononoirlorl rou enronemrdiou CTD r4q RNA no)upepooqq ll rord q UEToYpoQ1. 0rov q CTD qq RNA notrupepoonq ll ero6pXeror mo o0gntroxo npo6vopEnq 6ev e[vor Qooqoputrroudvn. H Qtr:otpoputr(oon rototrofnov oep(vnq dnptoupYe( O5oetg npdodeoqq t6oo yro ro 6v(upo ene[epyooioq Tou mRNA 6oo ror yro ng uvooeg nou rorotrUouv nspotTdpo tptootpoputr(tlon, dntoq neptYpdqtTot oIo oflUq. AvqTfntoorl on6 Trends Genet., vol.24,P. P. Gardner ror J. Vinther, Mutation of mRNA target sequences..., pp. 262-265. Copyright 2008, ge qv odeLo tou Elsevier [http//www.sciencedirect.com/science/iournal/01 689525]. Avqxurtroon rnq pol ll Enrp{xuvol on6 r4v pol ll 570 KEoAAAI0 20 H Meroypoqn orouq EuKopudTeq Y$prsts RNA Pol ll
l:= oi ono(eq pre q oetpd roug npoo66vowot orouq -::cyoweq ouppo<pr1g. Auq 11 6tepyoo(o (otog e(vot : :c rponog Ytq ro ouwovtopr6 tqg pieroypoqrlq ge rr1 - -:corpq. Tetrtr<d, rdnoto ouorortrd u1g ouoreuqg r<o- -".: :otruo6evutr(toorlq nou Xprtotponotoriwot xotd tov -::-orrogd rnq UeroYpq<pr1g eniorlg npoo66vowot oe l-) rporogopul,tcoprSvq orrl oep(vq 2. t-leptdpyoq, out6 - -*Soiver 11611 on6 rrlv 6vop[r1, ;rdMov 6tot trlore 1 R- ',: i:oluu0pdoq vo e[vot Srotprq Yto rtq owt6pdoetg . rlepyoolog tou 3' dr<pou pr6].tq [er<tvrloet q oJv0e- , - -t e[o6oq on6 tov nuplvo pt6oto ttrlv nuprlvtrolv n6- : - . enlor1g etrSyXetoL on6 tr; CTD ror nL0ov6v - -..ov((etot Ue rnv ene(epyoo(o rou 3' drpou O)'o --- : uno6qtrcbvouv drt 4 CTD iooq e(vot tivo yevtr6 . (.wpo yLo tq orjvdeoq dMtov 6tepyoottov Ut rn Ue- -:.:orpq. Itrlv nep[mcoon rou crXrlpotLoprof qg ro]'ti- :tr-( Kot rnq ouppoqnq, n CTD I'ettoupye( 5ppeoo, -: rtryowoq q ouvopprotr6Ynon rov nporttvtKov ouFl- -' : (rov nou ovol.optBdvouv ttg owt6pdoetg. Iqv nep[- *il rrlg 6qprtoupy(og tou 3' dr<pou, (oog ouprpret6leL r*.30 ornv ow(6Poor1. -o rplot;to orlpre(o 6tov r<oOop((etot ov (r<ot n6te, ' -,' nep(moon Utoq noLuprepdoqq oe eyprlyoporl) 0o ..:ooore[ 6vo yov(6to e(vot 11 xa)apo4 rou uftoKtvt]rtl, :-'.:5q 11 oneLeuOdpoorl rou ev([;rou on6 rov unoxt- - - Mdltq our6 ouprBe( xot oneLeuOepol0otiv ot nopd- :,.eg dvop[4g, enttp6nerot 11 petdBoorl mq qdoq : *iKUVonq. To pretoyporptx6 otiprntroxo onoteefuot - rcv on6 r1v RNA notrutrrepdoq ll, toug BootroUg no- :-.oweq TFrrE rot TFrrH, nopdYovreq enrprlxuvolg - .g o TFrrS) Yto vo e;-ino6(oouv rl'lv or<otdMr1trq noU- :* <ot d).o to dv(upro Kot rouq nopdyoweg nou e(vot -:cc5e6ep5vo oro CTD. Aut6 ro o0;lnl'oxo np6net trr;- : r 'vo UeroYpdqet 6vo er;.roye(o Xptoprot(vr1g, dtoptSoou '!v vouKleoorogot(ov. OL6r<i.qpo to yov(6to pnope( vo 3ploretot oe ovotXrrl Xptoprorivrl, t6(trlg ov 6ev evot no- .u ireydtro. EvoMorrtxd, oe ovotar4 lpcopror[vq ;rnope[ : bpioretoL pr6vo 11 neptoxrl yJpro ond rov.unortv4r4' ','eprr<d yov(6to, 6nolg to yov(6to tqg Mutr<r1q Auotpo- p(oq (Muscular Dystrophy - DMD),5Xouv g6ye0oq Ue- yoBdoeov ^o, yi' out6 'q preroyporprl. touq 6topre( noiL6s tipeq. To or<roprep( tcov totovrov np6net np6- ;"d uo -rponono,r1oo0v -optop6vo npoo<optvd ono- ouvoplloloyotivtot- rt 6netto vo ['ovoouvopUo- ilrnaJ;;o expove(o (BL5ne Keq6)'oro 10 r<ot 28)' To lO,o io orcro;iepriq e(vot 5to<popettx6' tiXowoq owtKoro- il;;,;;;;"ovtrr1 H3 ue rnv evotr)'o^tt^1 uopan H3.3"' - i,i1-,,grro p' 6vo notrot6tepo prow5lo' 11 npdrtq no- ),ru;;A;n no, eo cpUyer on6 rov urloKtvnrrl evepye(oq ouirurr-nc. K[pto ]'ettoupyto trlq e(vor vo.dtootpotr(oet J-i'iio*Lnpo to yov[6to 0o e(vot oe rotdmoon ovot- ;,i; il;u;.(vqq. MetocpSpet ;ro(( qs ev(uprtr<d orip- nl.oro yto vo -pot-,onoitloet rLq tot6veg Kot v' ouoO,o;rop.pOoet rn Xpopor(vq H rm6vr1 H2B uq(oro- roL OuroUtrn p,ouo-ou l,ntnoutrtv(tr;or1 (monoubiq u iti na- ii""i *ti" evepyd getoypo<p6prev-n. xpouor(vr1' Aut6 e(vot onopo(trlro Yto ro 6e0tepo gnuo' rn preou(oot1 rqq ror6vrlg H3, nou pre q oepd rrlg elvgt anopoiulrn y,o.r1u npoo6Lruoq ttrlv ovodtoptop<pcorriv rng Xpo)Ue itnnq G).ent Evor4req 10.4 t<ot Ev6t4ra 28'11)' Itilitpovo pe ro nto np6ocpoto Uow6I9, xdOe nolur' pepdoq' xprloigonote( 5vo otlprn)r,oxo ovo6tog6p9oonq ;t xdi,;.Gq ;ro(( pe uro tor6vn o1 ou1o66 npotet- "n y,; J oqotp5oet 6vo Etgepdq H2NH2B, oqrivowog 6vo'e(ogepSq (orn 06o'4 rou orroprepo0g), nou e(vot eurol,6tepo v; onoporpuvOe( npootoptvd' Aut6q ot rpo- nonotrloetg e(vor en(orlg onopofutlrcg Yto rnv enqvoou- uoplroi6y,'1orl rou ol(opepotiq tou vou rtreooulpot(ou mo'DNA 1.retd q 6tdLeuo4 t4q RNA noLuprepdorlg (BLdne Evoulta 10.10). 'Onog ovo<p6popre atry Ev6t4ta 20'7, to motle(o otrtrqtrouX(og tou DNA nou onotel'o0v tov rewptr6 unoo,uqtq' e;rqov(ouv orlprowtr<( erepoy6veto, 11 ono(o prnope( v' onotetre('rn Bdo1 Yto rnv et6u<6trpo rou uTIo- r<wrp{. 'Evo on6 out6 to cnotXe(o e(vot yvtoor6 Ue rov opo xoupn[ naioqq (pause button): np6renot Yto ptto nLotioto o"e GC otrtr1trou1(o nou tunu<d Bp(orerot ro- 0o6xd t4g oqeqpiog. Aur6 to orotXe(o 5Xet Bpe0e( o' 6vov onpoo66r<qto preydtro opt0pr6 ovomu(torriv Yovt- 6(cov qg DrosoPhila. H yevrrcl dtepyoo(o rrlg 6vop(qg e(vot nop6proto pe q 6tepyoo(o nou rqtol'ferot on6 q Bolcqprorrl RNA noLuprepdoq. H np6o6eorlqg RNA notru;repdo4q 6npt- oupye( 5vo rLetor6 o6pntroro, nou 6nemo peto'rp5nerot oe ovotXr6 oUgnLoro mo ono(o ot r.tr'cilvot rou DNA 6Xouv 6toXtoptore(. Itrl Borrcrlpror( ow(6poor1, o oxn- prolopr6q rou ovotxro0 ouprnL5xou oLoxl'r1pc0vet rnv onopofu4rrl 6oprri olloyri tou DNA. Mto 6torpopd orrlv europutottrc] ow(6poor1 e(vot 6tt pretd on6 out6 to ord- 6ro ononeftot nepomSprr: onoetr(rcrloq tou erpoye(ou' T6oo mo Bor<tripro 6oo rot orouq euxopuri:teg, undpXet dpeort o6vdeorl preto[ri qg RNA nouprepdorlq oo,.iq evepyonotrlorlq qq ent6r6p0roonq' Aur6 opXtrd 5ywe owtl.r1m6 enet6rl ro geroYpoutrr6vo yov(6to ent- 6rcplrisvowat xard nporfulor1. 'Eror, avaxaA0tp1qxe 6tt or6Xog e(vot pt6vo o xLc0voq erproye(o rou DNA, evri o xL<tvoi prq-er1-roye(o entdtopOrilvetot Ue rnv (6to to1itrl- ro 6n<og rot o r6ptoq 6yrog rou DNA' 'Orov r1 RNA no- LrU"pa'"n ouvow{oeu BI6Bn tou. DNA o-rov r}'ci'lvo il;y"(o, orogot6 enet6( 6a.gnope( vo xpqoqronoqoet rq npoBq;rotu<5g oMrl}'ouXbq oq erpoyeio,yto vo Kq- ili, e i"t, -d (euy6ptopo ttov ou prnr1 prrr;tonrcbv Bdoetov' nuiO tgnvei rnv ei6tr<6qto qg en(6poonq Yto rov-rtr<i- "" L^u"vtf" ftux6v gldgn orov ^I9Y9. prrl er<poye(o 6ev nopr1,,r,oOiq"i rrlv npo6l'oon rnq RNA.notruprepdonq)' jin o,"pvoo(o trlq enr6t6p0tr:or1g epntr6retor o yevtr6g uri"wt,fiOg nopdyo*og TFllH, o ono(og uq(orotot oi ruoLl.oo,o6q uopQ6q nou onorctro6wor on6 uto Kev- ip*n *p,oxn ouv6e8epr5vr1 pe dlleg unoprovd6eq' ' O fff ri-g1et rotvri )retroupy(o t9o9 mnv 6vop[r1 -nslrt-oYpo6ng 6oo rot orqv ent6r6p0toor1 tov Btro- BOv.'Ot iSteq unoprov66eq el'rdorlq fiPB xot XPD) 6n- lrroupyo,lv tqv opltrrl <PuooL(6ocrlltypoqqg rot ono6tfidooouv to pNA oe prq 06orl BLdBrlq' Ynoprov6- ;t; G;6vopo XP t<trldtronoto0wot on6 yov(6to trov ?S,S H 'Evop!r1 Axo)rou0e(rot on6 K(Oopon rou Ynorwqtrl rot Emg
ofloiov geroMd[erg npoxo].o6v to v6oqpro pelayypot- otxrj fr1polepp[a (Xeroderma pigmentosum), nou Xo- porcrrlpf(eror on6 npo6td0eorl yto ropr(vo. O p6].og rov unoUovddrrrv TFIIH orlv entdt6p0oorl tou DNA ovq.hie'rqr Lemopeprirg ornv Ev6rrya 16.4. H l,enoupy(o qg entdr6p0oroqg nr0ov6v onotte( rponono(r1or1 q ono66gnon rnq croproqpr5vqg RNA noLuprepdorlg. H peyd).r1 unogovd6o qq RNA nol.upe- pdonq 6rooildtor 6tqv ro 6v(upo mopord oe O5oetg BIdtsnS on6 unepuirdrl orcnvoBol,(o. H otiv6eorl pero[[ rrlg ouoxeurlg peroypogrlq/enrdr6pOtoorlq Kot rrlg 6td- oroonq r4q RNA nol,uprepdorlg nopopr5vet o6teuxp(vt- or4. l-h0ov6v q nol,uprepdoq np6ner v' onoporpuvOe[ pr6l,rg oroprorrloer. 2O.9 Or Evroxurdq flepr6xouv Apgi6popo Erorxeio nou YnoBon0o0v rnv'Evop[n e 'EvoS evtoXuqg evepyonole I rov unoxtvlrl nou pplorerot nto xovro rou ror pnopel vo pp[oreror oe onoto6tlnore onoorool elre ovo0rro e[re roOodrro ono rov unortvrlrfi. * Mro AUS (ovodrul evepyonorllrrl otrlntrouxio - upstream ac- tivating sequence) rou (u;ro1.ruKrlTo ouUneprQepeTor oov tvr- oxurng, otrtro treuoupye( povo ovo6rro tou unonvqrl. e 0t evLoxureq oppon(ouv ouprntroxo on6 evepyonou'lTsq n0u otrtrr1trenrbpouv opeoo ll egpeoo pe rov unorrvqrq. Irrq nporlyoUpreveg nopoypdtpoug, o unoKtvqrnq nopouordorrlKe oq onopovroprdv4 neproXrl uneU0u- vrl yro qv np6o6eon rnq RNA nouprepdonq. Om6- oo, or euKopuolrrro( unortvr;tdg 6ev evepyotiv ono- po(rr1ro on6 p6voL roug. Ttg neproo6tepeq eop5q, n evepy6rrlro ev6g unoxrvrlrrl ou[dvet ewun<ootord on6 qv nopouo(o ev6g evtoluq nou Bp(oxetot oe drd<popeg onoordoetg on6 tov rewptr6 unoxtvrlrq. Kdnoror evtoXur6g trettou pyotiv pr6ocr: oll.r1tr ent6pd- oeu:v pey6trou BeLqveroriq, qq rdqnq rrov Serd6tov kb. Aw(Oero, dLi,ot trettoupyotiv pSotr.l oLl.qLent- dpdoeov tr-ur<po0 Belqver<orig r<ot pnope( vo Bp(- oKowor opretd rowd mov rewpu<5 unortvqni. 'Evo on6 ro npdlro rowd ororle(o nou neptypdgrq- rov rowd orov unoKrwlui rpov 4 oMqLouX(o or4 06oq -75, 4 ono(o orlpepo ovoprd(eror r<our( CAAT, on6 tr1 ouvorvetrrrl ol,L4louX(o qg. Iupd Bp(oretot rowd mr1 06oq -80, oll.d ;rnope( vo tretroupy4oet on6 ono- ordoerg nou norxfi.ouv nolti on6 t4v oEerlpfo. Aet- roupye( Kor Trpoq lg 6rio roteuOUvoetg. Ot entmdloetg peroMd(eov mo rowi CAAT onoteLoriv 5v6et[q yto ro yeyov6g 6n ro0op((er onoqootortrd r4v onoteLe- o1lotx6rr1ro rou unoKrvrlr4, evcir 6ev enrlped(et rnv er- 6rr6qrd rou. 'Evo derirepo rorv6 ovodtrd morXeio e(vor ro r<ow( GC orrl 06o11 -90, nou nepdXet qv oMr;- LouX(o GGGCGG. Or evroXurdg ouXyd nepdXouv notr- l,onl.d owtypogo ourof rou ororXe(ou, efue orq pto efue orrlv dMq r<ore60uvor1. To rout( GC efuor en(oqg oXenrd row6 ororle(o xowd orov unor<rw1r1. H dnogr1 6tt o evrolutqg 6rorpdpet on6 tov unoKt- vqtrl owrrorontp((er 6Uo loportrlptmrrd. H 06or; rou evtoXurq oXelrd Ue rov unortvqtrl dev e(vot oro- 0epr1, olL6 prnope[ vq norKil,er no].0. H EIK0NA 20.15 6e(Xver 6rr o evtoXurlg pnope[ vo Bp(orerot qvo6txd, roOodrxd n U6oo o' 5vo yovi6to (runtrd mo rwp6vto). Enrnl.Sov, prnope( vo l,eroupye( ror npog ng 60o ro- reu0Uvoerg (our6 or1;ro(ver 6tr gnope( v' ovompogeQ oe oXSoq pe rov unomvrlrrl. Xerptopo[ tou DNA 6e[- ;3rouv 61 6vog evrolurr]g prnope( vo 6reye(pet oTlotov- 6qnore unoruvqrrl rono0erqOe( yUpco rou, or<6pr1 rot 6er<ddeg lrLrddeg Bdoerg trrorpud npog onotodqnote rore[0uvo4. 'On<og o unor<rvqtr1g, o evtolwrlg (rl o dMog eow6g ElK0tlA 20,15 'Evoq evtoXurrlq Unope( vo evepyonouloer Svov unoxrvrlrrl ono ovodurl n ro0odxrl 05o1 xot 1 otrtrr1trou1(o tou pnope[ v' ovoorpoqel oe oX6o1 pe rov unorrvqrrl. 0 ovodrxdq evrolurrlq evepyonoreI rov unoxrvl14 Ynouvrlqg 0 xo0oduoq evroXurrlg evepyonotei rov unoxtvrlrq 572 KEOMA|0 20 H Meroyporprl orouq Europu6reg ; dworeg
- - - r, cq onootconllrllq) e(vot 6vo nolupep5q oroLle(o - - - : .oreohot on6 Bpol6o orotle(o DNA Snou npoo- ,, -.:oi dtorpopot t[nor petoypotptrov nopoy6wt':v' - ., c-1utdg l.rnope( vo e(vot ontro( 11 otivOetot, ovdtro- r ..i :ov optOp6 rov orotXe[ov Kot rov t0no tolv peto- - . - .- *, nopoyowov. 3. *etoypogtr<o( nopdyoweg nou npoodtivowot o' , l, --i0g 6Lorp[vovtot oe OetLNoUg ror opvrittr<o0g - r: r, ovreq. Ot 0ertro( 6Leye(pouv tq pretoypotp4 .','EpYonotnr6g - activators), evco ot opvqttro( 11v . :-str.ouv (xorooroheig - repressors)' Ie roOe :.::*rvq ortYpl, dvo r<rirropo 0o nept6Xet 6vo prtypto . .-:.:ccptt<tr.lv nopoy6wr.ov tt<ovtrlv vo npoo6e0o[v o' , '.. t','toxurfi. Av npoo65vowot neptoo6tepor evsp- - -: -;eq on6 xotooroLeig, ro croLXe[o 0o eivot evt- , - -c Ie ow(Oeq nep(mr.':o1, 0o e(vot onooLonn- _. : I. cq dl).oq tp6nog dtdxptoqq ttllv preroypoqtrrltv - :: r . clrov Boo((etor orq Lettoupy(o toug' H npo-rq - :i-,apilo;lBdvet toug npaytrtatxolq uepyonoLqr{'q ",: activators), ot ono(ot Spouv t6oo pr6oo np6ode- r-: ti ouyxerptpr6veg O5oetq -rou DNA 6oo xot 6r1pt- - -: . ivroq enocp6q ;.re tq Bootr<r1 ouoreurl otov -:, .irn, e(re dpeoo (on6 ;r6vot roug) e[te, nro ou- r 'oou-) ouvepYoilotrlttrlv 6ncog o Mediator' ot no- - -.:, isq outrlg qq tolrlg trettoupyo[v e(ioou t<oLo , '. -cve[o DNA n Xpoprot(vqg. Arjo entnp6o0eteq td- :. : . . oOyonotntcilv eXouv ewel'trlq dtotpoperrxd pnxo- - ..l svspYonotnonq. H npoltrl nepLLopBdvet evep- :-:.nrsq nou snrorpoteUouv 6v(upro rponono(qo1g -- r,oopor(v1g t<ot otipntroro enovodtop6pQoong -- : ,.cLogor(vqq. l-1oMo( evepYonotnrSq orqv npoYpo- - . ::-rc Lertoupyotiv t6oo tr-lg npoyprortroi evepYo- - - -^:tq ooo xoL pr6oo npoodLxuo'r1q tpononotqtolv -- - r.c0JUor(vlq. H tpitrl td(r1 nepfioprBovet opxtre- .-, ..oouq UeroYpoQtKorjg nopdyovreq (archi- '.-'-'a transcription factors) , H provodtxrl Lertoupy(o -: - : iivor v' oMd(ouv rn 6oUn rou DNA, tuntrd vo to . :.'-r:ouv. 'EroL pnope( vo ep0ouv r<owd 60o oplrr<d r- - -trKpuop6vot geroyporpLr<of nopdyoweq, Steuxo- - , - .aog rl ouvepYoo(o toug, Itrlv en6pevq ev6rlro' : - ::ilooouue nto npooexttxd nirlg ouvepyd(owot o' - r, .',/loxurrq ot 6to<popetLr6q td(etq rov evepYonot- ---, (ct rcDV Kclroorotr6tov. Ito Kecpdtroto 28, 0o e(e- -:r:-.it rl ueroYpogrrrl pri0ptorl pre neptoodtepeg : . - JLPUTL' I:r (rj;rr1 undpXouv crrotleio ov6).oyo Ue rouq evt- . , - iq nou ovopld(owot ovo6tx6g evepyonorqrl169 ; ,-r lAouXieg (upstream activating sequences - UAS)' '-::.t vo Leroupyrloouv Kot ortq 6Uo roteu0fvoetg -. -.:tpopeg onoordoetg ovo6trd rou unoKtvltrl, 61t - --: (ot 6tov Bp[oxowoL ro0o6tt<o 'EXouv pu0prtortr6 : -' : Jiog rL ere( npoo65vowor ot puoptmtreg npolte'l- :: :cu evepyonoto[v ro ro0o6tt<d yovi6to' -iLpd;.roto ovoo0orootlq oro ono(o q oL)'q)'ouX(o - - - I rLo{urll otpotpe(rot on6 to DNA r<t 5netto evoopo- --,3.ct oe ronoto d).).r1 06o11 tou yovtdttill-totoq 6eG . - - ., orr rguototroyrr<q uetoYpo<prt prnope( vo 6torr1- pqOe( e<p6oov undpxeL evtoXurrlg, ove[oprlrc ] : :- ' or<pLBrl ew6ntorl tou (opr<e( vo Urlv undpXouv irc'';rii oro DNA nou peootr'oBe( B).6ne Evorqta 10'12) At i': yov(6Lo B-ogotp(vqq tonoOetq0e( o' tivo pdpLo DA no, ntpi6xti 6ro, er,oxur1, in vivo I ueroYpoQn rou O'ouEnde( ndvr,-l on6 200 <pop6g, toro Kt ov o evtoxurlq Bp(onerot opr<et6g kb ovobtr<d q r<oOobtrd rnq oQer!- pioq, o" onoto6rlnote npooovotol'Lo;r6 Ax6ptri 6ev 6Xet ovoxol.urp0e[ oe notd on6moorl netiet vo ]'ettoupye( o evLo'1urr1q. 2O.lO Ot Evtoxurdq nelroupYo0v AufidvovToq Tn IuYKdvTpoon rov Evepyonornrdv Kovrd oTov YnoKlvnTn * 0r evtoxursq ouvflOutg trettoupyo0v povo oe cis Stouoptptlorl 0t 0x00n ue Tov unoKtvllTn-oroxo * Ko0e evtoxurrlq unoypetirverot vo ),etroupyei p6vo orov ppe- Oet xovro oTov unoKtvllTll Or evtoXutdg Lettoupyoriv npoo6Svovtog ouv6uo- opro0q 1-reroypocptr<cov nopoy6wt :v, e(re. Oettxriv eire opvrlrir<cirv, nou etreylouv rov unoKtvqrq r<ot xot'en5- Kroon rrlv 5xgpoorl rolv yovt6(ov. O unortvqrrlg e(vot ri 06o11 6nou, srriv ovotXrrl Xptoprm(vq,.npoo65vowot or Boorr<oi peroYpoqtKo( nopoyoweg 6tot oore 11 R- NA no).upepdorl vo Bpe( tov unortvqtq l-lr'rrg dvog evtolutrlg 6teya(per trlv 6vop[q o' 5vov unoxtvrltrl nou'1-rnope( vo Bp(or<etot oe onotodrlnore ondorooq npog onotodfnote ntreuPo tou: H Lettoupy(o tou evtcnlurrl nepil.o;rBdvet otr-I'q)'ent- 6pdoetq pre rr1 Boorxq ouoreurl o--ro orotle(o rou Kev- rpu<o6 unoxtv4tq. Ot evtolut5q e(vot nol'upepq, 6no-tq Kor ot unoKtvrltdg. KdnoLo orotle(o Bp(or<owoL tdoo o' evroXur6g preydl'ou Bel'qver<o6q 6oo t<ot o' evtoXur6g nou.[p(orowot t<owd orouq unoKtvrlrSq' Kdnoto [eXo- p,*d -o,Xt(o nou Bp(oxowot t<owd oroug unortvqt5g 1-rotpd(owot Ue rouq onopoKpuop6voug evtolutdg rrlv rrov6qto vo Letroupyoiv on6 dtdqopeg onoordoeLg Kot npoq onoro6qnote npooovotoLto;-t6 'Etot q 6tdt<pt- o,1 1r"-oi,l evtcrlw6v preydLou t<ot ptr<poti Bel'rlver<orjq 6ev e(vot ooQnq. O ouotooru6q p6).oq tou evtolutq (ootg e(vot 11 o0- Enorl rnq ouyr<6wproorlq rou evepYonotrlrrl rowd otov ,noo,u,'l-tn oe dtopr6ptptrlorl crs. t-loMd netpdproto 66et- [ov drt to en(ne6o qq 6xqpoorlq tolv yovr6(ov (611]'o- O(, q tol:qro rrlq pretoypo<pqq) e(vot ovdtroYo ile rov xo0op6 opr0pr6 to:v 06oeov npdo6eo4q rou evepYo- noulrq. Ooo neptoo6tepot evepYonotnr5g e(vot npoo- 6e6epr5vot oe Uto 05or1 evtolurq, t6oo uqJn^'6repo ro enine6o tqq 6rcPPoorlg. To pLBootoprtr<6 RNA rou evtoxurq rou Xenopus :tt."i,; 0r EvLoxurdq Aeuoupyouv Au[ovovrog rr1 luyr6vrpolol rtrlv Evepyonorrlrtirv Kovro orov YnoxLv-l- 573
At#EvtoXurr'19 'Evog evtoyurrlg pnope(vo ),etroupyqoet rpepvovrog nporeiveg rovro orov unorrvqq.'Evog evroxurlg rr evog unoxrvlrnc oe Xoptoro xurtrlro DNA dev otrtrr1trentOpouv 6nog oro (f), oIIo pnope( v' otrtrrltrenrdpooouv orov ro 6uo popro elvor ouvdedeg6vo o),u0.- droro 6ntog oro (B). /aevls pnope( vo evepyonor{oer rn Ueroypog4 on6 rov unoKrvnrn rnq RNA no).uprepdonq L Aurrl 11 66- yepon e(vot oletrrd ove[dpqq on6 q 0dor1 r<r e[o- rolou0e( or6pro K[ ov o evroXurqg ogorpeOe( on6 ro Xpop6oro;ro ror tonoOerq0ei go(( Ur rov unoxrvqrq rou o' 6vo rurllr6 nLoopr(dro, evci ow[0ero, no6et 6rov o svroXuqq Kor o uTroKr.v4rrlg Bp(oxowor oe 6ro- gopettrd ntrootrl(6ro. Av 6prog o evroXurrig ronoOe-rr1- 0e( o' 6vo nLoopr(dro ouvde6ep6vo o),uor6ord pr, 6vo de6repo nLoop(6ro nou neptSXer rov unoxrvrlr4, 11 6vop(4 e(vor oXe66v 16oo qnore).eoproru<{ 6oo ror 6rqv o evrolutrlg Kor o unoKrvqtrig Bp(orowor oro (6ro r<ur),u<6 ;r6pro 6n<rlg go(vetor mrlv EIK0NA 20.16 (;roLov6rr, oe our4 r1v nep(nroon o evroxur]g ene- vepye( orov unoKrvrlui rou oe dropr6pqoorl trans). Au- t6 uno6rlLrirver 5rt ro x0pro XopoKrnprorrr6 e(vor 11 ew6nro4 r1g nporeivrlq nou e(vor npoo6e6epr6vrl fiov evroxur4, q ono(o ou(dver rlv nt0ov6tnro rou evroXurri vo 5).0er o' enorp4 Ue Uro npo:reiV4 npoode- 6epdvq orov unortwlr4. Eg6oov nparreiveg npoodedep6veg o' 6vov evroxu- 11 oe on6oroorl oprer6v kb on6 6vov unorrwlrl oMq- ).enrdpo0v dtrleoo pre npcoretveg npoo6e6e;rdveg rowd ornv oqernp(o, 11 opydvrrlon rou DNA np6ner vo e[vor opr<erd eu6Lu<rr1 rilore vo enmpSner orov evroXwq vo Bpe0e( rowd mov unorwqri. Aw6 npounoO5ret 6n to evdtdpeoo DNA oplpror((er pto Ueyqln nOrlfud". T6roeg 0r1Lr5g rilouv nopornpnee( oneuOe(og orqv nep(m<oor1 rtov Bomrlprorciv evrq4urriv. Tt neprop((er qv evepy6r4ro ev6g evrqXurl; Tunxd enevepye( orov nlqouiorepo unoKr.vrprj. Ie opropr6veg neprmc0oerg o evroXutrig Bp(oreror pero[6 60o unoru- vqrcbv oMd evepyonoref p6vo rov 5vov qn6 roug 613o Ue Bdon er6rrSq nprr:te'tvu<5g oM4lenrdpdoerg prero(ri r<ov ouprnl6rov nou e(vot npoode6e;r5vo oro 66o qror- Xe(o. H 6pdoq rou evroxurrl pnope( vo neprop[(etor on6 6vqv provorq - 6vo ororXe(o rou DNA flou nopep- no6(er qv en(6poon rou evroxur{ oe unorwqr6g ndpo on6ro govoq (BI5ne Ev1tqra 10.12). 20.l I H 'Ergpoon Tov fovt6iov lxeri(eror u€ Tnv AnopeouAioon o H onoge0utr(too4 oro 5' drpo rou yovr0(ou eivor onopoir4nl yro q per0yp0qrl. H pe0ul(cooq rou DNA e(vor 6vo on6 oprerd pu0pr- oru<d ouprBdwo nou enqped(ouv 14 6poor4pr6rqro ev6g unoruvrlrn. H preOuL[ooq orov ufloKrv4rq pnopei vq nopegno6[oer q peroypog4 xor q ogo(peorl rov peOuLoprd6cov e(vor onopo(rqrq npoun60eorl yro vo evepyonor40e[ o unoxrvqfig. Aur6 ro rporv6prevo dler ge).eq0e[ ro]d oe unorrwp5g 16oo qg RNA no].upre- pdonq I 6oo xqr qg RNA nol,uprepdoqg il. Irqv npoy- 1-rorrr<6rqro, q pre0uL(crloq e[vor 5vo ovrrorponr6 pu0gronr6 ouprBdv. Evepyonore(ror on6 rpononor- 4oerg rtrrv rorovciv, prero[ri dMcov qnooreru],(oon Kor pe0uL(oor1 (BI6ne Ev1nya 28.1 0). Me0u).(ooq en(o4q ougBo(ver Kor oq enryeverrx6 rporv6;revo. Irqv nporerpSv4 nep(moon, n rponono[- rloq gnope( vo ouprBe( er6xd oro oneptrroro(rodpro rl ro odpro, o6qyriwog oe 6togop6g prero[ri rrov 6rio oMrl- l,opr6pgov or4v en6prew'1 yevrd nou prnope( vo npoKo- L5oouv 6rogoprig mqv 5r<gpoon rov norprrriv ror pqrprxcbv oM4lopr6p<pcov (B),6ne Ev6trya 29.8). Ie our6 ro re<pdloto e[erd(oupre ro pdoo pre ro ono(o 11 preOu).(rooq en4ped(er rn Ueroypqgrl, ro ono[o e(vot ove[dprrlro on6 ro ov or pe0u].oprd6eg npom60q- 574 KEOAAAIO 20 H Meroyporp4 oroug Euxopuoteg Boorxeg evuoreq
ndqn us Mspl nam P8tho . r, n oeotp6Orlrov rog pu0prtortr6 q entyevertr6 ou;t- t-., i ;re0ul(oor1 oe unorwqt6q qq RNA notrupepooqq ,,,u[olur, oe dtvoureoti6to CG (ovoq5powrot rot 6t- ,."-ol!oiio,o "CpG,). H r<otovoprl tolv. pre0uoprd6tov .-l.orl vo e(etome( xpqotponotowoq riv(utrto neptopt- , -.u .o onoio n6mow b6o"tq-*6xoug nou neodxoyy -: 5rvoux.treot(6ro CG" Arjo r[not neptoptopou ouYKpt- .l,.roL ornv , '' To roocrtrt(oprepq nou Xpnot- -.:noLqOqxov oto ne(pogo elvot ev(upto to ono(o Ir.-oru tqv (6ro oLr1ou1(o-mo1o.oro DNA' o)'Ld .,cuv 6to.popertrd onotetrSo;toto ovdtroyo Ue rnv Ko- lf,Tool Pre0u(tooqq tou DNA' --i;;(rt- Hpall n6met trlv o)'rllouxto CCGG (vod- .rroi,l olLnLorXlo p6vo tou ev6q xtrtrlvou tou DNA)' :-Joo ov q 6e0tep1 C e(vot preOutrtcop6vrl' to 5v(upto :, irnoptl n6ov v' ovoyvop(oet q 06oq' Aw(0eto' to ,,lrJ" Mspl n5nter trlv (5ro 05or1-or6xo ove[dpqto , .. -n, xotdmoorl pe0ul'(rooqq orrl ouYKe-Kptp6vq C' :-'renrq, to Mspl lrnopei vo Xpnotponotn0e[ toore v' l, "vtrpi*"tv 6treq ot oMrlou{eq CCGG rot to Hpall ..: vo ro0optote( ov e(vot ge0utrttolt6Ye.S . - ev to ONA e(vot ope0u}'it''lto, to 6rjo 6v(upro 0o 6n- - ...lryqoouu ttg (6teq (oveg neproptotrrot'^Ito geOu- ' --.uo DNA, 6pcoq,'oi -pot'onotrl;reveq.06oetq 6ev -.-::cwoL on6 to Hpall' I-to r<d0e tStoto 06oq' 6vo pe- .'.-..po x).dopro Hpall owtro0tord 60o rldoproto M- :: -r ' 6(vet 6vo noPd6etYPto' -ctr'o yov(6to nopouotd(ouv 6vo np6tuno oto : t.c rl i<otdoroorl geOu)'(oor1q eivot oroOeprl ortg ne- , - ro.tptq O5oetq oM6 notril'et oe 6Meq' Kdnoteq 06- -'-, ;i;;, preoultrrrpr6veg q opreOuL(oteq oe 6trouq toug -.. ;. ALLeq O5oetq 1;rio' petov6trlto) e(vot pre0utto;r6- ': ce Loro0g orouq ono(ouq to yovl6to 6ev errppd(e- -: ovo e(vot opeOu}'(oteq oe toroUq 6nou to yov[6to . ..,-tutpVO. 'EtoL, 6vq evepy6 yov(6Lo prnope( vo nept- : lQe[ og unotrre0u].tco;r6vo' ffi Ztivn onorletmtxd mo HPall E ffi owxoOtmo rtq (tirveq tou ttlslt ffi ES- zOues qnor[etmtxd oto Msol | ] I I = Ueeultogdveq Odostq I L llr ZoVTI omv iDro 0eorl - L l =oue0u(orn06on .l 0r edoetc KoBowot oveqoprrlro ono n1 geOul[oon pi{aMg."ffi t Meouhtog6vo Mspl oxl [eoulrr,lu6vo[ ]lo ccdc -_-cgcq H oue0utr(olu 06on KoPeTot U npilrr r,rre I sa?fffraacffi';'-ffi;ffiGGCC lle H pe0utrttoP6vr1 0doq 0ev rdPerot To 6v(upo neptoprogotl Mspl n6nret oheq rLg otr- ,-ouxieq CCGG, e(re e[vot f-ie0u]'Ltogeveg oi1 deurepn C' e(re - ., ;vii,; Hpall nenret p6vo ro ope0utriolro rerpoueprl CCGG' E|K0NA20'lEToonotetr6ogotoflqn6UnqueMsplrotHpall ;;ffiil;i ue nlenporpdpnorl to:v rtroopdrov neptoptopot oe nr1xu1. I-letpdproto pte to 96pgot<o S-o(orund(vq nop6Xouv efrfr"on on66eiEn Yto ro yeyov6q 6tt 11 onopre0u)r(oor1 fi;;i; oonvno". oe 6rqpooq tov^vovt6(tov' To <pdp- Io^o ,uoruo.tivetor o'o oNR orq 06o11rqg r<ur6fur1q [;i;t";;ffi vo 1-reeumoee(' Aur6 o6qve( orqv e;re6vt- ililt6r"ra;-rr'e6o",u oro DNA pretd qv owrvpoq{ (pMne Etr<6vo 11.10)' Ot tpoworuntreq entmdroetq rnq 5-o(or<wt6(vr1q ne- prLoprBdvouv qv enovtov( oLl'ovtirv *ly ^:TT.::l b,o,poponohonq rov t<urtdp.tr:v' l-to nopd6etyuo' uurKq ^,:tiobo endyowot on6 np6Spopro pr1 purr6 ry11>: to qdppor<o evepyonoe( en(oqq yov(6.to oro otonnpo voou6otouo X, yeyov6q nou ougQove( ;re r1v t66o 6tt f,"dilffi ite,Jxr"ns oxet((etot ;re q 6pocnrlpt6- rnrorou xpouooouoroq' -_ !_' i*rOqind -o ,o e[etd(oupre qv xotdoroorl ueou- )r(toorlg ttr:v p6vt;ttov yovt6(tov, ;rnopo[ge v' owtnopo- |l.L" tir" ils' en,mooe,q rnq etoqY oYrlq peo uhtto;r6vo u [;;fudou DNA ot 'eo r0rropo [evtor6q' T6roto .,r,pouo* deftvouv pto ooqrl ox6oq: to-pre0utrtropdvo VouiOto e(vot ovevepy6, evr'il, ow(Oero' to otrle0u(toto yov(6to evePY6. ...!'- noia eivot 11 Srroorl qg unope0utrttopr6vlg nepto- fiq; Ito 60potopro youi6i" o-o"tpotpffiq rtov epuOpo- et6dlv r<urrdpov Tnq r6tog, q unopreOutr(tooq neptopf. (etot oe 06oerq nou ercrefvowot on6 -500 bp ovo6xd ;J.;il;;o ht o ro 60o evnr.ro o vovi6to 6tog -500 [p ^"e"ri,^a on6 to 6e0rcpo' oSoetq unope0u]'[<oonq ,Iapi" * tt o).6xlr1 pr1 tr1v neptoxq' ou ;rnepiloptBovo- uirJu' oo, tou otoorrl lroroq ovdpeoo crro 1?:i9:: I ::; i,oxn .ns uno ge0ul(trrorlq ou prnhtet. Fl.qv fl eptoxn ili}*,*a euotoorlo(oq orq Dndoq I (Blene Evot4ra io.l'l).'nrtO'to onot6l'eopro orrlp((et qv dnorl'rq 6tt q ,""lrlgriotr1 e(vor xopoKrnptoru<6 proq neptolrlq nou nrpi61r, 5vo pretoypoq6;revo yov(6ro n Yov(5ro' 'Onolq u"-"lilga ollov6q fin xpouqr(vq' cpo(vetot 6rt n [i"rol" ui,eulopo6tov gxet((etot pe tr1v xov6tnro ue- toypogf,q nopd ;re qv (6to rn UeroYpogn' To np6B)rr1po ornv eppqve(o tqq yevtr<dq oX6onq u".oi,i unoueeulioong oi t'"pyono(nonq ttov yovt6itov tiuo, b., oto tpow6pevo etr'tntr5retot p6vo pto petovorn- io tL" rrtgutuo;r5vtov OSoecov' l-lr0ov6v 11 rotdotoorl -is deli-"rlg e(vot r<p(otpr1 oe ouYKeKpu6veg 06oetg 11 oe pto neptoprog6vrl neptol4' En(onq' n el'dtcoon 20.11 H'Extppoorltov [ovtd(tov lxeti(eroL Ue nlv Anope0utrfoon 575
oro BoeU6 Ueeul(oong (ri or<5Un r<or 4 nLrlprlg oqo(pe- orlrov pe0ul,oprd6cov on6 rdnoro rgnpo DNA) e(vor ;r5- poq Uroq doprrrlg olloyrlg onopoirqtqg yto t4v e[5].r- En qq peroyporp4g. Iuyxerpt;r6vo, r1 onotrre0ul(oor1 mov unorrvqrrl [ocug efuor onopo(tr;q yro vo rov roromrioer 6ro0rior- Uo yro r4v Svop[q rqq Ueroypogrlg. l-to nopd6erypro, mo yov[6ro rnq y-oqorp[v4g 4 nopouo[o rcov ge0ul,o- prd6cov m4v neproXrl yripot on5 qv ogetrlp(o, peto{0 -200 Kqr +90, KoroqrdAler rq preroypoqrl. H o<po(pe- orl rcov rptc0v pe0ulopddtov nou Bp(or<owor ovo6rrd rnq qqrrnp(og, 11 rcov rpuiv ge0u).opd6ov nou BpG oKowor xo0o6rxd r4g, 6ev o(per r4v roromoLrl. H ogo(peor1 6ltrtv rtov pre0uloprdsov, 6prcog, ennp6net fiov unoKrv4rq vo ),enoupyrloer. Iuvenc0g, npoun6- 0eor1 yro rn Ueroypoqq iocog elvor q nopouo(o ptog opeOul,(ror4g neproplg orov ufloKrvrlq (B).5ne Ev6t4- ta 20.12). Qor6oo, undplouv e[orp5oerg oe outq tr1 yevrxrl oX6on. Meptrd yovidro er<rppd(owor or<6pr4 xor 6tov e(vot nolri trreOuLrcopr6vo. H 6norq oX5or1 perolti ;reOuL(oo4g ror 5r<<ppoo4g trov yovr6(rov 6ev e[vor ou<ouprevrrrl, oMd yevx6g rov6vog e(vor 6rt q ;re0uL(cooq onorp6ner qv 5r<<ppooq rcrlv yovr6itov, evri q ono;re0ul.(coor1 e[vor ono- pofuqr4 yro rrlv 5r<gpoor1. 2A.l2 Or Nnoi6eq CpG Eivor Pu0prorrroi Ir6xor * 0r vlor0eg CpG nou pplorovror orouq unoKrvrlTe g rotv rdLoou- ororo exqpo(o;revov yovrd[rrrv e(vor onope0u],urrpdveg. * N1o(6eq CpG pprorovror en[oqg Kor 0T0uq unonvlrdg opr- oprlvu.rv yovLdirov pe roroerdLrrl exqpool. * To ov0p6nrvo yovL6lropo nepreXer-29,000 vqo(6eq CpG. * H Ue0utrlooll luog vqo[dog CpG onorpener qv evepyonoqol rou oXel(opevou unoKrv1Tl. * Koroorotrrl nporotreiror ono nporelveg nou npoodevovror oe pe0utrLopevo (euyri CpG. H nopouo(o vqoidrov CpG (CpG islands) oug 5' ne- proX6g opropr6valv yovr6(ov ouv66eror pe rtg enrmri- oerq rnq peOul,(ooqq mrlv 6r<<ppooq rorv yovL6[ov. T6roreg v4o(6eg touronororiwor on6 r4v nopouoio ou lq pSvq g nu xv6ultog rr1 g 6tvo u r<}.eorr6rxrl g o).).r1- ou1(og, CpG (Cpc : 5'-CG-3'). To 6rvour<Leor(dro CpG ouvowdror oro DNA rotv onov6ui.rrtrrlrv oe oulv6rqro ;r6Lrg -20% ttlc, ovogev6- pevrlg ond qv ovoloyfo rotv (euycov Bdoerov G-C. (Au- td (orrrg ouprBo(ver enerdrl 6rov ro 6rvoux).eor(6ro CpG e(voL ;le0ul.rcopr6vo orr; C, 11 our6protrl onopr(vrrlorl trig prd0uLo-C q getorpdner oe T, 4 ono(o ov enr6top0o0e( eoqoLprdvo erodyet ;uo pretdMo(r1 nou ogorpe( ro 6r- vour}.eor(6Lo). Ie oproprdveg neptoldg otm6oo, 1 n_- rv6rrlto tclv 6rvoui<LeorrS(ov CpG grdver orrlv nc.- B),endprevrl trprl, dqAobrl ouorocnKd OexonLoord(et: ouyxpLttxd pe ro undl,orno yovr6(ropro. To brvourcLec:.. 6Lo CpG oe oureq rLg neproXdg yevrr<d e(voi ogeOu,L*- TO. Aurdg or nlorjoreg oe vrlo[6eg CpG SXouv ;r6ori r:i. prerlx6rrlro oe G-C -60%, nol0 preyotrilrepq ond :: yevtr6 DNA (pdoq ttprl 20%). l-lo(pvouv rq piop<pr1 r*- pdrrov DNA ;u'p<ouq 1-2 kb.Ito ovOprrtnrvo yovr6(o*: undpXouv -45,000 r5-roreg vrio(6eq. Opro;rdveg Bc- oKowor oro enovol.oprBov6pevo o-ror1e(o Alu, nL0o,,:, e[oLt(og rnq uqun].nq neprexlr6rlrdg toug se G-C, - oi.),q).ouX[o rou ovOprinrvou yovrdrcrtprorog entBepc . rrrveL 6rt, ov elorpeooriv or ouyKeKprp5veg vqo(6eq, or:3, pSvouv -29,000 vr;o[6eg. To yovr6(cr"lpo rou nowrKC- nepLdlsr fuydrepeg, -15,000. l-iep(nou 10,000 ond ..: npoB).endpreveg v4o(6eg Kor oro 6Uo e(6r1 qo(veroL c ewon((owor oro n).o(oro otrI.qAoulurtv ouwqprlpdv*. petol0 rrr.lv eL6crtv, yeyovdg nou unodr;Lrrtver 5-rr ou;:: (oog e(voL ot vrlo(6eg ;.re puOtrlrorrr<r1 orlpoo(o. H 6ol- rqg Xpropor(vnq onq ouyr<er<ptpr6veg nepLoXdg epec, - (et oMoydg nou oXer[_(owor pe rr]v dr<gpooq r<,DV yc. - 6(rov 6rov or vqo(6eg CpG e(voL ope0uA(roreg (BAo.. Ev6n1ra 28.11): lopor<r4p((eror ond 1o;rqL6repq oL- .. r<6wparor1 trlq Lordvqg H1 (nrOovdv oqpo(ver 6l 11 6c-- e(vot ).ry6repo oupnoy4g), trreydlo Bo0p6 oxetu).[cocr : tov dMotv torovciv (Xopor<rrlptorLx6 nou ouvrlOcoq qt r[(eroL Ue rrlv 5r<gpool rtrrv yovr6[ov) r<or nopouo(o e :- oov unepeuoioOrltirlv orq DNdo4 r1 oXeddv ono, . Loypr6.vorv on6 or<ropepfl rorovc6v (r<ord ro ovopevou:- vo yro evepyo0g unor<Lvrpdg). H nopouo(o peOuLro.rp,:- vorv 05oeir.lv CpG onor<tre(er on6 to vour<Leoocupror.: qv nopoMoyrl tor-6vqg N2A.Z. Aprerdg eop6q, v1o(6eg n),orioLeg oe CpG (errvo,, pr6}.rg ovo6u<d rou unoxLvrlrrl rL enercre(vowor xo0o6Lr: pSoa orq ;reroypo<p6trrevq neprolq dnou oro6tor: ,,OBrlvOUv". H ::..rr' .l . . : , Orgy6p(vet rqv nUXVdrrlrO r*. 6tvout<l.eol6(rov CpG or prcl <(yevlKr1" neproXrl rou yo,,'.. 6urr;rorog Kor or puo vr;o(6o CpG nou ovoyvc,-lp(mn<i ond qv oMrl).ou1io tou DNA. H vrlo[6o CpG nepLBo; et r1v 5' neptoXrl tou yovt6(ou APRT, ro ono(o extppc. (eroL 16too0ororo. O).o ro dLolerptorLr<d yov(6ro nou er<gpd(ovri. t6Loor-iotoro 6Xouv vrlo(6eg CpG, nou ovrrnpoocL- neUouv nep(nou ro 50% tov vqo(6otv. Ot un6),olnec Bp[or<owoL orouq unoKrvr;r6g yovr6(rrrv ;le roroetdLr<- ptiOptoq: nep(nou 50% outrrv rotv yovL6(ov dlouv vr. o(6e9. Irrg ouyrexprpr6veg nepLnro-loerg, or v4o(deg e.. vor otrle0u).(oreg ove[oprrlto on6 r4v rordorool rri 5xqpoorlg rou yovt6(ou. Iuvencbg, 11 pre0ul(cooq tcu. vrio[6ov CpG 6e oler((eror pe rn Ueroypo<pu<q r<orc. oroon rotv rmoeLdrr<otv yovr6(ov. H nopouo(o opeOt. L(ori,.rv vqo(6ov nLorjotcov oe CpG (oog e(vot ovoyKo.: oMd 6ev e(vor enopr<r1g ouv0r1<q yro Ueroypoqn. Apc 11 nopouo(o opreOuL(ortov vqo(6urv CpG pd).ov unc 6q).olver 6rt 5,vo yov(6ro e(vor 6uvqrrxd evepy6 nopc 6tt ovon6geurro 0o preroypo<pei. l-lo).L5g vrlo(6eg nc- 576 KE0AAAI0 20 H Meroypog1 mouq Euxopudrreg Boorxdg 6woreg
Y-0Q0tptvn iililiillllili 500 1000 bP APRT i ii lliiil liliilllllfillfillfrlilllilililliil l1 i I 5', E[rirvLo E[tivLo -:: .': H runtq [uKVoTr]To Yto 10 DtvouKtreoT[0to CpG rro DNA rov 0rlloolx6v ervot -1l1 00 bp, onoq Qo[veTot Yto T0 ,ovtDto qg y-orpotptvrlg. Ie pto ntrouoto 0e CpG vn0(00, 11 nurvo- .-ro ou[ovet oe > 1 0 6tvourieor(dto/1 00bp. H vrloLdo oro yov[6to :PRT (er<tvo -100 bp ovodtro 0n0 Tov unortvlq Kt tKTttvtTot ..100 bp Us00 0To yovtdto, Ko0e xo0eq ypopprl ovrtnpootdlleutt .,/o DrvouKtrtoT(bto CPG i[vor opre0u].(oreg oto (aro pre0u].ttovowot oe Kurrclpt- <eg oerpdq oe r<oMt6pyeto (r1 oe ronotouq xopx(voug)' Aurd 0o pnopo0oe vo oXer((eror Ue rnv ovtt<ov6-rqto :urci.lv rtov oeLprrtv vo ex<ppdoouv 6leq ttq Letroupy[eq rou XopoKrrlp((ouv rov tor6 on6 tov ono[o npo5plov- :oL VnopXer dvo oo<p6g nopo6etypro Yto rrlv toXupq qdoq peto(6 qg ;re0u).(oor1g tou unor<tvqrrl Kot rrlq 3,Kepoonq to:v yovt6[tov: ot vrlo(6eg CpG pre0u].L6vov- oL oro oSpovonotq;-rdvo Xpopr6oopro X ttlv 0r1troo-rt- <rrrv (B).dne Evoqta 29.5). H pre0ul.(roor1 qg vqo(6oq CpG pnope( vo enqped- Joi rrl Ueroypoprl. Or 60o nL0ovo( prqXovtopro( e(vot: . H peOu).(toon Utoq 05oqg np6odeoqq yto ronoLov nopdyowo prnope( vo epno6(oet rrlv npdodeorl tou' Aw6 oupBo(vet orrlv nepimolol qg np6odeo4g oe ;.tto puO;rtmtr<r1 nepropl 6togopettr<r1 on6 rov unort- vr1rrl (BL6ne Evoqta 29.9). . H Fe0u(roor1 trrnope( vo nporoltloet qv np6o6eoq eL6txtr.lv xotomotr6rov oro DNA. KotootoLrl nporoLe(tot on6 onotovdrlnote on6 :oug 6tio t0noug npoteivolv nou npoo66vowot oe pe- :utrLopdveg oMqLouX(eq CpG. H nprorelvq MeCPI yto ,'o npoo6e0e( oro DNA onoLre( tqv nopouoio opretolv -re0uLoprd6ov, evc0 q MeCP2 Kot pto ott<oySveto oXett- loprevo,tv npirtte[vcbv prnoporjv vo npoode0orjv oe 6vo *dvo pre0ulttrr;r6vo (erjyoq Bdoeov CpG. Aut6 e[r1ye( '.'Lor[ q 6vop[r1 rnq UeroYpo<pr1q npo0no0Stet pto ne- cLolii eLerjOepq on6 ;re0u)r.(coor1. H np6o6eor; nporei- ,'cov onotou6rlnote on6 rouq 60o rinoug eprno6((et r1 reroypo<p1 in vitro on6 6vo nuprlvtr<6 er<1itrtopro. H MeCP2, q ono(o t<otoot5Met dpeoo rrl UeroYpo- cr-1 oltrqLentdptrtwoq pe o[pntroxo orov unortvqrrl, in[or1q oMqtrentdpo Ue ro Koroorotrttx6 otiprn]'oxo Sin 3, nou nept5Xet evepy6rqreg onoqraTul[oonq srowir (BLene Eu<6vo 28.24). Auq q noporqpqoq ouvdeer ilio i ir,ouq rotooto.ttt<cilv tpononot{oeov: trre0u}titrurl tul DNA r<ot onooretutr(toorl rov tqrovtilv. Evdr or unoxtvqr6g nou nept6Xouv vrlo(6eq CpG (-60% CpG nur<v6trpo) 11 6ev SXouv epntrouttop6 oe CpG (-20% CpG nurv6tqto) yevtrd nopouotd(ouv qroxn ouoXdttotl ;reto[0 qq r<ordmooqg qq pe0u- L(toorlg rouq Kot rnq UeroYpqQnq, ot unorwqr5g pttog rpfur1g td(r1g <po(vetot 6tt pu0g((owot on6 tr1 pe0u}'ttrl- on rov CpG. I-lep(no u to 12o/o tcov yovtd(cov tou ovopri- nou nept6Xouv rtq xol,oti;reveg (qoeeve(q) vqo(6eq CpG ("weak CpG islands"), ortq onofeq 11 nurv6trlto tov CpG efvot -30o/o, evdtdpreorl peto[ti rolv 6rio dMtov rd[ecov unortvqrcilv. Autd to yov(dto Xoporcrrlp((owot on6 ro'Xupr1 ow(mpotp4 ouol6ttoq peto(0 qq peOu- L(rrlorlg tcov CpG rou unoKtvnrri r<or t4q np6odeorlq qq RNA no)ruprepdorlq ll. H onouo(o ;re0u}'opd6ov ouv66etot ;.re tr1 5x<ppoorl rcov yovt6(tov (ri touLdXtorov Ue rrl 6uvot6rpo yto 5x- qpoon). Qsr6oo, 11 r<otdoroorlrqq pre0ul[tooqg 6e gno- pe( vo 0eo:pr10e( cr.rq 5vo yevx6 pr5oo yro tov 5l'eyXo rqg 6r<rppoolq rov yovt6(rov. It4v Drosophilla melanogaster (ror dMo e(6r1 Arm6pov), to DNA e[vot etrdXroro peOu- tr rcopr5vo (nop6n 6Xet ovoyvroptoref pto pe0u].otpovo<pe- pdon, 11 Dnmt2, tqg ono(oq 11 oqproo(o 6ev e(vot ooAnq), evc0 oro vqgorrildrl Caenorhabditis e/egans xot orn (6pn 6ev e(vot xo06trou ;re0utrropr6vo' Ot dMeg 6to- qopdq prero[ri ovevepyoti n evepyo0 lproprotffiq qo(- vowor ow(orotleg pe ttg 6to9op6q ot ono(eg nopo- qpo0wot oe e(6q nou eprrpov((ouv preOutr(toor1. Iuve- nd,g, oe owo0g rouq opYovtopotig, 6trot ot p6trot rrlg pe0utr irrlorlq mq onovduLotd 6Xouv owtKqrooro0e( on6 rdnotov dMo Pr41ovtotrr6. Tpetg oMoySg nou ou;rBo(vouv mo evepyd yovbto efuot: . Mto unepeuoioOrFn 06or;(erq) eyroO(orotot t<owd orov unorwrltq. o H Xptouorivq groq neptoXqg, ouprnepil.ogBovop6vng Kor rnq petoyporp6prevrlg neproXrlg, y(vetot nto euo(o0r1t4 mr1 Dn6or1 l. o To DNA rng (6ro9 neptoX(g e(vot unoge0ul'ttrlp6vo' 'OLeg outSq ot oMoy6g e[vot onopofurlteq yto tq preroypo<9r1. 5'*r* e*$ E(rirvto E[titvto 12 2O.13 tlegirntlln An5 rg rpetq euKoputr:ttr6g RNA nol'utrrep6oeg, 11 R- NA nolupepdoq I pretoypdqet rRNA, q RNA no}'upe- p6o4 ll pretoyp6ger 6o;rtxd yovfdto yto mRNA xot nopouotd(et q preydt6teprl no u<t tr'o pto p<p(o npoi6wcov ror 4 RNA noluprepdoq lll peroypdqet ;rtrp6 RNA. To 6v(upro 6Xouv nop6proto 6opt(, pre 56o preydLeg unopo- vd6eg ror noMdq pxp6tepeg unoprov66eg. MeptrSg unopovd6eg e(vot row6g xot oro tp(o 5v(upo' : !-r':-l tll
Kop(o on6 nq rperq RNA nolu;repdoeg 6ev ovoyvo- p((er oneu0e(og toug unorrvqrSg qg. Mro r<orvr1 opXrl e(vor 6n or Ueroypoeu<o( nopdyoweg 6Xouv rrlv r<ripro eu0[v4 yro rrlv ovoyvri:proq rov ororXeiov Xopo]firlpr- orxtig otr},r1}.ou{og xd0e ouyrexprprdvou unorwqrl ror ge q oerpd roug npoo66vouv tqv RNA noLuprepdoq Kor rrlv ronoOerqoouv oosrd srnv oeernp[o. Ie r<d0e rUno unorrv4rrl, to orcrogep4 rrov rmovc0v np6ner v' orporpe0o6v ri vo geror<wr10oUv. 'Enemo ouvop1-roLoye(- tot ro o0pnloxo Svop(r1g on6 pro oepd owrdpdoeov or4v ono(o [eXtoproro[ nopdyoweg ouvdriowor (n oqn- vouv) ro orlpmLoro. O nopdyowog TBP ononekor yto tqv dvop[q ror on6 ng rperq RNA no].u;repdoeg. Ie rd- 0e nep(mcoor1, nopSXer pto unopovd6o ev5g Ueroypo- gtr<ori nopdyovro nou npoodSvetor rovrd orrlv ogeqp(o. 'Evog unoruvnrnq rnq RNA noLu;repdonq ll onore- i.e(ror on6 n).r1Oog BpoXS<ov morle[cov or4v nepropl ovo6rrd on6 rqv o<pen1p(o. Ie r<d0e orotle(o npoodd- vowor Svog 4 r1 neproo6repor Ueroypogrxo( nopdyov- teg. H Bootrf ouoreuri, nou onoreLe(ror on6 roug nopdyoweg TFII, ouvopprotroye(tor orqv oqerqp(o rr ennp6ner or4v RNA noLuprepdo4 vo npoo6e0e(. To rour( TATA (ov undpXeQ xowd orqv oger4p(o Kqr rl ne- ptoXrl Svop[qg (lnr) oxpgriq ornv oqerqp(o eu00vowor yro rrlv enil.oyrl qg ocoonlg ogerqp(og orouq unoKrvn- 169 yro r4v RNA noluprepdorl ll. H TBP npoo65vetor dpreoo oro row( TATA. Iroug unoruvqrdg Xcopig TATA Bpior<eror rowd orqv oger4p(o ;r6oal t4g np6o6eorlg rnq ornv lnr 11 ro0o6rrd oro DPE. Merd qv np6o6eoq rou TFllD, or 6Mor yevrro( UerqypogrKo( nopdyoweg yro rrlv RNA noLupepdoq ll ouvopl.roLoyoriv q Boou<r1 peroypogrxrl ouoreuq orov unoKrvrlq. 7U.o ororXe(o Tou unoKrvnrrj, nou Bp(oxowor qvo6rxd qn6 ro Kouri TATA, npoo66vouv evepyonor4r6g nou oLL4Lenrdporiv pe q Boorrrl ouoreurl. Or evepyonorqrdq xor or Boou<o[ nopdyoweg onel,euOepri:vowor 6rov q RNA nol,u;repd- or1 [exlqoa qv enrp4r<uvor1. H CTD rqg RNA nolu;repdoqg ll rpoogopulrcivetor rotd trlv ow(6poor1 6vop(49. l-1opri1er 5vo orlpe(o eno- qnq yro nporeiveg nou rpononoro0v ro RNA prerdypo- rpo, ;reto[ri d],].rrlv ro 6v(upro oxnponoUo6 14g 5' xo).Unrpog, nopdyoweg ouppogrlq Kqr ro oriprnl,oxo ene(epyoo(og rou 3' dxpou. Ko0c0g rl RNA noluprepd- or1 trleroruvekor 6ropr6oou rnq Ueroypocpu<rig provd6oq, ro orcroprepri rov torovdlv np6ner vo rpoTronorq0o0v yro vo enrp6qrouv rrl 6t5leuoq. Ot unorrvrlr6g pnope( vo 6reyep0orjv on6 evroXur6g, oMqLouX(eg rxov6g vo 6pouv on6 preydLq on6oroorl Kor fiouq 6Uo npooovoroLtoproUq, efue qn6 q pro efue on6 qv dMrl nleupd ev5g yovrdiou. Or evrolurSg en[- orlg onoreloUwor on6 opd6eg ororle(trlv, oMd e(vot nro ouUnoyr[q. Kdnoro orotXe[o Bp(orowor 16oo oe unoxwrlr6g 6oo ror oe porprvorig unor<rvqt6q. Ot evr- olurdg nr0ov6v Lenou pyo0v ouvopl.roLoyr6wog 5vo nporeivrr6 oUpnl,or<o nou otr).r1l,enr6pd pre npalrefveq npoo6e6epr6veq qrov unorw4r{, ovoyr<d(owoq ro no- pegBoM6pevo DNA vo oX4pror(oer uOq}.d". Or vqo(6eg CpG neptSlouv peydtro opr0pr6 6rvou- 578 KEOA Al0 20 H Meroypotprl oroug Euxopurirreg rLeorrd[rov CpG r<or oulvd neptBd].].ouv rouq uno(.- vqt6g ttrrv rdrooUoroto errppo(6pevo.lv yovr6(ov, n:- p6rr en(oqg Bpior<ovroL Kor orouq unorrvqrdg rL' , pu0;rr(6pevcov yovt6(tr;v, H vrlo'[6o nou nepfiopBo,r:. 6vov unorrv4q np6ner vo e(voL otr-re0utr(orr1 o-rme : unorrvrltqg vo pnope( vo (er<Lvrloer rl Ueroypoo- Mro et6tr<r1 npcorefvr1 npoo66veror oro pe0u].Lopo,: 6tvoux).eor(6ro CpG r<r epno6((er rqv Svop[r1 rlq -.- roypo<p49. BrBAroypogio 20.2 0r Europuorueg RNA l-lotrupepooeg Eivor l-lotrupepq 'Ev(ugo Avooxdnrloq Doi, R. H. and Wang, L.-F. (1986), Multiple prokaryotic RNA poly- merase sigma factors. Microbiol. Rev. 50,227-243. Young, R. A. (1991). RNA polymerase ll. Annu. Rev. 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P. and Tocchini-Valentini, G. P. (1988). Transcr:- tion by RNA polymerase lll. Annu. Rev. Biochem. 57, 873-91r Schramm, L. and Hernandez, N (2002). Recruitment of RNA po.- merase lll to its target promoters. Genes Dev. 16, 2593-2620 Bogenhagen, D. F, Sakonju, S., and Brown, D. D (1980). A co'. trol region in the center of the 5S RNA gene directs specific ir.:- ation of transcription: ll the 3 border ol the region. Cel/ l: Galli G Hofstetter, H., and Birnstiel, M. L (1981). Two conservE: sequence blocks within eukaryotic tRNA genes are major pr:- moter elements . Nature 294. 626-631.
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US/ 98. 5544-5549. Avurroryorl Maniatis, T., Falvo, J. V., Kim, T. H., Kim, T. K., Lin, C. H., Paretdr. B. S., and Wathelet, M. G. (1998). Structure and function of fie interleron-beta enhanceosome. Cold Spring Harbor Symp. QuanL Biol. 63, 609-620. Muller, M. M., Gerster, T., and Schaffner, W, (1988). Enhancer se- quences and the regulation of gene transcripti0n. Eur. J. Biochem.176, 485-495. Munshi, N., Yie, Y., Merika, M., Senger, K., Lomvardas, S., Agatioti. T., and Thanos, D. (1999). The IFN-beta enhancer: a paradigm for understanding activation and repression of inducible gene ex- pression. Cold Spring Harbor Symp. Quant. Biol. 64, 149-1 59. 'Ipeuva Banerji, J., Rusconi, S., and Schaffner, W, (1981). Expression of p-globin gene is enhanced by remote SV40 DNA sequences. Cell 27,299-308. 20.10 0r EvroXureg Aeuoupyouv Au[ovovrog q Iuyrrivrporm rov Evepyofiorrptov Kovro orov Ynorrvrlrq Avordnqoq Blackwood, E. M. and Kadonaga, J. T. (1 998). Going the distance: a current view o{ enhancer action. Science 281, 60-63. 'Ep*uvo Mueller-Storm, H. P., Sogo, J. M., and Schaffner, W. (1989). An enhancer stimulates transcription in trans when attached to the promoter via a protein bridge. Cel/ 58,767-777. Zenke, M, et al. (1986). Multiple sequence motifs are involved in SV40 enhancer function. EMB} J.5, 387-397. 20.12 0t NqoiDeq CpG Eivor Pu0gtolro[Ir6Xot r ill llil.l::!lr l: : Bird, A. (2002). DNA methylation patterns and epigenetic memory Genes Dev. 1 6, 6-21.
:a ..,. , ii Antequera, F. and Bird, A. (1993). Number of CpG islands and genes in human and mouse. Proc. Natl. Acad. Scl. US,4 90, 11995-11999. Bird A. et al. (1985). A fraction of the mouse genome that is derived f rom islands of nonmethylated, Cp-G-rich DNA. Cel/ 40, 91-99. Boyes, J. and Bird, A. (1991). DNA methylation inhibits irais:':- tion indirectly via a methyl- CpG binding protein, Cel/ 64 l'2r- 1134. Zilberman, D., Coleman-Derr, D., Ballinger, T., and Henikoff S (2008). Histone HZA.Zand DNA methylation are mutually antag' onistic chromatin marks. Nature 456,125-130. Br9trroypotpLo 581
Euppogri Kr EneEepyooio Tou RNA ffi eLooyoyrt ffi fo 5'Arpo rou Europuarnrou mRNA 06pet Kotrunrpo * Mto 5' rotrunrpo oXrlporr(eror npooOerovrog pro G or4v retrrrrl pool rou peroypotpou Ueoo evdq deopou 5',-5',. e H dtepyoo[o oXlportopou rotrunrpog ouppo(veL roro 11 peroyporpl, ron nou rooq ent1peo(er rrlv enov6vop[r1 rrlg peroypotprlg. * H 5' rotr0nrpo tlv neproooreptov mRNA e(vor lrovope- 0utrulpevl, evtir opropevo Utrpo Un rtodtronolqlro R_ NA e(vor rprpe0utrrurpdvo. + H doilrl rotrunrpog ovoyvop((eror onri nporeivrroug nopoyovreg nou enrlpeo(ouv q oroOepdr4ro, Trl oup_ potp1, rr;v eloVtoyn rorq gerogpool rou mRNA. ffi O, Iluprlvrreg Iuppotreq Iuppoqnq ervot BpoXereg Atrtrq- trou1(eg * 06oerq oupporprlg eivor or otrtrqtrou1reg nou pp(orovrot opeotog Otnlo ono ro dpro e[ov(ou-rvrpov(ou. l'lotp- vouv T0 ovopo roug ono rq 06oq roug ouyxpurro pe ro rvrpovro. 582 KEOAAAI0 21 lupporp1 rr Ene(epyoo(o rou RNA @ Kenneth Eward/Photo Researchers, lnc * H 5' 06q oupporpnq oro 5' (oprorepd) orpo Tou rvTpo- viou nepiloppovrr ul ouvorvrTrrr1 otrtrr1trouX(o GU. e H 3' 0eon ouppoqriq oro 3' (6e(ro) orpo rou rvrpoviou nepiloppover rr1 ouvotvelq otrtr4trouXio AG. * 0 rovovoq GU-AG (nou opyrro ovopo(drov rovovoc GT-AG ovorpoprro pe qv otrtrr1trouX(o rou DNA) opr(eL on ouro ro oro0epo 6rvourtreor(Oto e(vor onopo[qro ong 6uo nptireg ror 6uo reLeuro(eg Oeoerg Tov rvTpo- v[tr-rv oro npodpogo mRNA. e Ynopxouv etroooovo rvrp6vro ouyxprnxo pe ro pei(ovo lvrpovro nou orol,ouOouv rov rovovo GU-AG, * To etroooovo rvrpdvro orotrouOouv rivo yevrro rovovo AU-AC pe pro 6rorpopelx4 opo6o ouvorverxtirv otr),1- trouXuirv oro 6pro e{tov[ou-rvrpoviou. 0L Iuppotr6g Iupporpr.lg Aropo(ovrot ovo Zeuyrl * H ouppoqrl e[oproror ;tovo ono qv ovoyvdrprorl (eu- ytirv ouppotrrirv oupporprl g. e '0treS ot 5'xor ot 3'0eoetg oupporprlg e(vor tooduvopec treuoupyrro. * Enrnprio0ereg ouvrrlp4g6veg otrtrr.1trouXreq onq 5, xoi ffi
3' 0doetg ouppoqnq drorplvouv ng treuoupyrrdg 0e- oerg oupporpr1q gero[u oprer(rv otrtrrrlv nr0ovdrv Oeoe- tov oro pre-mRNA. ffi U Iuppoqrl rou pre-mRNA l-lpoXrrlpeiAropeoou Evog Ao- o0u e H ouppoqrl onoue[ tLg 5' ror 3' 06oerg oupporpflg ror pro 06ori dtortrddoolg ovodrro rqg 3' 06orig ouppo- arls r H otrtrlloux(o drortro0toolg elvor ouvrrlpnU6vn or1 (upq oIIo dXr rooo oroug notrur0rropoug europutir- Teq. * Orov ro rvrp6vto rone( orqv 5' 0eoq ouppoqrlg, oXrl- po((erot gro doprl oe oXrlpo Ldoou xot ro 5' oxpo ouv- dderor orq 2' 0eoq prog A mr1 0rlo1 drortrodroorlg g6oo oro lvrprivro. * To rvrp6vto onetreu0epdrveror und poptpr; trooou drov ronei oqv 3' Orior1 oupporpflg u 6neuo ro oprorepo ror ro 6e[r6 e[rivro ouvdeOouv pero[u roug. ffi fo snRNA ELvorAnopolrr1ro yro n1 lupporprl * To n6vre snRNP nou epntr6xovror orrl ouppoqrl eivot ro Ul, Ul, U5, U4 rot U6. * To snRNP po(r pe optog6veg otrtreg nporeiveg oX11ro- r((ouv ro otogor[dro oupporpnq. * Otro ro snRNP errog on6 ro U6 nepr6Xouv Uro ouvTrl- pqp6vrl otrtrqtrouX(o nou npoodever lq npoTeivrq Sm or ono[eg ovoyvrrlpl(ovror ono ovlodrporo nou nopo- YovTor 0e ouroovooo voorlporo. €& leopeuon Tou llpodpogou mRNA orrlv 0d6Iupporprlg * To Ul snRNP nporotret rrlv 6vop(r1 qg oupporprlg pd- trtg npoode0e[orrlv 5' 0eori ouppoqrlg peoo_r prog ovl- 6pooqg (euyoptirporog RNA-RNA. * To oupntroro 66opeuorlq neprdXet ro Ul snRNp npoo- dedegevo oqv 5' 06011 ouppoqrlq Kor Trlv rpoTeivl U2AF, 11 ono[o npoodrlvsTor 0e Uro ouororX(o nupiprdr- vov pero[u rou orlgelou drortrodoiorlg ror qg 3, Oeor1g oupporprlg. & Irouq notruxurropoug europu6reg, or npotreiveg SR no((ouv onopolqro p6tro orrlv tivop[r1 rou oXnUoTr- opo0 rou ougntrrlxou ddopeuolg. " To (euyopogo rov 0eoeov ouppoq1g enuuyyoveror g6oto opropou rov rvrpov(ov 11 rotv e[o.rvrov. ffi n 06oq lXqpolo;lorj rou ltogoldtou Iuppoqnq e To oUUntroro 66ogeuorlg e[etrfooerot oe np60popo oo;ror[dro oupporprlg (oupntroro A) nopouo(o ATp. * H npoodeon rov U5 ror U4lU6 snRNp perorpdner ro np66popo oroporidro oupporprlq oe tirptpo orrrportdro oupporprlg (ougntroro Bl ). e To oupntroro 81 enerro perorp6nerot oro otipn),oro 82, oro onolo ro Ul snRNp onetreu0eptiveTor yro v0 enttpdtl.tet oro U6 snRNA v', otrtrntrentbpooet ge rrlv 5' 0eor1 oupporprlg. * H U4 onoporpuveror on6 ro U6 snRNp enupenovrog oto U6 snRNA vo (euyopooer ge ro U2 snRNA yto vo oX4gorfoer ro roro],ulrrj r6vrpo yro q ouppoqrl. e Kor or dtlo ovrLdpooerq trans-soTrponoulorlq ougpoG vouv 0T0 evepyonorlpevo oroporblo ouppoqnq (ouU- ntroxo C). * H ovr[dpoorl oupporplg e(vor ovnorpenrrl oe otro ro 0nUoro. *q&3 'Evo EvotrLornrd lorpor0ro Iuppotprlg XprlorponoLei ALo- gopenro snRNP yto vo Ene[epyoore( r0 ,,E],oooovo,, lv- rpdvro * Mro evo),trorrur1 odr5g oupporpnq Xplotponorei pro otr- trq opo6o snRNP, q ono(o 6Xer rorvo pe ro rupro orolro- 16ro ouppotprlg ;ulvo ro U5 snRNP. * To rvrprivro orriXor eXouv geyotrtirepeg ouvorvenr6g otrtrqtrouX(eg ong ougpotreg oupporprlq, Xopig v' oro- trouOorlv ntoro roug xov6veg GU-AG n AU-AC. * To Uei(ovo ror ro eLdooovo oropor0ro oupporprlg por- pd(ovror olgovlro0g nporeivrroug nopdyovreg, ge- rolu otrtrrov rtg npolreiveg SR. ffi n Iuppoqn rou pre-mRNA llt0ovov'EXer Kotv6 Mryovr- op6 pe r' AurororoLulro lvrprivro rqq 0po6og ll * To rvrprivro qq 0po6og ll err6pvovrot on6 povo roug onri ro RNA p6oto evog ouppdvrog ourorototruurrlg ouppoqnq. * 0r ouppotr6g ouppog1q Kor o U1x0vrop6g oupporpqg rt0v rvrpovlrov qg opodog ll e[vot nopolroro Ue Tr.l oup- poqrl rtrlv nupqvxcirv rvrpovltrrv. * 'Evo rvrpdvto qg ogodoq Il drntrtirveror oe pro 6eurepo- royrl 6opr1 nou d4proupye( evo rorotrulro r6vrpo ro ono(o 0upr(er q Dogr'1 rou nuplvlrou rvrpovrou U6-U2. ffi U lupporprl e(vor Xpovrrd ror Aeuoupyrro lu(euypevl ge l-lotrtro Bqporo oqv'Erqpooq rolv l-ovldurlv * Iuppoql gnope[ vo npoyporonorlOer xoro q 0ropxero qq ueroYpoQnq n uero q peroyporpl a 0r UnxovroUoi qg peroyporpqg roL qg ouppoprlg ervor quotro ror tretroupyrxo evonollpevor, * H oupporpl ouv6derol pe qv e(oyu.ryr1 rou mRNA xor rov 6treyXo qq moOepoqrog. r H ouppoQrl orov nupilvo pnopeI vo enrlpeooeL q gero- Qpoorl Tou mRNA mo runopontroopo. @ H Evotrtrorlrrl Iuppoqn E(vor Kovdvog,'OXr E(orpeol, oroug llotrururropoug Europudueg * Erdrro e[covto 11 otrtr4trouXieg e[ovulv pnope( v' ono- rtrerorotiv 11 vo oupnepilrlq0oUv oro mRNA npotovro Xprlorponouivrog evotr),orrt16q 0eoerg oupporpdg. e H evo),),ornrr1 oupporprl ouvetoq6pet oq doprrrl r<or treuoupyrrrl norril,[o rtrlv npoiovrov rov yov16(tov. * 0 ro0optoprjg rou rp0trou oq Drosophila nepilogpover pt0 oepo ouppdvrtov evotrtrornrrlg oupporprlg oe yov(- 6to nou roduonorouv 6lo0oXuo npoiovro ;uog o6ou. -ffi H Iupporprl Pu0p(eror ond EvtoXureg ror Anooronlrrlg nou Bploxovror Tooo oro E[drvto'0oo ror oro lvrp6vro e H evotrtrorrun ouppoqn ouXvo oXer(erot pe oo0eve[g 0doerg ouppogr.lg * 0t otrtrntroux[eg nou neprpotrtrouv evo],],oxlro e[tirvro ouXvo e(vor nto ouvrrlpqp6veg e[etrulro on6 ng 0trtrr1- trouXieg nou neprpotrtrouv ro rdroouorolro e[tirvro. e ErdLxeS otrtr4trouX(eg e[ov(ov ror tvrpov(rrlv evroXuouv 11 xoroore),trouv rrlv eniloy( qg 06oqq ouppoqrlq. e H en[0poo1 rcov evroXurtiv Kor Tov onooro:nlrtirv qg ouppoq1g DropeootropeLrot onti er6Lr6g yro rrlv otrtrq- trouXlo npolreiveq nprlodeolg oro RNA, notrtr6g ono ng onoleg pnope[ vo puOp((ovror ovonru[roro ro/r1 vo erqpo(ovror roro-erdrro. e H roxurnro peroypoqrlg enrlpeo(er ogeoo rrlv drpooq qg evo),),orlr1g oupporpfl g. ouvey[(eru or4v enopevq oeAido 583
dffi" &lxtj 'w 0r Avndpooerg frans-ouppoqrlg Xplorponorouv Mrrpo R- NA * 0r ovrLDpooerq ouppoQnq ouvrl0oq npoypcronorouvror ;-rovo in cis perolu ou;rpotrrirv oupporpqg oro [dto ;ropLo RNA. e Trans-oupporpll ouUBoiveL oro rpunovooriruoro Kor T0 orouLrlrro onou puo ppoXero otrtrqlouXio (SL RNA) oupponrerol oro 5' orpo notrtrtirv npodpoprrlv mRNA. * To SL RNA 6Xouv doprl nou poro(et ge q 05o1 npoo- deoqq Sm rov U snRNA. To 3' Arpo rorv mRNA Alptoupyo0vror pe l-leg1 ror l-lo- truoDevutr[oor1 * H otrtrltrouxio AAUAAA e(vor 6vo onuo ndUnq yto rl drlgroupy(o evoq 3' notruodevutrulprlvou oxpou mRNA. * H ovr[dpoon onoue( 6vo nporeivrrrl oupntroro nou ne- preXer evov nopdyovro etOrrdrrlroq, gro evdovourtreo- oq rot notru-A notrupepooq. o 0 nopoyovrog erdrxorr;rog ror q evdovourtreool n6- nTouv To RNA ro0odrro qg AAUAAA. * 0 nopoyovrog eL6trorrlrog rot 11 notru-A notrupepoorl npoo06rouv - 200 rordtrorno A oro 3' drpo. * H oupo no)tu-A etreyXer q oroOeporrlro rou mRNA xr enlpeo(et q peroqpoorl. * H noLuodevutr[roor1 oro xurropon],oo1ro noi(er potro oqv epppuirq ovomu[1 rou Xenopus. H Ene[epyooio rou 3' Arpou rou mRNA Ervor Kpiorlrl yLo rov Tepponop6 11g Meroyporprlq * Yndpxouv drorpopot rponor repponouou rnq usToypu- qflg ono dLotpopenr6g RNA notrupepooeq. * 0 0XIU0TtoU6g rou 3' orpou rou mRNA oqpotodore[ rov reppoloiui r1g peroypotpflg ond rrlv Pol ll. 0 lxrtfrorropdq rou 3' Arpou rou mRNA tov lorovrilv Anouef ro U7 snRNA & H 6KQp0on rcov mRNA rtov rorovdrv e[oprorot on6 rr1 peroypoqrl xor puOp((erot roro rr1 droprero Tou KUTTo- prrou rurtrou. o To mRNA rolv lorovtirv dev eivor notruoOevutrrrol.rtlvo: ro 3' orpo rouq drlptoupyelror gdoro luog ovr[dpoolg n6fig nou e[optorot on6 q 6o91 tou mRNA. a H ovr(dpoorl neQrlg npouno06reL npoodeol rou SLBP oe pro doprl proXou-0r1trroq ror (euyoprrrlro rou U7 snR- NA pe pro yeuovrrl povortrovrl neproXrj. * H ovrldpoon ndQqq rorotrueror ono 6vov nopdyovro rorvo ;le ro oupntroro notruodevutr(trlor1g. H luppotprl rou tRNA llepilol.rpovet Konrl rr Enuvoouvde- o1 oe :exolproreg Avldpooerq * H RNA notrupepoorl lll repuort(er rr1 geroypotpq oqv otrtrntrouxto notru(U)4 nou pp(orerot geoo oe Uro otrtrn- trouXio ntrouoto oe GC. c H ouppoQrl rou IRNA npoyporonotelrot ;le 6to6oXLxe: ovnOpooerg ronqg rot ouv6eoqg, * Mro evDovourtreoorl rdper ro npodpopo IRNA roL c.: 6uo oxpo rou rvrpovfou. * H onetreu06po0rl Tou rvrpov(ou dnuroupyei 6tjo ptol- IRNA pe oouvrlOtoro oxpo nou nept6Xouv 5'-0H roL 2 - 3' ruxtrrro qtoorpop116. * To 5' -0H drpo qootpoputrtoverot on6 pto notruvo,- r),eoldxfl rrvool, q ogddo rou rurtrxo0 QooQOptK:, ovo[yet on6 pro tproorpodreotepoorl onodtdovroq 2'-q.:- oqoptro tetrtro orpo rot 3'-0H opddo, ro drpo i,-' e[ovlov ouv66ovror ono pro RNA trryrooq xor ro 2 - rpulorpopuo otpoLpelror ond pto tprrrorpotootl. H Anovqoq orrlv llopouofo proq iedtntrtop6vlg l-lporra.- v1g IXer((eror Ue rn Iuppoqrl rou IRNA o H lrel e[vot ;uo eotorepxrl nupqvrrrl UeUBpovrx! np';- Tervn Us rrlv N-retrrrr1 neproXl T1q oTov outro rou :: rot q C-retrtr1 neptoxrl Tlq oTov nuprlvo. H C-re),,1<: neproX1 6Xer evepyoqreg rLvoolg n evdovouxtreoorl * H npoodeorl ptoq [edrntrotp6v4g nporeivqg oqv N-."- tru1 neptoXrl evepyonotel rrlv C-retrrrr1 evdovourtreo:- Ur ouToqtrtoQoputr(oor1. * H evepyonotqptlvl evDovourtreooq ropet ro HACl rl- NA (Xbpl oro onovduLroro) onetreu0ep6vovrog e,: rvrpovro ror 0rlgroupyrovrog e(6vto nou ouvdtlo','ri. pero[u roug ono pro IRNA trryrooq. * Movo ro ouppoprlvo HICl mRNA ;inope[ vo USToQC]- orel o' evov peroypotprxo nopoyovro o ono[og evep].- nore( yov[6Lo yLo ouvodoug nporretveg nou poq0o-, oro 6tntropo rov [edtntro;r6vtrlv nptoreivtirv. * H evepyonotluevn lrel endyet on6nrooq orov ro rL.- ropo nepr6Xer nopo notrtreq [eDrntropeveg nporeivec H l'lopoyoyrl rRNA l-lpoUno0ereL lupBovro Kon1g ror f :- prtroppovet prrpo RNA * H RNA notrupepool I repuon(er q peroyporprl o evr, reppolorfl 1 8 pooeorv. s T0 Ueyotro rot ro ptrpo RNA onetreuOep6vovrot pe x:- n1 on6 6vo rorv6 npridpo;ro rRNA. To 55 rRNA pera- ypoqerot IeXtoproro. * H ouodo C/D rov snoRNA elvor onopolrr1Tl yro T'i rpononoqorl qg 2' Oeoqq qq prp6(qg pe npooOq<- lrLog pe0utro;ro6og. a H ouo0o H/ACA rov snoRNA e[vor onopo[rr1Tl yro T" perorponl rrlg ouptd(vr1g oe qeu6ooupt6[v1. * Ie ro0e nep[nroor1, ro snoRNA (euyoptirver pe pto o,- tr1trouX(o RNA nou nepreXer q pooq-oroXo tlore l'c drigroupynoer pro runrrrl 6oprl nou elvor ro un6orpogc yro rqv rponono[qor1. llepilqtpq iasE{ YicfJt! ffii 21.1 Erooyuryri To RNA no[(et rewprr6 p6].o orrlv 6r<gpooq rov yovr- 6(tov. Aplrrd 1oporcr4p(ornKe oq 6vo ev6r6preoo rnq npolreivoo0vOeorlq, qA d 5lftore ovoroL6rp0qxov nol.- Ld dMo RNA pe 6opuxoUg rj Leroupyu<oUg p6Aoug oe 584 lGOAllAl0 21 Iupporprl rr Ene[epyoo(o rou RNA 6rd<popo ord6to qq Srrppoolq rov yovr6(u:v. H ou-- preroXrl rou RNA oe noMdg Aenoupy(eg nou oeopor. orrlv 5r<<ppoorl rov yovr6(iov unomrlp((er rrl yevtKr dno!.rq 6l q (or1 prnope( vo SXer e[e].rX0e[ ond dvc', "K6oUo RNA" orov ono(o ro RNA opXLKd rlrov tc evepy6 ouorotLr6 yto rr1 6roqpqorl Kor 5r<rppooq rcu,, yeverrrcbv nl.qporpopttrrv. Itq ouv5Xero no).).59 onc
ourdq rrq Leroupy(eg unoBorl0ri0qxov ri ovoLrirp0rl- xov qn6 npoteiveg, p' enox6trou0o trlv otifioq rqg euell((og r<or pdMov Kot. rnq onotetreogotx6rrltog. 'OLo rq RNA nou ;retoypdtpowot on6 r' ow(orotXo yov(6ro yro vo y(vouv ti:ptpro r<ot Lemoupytt<d Xpetd(ov- rot nepon6pco ene[epyoo(o. Atorer<otrrp5vo yov(6to undplouv oe 5Leg ng KorrlYop(eg ttrtv euxopurrlttrdlv opyovtopd:v. Awtnpooo:neUouv 5vo noLrJ prxp6 p6pog trov yovrd(tov trov ontroUorepov euKoputotcilv, oLtrd rr1 Ueyoll nlerov6rqto ttov yovtd(cov oro yovt6triproro rov ovtirepctlv euxopuror6v. To yov(6to notril,ouv noAU orip<pcovo Ue rov opt0pr6 xot to ;rrlxog rov twpov(tr:v, 6pr<og 6vo runrr6 yov(6to ttov 04loonrri:v 6Xer 7-8 e[c0- vro nou eme(vowot oe Uto neptoX4 -16 kb. To e[6vto e[vor oletrd ;ru<p6 (-100-200 bp) evtil to twp6vto q4e- rrr<d;reydl,o (>1 kb) (06ne Ev6tr1ra4.7). H ooul.tEcovio pero[U rlg dtoxeroUpdvnq opydvro- onq rou yovt6(ou Kot rnq 6tor<exopr;r6vqg opydvooqg rou ow(qrorXou mRNA npouno06tet enefepyoo(o tou npcotoyevoUg npoi6woq rnq UeroYpoqnq. To Tlporo- yev6g pretdypocpo SXet rqv (6to opydvoor'1 pre to yov(dro xot rolefuor pre-mRNA. H o<po(peor1 to:v twpov(tov on6 ro pre-mRNA og(vet 6vo tuntx5 oyyeAtorp6po - 2.2 kb. H 6repyoo(o ogo(peo4g ttov twpov(ov rolekot ouppo- gri rou RNA (RNA splicing). H ogo(peoq rov twpo- v[cov e(vqr r<[pro pr5pog rrlg nopqyoyrlq RNA oe 6trouq rouq euKopurireq. I-lop6tt to dtoreroppr6vo yov(6to e(- vor olerrrd ondvto orouq neptoo6tepoug l.tovot<6rro- poug/otrryor<6ttopoug europuriteg (6ncoq n (6pn Saccharomyces cerevisiae), 11 ouvoLt- r<q ovotroy(o unoerttpd rq oqproo(o rc,rv rwpov(ov eneL6rl ro neptoo6tepo 6Lorcer<oprp6vo yov(6to xro6Lronototjv oletri<d oqOoveq npotelveg. Iuveno.rg, 1 ouppo<pr1 epn5r<eroL orrlv nopoYoJ- yr1 peyolrjteprlq ovoloy[og otrLxoi m- RNA ond ro ovogev6pevo pe Bdoq qv ovd).uot1 tou yovtdtcrtprotog, (otog 50%. Mro on6 rtg npu:teg ev6e([etq o1e- iLr<o pe q gUorl rnq ooupeov(og og npoq ro p6ye0og preto(ti rolv nuplvt- <r.,-rv yovt6(r.,.lv KClt Tov npoi6wtrlv toug ;roug ovrr.lrepouq euKoputr.lteg 6601- <e on6 rrg rdL6tqteg rou nuprlvtt<oti RNA. To pr6oo prdye06g tou e(voL noL0 geyorirepo on6 to prdyeOog tou mR- NA, eivor no).ti oo-roOdg r<t 6Xet noLti rotr un).or<6tepr1 otrtrqlou1(o. Awtr<o- To RNA rpononote(tot orov ruprlvo pe npoo0ilreg oro 5' rot 3' orpo roL ueoor ouppoqrlq yto Tlv oqo(peo1 Tolv tvTpo- iLov, To ouUBov oupporptlq npouno06ret 3oouol rov oup$otrtlv e[ov[ou-tvrpovlou rot ruvOeol rorv oxptrlv rtov e[ov[tov, To titpLpo nRNA UerotpepeTor Utoo rov nuplvtro.tv no- J0lv oT0 xurropontroopo, onou pteroqpo(eroL. Eqd)vlo1 lwp6vrol Eqovto2lwp6vto2Eq6vtoSlwp6vto3Eqovro4 lwp6vto4 E(civto5 a I ut.oypo,ln Y I Tpononoiqorl tou oxpou l-lotru-A mo 3 oxpo V tAzoo {:uppo,pr1 -,*g!!9-l*[0,:,= I 0r ouu0otreq e(oviou-tvrpov[ou onove Y _-----_ 't- j I To eidrvLo ouvd6ovrot I A h200 nYPHNAI lVletorroPd romp((owoq rlv eupe(o rorovoprl r<ov peye0dlv rou, rotre(ror erepoyevdg nup4vrx<i RNA (hRNA) (hetetu geneous nuclear RNA). H quou<q Uopeti rou hnRNA e(vot 5vo ptBovouxleo- npoteivx6 otopot(6to (hnRNP), 6nou to hnRNA efuot npoode6e;r5vo oe Uro oprddo dgOovtov nporeivrirv. Opt- opr5veg npotetveg prnope( vo 61ouv 6opru<6 p6Lo orq ouoreuoo(o tou hnRNA. Apretdg e(vor yvcoor6 6tt en4- ped(ouv qv ene(epyoo(o rou RNA r] 6reuxotrUvouv rqv e[oy<oyr1 rou qn6 rov nup{vo. H ouppogrl npoypororoe(rot pr6oo orov nuprlvo, po(( pre ng 6trl.eg tpononott'1oetq nou ouptBo(vouv oro veoouwe0erpr6vo RNA. H 6repyoo(o tqg 5rqpoorlg ev6g 6roreroprpdvou yovt6(ou nopouotd(etot orqv EIKO' NA 21.1. To ;retdyporpo onorcrd r<olr3mpo oto 5' dr<po, ro rwp6vro o<potpo0wot t<ot ro 3' dt<po nol'uov6evu- hr.6vetor. 'Enero, to RNA petorp6petot ;t6oto ttov nupq- vxriv n6pov oro rurtop6ntroopto, 6nou e(vot 6tq06ot- pro yro gerdgpooq. Avo<poprd pre rtg 6tdcpopeg owtdpdoetg ene[epyo- o(og nou oupBo(vouv orov nuprlvo, 0o O5tropre vo yvcopG (oupre n6te or<prBriq npoYUoronote(tot q ouppotp4 oXenxd pe ng dltreg tpononot4oetg tou RNA. Apoye, o'uppoqri ouprBo(vet oe pto ouyxer<ptpt6vq 05or1 pr6oo orov nupqvo r<ot ouv66etot pe dMo ou;rBdwo - fl.X. 'rrl pretoypoEq r<o/q q preto<popd on6 tov nup{vo oro KUr- rop6ntroqro; En(oqq, rt ow(rcruno orrlv 5rrppoorl tcov prq dtoreroprprdvcov yovt6(ov 61et 4 6lstr!4 ouppotptig; Avorpopu<d pe rnv (6to qv ow6pooq rnq ouppo- E[r0vto 1r- Y- I -l- lMerooooon V" :a:_--:: KYTTAPONAA'[,1A Kotr0ffipo oro 5' oKpo Azoo 585
tptig, 6vo on6 ro x[pro eporrlporo e(vor ndlg eMyleror q et6rr6rrlrd rqg. Tr 6toogol.i(er 5rr ro drpo rd0e rv- tpov(ou ovoyvop[(owot ovd (eUy4 Sror cilore on6 ro RNA v' orporpe[ror q ocr:orr1 o].].q].ou1(o; To twp6vro r6Bowor on6 5vo np6Spogo ;r6pro pre r<o0opropr6vr1 oerpd; H rop[provoq rou RNA XpnorUonore(ror yro rq p60proq qg 6r<cppooqg ro:v yovr6(ov prSoco 6rdr<prorlg peto(0 rrov 6to05or;rolv npodp6prov ri oLLoyrlg rou nporUnou oupporpfg; Emdg on6 rr1 ouppogrlrou RNA yro rnv ogo(peor1 rov twpov(o:v, ene(epyoo(o yro vo rrtprprdoouv Xper- d(owot en(or1g noMd pr4 ro6xonorqru<d RNA nou no(- (ouv p6l,oug oe noxll,eg muX6g qg 6r<gpoorlq rov yo- vr6[ov. 21.2 To 5' Arpo Tou EuKopuoTtKo0 mRNA Odper KoA0nrpo * Mro 5' xotrunrpo oXrlporl(eror npoo0drovrog pro G oqv retrr- rr1 pool rou geroyporpou ;.uloo evog deogou 5'-5'. e H 0repyoo(o oxlpologou rotr0nrpog ou;rpo(ver roro q pero- yporprt, rort nou (otog enqpeo(er qv enovdvop[n Tnq UeTo- Yp0arlq * H 5' rotrunrpo rtov neptooorepcov mRNA e(vor povopeOutruo- gev1, ev6 oprol.ulvo pxpo 1ui rodrronorlrxo RNA e(vor rpr- pe0utrulgevo. * H 60[rl rotrunrpog ovoyvrrlpl(erot ono npotreivrrouq nopd- yovTeq nou enqpeo(ouv q oro0epo-rr1ro, rq ouppoqrl, qv e[oyrrryrl rorq perorppool rou mRNA. H peroypogrl (er<rvd pr' 5vo rpr<proorpoptx6 voux).eoo(- 6ro (ouvr1Oog noup(vr1, A n G). To npcoro vouxl.eor(- 6to 6Lorqpe( tqv 5' rpretrtoeoprr<r1 opiddo rou Kor 6q prou pye[ to ouvrl0topr6vo <proo<po6reoreprr6 6eopr6 on6 rqv 3' 05o4 rou ornv 5' 05or1 rou en6gevou vou- Bp[orerot oe dtreq rtq r<otrrlnrpeg I Bp(oreror oqv ro)Unrpo 1 Bploxeror oqv xotrUnrpo 2 rheol6(ou. H opXrrq oALqLoul(o rou preroypdgou gnope( v' ovonopooroOe( og: spppYopNpNpNp . . . MoLoro0ro, 5rov to r6prpro mRNA rorepyoore( rn vitro $e 6v(upro nou ro ono6opro0v oe (eXoprord vou- xLeor(6ro, ro 5' dxpo 5ev ono6(6er ro ovopev6pevo rpr- rpcoorpop116 vourleoo(6ro. Avr[0ero, neprSler 60o vouxLeor[6ro nou ouv66owor 1.r' 6vov 5'-5'rpreoogopr- 15 6eopr6 n en(onq 6ro05touv puo pre0u).oprd6o. H refu- x4 Bdoq efuor ndwo plo youov(vq nou npoor[Oeror crro ou0ewtx6 g6pto RNA ;rerd rq geroyporp4. H npooOrlrq qg 5'rcLxr1g G rorol0eror qn6 5vo nupqvx6 5v(u;ro, qv youovuLuo-rpovogepdon (GT). Ito 04l,oonrd, r1 GT 61er 6Uo ev(upur<5q evepy6rqreg. Aenoupye( 16oo rog tprEcoorpordoq o<porpcirwog 60o groogoprr6g ogd6eg on6 to GTP 6oo Kor oq youovrr l,u).o-rpovorpepdorl ouwrlrowoq q youov(vr1 pe ro ou- 0ewrx6 5'rpr<prrlogoprr5 dxpo tou RNA. Ir4 (rip4, our6g or 60o evepy6rqreg ov4Louv oe 6rorpoperrd 6v- (upo. To v6o xotdlorno G npoor(Oeror oro 6xpo rou RNA ge ow(orpogo npooovoroLrop6 on' 6Lq t' 6IAq vour<Leor(61o. SGppp + 5 pppApNpNp. . .-+ GpppS-SApNpNp. . . + pp + p Aurn n 6opr{ r<o}.efuor xotrrlmpo (cap). E(vor ro uno orpoUo yr' opxerd ouprBdwo ;re0uL(oo4q. H ElK0tlA 212 6efiyer r4v n^,npn 6oUri puog xoA0mpog og6rou npoorE- Oorjv 6l,eg or nr0ovSg peOuLopddeg. To nro oqprowx6 ouprBdv efuor q npooOrlxrl puog provo6rrrlg pre0u).otrr66og orn e6on 7 rqg reLx4g youov(vr1g, q ono(o roro.l,feror on6 rrlv 7-peOuLorpovogepdorl rnq youovfuqs (MT). flop6rr q 6ro6roo(o rqg r<dLurlqg ;rnopef vo flptry- poronot40e( in vitro pre ro0opd 5v(upo, rovovrxd n owi- 6poon npoyporono€kor rqrd rn Ueroypoerl. Ap6orog prerd qv 6vop(q rnq Ueroypoens, n Pol ll oroprotd -30 vouxl.eoil6Lo roOodtrd rqg ogeqp(og, nept;r6vowoq vq enrorporeu0orjv ro 5v(upro r<dLur[4q nou 0o npoo06oouv rnv r<otrrinrpo mo 5' drpo tou ovo6u6pevou RNA. Xrop(g our4v rrlv npe oroo(o, ro ovo6u6pevo RNA e(vor eudltoro d en(0eoq on6 5'-3' e[rovour].edoeg; 5vo rSroro rfoL(dropo prnope( vo rdva ro oriprnl,oro Pol ll vo n6oer on5 ro DNA ergoye(o. Iuven<iq, 11 dtodtxoo(o qg rdi.ur.f4g efvor orlgowxrj rilore 4 Pol ll vo nepdoer oe nopoyoyu<r1 enrprlruv- orl yro vo peroypdtler ro un6lorno yov(6ro. An6 oufi qv dnorfrrl, 11 el5tu[q rou prlxovr- oproU norio4q yro rnv npoo0r]r<q r4g 5' xoA0- nrpoq owrnpoooneUer 6vo orlpre(o eL6yXou yro rrlv enov6vop(r1 rnq Feroypogqg on6 rqv oplu<ri 05oq noUorlg. ElKOllA 21.2 H xotr0nrpo (0noqpdootr,, ro 5' drpo rou mRNA ror eivor pe0uhuop6vl oe opxer6g 06oetg. 586 KE0AAAI0 21 Iuppoqrl r<t Ene[epyooio rou RNA Boorxeg eworeg
I' 6vov ntrq0uopr6 europutottxc0v mRNA, r<d0e pr6pto nepr6Xer ;r6vo puo ;reOutroprd6o orqv retrtni youov(vr1, nou yevxd ovoE6petot trtg provopreOutrttopr6vq r<oL0mpo. Aw[Oero, optopt5vo pupd prrl rodxonoqnrd RNA, n.X. exefuo nou epdSrowot oq ouppog4 rou RNA oro oo- pcn(6ro ouppoqqg (BI6ne Ev6rrya 27.6) preOuLttirvowot nepordp<o t<ot nepdlouv tperg preOutroprddeg orrlv rcLrq youov(vr1. Aurri q 6oprri r<otrekot tptpeOul,trog6vq rol[- mpo. To 6v(upo yt' out5g ttg entnp6oOeteg preroEop6q peOutrop66tov Bp(or<owot oro Kurrop6ntroopto. Aur6 (ooq drooqoA(er 6rt p6vo optopr6vo e(et6u<eug6vo RNA rpononoro0wot nepor6pco ortg xo}Umpeq rouq. Mro on6 lg r<Upteg l.eroupy(eg rou oXrlgottol.to0 xol.Umpog e[vot 4 npootoo(o tou mRNA on6 ono66- pqoq. Iqv npoy;rorx6qto, q ev(upxti ogo(peor1 qg xol,0mpog e(vot 6vog on6 roug rfptoug pr4XovtqroUg yro q p0Optorl rnq ovorfxtroorlg tou mRNA oro euKo- puortrd r<[rropo (F],5ne Ev6tqta 21.11).Itov nupfvo, ro erepodrpep6g np6o6eonq ornv rotrUmpo CBP20/80 ovoyvrop((er qv xolUmpo rot npoo66vetqt oe ourqv. Aur6 6reyeQet r4v o<po(peorl rou npd:rou twpov(ou rot, prSoco dpreorlq otrl.ql,en(6poonq Ue rn Unxovn e[oytoy{q rou mRNA (otiprnLoro TREX), 6teuxolUvet qv 5[o6o rou mRNA on6 tov nup4vo. 'Orov to mRNA gtdoet oro xurrop5ntroopo, pto 6toEopertrq oprd6o npulteivdtv (elF4F) npoo66vowor offlv rotrtintpo nporotrri:woq rqv 6vop(4 rqq perd<ppooqg. 2'1.3 Or llupnvrrdq IuUBoAdq lupporphq Eivor Bpoxeieq AAAnAouxieq a 06oetq oupporprtq e[vot ot otrtrrltrouxteq nou pp(orovrot optl- oog drntro ono ra opLo e[otv[ou-tvrpov[ou. 11o(pvouv ro ovol.lo roug on6 q 05011 rouq ouyrptrtro pe ro tvrpovto. * H 5' 06on oupporprtq oro 5' (optorepo) orpo rou tvrpovlou neprtroppovet rr1 ouvotvettq otrtr1trouXio GU. * H 3' 06oI ouppoQnq oro 3' (de[t6) drpo rou rvrpov[ou nepr- troppover rr1 ouvotvertrtl otrtrltroux(o AG. u 0 xovovoq GU-AG (nou opxtro ovopo(orov rovovog GT-AG ovorpoprro pe rrlv otrtrr1trouXio rou DNA) op((et ort ouro to oro- 0epo Drvouxtreor[Dto elvot onopo[qro ortg duo nptitreg rot 6uo retreuro(eg 06oetq tu-tv tvrpov[tlv oro np6dpopo mRNA. * Ynopxouv etroooovo tvrpovto ouyrptnro pe ro ge(ovo tvrpd- vto nou orotrouOouv rov rovovo GU-AG. * To etrdooovo lvrp6vto orotrou0ouv 6vo yevtro rovovo AU-AC pe gro 6rorpoperrrrl opo6o ouvotvertrtlv otrtr1trouXttilv oto opto eItov[ou-rvtpov[ou. l-ro vo eortdoouUe oro proproxd ouprBdwo nou e;intr5- KOVTor orn ouppoQl ttr;v nuplvtr<c0v twpov(olv, np6- ner vo e[erdooupre q gUorl ttov 06oeav ouppaEt]q, 6q).o6rl rov 6tjo op(ov e(cov(ou-twpov(ou nou nept- LoprBdvouv lg 06oerg OpoUorlg xt enov6vtoonq. Ot ouprBoldg peto[ti e[ov(ov xot twpovf<ov pnope( vo r<o0opLoroUv ouyrp(vowog qv oMlLouX(o voux.treo- rL6(rov rou mRNA Kot rou dogtr<otl yovt6(ou. Aev undpla ercretopt6vq o;rol'oy[o rl oupnA4potr rrr6rlro ;reto[rj tcov 6Uo drpov evSq twpov(ou. Ot oupBol.5q, tr:01600, 6Xouv ro}'d ouvrrlpqprSveq, clv Ktrl oXenr6 prxp5g, ouvowerr6g oMql,oulieg. E(vot egu<r6 vo npoodrop(ooupe 6vo et6tr6 dr<po oe xd0e rwp6vto Ue Bdon q ouwrip4crq tov ouprBoLcirv e(ovfou-twpovG ou. 'OLeg or oupBol,6g e[cov(ou-twpov(ou efuot nopotr- ).oy5g r4g ouvotverxqg oMrlLouXfoq nou dfuetot oro ndvco pr6pog rnq EIK0NAI21.3. To rjryog r<dOe ypd;r;rotog unod4trdrvet rrl ou1y6r1- ro proq ouyr<erptpt6vns B6onS oe xdOe ouvotveurcl 05- orl. YqrrlLi ouwripqorl nopoqpe(tot pr6vo pt6oo mo rwpSvro orrq unonO6preveg ou;rBol6g. Aur5 op(er rqv otr,r1trou1(o ev6g yevtrori twpov[ou rrlg: GU...,.. AG To rwp6vro nou op((etot rqt' out6v rov tp6no [ert- vd pe ro 6tvourtreot(dto GU rot teletrilvet pe ro 6wou- rLeor(dro AG. 'Etot, ouXvd l,6yerot 6tt ot ouprBoldg orol.ou0o(v tov xov6vo GU-AG (GU'AG rule). Iqprerdrme 6tt ot 6uo O5oetg 5Xouv dtogopertr5g oMrltrouX(eg on6te op((ouv rov npooovototrtotrl6 rcov drp<ov rou rwpov(ou. Ovotrrd(owot on6 t' optmepd npoq ro 6eltd xotd prrirog tou twpov(ou rog 5' 06o11 ouppogriq (peprr6g Eop6q rotre[tot oproreprl 06on n 06o11 d5qq) ror 3' 06oq ouppogrlg (en(or1g rotre(tot 5e[rd 06oq ri 06on 6611119). Erlpretox6q preto]']'d(etg orrq ouvqtvertr6q oMqouX(eq nopeprno6((ouv rrl oup- pog{ 16oo in vivo ooo rot in vitro. To neptoo6tepo twp6vto ot<olou0oriv tov rov6vo GU-AG. Oar6oo, 5vo ;lrp6 xLdopro twpov(tov 6ro05tet puo 6togopetu<n opddo ouvowertrcilv oMrltrouXtrilv mo 6pro e[roviou-twpov(ou, 6ncog <po(vetot oro r<dtto pr5- poq rnq Erx6vog 21.3. Autd to twp6vto oplrxd nept- ypdrpqr<ov crlg etrdooovo twp6vto nou orotrouOotiv tov rqv6vo AU-AC L6yco rcov ouwqpqprdvov 6tvoux.treott- 6(rov AU-AC Kot oro 6Uo drpo xd0e twpov(ou, 6ncog qo(verot oro p6oov rrlg Etx6vog 21.3. Tq pe((ovo t<ot eldooovo rwp6vto neptypdgowot rotrUtepo og rwp6- vro rfnou U2 r<or U12 ;re Bdoq rtg 6torpn6g pqXov6g oupporp4g nou XpnotUonoto[wqt yro rrlv ene[epyoo(o roug (Btr5ne Ev6tryra 21.9).'Ero; evri: optopr5vo twp6- vto qo(vetor v' qxolou0oUv tov rov6vo GU-AG, orqv npoyprorx6rqro ene(epyd(owor 6nog to twp6vto tti- nou U 12 (pMne r<fuo pr6pog r4g Etx6vog 21 .3). 21.4 Or lupBoAdq luppogriq AroBd(ovror Avd Ze0yn e H ouppoQn e[oprorot povo ono rrlv ovoyvdrptol (euY6v ouu- Botrtitv ouppotplq. c'0trsS or 5'ror oL 3'06oetg ouppoQnq elvot tooduvopleg tret- roupyrro. a Enrnpoo0ereg ouvqplp6veq otrtrltrouxieq ortq 5' rot 3' 0d- oeLq oupporpnq 6rorp[vouv rtg tretroupytt<6q 0eoetq 0uppoQ]lc pero[rj oprerdrv otrtrtov ntOovtirv 06oeov oro pre-mRNA. 21.4 0r Iuppotreg Iupporpnq Atopd(owoL Avo ZeuYTl 587 Boorxeq eworeg Boorx6g eworeg
Aproreprl (5') 0eon Irlpoto ouppoq1g yto ge((ovo rvrprivto (rUnou US n GU-AG) Ilpe(o 6LoxtrdDooqg o -74--54 3 2 1 1 2 3 4 5 Ae[td (3') 06o1 IrlUqTo ouppoQrlq yto e)dooovo rvrprivro (ilinou U1 2 n AU-AC) Iqgeio 6tortrd0rrlo4g o I C. Aeird (3') 06011 IrlUoTo ouppoQnq yro eJdooovo twprivro (runou U1 2) nou flep0dLovror on6 GU rot AG mo dxpo Irlpe(o drortrddroorlg o Ae{ro (3') 06ort To orpo rov nuprlvtKov tvTpovll]v Koeopl(ovTor on6 rov K0v0v0 GU-AG. To etroooovo rvTpovro Koeopi(ovTor U60o 6r0Qops- TrKd)v 0uv0rveTrK6v otrtrntrouxuilv oqv 5 Oeoq ouppotplg, rq Oeoq dLoxtrodrrrolg xot qv 3' 0do1 oupporpqq. 'Evo runx6 mRNA Or1).omrroU 6ro06rer noMd rwp6- vro. To Boorr6 np6Blqpo rnq ouppogrjg rou pre-mR- NA onoppder on6 rrlv onl.6rnro rov 06oecov ouppogrlg Kor nopouor6(eror mrlv EIK0NA 21.4. Me de6op6vq rqv nLrlOcirpq oMnlouxrcbv rou rwpov[ou nou rorprd(ouv pe rn ouvorvelrr; oll,ql,ouX[o rrov yvtiorcov 06oerov ouppognq tr 6roo<pol.[(er 6n 0' ovo- yvrop((owor xor 0o ouppdnrowor ro ocomd (e[yr; 06oeo:v; IlpSner vo ouv6e0oriv rordMrll.o (eUVn GU- AG nou on6louv noL6 (opro;r6vo rwp6vro e(vqr > 100 kb). A0o trjnor prrlXovrol.rcilv 0o ;rnopotioe vo e(vor une(Ouvor yro ro (euydpropro rcov xotdMrllov 5' xor 3'06oetrlv: o H oriv6eorl rorv O5oeov or' dr<po ev6g ouyxer<prpr6- vou rwpov(ou 0o pnopofoe vo e[vor 1.uo eyyev7g tdt6rqta rou RNA. Aur6 0o npoun60ere to ro(pto- opo er6trriv otrtrr1trouXr6v 4 6oprirv, rdtr nou 61er nopotqp4Oe( oe opropr6vo yov(6ro ew6purlv, oMd ''t.::i:::-. .-..----x 1 i; :.i:r!r;ir*ili; ,r,:;ili;rl :;ti.tii:t,r:;ty l] tt]i-ti!l+tir,rlr ait.i;t':r.t 0t ou1tBotr6g oupporplq ovoyvtr.tpf(ovrot p6vo 6rov 6Xouv rov xorotrtr1tro ouvduoolro (euyopo-rporog, 588 KEOAAAI0 21 luppoQn xr Ene[epyoolo rou RNA Aptorepq (5') eeon Aprorepn (5') 06on
6e go(vetot vo toX0et Yto ro neptoo6tepo euKopuoJ- uxd yovbto. o'OLeS ot 5' Kot ot 3' 06oerg 0o prnopo0oe pev vo e(- vot Lemoupytxd too6tivopreq otrI'd 11 ouppo<pf oro- )touOe( xav6veg nou 6toogol'((ouv 6tt ptto 5' 06o11 n6wo ouv66erot Ue rnv en6;revr1 3' 06oq oro RNA. Orjte ot 06oetg ouppogqq oUte ot neptBdMouo'eg neprox6q 51ouv rdnoto ouprnl,4ptrlpotx6trpo toq flpoq rqv oMqLouX(o, yeyov6q nou onoxtre(et to 1.tow6tr'o nou unooq p(ouv ou prntr 4 p<oprorx6 (euydpcopo prero(U tov dxprov rov twpov(tov. l-letp6proro nou Xprlottrronotorlv np66popro RNA 6e(1youv 6n onotodrinote 5' 06oq oup- poqns prnopei Oeoprltxd vo ouv6e0e( pre onoto6qnore 3' 06o11 ouppogrlg. l-to nopd6erypro, 6tov to npcilro e{6vro rqg nprirrtrtrlg preroypoqtrulg govddog UeroYpo- grlg rou SV40 ouv66etot pe ro rpiro e[cirvto rqq B oqot- pivr;g rou nowu<oU, to uBpt6rr<6 twp6vto ogopekot yto vo 64puoupyr10e( puo t5treto otiv6eorl ptero[6 rou SV40 e(ov(ou Kot rou e(ov(ou rqq B orpotp(vr1g. flpdyprott, ourn n owotrLo[tpr6t4to e(vot n gdon Ylo rrlv telytr<r1 nayi6euoq g e[cov[cov no u neptypdqn Ke nponYo u p5vtog orqv Eu<6vq 5.8. Tdtoto nepdptoto ovodetxvUouv 60o yevxd onpe(o: o OL 06oeq ouppaEriq e(vat yevxiq: Aev 6Xouv etdrx6- rnro yro (eXroptord np66pogo RNA, ofte to [eXor- prord np66pogo RNA g5pouv et6rrdg ntrqpoqop(eg (n.X. 6ewepotoVri 6oUri) onopofurlteg Yto rn oup- poen. Om6oo, oe optop5veg neptntrioetg 6Xet ono6etX0e( 6tt et6tr<5g nprore[veg nou npoo6Svowot mo RNA (n.1. hnRNP A1) npo6youv to (euydptol.to mrg Odoetq ouppog{q prSoro np6odeonq oe Yetrovt- x6g nr0ov5q O5oetg ouPPorP{q. o H ouoxeui yrc rn ouppaEn 6ev e(vu torc-etdmfi: 'Evo RNA ouvrlOog ouppdmetot oo:md oe onot- o6rinote r0rropo, ove[dptqto on6 to ov ouvrlOtrrq ouw[0erqt pr5oo oro ouyrer<ptptdvo r<0rropo. (Itqv Ev6t4ra 21.12 0o e(erdooupe rtg e[orp5oetq ortq ono(eg nopoulpotiwot toro-et6trd pot(Bo evoMq- rcttruig ouppogrlg). Mol.ov6tt ot 5' t<ot 3' 06oerg ouppoqrlg go(vowot 6poeg on1 ouoreurl ouppoq(q, notot rov6veg 6too<po- Ai(ouv 6l 11 ovoyvrilpton rov Odoetov ouppo<prlg nepto- p((eror 5rot dlme vo ouppdmowot pt6vo ot 5' rot 3' fuoerg tou [6tou twpov(oui To twp6vto orpopotiwot pe xdnoto et6u<r1 oetpd on6 5vo ouyrexptp6vo RNA; H ouppo<prj e(vot lpovtrd ou(euyp6vq Ue rn Usro- ypqqn ft.1. notrl.d ou;rBdwo ouppo<prig oLoxtrrlpdrvov- ror nporoU q RNA notruprepdorl rptdoet oro t5Log tou yovt6iou). Iuvendlg, efvor e6)loyo vo uno0Sooupe 6tt r1 trEf,cypogn nop6ler ;lo o6pri oepd ouppoqnq oe ro- teri0uvorl 5'-3' (6rllo6n, rqpefuot xdnoto noepd npore- por6qtog"). Aeftepov, 1-tto Leroupyxq 05oq oupporp{q orry1vd neptBdlletot on6 pto oerpd orotle(cov oM4lou- foq nou evtopjouv ri r<otoorS].].ouv rrl 05or1 (B].6ne Ev6r4ta 21.13). 'Etot, oMrltrou{eg 16oo rov e[cov[trrv 6oo xot ttov twpov(tov treroupyo0v en(oqg og pu0ptort- xd orotle(o yto rnv enttroyri tov 06oetov ouppog(g' l-to v' ovoyvtrlptore( onoteleoporLrd ono rc -- ,.1' .- opr6 oupporpnq, Uto ).ettoupyLxq 05or1 ouppoQrc ::- ntr vo 6to06ter q oomrl oMrll'ouX(o, peto[u c)-'*. er6Lxdg ouvotvertrdg oMrlLoulieq rot ototlelo flou t!c- 6civouv q ouppoprl to ono(o r<uptoplotiv 6vowL o)'- Lrov ororxe(ulv nou rlv t<otom6Mouv. Iutrtroytr<d, ouroi or unxovtopoi 6rooqol((ouv 6tt to oriproro ouppocpqg 6roBo(owot ovd (e0yr1 Ue pto o{ertKd ypop;rtr<r1 oetpd. 2l "5 H Xuppqqr'r Tou pre-mRNA llpoxopei Arqudoou Evdg Adoou a H ouppqQn onotre( rtq 5' rot 3' 06oetq ouppqQrlq rot gto 06- or1 0Lortro6oor1g ovodtrd rnq 3' 0eonq oupporpflg. * H otrtrntroux(o otododtoolg e(vot ouvqprlg6vq ott'1 (uUn oIIo 6Xr 16oo otoug notrux0uopoug euxoputlteg. . 0T0v ro wrp6vro xone( oqv 5'06o11 ouppoQnq, oplpor((etot Uro 0opn oe oXflpo trdoou xoL to 5' orpo ouvD6etot oq 2' 0e- on Uroq A mn 06on 6toxhd6oonq p6oo oto tvtptivto. . To tvrp6vto one)eu0ep6vetot un6 poprprl trdoou 6tov ronef aqv 3' 06orl oupporpnq rt 6netro to optotepri roL to de[t6 e[tirvro ouvde0ouv peto[U roug. O prrlXovropr6q qq ouppo<prlg 6Xer loporctqprore( rn vitro Xpqor;ronotcilwoq ouorqpoto el'eU0epo xurrd- pov oro ono(o twp6vto pnope( v' ogotpeOoriv on6 np66po;ro RNA. I-lupr1vu<d exluA(opoto ouppdmouv roOopd np66popro RNA. Aur6 6e(lyet 6tt 4 ouppogfl 6e ouvS6etot uno1ceortKd ;re tr1 6repyoo(o rrlq Uero- ypoqns. Iuppogq prnope[ vo ouprBe( oe RNA nou 6ev e(vor o0re roLugprdvo o[te no].uodevu].ttr:p5vo, nop6- r. oro rUropo owd to ouprBdwo t<ovovtr6 npoYpo- ronorotivtot pe ouvrovtopt6vo tp6no Kot n onotetreotrtottr6rrlto rnq ouppoqrlg enqped(etot qn6 rn Ueroypoqri xr dMo ou;rBdwo ene[epyoo(oq (B16- ne Ev6tryo 21.11). Errlv o66 r4g EIK0NAI21.5 nopouotd(owot tq ord- 6ro r4g ouppogrlg in vitro. Oo e[etdooupe rrlv ow(6poor1 on6 rq orontd rcov feloprmdrv RNA nou pnope( v' ovoyvtoptororiv. Qor6- oo, in vivo, RNA nou nept6Xouv e[drvto 6ev uqiorowot og eLeriOepo p6pto oMd ouyt<pototiwot on6 rn ou- oxeurj oupporpqq. H EIK0NA 21.6 defiyet 6tt to npcino B4go qg ow(- dpqonq ouppogrlg e(vot 1.tto nupqv6Eil.q en(Oeo4 tou 2'-OH mqv 5' 06o11 ouppoqrig. To optmep6 e[cirvro no[pver rn Uopqn ev6q ypogpxotl prop(ou. To 6e[t6 pt6- pto rwpoviou-e[ov[ou oXqgot(et prto 6ogt1 oe oplpo tr6oou (lariat), 6nou to 5' dxpo nou dq;rtoupyefuot qro drpo tou twpov(ou tout6Xpovo trans-emeponote(rot drors vo ouv6e0e( pr5oco ev6q deopro0 2'-5' ge gto Bdoq ;16oo oro rwp6vto. H Bdorl-m6Xog e(vot ;rto A nou Bpi- oKeror gdoo oe puo oMqlouX(o yvooni toq 06o4 6lo' xtr66toot1g (branch site). 21.5 H Iupporprt tou pre-mRNA l-lpoXopeiAropdoou Evdq A6out 589 Boorxeg 6vvocg
5'r3' 5'0dq 3'060n GU UACUMC AG Pyr6 N Py66 PysT Pu75 A Pye5 ffiffii[I-0H :li -0 'lP :* i ,rj-ir'il E{rirvro 2 :f!dii0l-O:' r' "fi E{(rvro 2 ElK0llA 21.6 H nupqvxrl oupporprl npoypotonou[rur p6otrl 6uo ovrtdpdoeov trans-eoreponoqonq otq ono[eg pro oUd6o 0H enr- r[Oerot o' 6vo rproorpodreorepxo 6eopd. ).erQr1 ouvqpnonq ornv oMrlLouX(o rnq e6o'nq 6toxL6- dcoonq enrpdner rn Xpnorpono(qor1 nopepgepcbv oAtrq- LouXrdrv (xoloriwot r<purpdg 06oerg, cryptic sites) ov 4 ouOewx4 6tor<ld6ooq onoLor<pe[ r1 ;reroLl.o10ei. H ey- ywrpo ornv 3' 05or1 ouppo<pr1g rpo(vetor 5r e(vfl snpov- rrrl, ener6{ n Kpugri 05or1 Bp(or<eror ndwore rowd orqv ouOewu<r1 06o4. Mto rpugrj 06o11 1p4oq-ronoefuor g6vo 6rov 11 05oq droxld6tooqq 6Xer odpovonorqOe(. 'Orov pno Kpugn oMqlou{o 6ror<}.ddroorlg Xpnorponorekor xcn' ow6v rov rp6no, 11 ouppogrj ouve((eror xovovrrd ror ro e[ri:vto ono6[6ouv ro (6ro npo't6wo pe rov dypro ri- no. Zuventiq, o p6Log qq 06or7q SLaxAddaor1q e[vat v' avayvupi(a qv nA1o6areprl 3' 06o4 ouppaE4q aq otG Xo y@ o0vdeoq pe rnv 5' 06o17 ouppacprTq. Aur5 pnopei vo eppqveuOe( on6 ro yeyov6g 6n npoyproronoefuor puo oMqleni6poorl ;rero[ri rolv nporcivu<riv oupn],6rov nou npoo65vowor oe out6q ng 6Uo 06oerg. de5'@ 3, L==.lL ' E.- ., UACUAAC AG I2'5', AnoDtortrd6oon tvrpov[ou 5',Gr-i 'u_*UACUMC AG 3' . : :r.:. H oupporprl npoyporonoLelrot oe 6uo oroDLo. l-lpdr- ro roperoL ro 5'-e[drvro rr enerro ouvdeeror pe ro 3'-e[ovto. Ito 6efrepo Brlpo, to e].e[0epo 3'-OH rou e[rov(ou nou oneLeu0eputOrlxe on6 qv npotq ow(6poor1 enn(- Oetot oro 6eopr6 orrlv 3' 06o11 ouppo<prlg. nopoqp(- ote drr o opL0;r6q rov qotoqodreoreprrolv 6eoprriv ouwqpefuot. Aplrxd unqpXov 6rjo 6eoproi 5'-3' ong 05- oerq ouppog4q e[cov(ou-rwpov(ou. O 6vog 61er owrr<o- rosro0e( on6 ro 6eop5 5'-3' pero[0 totv e[o:v(o:v, evot o dl),og on6 to 6eopr6 2'-S'nou olrlprot((er to Ldoo. 'Eneno, ro Ldoo "ono6[oKlo6((eror,,, ono6(6owog 6vo ypoprpu<6 rwp6vro ro ono(o ypnyopo ono6ope(ror. Anopofurlteg otr-I.r1).ou1(eg yro rn ouppogq e(voL or Bpole(eg oMlloul(eq olq 5' r<or 3' O5oerg ouppo<pr1g Kor orn 05oq 6tor<)d6roor1q. Ko0rrtg ro peyol.rirepo U6- poq rnq oMrlLouX[og ev6g rwpov(ou pnopei v' qno].or- <pe( Xop(g ow(rruno orn ouppoerl, ouvdyeror 6tr 6ev onorefuor rdnoro er6u<4 6ropr6pqoon ;rdoo oro rwp6- vLo (q to e[otvto). H 06oq 6rori.d6oor1q no((eL o4prowrr6 p6).o mqv ovoyvtrrpto4 qq 3' 05or1q ouppocprlg. Ir4 (Upq, 11 05or1 6ror<Ld6oroqg e(vor e[otperu<d ouwrlprlpr6vrl r<or 6to05- rer rl ouvorverrr<r1 o).).r1).ou1(o UACUAAC. Aw(Oeto, oroug nolurUrropoug eur<opurrtreg 6ev e[vqr r<o],d ouv- qprlprdvr;, oMd 6e(Xvet ;uo npot(pu1oq yro noup(veg r1 nuptprt6(veg oe 6torpoperLr<6,9 Odoetg r<or 6rorrlpei ro vourd.eot(6to-or6Xo A (B).6ne Eu<dvo 21.5). H Odq 6ror).d6oor1g Bp(oxeror 18 6cog 40 vour<).eot[- 6ro ovodu<d qg 3' 06oqg ouppocp4q. Irn ([Un, peroMd- [etg r1 ono].oupdg orrl 06o11 6rox].d6oonq nopepno6((ouv q ouppo<pr1. Ztoug no).ur<Urropoug eur<opuroreg, q 6L- 590 KEOAAAI0 21 Iupporprl rt Ene[epyoo(o rou RNA 2 1.6 To snRNA Eivor Anopqirnro ylo Tn Iuppoqi a To n6vre snRNP nou epntrdrovror orq ouppoqrl e(vor ro U1 U1, U5, U4 xor U6. a To snRNP Uo(r Ue oproUeveg otrtreg npotrelveg oXrlpon(ouv ro oopor[dro oupporp1g. * '0tr0 ro snRNP errriq onri ro U6 nepr6Xouv 1uo ouvqpqpevl 0trtrntroux(o nou npoodever Trq rpoTeiveq Sm or onoleg ovo- yvropl(ovror ond ovnotirporo nou nopoyovror oe ouroovooo voorlporo. 3', {e ls' Boorr6g eworcg I
Or 5' ror 3' 06oerg ouppo<priq r<ot q oM4l,ouX(o 6ro- x,l,66oor1g ovoyv<op((owqr on6 ouqrorrrd rnq ou- oxeurlg oupporp4g nou ouvoOpo[(ovrot oXrlpror(- (owog rivo peydLo o0pnloxo. Aw6 ro orjpnloxo g6pva rowd rrg 5' r<or 3' O5oerg ouppogqg nporori oupBe( onoto6{nore ow(6poor1. To oUpn}.oro ouvqp- poloye[ror 6ro6o1u<d ndvo oro pre-mRNA. Apr<erd ev6rdgeoo prnope[ vo rouroTror.4Ootiv prSoo qg x].o- oporono(4 o4 g ou prnl5xrov 6rogopetrxo 11 preydOou g. Iuppog1 ouprBo(ver ;r6vo 6rov ouvoptr_roLoyqOo0v 6)to ro ouorolrd. H ouoreurl oupporpqg 6ro06ter 16oo npcoreiVeg 6oo xor RNA (exc6g rou pre-mRNA). To RNA no(pvouv r1 popei pru<p<itv 1.top(<ov nou ug(orowor rog prBovoux.treo- npcoreivu<d oo;rot[6to. T6oo o nup4vog 6oo xor ro KUr- rop6nl,oo;ro rov euKopuonr<cilv xurrdpov nepr6louv noMd 6ror<prd prxpd RNA. To p5ye06g rouq xu;ro(ve- ror on6 100 6ro9 300 Bdoerg oroug ovrirepouq euKo- puri:reg, evri m4 (rip4 ercre(vowor oe -1000 Bdoerg. Arogdpouv noLrj og npoq rrlv og0ov(o toug, on6 10s irog 106 ovd r<Urropo Srog ouyxewprioerg 16oo 1oprr1- ).eq rriore vo pn Unope( v' ovrXyeu0oriv d;reoo. To RNA nou neprop((owot orov nup4vo r<ol.oUwor prxp6 nupqvrxd RNA (small nuclear RNAs, snRNA), evri exe(vo nou Bp(or<owor oro xu-rrop6nAoopo plxp6 xurropontroopolx6 RNA (smalt cytoplasmic RNAs, scRNA). Kovovu<d ug(orowor cog prBovouxleonporei- vrx6 oor;.rori6to (snRNP r<or scRNp). yndpXer en(o4g pro xorqyop(o pru<priv RNA nou Bp(orowor orov nuprl- doxo r<or rqloUwor snoRNA (plxpri RNA rou nupqvi. oxou, small nucleolar RNA), ro ono(o eprnldKowor orqv enelepyoo(o rou prBooropu<ori RNA (B).6ne Ev6t4- ru21.20). To snRNP nou etrmfuirowqr orrl ouppogri, pro(( ;re noA.69 dMeg npolreiveg, q4qprot((ouv dvo ;reydAo oro- pmrdror<6 orjgnLor<o yv<rror6 <og oopori6lo ouppog{q (spliceosome). 'E1er onopovco0e( ond oum4proro rn vrlro ou p porpn q coq p rBovo u r<Leonproreivu<6 oco 1_ror(6 ro 50-605. To otol.rot(6ro oupporprjg nr0ov6v oXqtrror((etor mo6rord xoOdrg ouv66owor rq snRNp. E(vor peydLo orol.rorf6ro, pre preyo].Urepn Ud(o on6 ro ptBoorr:gduo. H EIK0NA 21.7 ouvotp(er ro ouorctrrKd rou ocol.rorrd[- ou oupporprlg. To ndwe snRNA onoreloUv ndvo on6 ro 6vq r6ropro qg pd(og rou ocopon6(ou ouppogrlg. Mo(i pe rtg 41 nporefveq Ue rrq ono(eg ouv66owor, ov- nnpooone0ouv oXe66v ro 1.uo6 qg pd(og rou. l-tep[- nou 70 dtreg npcoreiveq nou Bp(or<owor oro oopor(dro ouppogrlg ovogSpowor <og nop6yoweg ouppogfq (splicing factors). l-'lepd.oprBdvouv nporetveg onopof- rryreq yro rr1 ouvopl.rol,6ynon rou ocotrrorrd(ou ouppo- gnq, rnv np6odeori rou oro RNA un6orpogcl Kor rnv xcnooreu4 ev6g r<5wpou pre Bdoq ro RNA yt, owr6pd- oerq trans-eoreponoi4o4g (transesterification). Ercr6g on6 owSg ng npcoteiVeg, nep(nou S0 enrnp6oOereg nprrlreiveg nou ouv65owo. oro orrrprot(dro ouppogrlg Oecopefuor 6rr evepyoriv oe dlLo ord6ro rrlg dr<rppoo.qg rov yovr6(rov, yeyov6q nou unodrltrc6ver 6n q ouppogrj (oog ouv6detor pe dMo Bri;roro or4v dr<gpooq rov yo- vr6(cov (BMne Ev6rrya 21.11). EIKONA 21.7 To otoporidro oupporprlg eivoL -12 MDa. ll6we s- nRNP onoretrouv oXedov ro pro6 Tlq Ud(oS rou. 0r undi,orneg nptrlrelVeg nepiloppdvouv yvtrlmoUg nopdyovreg oupporp(g, ro- Otig en(o4g Kor rrpoTelveq nou epni,6rovrot oe dUo mddro qg 6rgpoorl g rov yovr6[rrrv. To oropor(6to ouppog4g 6rlgroupye(ror ndvo mo or6,poro np66popo RNA ror nepvd prdoo puog evdrdtrre- orlq rordoroonq ornv ono(o nept6Xer ro ypogptr6 ;16- pro rou 5' e(ov[ou ror to 6e(t5 ].doo rwpdvro-e[rivro. MSoo oro orjprnLor<o Bp(oreror pr6vo 5vo ptxp5 pdpoq rou ouppopgdvou npoi6vrog, 5v6er[r1 6rt ouvrl0rug oneAeu0epcbveror opr6ooq prerd t4v ronrl r;g 3, 06o119 r<or 14 oUvdeorl rov e(cov[<ov. To orrlpro{6ro snRNP eprnfuixowor oqv rorooreu4 rrlg 6ogng rou orrlproldiou ouppogrlq. 'Onog Kcu ro pr- Booco;rdrro, to oropor(6ro ouppo<pr]g e[optdror qn6 oI- trrlAenrdpdoerg R NA- R NA r<or o).l,r1Asnr6 pdoerg npo- reivqg - npolretv4g. Opro;r6veg owr6pdoerg nou nepr- Lo;rBdvouv ro snRNP npounoOdrouv dpeoo (euydpogo pero([ rou RNA rouq Kor. rou RNA nou ouppdmetor. lMeg owtd pdoerg npo uno0Srouv ovoyvrilproq prero[ri ttov RNP { pero(rj rov npcoreivdlv toug r<ot dMrov ou- ororxriv tou orrlpold(ou oupporprjg. H orlpoo(o rov snRNA prnope( vo eLeylOe( dpreoo orn (UUn p5oo qg np6x).r1oqq preroMd[eo:v oro yov! 6ro roug 11 oe owr6p6oerg rn vftro ouppogrig ;16oo qq oroXeuprSvrlg ono66;rqoqg [eXroprorci:v snRNA oro nu- pqvx6 erlUtrropro. H odpovono(4or1 tov nriwe snRNA, [eyoprord r1 oe ouv6uoqr6, onorprinet q ouppocprl. OLo ro snRNA nou epnl,6rowor orrl ouppo<pr1 ovo- yvrop((owor oe ouwrlpnUdvq popgrl oe 6l,oug toug euxopucirreg, ou gnepil,opBovoprdvov r<ov rputcirv. Irrl (6Un r' ow(orotlo RNA ouyyd e[vor preyo],rirepo, oMd or ouwnprlpdveg neptoXSg nepil,oprBdvouv XopoKrrlpr- mxd nop6proro Ue ro snRNA tov nolur<Urropov euKo- purorciv. Tq snRNP nou epnl,6rowor qrrl ouppog4 e(vor ro U1, U2, U5, U4 ror U6. Ovoprd(owor orjpgcovo Ue rq s- nRNA nou nept6louv. Kd0e snRNp nepr6let dvo provo- 6116 snRNA ror oprer6g (<20) nporeiveg. To U4 ror U6 snRNP ouvqOcog Bp(or<owor go(( oq dvo ocopro{dro 6r-snRNP (U4IUO). O xorv6g 6opur<6g nuprlvog xd0e s- Ailteq 30 rtporeiveq, l7% mc uotoc 5 snRNA, aao/ *^ ., ,o trg,, 1td(0! 70 nqoqvowec ouppoqns,sai. ;jXf#ffi*'nlq [o(oq iar" irlC ul(.s 5E1
nRNP ofloretrefuot on6 ;uo o;rd6o orcr6 nporeivdlv, or ono(eg ovoyvco p((owor ro ouroclwrorirproto qwr-S m (anti-Sm). Iuwqprlprdveq oMqLou{eg orrg nporteiVeg onoteloriv ro or6Xo owrilv roJV owroopdrrov, Or un6- Lorneg nporelVeg e(vot llovo6rxdg yro rd0e snRNP. Or npo:teiveg Sm npoodSvowor orrl ouwrlp4;r6vr1 oMq- LouX(o PuAUg-oGpu 11 ono(o undpXer oe 6Lq to snRNA ercr6g rou U6. To U6 snRNP neprSXer pro opd6o npo- reivdlv nop6pouov pre ng Sm (Lsm). Opropdveg nporetveg rov snRNP nr0ov6v epntr5r<ov- ror dpeoo orn ouppqqn. iMeq (oo4 SXouv 6oprrorjg p5louq 11 e[vot onopo(r4reg yro rrl ouvopprol6yrlon n yr' oMqlentdpdoerg pero[0 rov ocol-ron6(ov snRNP. l-1epfuou ro tivo rp(ro rov npoteiv6v nou epnfuixowor orn ouppoq4 efuot ouoronxd rcov snRNP. 'OLo r<or ne- ptoo6repeg evde[(erg uno64],d:vouv 6u to RNA 6Xer dpeoo p6Lo or4v r<otdtruor1: ouvend:q, or neproo6repor nopdyoweq ouppoEr1q Unope( vo 51ouv dopur<oUg p6- ).ouq n vo oupr;rer51ouv fin ouvopprol.6yrlorl rou oo1.to- n6(ou ouppogfq. 2 L7 Adop€uon Tou fip66popou mRNA oTnv 006 Iupporpfiq " To U1 snRNP nporotre( qv evop[1 11q oupporprlg pdtrLq npoo0e0er oqv 5' 06oq oupporpqq U6orrl luog ovr(6poo1g (euyoptirporog RNA-RNA. u* To ouUnl,oro 66opeuoqg nepL6yer ro U1 snRNP npooDe6epS- vo orrlv 5' Oeoq ouppoQnq rol rrlv nporelvl U2AF, 1 ono[o npooddveror 0e Uro ouoTorxlo nuprprdrvdrv pero[u rou orlpe! ou 6ror),o0cooqq Kor rnq 3' 0eo1g oupporprlq. e Iroug notrururopouq europutlreg, or nporeiveg SR nor(ouv onopo(qro p6lo oqv evopfr1 rou oXr11-rolopott rou oupntrri- rou deopreuorlg. * To (euyoptolro rorv 0doeov ouppoqlq enuuyXdveror pulotrt oproliou rov rvrpovlov r1 ralv e[trlvirrtv. H ovoyvrrrpron rcov ouvorvelrcbv orlpdro.lv ouppogrlg nepil.oprBdver 16oo RNA 6oo rot nporelveq. Opropr6- vo snRNA 61ouv oMrl).ouXieg nou e(vor ouprn).r1popro- nr6g trrrv ouvqtverrKcbv oL).q).ouXrov 11 prero[rj roug r<or to (euydpc,.rpo ;rerofri tou snRNA Kor rou pre-mR- NA r1 peto[ri rov snRNA no[(er orlprowrx6 p6].o or4 oupporprl. To U'1 snRNP tou ovOptonou neprdXer trg r<ewptxdg nporeiveg Sm, rperg er6u<69 yro ro U1 npurrelveg (U1- 70k, U1A r<or U1C) xor U1 snRNA. H deuteporoyrlq 6o- prrq rou U1 snRNA qo(veror orrlv . l-1ept61et opxetSg neproX6g. H 06oq np6odeoqg Sm e(vor ono- pofuqrq yro rnv otrLr;l.eni6poorl ps rrq r<orv6g npcoret- veg snRNP. Or neptoXdg nou ovoyvop[(owor on6 ttg [eXoprm5g 6o;rdg p(oXou-0r1Lrdg nop6Xouv 06oerg np6o6eoqg yto nporeiveg nou e(vor provodrx5g yro ro Ul snRNP. H np6odeorl rou U1 snRNP orqv 5' 05or1 592 KE0AAAI0 21 Iupporpn xr Ene[epyooto rou RNA ouppoqqg e(vot ro npri.lro prtpo o-cn oupporprl. Tc -- RNA oMq).enL6pd pe qv 5' 0dor1 oupporprlg pe (e". :- popo pero[0 tou pov6r.trovou 5' dr<pou rou Kor 4-6 3:- oeov rrlq 5' 06oqg oupporplq. MeroMd(etg orqv 5' 06o4 ouppo<pqe r<oL oro U' -- nRNA gnopei vo lplorpronoLrl0otiv c,rryre vo eley):: dpeoo ov e(voL onopo(rr1ro ro (euydprop-ro rouc - onore).dopioro ev6g rdrotou nerpdprotog nopouoro(:, - ror ornv H oMrl).ou1[o oyprou t0nou :-: Odor1g ouppogqg mo pre-mRNA rou 125 odevo.:- (euyoptrrver oe ndvte ond rrg 6(L O5oerq pr ro ;' snRNA. 'Evo pretdL).oyUo rou 12S RNA nou 6e oupc:- ftrtror dler 6uo oMoySq orqv oMrl].ouX(o: to r<ord.: - no GG orLg 0eoeLg 5 r<ot 6 rou twpov(ou 5.Xouv oMcl: oe AU. Av eLooX0e[ puo prerd].].oEn mo U1 snRNA * ono(o onoro0Lmd ro (euydpropro mq 05oq 5, q oupc:. qq pnope( vo ouprBe( xovovLrd. ALleg nrprnroJCs.: mrg ono(eg ow(morleg preroLi.dleLg drlptoupyou, : oro U1 snRNA rrrore vo 6ronLoro.l0e( ov e(vot rKovrq .l rotooreil.ouv q prerdMo[r1 orrl 06o11 ouppocprlg, c,.:- detrvrjouv ro yevrrd rov6vo: q ou;rnLqpcr;1lorrrorr:: piero(U tou U'1 snRNA Kor rnq 5' 06o119 oupporprlg eL',: onopofuqtrl yro rrl ouppogrl, oMd q onoreLeoporL<c- rlro rrlq o'upporprlq 6ev r<o0op((eroL onox.l.eLorLxd cr : rov opr0;rd trov (euyrirv Bdoeotv nou oXq;ro-r[(owor, H ow(6pooq (euyoprrtl.ioroq rou U1 snRNA ;-re r-. 5'06o11 ouppogrlg oro0eponore(toL ond nporeiveq. - nporelvq nou npoo6S.veror orrl 05or1 6Lor<Lddcoor : (branchpoint binding protein, BBP, yvroorq r<oL og SF' oM4).enL6pd pe rrlv oM4).ouX(o rou orlpe(ou 6rontrc- 6oro119, evo q U2AF (5vo e-repodrprep6g nou onore,s.- ror ond rqv U2AF65 r<or U2AF35 oe noLux0rtoc: europurotrr<d rUrropo q rqv Mud2 orn (6Un Saccha- romyces cerevisiae) npoo6SvetoL orq oumorX(o nupt! - 6rvcrrv prerolU rqq otr).r1Lou1(og rou oqpe(ou 6ror<Lc- 6ooqq Kor rou mo0eporj 6rvour<LeotL6(ou AG oro :8- Log xdOe rwpov(ou. An6 pr6vo-rou, r<ov6vo on6 ourd i: ouprBdwo np6o6eoqg 6ev e(voL opr<erd ro1up6, r,lmc- oo rl ouvepyoo(o roug o6r1ye( oro oxnUorrop6 evoc oleru<d oro0epo6 oupnl.6rou nou ro).e[tor ot-ipn].ox: 6do;reuoqg (commitment complex). Iro rrjmopo rotv 0qLomLrov, ro orjprnl.or<o -:,- opeuorlq e(vor en(or1g yvoor6 otg oripntroxo E (,,E,, y:c ,,e?rly>, npt,ttpro). O ol4porLo;r6g rou 6ev ononel ATP (ow(Oeto on' 6Lo ro dtl.rtpro Brlporo xord q ouvoppic- ).6yqor1 rou ooUor.6[ou ouppo<pr'19, B],6ne Evotqrc 21 .8). Aorcoo, or ouvorverrr<5q o).).r1LouX(eg orLg 0e- oerq ouppognq oro yov(6Lo rrov 0lAoortrurv e(vor 1c- Lopd ouwqpqprdveq xot, ouvencog, yro ro oXnUoloUC rou ougnL6rou E onoLrorjwoL enrnp6oOetor nporetvt- r<o( nopdyoweg. Or nopoyoweq nou no((ouv rewpu<6 p6lo 16oo oe ourqv 6oo Kor oe dMeg 6repyoo(eg ouvopl-ro).dyqo1c rou oro;rotr6(ou ouppogqg efvor ot nporeiveg SR, prrc ou<oydveto nopoy6wolv oupporpqg nou nepL6louv dvc rq 6Uo goriBo ovoyvrllpLorlg RNA oro N-reLrr<6 drpo xo. pto neptopl "unoYpogri" nLoUoLo oe noMonLSq enc- voLrlqUetg rou 6rnenrr6(ou Arg/Ser (r<o).e(ror neproxr RS) oro C-re).Lr<d dr<po. Aurd ro ;ror(Bo ovoyvrirproqc Boorxfq 6womg
. To U1 snRNA 6Xet pLo (euYoptlp6vrl 5opri nou di'lptoupyer oprer6g neptoXeq. To 5' orpo iopo;t6vel povortrovo xot (euyoptivet pe rqv 5' Otlot1 u ppoQIlq. RNA e(vot uneU0uvo yto etdtrrl yto qv otrr1- LouX(o np6o6eoq mo RNA. En(or1q, n flepto- Xrl RS ;rnope( vo npoo6e0e( t6oo oe RNA 6oo xot oe dMoug nop6yoweg oupporprlq prSoo otrrlLenr6pdoeov preto(ti npotetv6v, nop6lowoq 6tot np6o0etrl ur6Mo" yro 6td- rpopo p5p4 tou oupnl'6rou E. 'Onog nopouotd(erot orrlv EIK0NA 21.10, ot npu:tefveq SR npoo66vowot oro ouororx6 70 kD rou U1 snRNP (r1 Ul np<otetvq 70 kD neprsXet en(or1g prto neptoXrl RS, ol d 6e Oeo- pefuor rog runu<r1 npcotetvq SR) yro vo evtg4ti- oouv rl vo oroOeponot{oouv to (euydpogd 'rnq Ue rrlv 5' 05or1 oupporpqq. Ot npcotetveg SR npooS6vowot enbqg ornv U2AF nou efuot 6eopreupr5vq ornv 3' 06o11 ouppotpr]g (pto ne- proXq RS undpXet entonq t6oo qrnv U2AF65 6oo rot orlv U2AF35). Autd ro 6(rruo npto- rcivrrdlv oMrllentdpdoecov 0eolporiwot rp(ot- Uq yro ro oxrlpqrtopr6 rou oupntr6rou E. Ot npoteiVeq SR onopovdrvowot pro(( pe to o6pr- nloro Pol Il t<t 6Xouv rqv rov6rrpo xrvrltu<tig 65o1-reuoqg tou RNA ornv o66 ouppotprlg. Iu- vencig, nt0ov6v l.eroupyofv cog evoprcrrlptot nopdyoweg rnq ouppoqriq oto rfrropo ttov notrur<Urropolv euroPuotd:v. nepLoxn B ^A^ HH JY IbrfiD $B-H$' ffiH HF.B s&E npoooeontrsf. t&ftKg #"# "g sfry ocu ccu'oii uuo cuGAccc " rFffi ,,^^*' % r rnc c un u u" oooo"oo oot^ EETiS:l AC ^dAdcouccA u u cA u ApppGme 5' c E s'E[rr]vto GU A A GU A lvrPovto 3' cG I UA I UG I UA I , H ZeuYdptouu tvtPov(ou GC GC nu'oru Tunu<5q npcoretveq SR 6ev rco6tronotofwot sro Yo- vt6(orpto tou S. cerevisiae o0te e(vqt ovoyr<o(eg Yto rn ouppogrl oe ow6v rov opYovtopr6 6nou ro origoto ouppo<pr1q e(vot oXelrd opretdBtrrlto' Aw(Oeto, e(vot on6Luto onopo(tqteq Yto rn ouppoqrl oe 6).oug toug notruxfrropoug europucireg 6nou to or]proto ouppo- griq onoxL[vouv notr0. H e(5].t[4 tov npu:teivriv SR oroug notruxrirropoug europutiteg nt0ov6v ouvetorp5- per oe ouppoErl uUnInS onotel'eopotr6tqrog Kot nt- m6tqtog oe Xol.opd ouwrlpqpSveg 06oerq ouppotp4g' H ovoyvrirptorl tretroupytr<drv 06oeov ouppoEriq r<otd rq dtdpr<eto rou oxrlporl.opo6 tou oupntr6rou E qxo- )rou0ei 66o nope(eg, 5nog <po(verot orrlv EIKONA 21'11' Erov S. cerevisiae,6nou oXe66v 6Lq to yov(6to nou ne- pdXouv twp6vto 6tox6mowot on6 6vo provo6tr6 pxp6 rwp6vto (prlroug -100-300 vouxtreond(rov), or 5' rot 3' O5oetg ouppqgnq ovoyvop((owot tout5lpovo on6 to Ut snnNp, BBP rot Mud2, 6n<oq npoovorp5pO4r<e' Auti q 6repyoo(o ovoq6petot oq oprop6g Iwpovitov (lntron definition), 6ncog <gofvetot oro optorep6 pdpoq qg Et- x6vog 21.11. (l-lopoqprlore 6tt o prqXovropr6q optogoti rwpov[tov toXUet yto prtrpd rwp6vto oe nol'ur6rrqpo eu- ropuo:ru<d r<rirropo rt 6tot q etx6vo lpqorpronote( r4v opol.oy(o rov nopoy6wtov ouppoqdg tcov 04).oortr<drv nou egntrSrowot srrl 6tePYoo(o). Aw(0eto, mq notruK0rropo euKopucottx6 yovt6ttil- AAoo"oa' nePoXn n poro ro twp6vto e(vot preydtro n 61ouv nol'0 getoBl'r1- 16 p6ye0oq. En(or1g, nepr5Xouv noM6g ol'I'r1}'ou1(eg nou pot6(ouv Ue npoYportxdg Odoetq oupporprlq' Aur6 duoXepo(vet ri rot r<oOtmd ov6qtrtq q ou(euy;r6vr1 ovoyvtirptorl rov 5' t<ot 3' 06oeov ouppogriq. H )'Uorl owori tou npoBldpotog e(vor 11 dtepyoo(o rou opl- opof e(oviov, 11 ono(o o(tonote( to ;uxp6 p5yeOoq tcov eiov(ov oro rcittopo tcov ovrilteprov europutrltcirv (pui- rog - 1 00-300 voux).eot(6to). 'Ontog cpo(vetor oT o 6e(t6 p6pog qq Erx6vog 2'l ' 1 1, xotd tov oprog6 tov efcov(ov, to etepo6tptep6g U2AF npoo6Svetot orrlv 3' 05oq oupporpdg xotto U1 snRNP (euyopcirvet pe rnv 5' 06oq oupporprlq ro0o6tr6 qg oi'- l,rllou{og rou e(tov(ou. Ir1 ouyxerptp6vrl 6tepyoo(o ouverog6pouv nptotetveg SR nou npoo66vowot oe et- 6u<5g oMqlouX(eg tov e[tov(tov preto[rj rcov 3' Kctt rov xo0o6xrilv 5' 05oerov oupporpriq. M5ool ev6g dyvtrlorou prrlXovropoti, ro oUpmtroro nou oXrlprotf(owot r<otd grl- xog tou e[ov(ou peton(mouv oo oUpntror<o nou ouv65- ouv rrlv 3' 06o11 ouppoqriq otrlv ovo6trri 5' 06o4 ouppogrlq Kot rrlv roOodtr<r1 5' 06o4 ouppoqrlq ottg en6lreveg ro0o6tr<5q 3' 06oerg ouppoqrlg 6top6oou rolv twpov(tov. Aw6 eyro0tord tqv uenttpemtr{" 6rqp6p- qoorl nou enup5net vo ouptBotiv to peroyevSorepo Bn- Uoro ouvopUoL6yrloqg rou oto gon6(ou ou ppoqf g' H ovooro)rri ourlg qg ;retdmoorlg e(vot 5vo p6oo yro rn pU0prto4 qg enrloyriq ouYKeKptgdvtr:v e(tov(tov rord tr1 pu0g(6prevq oupporprl (B).6ne Ev6t4ru 21'13)' 3'1"? A6oueusn rou [lp66poUou mRNA otlv 066 ttnpoqrts 5S lleproxn C
Ul snRNA dyprou flinou rqt 'l 25 pre-mRNA h. : ... i,. ' .:.J5[ ,: :, flH g3' flfl fgqr JIH Y ,:R,F;@FrurubeFkut-5 5' Ep$vro G U G A G G lwp6vro 3'Hq $l 06on oupporpftq 125 qdevolori U 1 snRNA dyptou rUnou xor gerdUoygrivo 1 25 pre-mRNA i1r '. "',rCSl 3', 5' E{<0vro G U G AA iJ lwp6vro 3' irl g: gli :riii*?:t *ir;1 f rrflf *{ir: -t MeroMoypdvo U1 snRNA (0r Uer0ll0yU6v0 125 RNA ;'':.i:::. 3', iiiiti,lflg:: iiit i"il 5rRliil ' : .,.C.A.U,,Ui-r..AU 5' 5' E(tirvro G U G A A u lwp6vLo 3 tlK0NA 21.9 MeroMd(etq nou KoTopyodv rn treroupy(o rnq 5' 06o4q oupporprlq KoTqor6trtrowor U6oo ovTrppoflrorrKd)v UeTotrIo- [eov mo U1 snRNA ftou orroKoeroroUv ro (euydptolro Tov Pdoeov. E[tivto reuu Te).u<d, o pu'11ovrop6q opropoJ e[ov(ov nou 6roLieoolc- Befuor on6lq npore(veq SR nopSler en(onq 5vo prlXc- vrou6 nou enrrp6ner (euydprr;pro Kor oUvdeorl U6oo r.( ouppoenq pi6vo peto[rl yerrovrrov 5' ror 3' 05oea, ouppoQriq. 21.8 H O66q Ixnpono;"ro0 rou Etopon6iou Iuppogfiq * T0 ouuntroKo dSoueuonq e€rtr(ooeTor 0e np00p0u0 o0ll.10:- Dro ouppoqqg (oupntroro A) nopouo(o ATP * H npoodeon rov U5 ror U4lU6 snRNP perorpSrer To npur- 6popo otolrorrdro ouppoQrlq oe dlprUo otopor(6ro 0uppo0-i (o0Untroxo B1). e To ouUn),oro B1 6neuo perorpeneror oro oupntroxo 82, c.: onolo ro Ul snRNP one],eu0ep6veT0r yro v0 enrTpriqrer 0.: U6 snRNA v' otrtrltrenrdpdoer Ue qv 5' Oeol ouppoqrtq * H U4 onouorpuveror ono ro U6 snRNP enupenovrog oro Uc snRNA vo (euyoptioer pe ro U2 snRNA yto v0 oxlpolosr:: rorotrunro revrpo yro tr1 oupporprl. * Kor or 611o ovldpooerq lrans-eoTeponoinqq ouptpolvouv o.: evepyonorlp6vo orol-ror[0ro ouppoqnq (oupntroro C). * H 0vT[6p00rl oupp0Qnq tlvor ovTroTprnrrl oe otro ro BrlUorc M6Lrq oplpotrme( ro oriprntror<o E, r' dl,o snRNP ror or nopdyoweq nou eprntr5r<owor fir1 ouppoe- oL).qen16poriv pre ro oripnlor<o pe ro0oproprdv'- oetpd. H llil,ri:l;i ;:= l,: 6e(Xvet To oUoToTtKd ttlv ou1-- ni.6rc,rv nou ovoyvop((owot xo0rrrg npoXrope( q c,,- r(dpooq. Iro npcoro Brlpro, to ono(o e[optdtor on6 ro ATF Eltirvro -ry -Au 06on luorory(q Pyg' 6uddd0onq Odoq ouPPotPtlg sPyssAGFm lvrpdvto 5', 0dor1 ouppoqrlgH Ul ror ot nporetveg SR xoOop((ouv qv 5' 06o1 ouppoqrlq '- ':il, SR :'. H U2Af npoo66veror oqv oupri Py roL qv 3' 0dor1 oupporpr'1q, gto etbtrrl npoTsivn ovoyvrop((er m Oeon drortroDoonc SR" 0t nptoreiveg SR ouvd6ouv ro oUpntroro mrg 5' rot 3' 06serq ouppo9ng 6me vq oxnuqrrme( ro oUpntroro d6opeuorlq (E) BBP ,U2AF65i35 GU ::UACUMC --Py =AG I SR *"U24ffi,/35:- -, ------t {@_UACUMCI EIK0NA 21"10 To oupntroKo d6opeuoqg (E) oXrlpori(eror pe q drodoXxrl npooerlKn rou U1 snRNP orrlv 5' 06o1 oupporpqg, qg U2AF m1 ouororX(o notrunuprprdrvrirv/3' 0604 ouppoQnq ror qg nporelVlq ouv6vrrlolg SF1/BBP. 594 KEOA Al0 21 Iupporprl rr Ene[epyoolo rou RNA Boorxeg eworeq
I o*r^rn,uouor,, V 0tou6oou rou tvrpov(ou 0propdg rvrpov[tov Elrivto lvrpovro E(rivro wGUwUACUAACT PyxAGrw 5' 06o1 Odoq DrortrdDoorlg 3' 06011 0propdq elov(trrv E[tivro lvrpdvro E[rirvro xAG wq gg w utrQLJMC x Pya AGwsslE GU = 5' 0do4 06oq Dtortrd6ooqq 3'0dor1 5' 06o11 I or,,nrrr,,oooor,, V 6ro[doou rou e[ovfou UACUAACE Merdnroo4 ond d1,flentbpdoerg 6ro96oou tou e[orv(ou oe otrtrltrentdpdoerg 0rog6oou rou tvrpov[ou YnopXouv 6uo odot yto qv opXtrrl ovoyvtirprol Ttrv 5' Kor 3' 06oeov ouppoQnq, e(Ts Us opropd rvrpov[rov e(Te Ue oproUo ,:-,.iJV. -: .12 snRNP ouv66etot pe ro U1 s- - : ' P ndvco oro np66popo mRNA 95- :* rpdodeonq ornv oMq).ou1(o rou r-..erou 6ror).d6coo4q. Aur6 en(onq -i:ri.opiBdveL (euydpopro UeroqU rnq :'-'.-ou1(og rou U2 snRNA Kor rrlq : .'.1),ouX(og rou onpe[ou 6rox].d6tr:- -'.: To or-iprnloxo E petotpdneror - ..3v oro np66po;ro ocopor[6ro oup- : ri-q nou e(voL yvcoor6 oq oUp[tro- .3 A xor our6 ro Bqpo onorre( - I:c,n uor1 tou ATP. -c orlpn).oro Bl oXqpor((eror p6- : t',,o rprpep6g oUprnLoro on6 ro -: <eL U4lU6 snRNP npoo6ege( oro - --:^oKo A. To 81 Oeope(tor drr e! : :'"rpor[6Lo ouppo(pnq enerdrl ne- : I :.ai To ouotottrd nou onotrofwor : .1v ovr(6poon rnq ouppqerlq. -,:J oneleuoepooe( ro U1, ro oUp- - :(c ourd perotpS,neror mo o0pr- - :<c E2. H o<po(peoq rou U1 e(vot :-::o[rqr1 otote ro und].orno ou- . H ovT[dpoon ouppoglg npo- -,. ir Storpuo orodro roro T0 oilolo o . --: :1.rog rou otrllrofldlou 0uppoQnq ne- - :-i:','er qv otrtrqtreni6p00rl Tov ouoToTr- , l- ovoyvop((ouv Trg ouvolverrreg otr- - . ,.. IUpntroro E IU[ntroro A IUpiltoxo 81 Itlpntroro B2 Idpntroxo C1 1 U2 IXqporLoprlg tou ougntr6xou deoueuonq oro ono(o n U1 eivor (euyopouevn ge qv 5 0€orl ouppoQng tlpoo0nKn U2 yro v0 (euyoptioer pe rn 060r1 6rox),odooqg nopouofo ATP ' ^46t!n@ *AU" + Iuvdeoq tov U4,6/U5 rpL-snRNP Anetreu0dproorl rov U1 ror U4 IXrlpouopdg rou rorotrurrxorj Kdvrpou oro onolo ro U6 (euyopriver Ue rnv 5' 0don oupporprlg, ro U6 (euyopriveL en[o1q ge ro U2, ro U2 nopogdver (euyopopevo pe q 0dor1* ^F@ - ^u-- 6rortro6oror1q. ro U5 o),trr1trenr6po ror ge ro 6Uo e(6vro U6oo rnq Onroq rou & , # To npdrro gnUo rnq trans-eoreponoqorlq ' ffi; xlBe-0,,'r5'06o11ouppogr1s,o1r1pori(eror U2 Gqw "" To trooo UACUMC:PycEAGr?Em To 6eurepo BnUo tr1S trans-eoTeponorn0nq Krlperot 11 3' 0eor1 oupporprlg, ouvddovror ro eIdrvro m.'5AGu IUgntroxo C2 ?:.fi H Odog IXr;poloUOU TOU IorgorLd[ou Iuoorc.t 595
srcntx6, 16(ro9 ro U6 snRNA, vo Bpe0oUv 5hLo orrlv 5' 05or1ouppoqrlg. H rorol.urxrl ow6poo4 nupodotekot on6 r4v one- Aeu06pooq rou U4 nou en(oqg ouprBofuer rord q prerd- Booq on6 ro oUpnloro B'l oro B2.To U4 snRNA (orog "eyr<Ao9((er,,to U6 snRNA 6cog 6tou ttotoore( onopokq- ro. H EIK0NA 21.13 6e(yyer rg oLl,oy6g nou ougBo(vouv mrg oLtrrllenr6pdoerq (euyopdrporog Bdoeov gero(U rov snRNA xord 11 ouppogrl. Iro U6/U4 snRNP, puo ouveXriq 6rcrooq 26 Bdoeov rou U6 (euyopdrver pre 6uo [eXopror6g neptoXdq rou U4. M6].rg onoproxpuv0e( ro U4, 11 neproXrl rnq UO nou onetreu0ep<ilveror prnope( nL6- ov vo uro0erqoer dMq 6opq. To nprito pr6pog rqg (eu- yopcirver ;re ro U2, evtil ro 6er3repo oXqprot((er puo evSotrroprorrl qoupr5ro. H otrl,qLenbpooq prero(ri trov U4 r<or U6 efuot ol.rorBo[o ootiprBoq pe rqv oMqlen(6po- o1 pero[ri trov U2 Kqt U6, on6te 4 one].eu0dpoor1 rrlg U4 eMyXet rqv xov6r4ro rou ooUorr6(ou ouppo<ptjg vo npoXcoprloer ornv evepyonorngdvn rordorooq. l-ro L6youq oo<pfvetog, q err6vo 6e(1vet ro RNA un6orptol.ro o' errerop6vrl poperl. Iqv npoyponr6q- ro, 6prcog, r1 5' 06oq ouppog(q Bp(orerot xowd orqv oM4lou{o U6 oprdoog 6(n},o orqv 5' ntreupd rnq oI- LrlLouX(og nou e(vor npoo6e6epr6vq oro U2. Auuj q o),- LqLouX(o rou U6 snRNA (euyoprirver pre oAlr1trou1(eg 5'(opLmepfl) 06on ouppoQnq EIK0NA 21.'13 To (euydpogo U6-U4 eivor ooripporo pe ro (euyo- proUo U6-U2. Orov ro U6 ouvdeOei mo ourl.roridro ouppoQnq, (eu- yoprlver pe ro U4. H onetreu06proon rou U4 enrp6net ULo dttroyrl orn druu6pQoon rou U6. 'Evo u6poq rnq otrtrrltroux(oq nou onetreu- 0eptir0nre oxnUor(eL pro rpoupr6ro (yrpr) evdl ro dtrtro (poQ (euyo- pdlver pe r1v U2. Mro yerrovrrrl neproxn rou U2 eivor 11611 (euyopop6vrl ge q 0eoq dtodddooqg, q6pvowog ro U6 ovnrpu- or0 oq 6rortro6oor1. l-lopoqprlme 6r ro RNA unomptrllto 6ev 6Xer ro ouvl0top6vo npooovorotrropr6 ror qofueror oe rore00uvorl 3'-5'. To Ul (euyoptiver pe qv 5' 06srl ouppoqnq 5' 0dorl ouppqqrlq 3'AU'GAU..5' 5L-U A C U AC ------------AG--- 3', 06011 0Lortro0roor1g n To U6 (euyopdrver ge qv 5' 6dor1 oupporprlg U6 3'GAGACA.5' 5,AGGUAUGU 3, 5' Odon ouppoQnq E[tirvro 1 at +1 Eqdvro 2 at +1 H oupporprl Xplorponorel pro oerpo ovrr6pooa-, (e uyoptirporog pero[0 rtov snRNA ror rorv 06oetov 0uppoQrlc rou twpov(ou nou Bp(or<owor r<o0o6rxd qg ouwrlp-- p6vqg GU mqv 5'0rior1 ouppo<pr1g (prerol.LdleLg nc- evroXriouv 6vq rSroro (euydpulpro Betrtr<r-lvouv rnv or:c- re).eopotrrdrnro rnq ouppocp4g). Iuveno-rg, xord rqv nope(o rqq ouppogrlq, npoy;-c' ronoLorjwor op rer6g owrS pooerg (euyopr.,.r ;roroq [rer c- (U turv snRNA Kor rou RNA unoorpcirprorog. O. owrbpdoeLg ourdg ouvotlr[(owot orrlv C. snRNP dXouv oMrlLoul(eg nou (euyoprrrvouv tdoo -a ro un6mpull-ro 6oo xot prero[ti toug. En(oqg, 51ouv ;-rc- vdxi.toveg neprol6g oe Oqlldg nou Bp(or<owot t<owd o: oL).q).ouX(eq tou unoorproporoq Kor no((ouv onUowrK. p6).o, 6norq ouvdyeroL on6 ro yeyov6q 6rr preroMd(e.c ortg Oqlteg roropyorlv q ouppogq. To (euydptopro preto[rj rou U2 Kor rou otlpe[ou 6Lc- r<),d6oror1g xor prero[ri rov U2 r<ot U6 6rlprroupye( pric 6oprr1 nou por6(er pre ro evepy6 r6wpo rov ouro-oup- pont6pevorv rwpov(tov 11g opd6og ll (B).5ne ELrovc 596 KE0AAAI0 21 IuppoQrl rr Enelepyooro rou RNA J 5', Korotrurtxd
2 1 15 oillv Evorrlra 21 '9). Auro eye(pet rlv nt0ov6trlto :c rorotrurrr6 oustottx6 vq e(vot Utq 6ogn RNA nou :opoyetot on6 qv oMrlleni6poorl U2-U6 To U6 (eu- ,iopover Us rnv 5' 05oq ouppotpqq Kot Pto 0r1l'rd mo U5 snRNA Bpioxetot or<ptBtog 6(nLo on6 ttg npolteg 3ooerg Kot rov 60o e(irlv(tov. Motrov6tt ot 6ro0Sotpeq avbe([etg unodetrvJouv 6tt o prllovtopr6g r<ordtruorlg reoo oro otrlpor(6to ouppoqrlq Boo((etot mo RNA, q :uveto<popd tov npotte[vtiv 6e pnope( v' onox]'etore(' ',lio uno{.rq<pto npote(vr1 e(vot t1 Prp8, ;rto prey6trq npo- :eLvr1 trptoprqroq nou 6rlpuoupye( dpeoeg enorp6g rot -a rLq 6rio O5oetg ouppqqrlq, 5' t<ot 3', pr6oo oro ooJpo- :5ro oupporpqg. Kot ot 6[o owtdpdoerg trans-eoreponoiqorlq npoy- - oronoto Uwoi oro svepYoTlo tn Udvo otr: prot(6to ou ppo- :11g (o0prnLoro C) p6LLq olorLqpo0e( pto oetpd :ro'6toro(eov tou RNA. O oXrlpiottop6g tou Ldoou -q 06oq dtortrddororlg roOop((er q lplotprono(qoq :'^c 3 06onq ouppotp4g, enetdrl ri 3' ouvotvettr<4 oMq- ':.X[o nou Bp[oxetot nto xowo oqv 3' n]'eupd tqq :.:nd6too4q y(vetoL o or6Xog Yto rrl 6eUrepq trans- . -apono(qoq, To orlprowtr<6 oupndpoo;.to oro ono(o xotoLqyouge :.. r. ort to snRNA rqq ouoKeuqq ouppaqnq aM4Lent- - :: -tv t6oo peralu rcuq 6oo Kot pe ro RNA unootpupa ,.: -t (euyapds;tarcq Baoeotv xat auttq ot aMrllent6pa- -. : €nLTpenouv aMayiq or4 5opr1 ot onobq pnope[ va -..auv Kowa rLq oilddeg nou awtdpolv 4 ax6pa xat va :' -.o u pytloouv xata)'urLxa xiwPa. -op6rt to ocopot[6to ouppo<pr1g prd].]'ov efvot f-ito -..:q Ulxovn RNA, 6noq to ptBooto;rdtto, yto vo Let- -: -:v(oet lpetd(erot noMo0g npoteivtr<otlg nopdyov- -:: Extetop6veg ovoL0oetg pretoLl'o[eov nou -ttruorollotrloqrov orn E[un ovoyvtrlptoov t600 to :',- ooo Kot ro npoJTeivtxd ouoroflr<d (to ono(o ovo- -.rc(oi Ue ro oKpovriprto PrP: pre-mRNA processing, ,-rieoyoo(o pre-mRNA). Apr<etd on6 to npoi6wo ou- - - , :Lov yovt6(tlv SXouv prot(Bo nou ro rowonototiv trlg : - :', eveLo RNA eltrooov e(opttiprevcov ono to ATP, ot :-:.eq e(vot rp(otpreq Yto Uto oetpd ovo6tord[eotv mo - -*:;i6Lo oupPotPrlq. + Prp5 e(vot rp(otpr1 Yto rnv np6o6eor1 tou U2 m1 :: ;- Sror<.l.d6coor1g xotd q pretdBooq on6 to o0;rnLo- . - f mo A. H Brr2 6teurol0vet trlv onel'euO5ptr-loq - - , J 1 roL U4 xotd tn pretdBoorl on6 to orlprntroro 81 - : 32, H Prp2 e(vot uneJOuvn Yto tqv evepyono(1or1 -:, :.1;.tott6(ou ouppoqrlg r<otd tr1 trletotponq rou , -- .i.o<ou 82 oe o[prn}'or<o C rot 11 Prp22 Forl0d . -., :neleuOdpooq tou r.,-lpt;'rou mRNA on6 to oopo- - : : ouppotpqq. Entnl,6ov, opropr6veg RNA etr<doeg -: -:-v pd).o orrlv ovoxtit<trtooq tov snRNP Ytq rov : -: -i,/o r<Urtro ouvoppro).6yrlonq rou otr-11-'tott6(ou ---*-vll, : -:o to euprl;roto e[qyoriv yrot( xpetd(erot udp6- - r- :ou ATP oe 6td<popo orddto rrlg ovt(6pooqg - -:::c.cqq, polov6tt ot (6teg ot ovtt6pdoetq trans- , - .:cno(qonq 6ev onottoUv ATP. I-1op6tt oro ooUo- - - : ;..rppogr1q ouprBo(vouv 6to6oXtrdq ovodtotd(etg rou RNA, e(vot o[tooqtrle(oto 6rt q 6to6troclo ctr ''' Tot noq e(vot owtorpentrl prerd rqv nprr-ltt1 roL 5a-::- pr1 ow(6poorl trans-effiepono(r1or1q. 21.9'Evo EvqA orrtro Ecouori6ro lupporpfiq Xpnotponolei AlogopeTlKd snRNP Ylo vo EneEepyoorei To <tEAdooovorl lvrp6vto o Mto evotr)ronxn o66q ouppuQnq Xplotponore[ Uto dt]rn oud- dq snRNP, 1 ono(o 6Xet xotv6 pe to rupto otoporidto oupp0- qrlg privo ro U5 snRNP. . io tvtp6vto ot6Xot 6Xouv ueYolUtepeq ouvotvetxdq olJrrl- trouXieg oilq ouUBoI6q ouppqqnq, Xolpig v' orolou0ouv nt- md roug rovdveg GU-AG n AU-AC. . T0 uei(ovo rot to eldooovo otrlpor6to ouppoqrlq uopd(owot orlpovrxoug nptrlreivtrofg nopdyovreg, gero[6 dtrtrtov ttg nPtrlreiveq SR. To twp6vto GU-AG enu<potoUv (>98olo rov ougBol'ti:v ouppoq(g oro ov0pdrnwo yovr6(topto)' E(orp6oerg e[- ,oi o, 1r4 t<ovovtxSg ouprBoL6g ouppog(q AU-AC rt dMeg nopohLoy6g. l-1ol,ot6tepo, ro twp6vto out(g -qg "dooorog r<otrlyop(og ovoq6powov oq twp6vto AU-AC oe owtdtomotr'n npoq to "ge((ovo" tvrp6vtq nou qrolouOo0v tov t<ov6vo GU-AG rot6 q ouppo- <p(. I(;repo nou 6tol,eurdv0qre n pnxov{ ene(epyo- oloq tOoo t<ov pret(5vtov 6oo Kot rov etrooo6vtov rwpov(ov e(vot ntr6ov oo<p6g 6r ourrl q opotroy(o yto tr1v etr6ooovo rorqyop[o twpov[tov 6ev e(vot on6truto orptBrlg. Me o6nV6 trlv epnetp[o nou onorcrf04re on6 tqv notr01povr16peuvo pe owxe(pevo to pre[(ov otopot(6to ouppo<priq, n Unxovn ene[epyoo(oq tov e]'ooo5vcov w- tpov(tov 6ioLeur<dv0r1re yp(yopo. AnoreLefuot on6 U11 r<ot U12 (q4eil(owot pe to U'l rot U2, owtorofioq), 5vo r<orv6 U5 nou propd(etol pe ro pre((ov oopotfdto oup- pqenq Kqt ro U4atac Kot U6atac snRNA' H ow(6poor1 buppoEnq ouotocnxd e(vot nop6ptoto ge rqv ow(6po- on ytq to pre((ovo rwp6vto Kqt to snRNA 61ouv ovdtro- youg p6trouq: to U11 (euyopdrvet Ue rtq 5' 06oetg oupporprlg, to U12 Ue rrlv oMrlLouX(o rrlg 05oqg 6to- drddooqq rowd orrlv 3' 06on oupporpqq Kot ro U4atac Kor U6rt"c nop5louv ovo)ioyeg Leroupy(eq rotd tr1 ou- vop;rol6yr1or1 Kot rrlv evepyono[r1or1 tou otrll'ton6(ou ouppogrlq. 'Onoq ono6e(X0nre, q enil'oyr1 tou rtinou rou oo- pron6(ou ouppo<prlg en4pe6(etot en(or1g on6 ng oMrl- ioul(eq dLl'tov rprlprdrov rou tvrpov(ou, on6re un6pXouv optop6vo twp6vto GU-AG nou ouppdmov- ror on6 to otopor(6to ouppoqriq rfnou U12. Mto tolu- pr1 ouvotvettrri oLLqLoul(o oto optorep6 drpo r<oOop((er rov U12-efoptdrprevo tUno tvtpov(ou: 5' ,i:,.j 'Ev0 Evotrtroxlr6 loporioro luppoqnq Xplotgonote(Atorpopertxo snRNP Yto vo Ene[epyoore(ro "E],com"l ' :: ' I 5Ei
GnUAUCCUUU,..PyAGc 3'. Itrlv npoYportK6rnro, ro ne- proo6repo U12-e[optrirgevo twp6vto 6Xouv oro 6rpo rouq rnv oMqLouX(o GU...AG. Entonq, 6Xouv pto el,ot- pertrd ouvrrlprlpr6vq 06oq 6tortrddtootlq (UCCU- UPuAPy), 11 ono(o (euyopcilvet Ue ro U12 (BL6ne Etx6vo 21.3). Aurq 11 6togopd mtg oLl4LouX[eg qg 05or1g 6rox).66oo49 e(vot 4 rripto Srdxptoq preto[ti Uet(5vcov xor eLooo6vrov rwpov[rov. l-t' our6 to tr6yo, to pe(ovo rwp6vro r<otroUwot U2-eaptdpeva, wti) ta eAdooova U 1 2-eaprdpeva, aw( yn tw p6vLa AU -AC. Or 6uo r6nor twpov(tov ouvundpxouv oe notrfi,o yo' vrdtcill.roto Kor rrq neptoo6tepeq gop6q Bp(or<owot oro (6ro yovi6to, To U12-e(optcirpevo twp6vto ouv4Oog ne- prBdMowot on6 U2-e[oprdrpevo twp6vto. H <pu].oyo- v(o ourrilv rrrlv rwpov(cov uno64Ldrvet 6tt (ooq xdnote rupropXotioov ro AU-AC U12-e(opt<irprevo twp6vto, oL- trd oe peroyev6orepo orddro qq eE6LtEnq enxpdrqoov ro rwp6vro pre drpo GU-AG nou e(optriwot on6 ro U2. H rorvrl e[6r[q ttov ouorqpdrov unoypoppr(etot on6 ro yeyov6g 6tr XpqoqonotoUv ovdloyeq op66eq (eu- yopropr5vov Bdoeov peto[0 trov snRNA Kot rou pre-m- RNA unompcbprotog. EntnLSov, 6tror ot onopo(rrltot nopdyoweg ouppoqrlq (n.X. npoteiveq SR) nou petre- rjOqrov p5Xpr rcilpo e(vot ovoyro[ot yto t4v ene[epyo- o(o r5oo rov rvrpov(rov t6nou U2 ooo Kot rov rwpov[olvilnou U12. Mro o[roorlpre(toq dtogopd peto[ti ttov U2 xot U12 ninov rwpov(tov e(vor 6tr to U1 rot U2 go[verot v' ovo- yvrop((ouv ove[dpqru ug 5' rot 3' 06oerg ouppoqrlg oro pre(ovo rwp6vro xffid ro crXrlprottopr6 tov oupntr6- r<ov E ror A, evdr ro U11 rot U12 npcilto oXqpror((ouv 6vo oriprnLor<o ru 6nero 6pXowot 1-to(i o' enorpq pe ng 5' ror 3' 06oerg oupporpfg yto rrlv 6vop(r1 tqq ene(ep- yoo'(og rrov eloooSvcov twpov[tov. Aw6 6tootpotr[(er 6rt or 06oerg ouppo<prjg oro eldooovo twp5vto ovoyvcop[- (owor rout6lpovo on6 to prqlovtopr6 optopo6 twpo- v(ov. En(oqq, 6rot onogefyetot I "otiYxuol, ttov pr4Xovrilv ouppoqrig rord r1 gerdBoorl on6 rov optopt6 efov(rov orov opropr6 rwpov(cov yto rrlv ene[epyoo(o rov prer(6vtov rot etrooo6vtov twpov(ov nou undplouv oro (6ro yovbro. 2 l .l O H luppocpri Tou pre-mRNA llr0ov6v 'Exer Korv6 Mnxovrop6 U€ T' AuToKoToAuTtKd lvrp6vro Tnq OU66oq ll * To rvrpovro uq 0uo6oq ll err6;tvovrot on6 p6vo roug ond ro RNA peoo evdg ougpovrog ourororotrurtrlq ouppoqnq. * 0r ouu0otr6q oupporprlq Kot o pnx0vtou6g ouppoqqg rov tv- rpov[rov rnq opodoq ll eivot nopo;toto Ue Trl ouppotptl Tov ru- pqvrr6v rvrpovlrov. e 'Evo tvrpovto rqg ogoDoq ll Stntrdrverot oe gto deureporoytl 6oprl nou dlUroupyei 6vo rorotrurtxo r6vrpo ro onolo Oupi(et q 6oUrt rou nupqvtrou tvrpov[ou U6-U2. To rwp6vto rcov yovt6[cov nou rtodtronotofv nporct- veg (orr1v npoyl.tottr6tnro, 6Lcov ttov yovt6[tov erc6q rov Truprlvrrrirv yovtS[tov TIou rodtronoto0v IRNA) droxp(vowor oe rpetq yevrxSg r<otrlyop(eg. To twp6vto rov nupnvrKdrv pre-mRNA ovoyvop((owot g6vo on6 rnv nopouo(o ttov dtvourLsott6(trlv GU...AG mo 5' ror 3' dr<po Kot rnq 06onq 6roxld6<ooqg/ouororXioq nup4.u6rvrirv rowd mo 3' dxpo. Aev 61ouv row61+ porrrlprorxd 6ewepotoyoUq 6oUiq. Aw(Oeto, tq u- rp6vro qg oprdSog I xot qg oprddog ll nou Bp(orowcu oe opyov[dro xor Borcrrjpro (rwp6vto otrrddog I undg louv en(or1g mov nupf vo ptovorUrropo:v/otrtYorfrre pov euKoputotc0v) to[tvoproUwot oigqrovo Ue rnv eororeptrrl opydvtoo4 toug. Ko05vq 6tnl,c0vetot oE 1.uo tunrxq 6eurepotoyrj 6oprr1. To rwp6vto oprd6og I rot oprd6og ll 6Xouv rrlv ofie oqpre(coq trov6rlto vo err5gvowot on6 p6vo roug on6 ro RNA. Aut6 xoLefuot ouroouppog{ (autosplie ing rl self splicing). To twp6vto optd6og I efuot nto xor- vd on6 ro rwp5vto oprd6og ll. Yndplet ptrpri oX6orl peto[( rcov 6tio rd[ecov, oMd oe r<dOe nep(moorl ro RNA npoyproronore[ to (6to qv ow(6pooq ouppoqrlq in vitro, Xrrlp(q vo Xpetd(etor ev(upru<69 evepy6rqteg nou nop6lowor on6 npcorefveg. 0o16oo, e(vot oXe66v B6- Boro 6rt ot npoteiVeg e(vot onopofuqteq in vrVo roOtirg Borl0oriv oro 6(nLopro (B).6ne Keq0roto 23). H EIK0NA 21.15 defivet 6tt to twp6vto rot tov tpttil rd(ecov emdgvowot pr6oo 6uo StoboXtt<triv trans-eore- ponorrloecov (6ntoq ebopre npo4youpr6voq Yto ro nuprF vrrd rwp6vro ornv Etr6vo 21.6). Inlv nprirrl owL 6poon, n 5' ouUpolri e[ovfou-twpov[ou 66Xerot en(Oe- oq on5 puo eLerl0eprl oprddo udpo(uL(ou (nop6Xelot on6 puo eocoreptq 06oq 2'-OH oro nupnvtKd twp6vto xor ro rwp6vro oprd6og ll, f on6 6vo eLeri0epo vourle- ot(6ro youov(vr1g oro twp6vto opd6og l). Itrl 6ewep4 ow(6pooq, ro eLe0Oepo 3'-OH oro drpo rou one),eu 0epcopr6vou e[ov(ou enm(Oetot or4v 3' oupBolrl rwpov{- ou-e(ov(ou. Yndplouv o1.tot6tr1teg preto[ri tov twpov(ov rqg oprd6oq ll rot rqq ouppogtig rou pre-mRNA. To pme Xov6pror<d twp6vto oprddog ll err6gvowot p6oo rou (6rou 1-nlXovtopro6 pe ro nupqvtrd pre-mRNA dtoptSoou ev6g trdoou nou ouyKpore(tot 1.t' Svov 6eop6 2'-5'. 'Orov 6vo pregovtopr6vo RNA opd6oq ll enod(erot ln vtf- ro ltop(q entnp6o0eto ouoronrd, prnope[ vo npoYpcnG notrloer qv ow(6pooq ouppogrlg. Aw6 oqpro[vet 6rt ot 6uo owrdpdoetq trans-eorepono(qoqg nou go(vowor orrlv Ex6vo 21.15 npoypqtonototiwot on6 rrlv 6to rqv oM4l,oufo RNA tou twpov(ou opr66og ll. O optOgoq rrov Etootpo6teoreptrciv deqrtirv ouwqpefuot ornv owi- dpoorl rr 6tot 6ev onome(tot e[roteptni nopoXq ev6p yerog. Aur6 0o prnopo[oe vo e(vot oqpovttr6 XopoKrnprortr6 orqv e[#rt[q trlq ouppo<pr1g. 'Evo rwp6vto ogd6og ll onor,cd 6ewepotoyq 6o94 nou nepdler opr<et5g neprol6q ot ono(eg oXq;rot(ov- ror pr5oro (euyoptop6vov ;t(oXtov xot l.tov6xtrtovtov 0q- l,u6v. H neproXq 5 6toXrr:p(etot on6 56o Bdoetg on6 rrlv neprol4 6, 11 ono(o nept6Xet 6vo rotdtrotno A nou 6olp((et qv oprddo 2'-OH yto tqv npdnrl trans-eorepo- 598 KEOAAAI0 21 Iuppotpn rt Ene[epyooio rou RNA Boorxcg eworcg
rov o opLep6q rov 6uvrlnKov unoorpoprdtcov), or npo- re'iveg 6nor(ov 6Lo r<ot Trro orlpowrK6 p6Lo, 21.11 H luppogli eivor XpovrKd Ko! Aetroupyrrd Eu(euypdyn p€ tloAAd Bfiporo ornv'Ergpoon Tov fovr6iov o IuppoQrl gnope[ vo npoyporonoul0ef rord rr1 dtdprero u1g petoypoqrlg d pero q peroypoqrl. . 0t unxovrouo(qg peroyporpfig ror qq oupporpnq e[vor guoxd xor treuoupyrd evonorrlp6vor. o H ouppoQtl ouvd6eror ge qv e[oytrlyrl tou mRNA Kor rov dtreyxo rnq moOepdqrog. r H ouppoQn orov nup4vo pnope( vo enqpedoer q perdqpoorl rou mRNA oro xurrop6n),oopo. An6 xop6 ovoyvrop[orqre 6tt q ouppo<prj rou pre-mR- NA ouprBo[ver row6lpovo Ue rn lreroypo<pri, nop6rr or 6uo owt6p6oerg prnope( vcr Trpoypqronorrl0o0v [elcopr- oro, in vitro ru 5Xouv peLerrlOe( og (eXo:ptorSg 6repyo- o(eg crrqv 5r<gpooq r<ov yovr6(cov. Kripteg nepopcnu<69 ev6ef(erg nou unoornp(ouv our6 to yeyov6g npoq].- Oqv on6 rrq flopcnnp{oetg 6tr notrtrd oupBdwo ouppq- gqg oLoxLqprivovror nprv rrlv o).orl,rlptr:on rnq peroypogrlq. l-evrrd, ro rwp6vro nou Bp(orowfl xowd oro 5' dxpo rou yovr6(ou oEorporiwor rord q Srdp- Kero rrlq peroypoqrlg, evd: to rwp6vro nou Bp(oxowor xowd oto 16l,o9 rou yovrS(ou ene[epyd(owot xord rr'; drdprero rrlg preroypoqrlg 11 r<or prerd q pretoypogrj. Ercr6g on6 q Xpovxri o6(eu[r1 prero(ri peroypotp4g xot ouppogrjg, nt0ov6v undplouv rt dll.or L6yor yro rq tremoupyu<q oUvdeorl outcbv rov Boou<tilv Stepyooui:v. 'Owoq, ot prlXovSg yro rnv 5'- rdAuQrl, qv oqo(peoq rwpovirov, or<6pro Kor rnv noluo6evuL(toorl oro 3' drpo (BL6ns Ev6trya 21.16) ep<pov((ouv rpuotr5q oMrl).ent- dpdoerg Ue rrlv r<evrpu<r1 pnxovn r4g peroyporprlq. 'Evog r<ow6g prqXovtotrr6g efuor vq Xprlorponororiv 14 pe- yd).q C-reLu<q neptoplg qg peyotrrjrepqg unopovd6og rqg Pol Il (yvoorrl og CTD) rog 06o4 yro ro <p6prco1-ro 6rdgopulv nopoy6wrov ene(epyoo[og tou RNA, nop6rr orrg neptoo6repeg neptmdloerg 6ev 6Xet ro0optore( ov r; np6odeorl e(vor d;ieoq 11 dropeool,oBekor on6 opr- opr6voug rorvo6g npcoreivrxoiq i RNA nopdyoweg (BL6ne Ev6trya 20.8). Mro r6roro rpuou<rl evono(qor1 droo<potr(er rrlv ono- tel,eoprotxrl ovoyvcirproq ovo6u6pevov o4 prdttov oup poeiq yro ro (euydptopo yetrovrrciv l.enoupyrxri:v 06oeov ouppo<p{q xord 14 ;reroypo<ptj, 6torqp6wog 6ro1 1.uo o6prl oepd ouppog4g oe rore60uvoq 5'npog 3'. H ovoyvcOprorl rov ovo6u5pevov oqprdrcov ouppo- tp4g on6 roug nopdyoweg ene[epyoo[og rou RNA ror ro 5v(upro nou ouv66owqr pe ro oriprntroro enrpqruv- orlg Pol ll enup6nouv en(or1g oe ourorig roug nopdyov- req v' owoyovroroUv onoreLeoporxd dMeg prq er6xdg nponelVeg np6odeo4g oro RNA, 6nog or npoter'veg h- nRNP, nou Bp(or<owor nowof orov nuprlvo yro rrl ou- oreuqo(o rou RNA. Av 4 ouppoEri tou RNA encogel,e[ror on6 q pero- ypoqn, ytot( vo prrl ou;rBower ror to ow(orpogo; l-1pdy- 1.rorr, ou(ov5;reveg ev6e([erg unomrlp((ouv 6rr rdrr r6roro 6wtog ouprBoiver. 'Oncog nopouord(eror qrnv EI- K0NA 21.17, ro 6v(upro qg 5' rdlur,l.rrlq rpo(veror 6l Boq- OoUv vo [enepome( n opXrxri UeroypoerKq nofor1 : .',,:. ... I Iu(eu[1 rrlq peroypotprlq [0 r-. ovridpoorl 5' Kotrurflq. H UeroypoQn p5oo q-q P: ll opXrro oroporo rovro oqv oqerqplo Tlq Usrc- ypoqrlq. H youovutro-rpovoqepoorl (GT) rot q - pe0utrorpovorpepool (MT) enrorporeuovror o : o0pntroro rqq Pol ll yro vo rorotruoouv q 61proi"-- yio luog xotrunrpog oro 5' oxpo rou veooXlpori.:- pevou peroypotpou onou npood6verot 6vo etDii: oupntroro. Aur6g or ovrr6pooerq enupenouv o:-. Pol ll vo nepooer oe nopoyr,lyrrl enrprlruvol, 600 KE0MA|O 21 lupporp1 rr Ene[epyoo(o rou RNA Boorxeg 6wong
|:rppo,p,l I H nporeivn nPoobSvetoL { oto ouuntroro oupPoQnq .*+F*".,:, ---- i- I H nPulteivrl noPogdvet on1 t ouUgoIn e[ov[ou-e[ov[ou .....,"..,,' ; To oripnhoxo (EJC) * ryvryl*qv:lll , RNA ot oilofeq epnL6rowot ornv eEqYoY( tou mRNA l-lporpov6q, outo( or ;rllovtopro( ;rnopeivo eveplofv or vepyortrd npodyowoq qv e[oytrlyn tou mRNA nou npodpleto, on6 rr1 peroypoqrl rot tr1 oUylpovn Fetn petoVplqri depyoolo rrlq ouppoqriq' Aw6 forr:q [erv6 vop(q r<otd rn UeroYpoqrl. To oign)'oro rp6odeoqq orqv rol0mpo CBP20/80 go(vetot 6r npoo6Svetot dge- oo orn pnxovn e[oytr:yr1g mRNA (ori;rntroro TREX) r<otd tp6no'nou'e[optoror on6 rq ouppoqq cilsre v' orporpS- oer to npdlro twp6vto xowd sto 5' dr<po yto vo 6euro- l.rjvet rqv e(oyoyrl rou mRNA. 'Evog Bootr6q nopd- yowoq t o, 6,opeooloBefuot orqv e(oycoyrl tou mRNA e[vot o REE (orrf (6Pn rotrefuot Aly, Yralp), o ono(og ono- reLe( pSpog tou EJC t<ot oMrllenrdpd dge.oo Ue rnv nprtdtun TAP (orq (6Un, Mex67p) nou peto<p6petto mR- NA 6nog rpofuetot orqv EIK0NA 21.19' To otiprntroro EJC 61et 5vo np6o0eto p6Lo oq ouvo- 669 rou mRNA ercr6g tou nup(vo, YeYov6q ;re preydtro o*(,.rrr,o orrl oro0ep6tnro rou mRNA oro rurrop6- ntroqto. Aut6 ouprBo(vet enet6q 5vo EJC nou SXet ouy- r<por4oer optoptSvo ov6prol,o mRNA. prnope( vo enrorporcUoet 6Moug nopdyoweq ot ono[ot npodyouv rrlv o<po(peo4 rnq npoororeutrrlg r<oLrimpoq on6 ro 5' 6xpo rou mRNA pe q 6pdoq ev(0prov onoxdLu$r1g' 'Onog qo(vetot ornv EIK0NA 21.20,to EJC rovovwd ono- 1ro^p,irttot on6 to gerorloUgevo ptBooto;r6tto rotd u1 btdpr<eto tou npcitou r0r<trou gerd<ppooqg .oto rurrop6- ntroopto. Qor6oo, ov o' 5vo ene(epyoopr6vo mRNA et- orn ou$oln e(ov[ou-eqov[ou I I o EJC npoo66ver npoteiveg nou epntr6rovroL ornv eqoYurYn' evrontol, onoddpnon rou RNA : To EJC (oupntroro ouUpotrnq e(ovitlv) evonorl- :,-r r0VT0 orn ouu0otrn ouppoQrlq olq ouveneto rnq ovriopoolq ,: litlq, . : , .l orov unor<wr1rr1. Ot nopdyoweg ouppo<pr1q 9o(- . :. ort no((ouv rdnoto p6l'o oio vo 6teurol'Uvouv -- . inL;.rrlruvon rnq ptetoyporprlq Kot o oxrlUottopr6g -, 3 oxpou tou mRNA e[vot ooqrog ro0optmtr<69 yto - -. :epUorLop6 trlg pretoypo<pqq (BL6ne Ev6trlta -- -5r Iuvenc,:q, 11 petoypoq4 r<ot 11 ene(epyoo(o tou :',: e[vot e(orpettxd ouwovtopdveg oro r<[rropo rov - :'...,r <urropolv euxoputotrov. - ene[epyoo(o tou RNA ouv66etot ]'ettoupytxd 6Xt --,3 ue rlovo6trd preroypoqtr<d ouprBdwo, oMd eni ---: Jt ro0o6tr6 Bqpoto, 6n<oq q e[oycoyrl tou mRNA . : : eleyXoq qq oro0ep6qtog. An6 rotp6 yvorp((ou- -. .., .o RNA nou 6Xet unocne( otelq ene[epyoo(o t<t .1:<oou0e( vo neptdXet oprop5vo twp6vto 5e ;rnope( : "!o10ei onoteLeopottxd, yeyov6q nou (otog ocpeil'e- -: :ro cpotv6pevo KoroKpftnongtou otollott6lou oup- :::qg 96oo mov nupqvo. H eu66oor1 rqq e[oytoyqq -, - "nRNA on6 q oupporprl ovodetrv0etot pe tqv 6ve- r'- trrov nuprlvo RNA Xop(q twp6vto nou npo6pXetot --: oDNA n pre-mRNA nou 0' ono6oloet tow6orlpio : "q xotd q ouppotprl' To RNA nou 6xet nepdoet pr5o<l _-: 5Lo6troo(og qg ouppo<pqg e(dyetot nto onotee- - -:ttrco on6 to RNA nou npo6pXetot on6 to cDNA, ye- :.cq rou unodrlltovet 6rt q dtepyoo(o trlg ouppoqqq ::-0o r1v e[oYtoYq rou mRNA. Onrr.lg <powetot orrlv :.:".1r:ri ':-r ':, ndvo'l orrl oupBoLq . : lviou-eEoviou evonor(Oerot 6vo et6tr6 o0prnLoro, yvol- r : oq oripnhoxo oupBoA{q e[tovitov (exon iunction :omplex, EJC). Aw6 to otiptnoro qo[verot 6tt entorpo- -.-.i dp,.oo opto;rdveg nporelVeg nou npoo66vowot fio fl I KyTTApon^ArMA I @ l#r#lX.**, Mto npotreivrl REF npoo66verol o' dvov nopoyov- ro ouppoQrlq rot nopol.tdvet Uo(i Ue ro ouppoltp6vo npoiov RNA H REF npoo06veroL o' tivov nopoyovro eEoytoVrtq nou npoo6eve- T0r 0T0v nuplvtro nopo. H rporsivn REF (Aly) e(vor p6pog rou EJC 0 nopdyowoq Uerogopdq TAP/Mex npood6vetor cqv REF tou nupqvtro! ndPou ?,t :t H lupporpn e[vot Xpovu<o rot Aerroupytro lu(euypdvrl ue l-lotrtro Brlporo orqv'Errppoon roi' [:' i *' 601 E -&ilvto lvrPdvto EEOvto,
Anetreu06poor1 prpootogor[ou ond ro npoopo xrodrrovro repgolo;rorj EJC ; Entmpdreuorl ev(upou ogo(peor1 + Trlq Kotrt1flTpoq Ano66pr1or1 rou mRNA EI(0NA 21.20 To o[UntroKo EJC ouvd6erq ouppogn Ue Tnv NMD. To EJC enrmpoTeuer ettionq rporefveq Upr ov nopoge(ver oro s[o- ydpevo mRNA. Merd qv eqoyoyn ono rov rupnvo, ro EJC np6ner v' onogorpuvOef U6oo Tou UeToKrvougevou prpootogor(ou orov flpdlro yupo qg perorppoqq. Av 6v0 EJC nopogeiveL mo mRNA trdyo rqg nopouo(og evdg npdopou rtodrxovfou repponoporj, ro onofo onetreuOep6ver ro prpoorrlpdlo, ro EJC 0o enrorporeUoer enmpdoOereg npoTetveq, rt.X. rr1v Upf, or onofeg pe q oepo toug 0o enrorporet1oouv ro 6v(upo ogoipeoqg qg xotrunrpoq (DCp). Aurd endyeL qv onopdrpuvoq qq xotrunrpog mo 5' drpo xor n1v onoddprpoq rou mRNA oe roreu0uvorl 5'-3' mo runop6doogo. oo10e( 6vo np6opo rrodx6vro repporroproU e[or(og oq- perorqg perdMo[4q 11 evoMorcnruiq ouppogrlg (BI6ne Ev6tryta 21.12), to prBoo'<o;rdrro 0' onoonoqre( on6ro mRNA erpoye(o nporof <prdoer oro xovovx6 rto6rr6vro repporropro0, ro ono[o ouvrl0og Bp(oreror oro ts^.eu- rofo e(c0vro. H o6uvopr(o rou prBoo<o1.ror[ou v' onoB6.tret ro ofpnloxo EJC nou Bp(oxeror gerd ro np5opo xo6r- x6vto repporroproU enrp6ner vo enrorporeu0orjv 6v(upro nou orpoporiv rqv xoLUmpo, endyowog roXe(o ono66- pnon rou mRNA. Aurn n 6repyoo(o rolefuor g0op6 rou mRNA 6ropeootropo0pev4 on6 oouv0enx6q pl6Beg (nonsense-mediated mRNA decay, NMD). Awrnpooro- ne0er dvo prqlovrop6 enmrlprlorlg rou mRNA o ono(og onorpdna q perdrppooq roloBtirv npu:reivdrv on6 mR- NA nou rp6per rivo np6trrpo rodx6vro tepprorroprori. H NMD 0o nopouoroore( tremoprepc0g atlv Evotrpa 22.g. 21.12 H EvoAAomtKr{ Euppoeri Elvor Kov6voq, Oxr E[oipeon, oTouq lloAur0rropouq EuKopu6Teq * Erdrro e[tirvro 11 otrtrr1trouX(eg e[trlv(ov gnope[ v' onoxtrs . orouv r1 vo ougnepilrltp0ouv oro mRNA npoiovro Xprlorpot .: - tilvrog evotrtrorlreq 0eoerg ouppoQ1q * H evoLl,orlrl ouppoqn ouverorp6pet oq dopxrl rqr ),euo::- yrrrl norrr),[o rov npoiovrov rov yovr6(rov. s 0 xoOoprogdg rou tprltrou oq Drosophila nepd,oppdvaL ;.: oepo oulrBovrov evotrtrorrtxrlg oupporpqg oe yovldro nou r:- 6uonorouv dLodoXrro npoiovro proq odo0. Orov 6vo 6Lorer<oprprSvo yov(6to ;retoypoge( o' dvo F- NA nou ono6(deL 5vo provo6u<6 tUno ouppoll;rdvou r- RNA, 6ev undpler oodgero orrlv ovoyvtiproq rc, e[,orv(ov Kol rov rwpov(rov. To RNA rov neproodrepa. yovr6(ov, toor6oo, or<olou0oUv np6runo evoA,lorrr. x{g oupporp{q (alternative spticing), q ono(o oup_rBc.. ver 6rov 6vo ;rovo6u<6 yov(6ro ono6(deL neptoodrepe: on6 pLo otri.ql.ou1(eg mRNA. Me ng tey3rLr5g rnq cD: x).rovono(qoqg peydqg xi.(proxog Kor rnq oMq).oLJX- oq, Syrve oogdg 6tr ndvo on6 ro g0% trov yovr6il-, nou erqpd(owor oro O4looru<d ouppdmowor evci.- Aornr<d. Iuvenog, 11 evoMornrrl oupporp4 6ev eii,c. on).c69 ro onotS,l.eo;ro LoOri:v nou npoyporoflora - Unxovn ouppo<pr1q oMd pr6pog rou npoypd;r[-loroQ gr- epoolq rov yovr6(ov nou o64ye( oe noMon).d yo,;. 6rord npoi6wo ond rov (6ro yeveru<6 r6no. YndpXouv drd<popot rUnor evoMor<nxr1g ouppogr : peto(0 d).).rr;v 11 ouyr<pdrr1o4 rwpovirrtv (intron rete^- tion), r; enil,oyq evotr,orcrtrrrlv 5' 06oeotv ouppogrlc * 6wo(q r1 nopdLerrpq e(olv(rov (exon inclusion or sk c- ping) ror 4 oporBo(o onor<).er6gew1 enil.oyr1 totv evoMc. t<ttt<tr.lv e[olv(ov t::]i;...,:;,i :r r ',, To (6to npoToyeve i gerdyporpo ;rnope( vo unoore( nepLoo6tepoug ond dvc, rUnoug evoMorcrrr<r1g ouppoqrlg. T' o;rotBo(o onox,,e.. 6pevo e(trrvro r<ovovrrd pu0pr((owoL ge roro-er6rx6 rpc- no. Meprrdg eop5q, oe ourrlv rqv nolunLor6rq:: npoor(Oetor to refur6 np6runo 5r<ppoorlg TroU KorcUgc.- vetoL on6 rn Xpnon 6rogopetrrcov orlpre(rov 5vop(qg r: geroypogr;g 11 qv nopoyrrtyq evotr),orcru<ov 3', dr<po.lv H evoMomLrq ouppo<pr1 prnope( vo enrlpedoer r", 6r<rppoorl rov yovr6(otv pre 6Uo rou).d1rorov rp6noug, C Svog e(voL q 6r1;rroupy(o 6optrrlg norxL).(og oro npoic,- ro rov yovr6(o.lv oupnepil.oprBdvowog r1 nopotre(no.- rog optop6veg rodrr<onorqrrr<d,g o).).r1Lou1(eq - 6lprroupyrrtwog evoLLomrxd nLo(oro ovdyvororlg v,: 5vo rprrlpo rou yovr6(ou. Aur6 ouXvd rpononore( q ),e.- roupyrxq rdr6qto trlv r<rr.ldrronoroUpevov nporeivo, FLo nopd6etypo, ro yov(6ro CaMKlld nepr6,1er rp(o eE.- vro nou ouppdnrovtot evq^.^.oKtLxd, 6nrog <po(ver: oTllv rr ':i.l .i ,l- ,.,'. TO yOv(6tO er<rppd(erOt oe 6).Oug o1:. cap Ptpooopdlo EJC I cap 602 KEOA AI0 21 Iupporpn rr Ene{epyooio rou RNA llprirtog y0pog gerdqpoolg Boolxeq evvoreg EJC I
KorqKpoTnorl rvTpov(ov Evotrtrorrudg 3' 06oeLg ouppoQnq 'Evroqr/ropdtrerq,1 e[tovltov :lK0NA 21,21 Arorpoperlro(rUror rvotrtroKTrKr1g ouppotpqg. 1:.' roug xurtoprro[g r0noug r<ot toro[g rrrlv Oq].oon- . *, Av nopo).er<p0otiv Kor ro rp(o evoMorrrr<d e[utvro, -: rRNA xro6xonore[ Uro Kurropon].ool-torrrrl rrvdorl -:, eooeopulr<r.lveL nol d nporeivtrd unoorpo.lproro. 3 :,,, oupnepd.oprBdveror ro eqovro 14, q r<rvdorl prero- :.:.ror orov nuprlvo eneL6q ro e(utvro 14 neptdlet 6vo l-*o nupnvrKoU ewonroproU. Aw6 ennp5ner orqv rrvd- - .: pu0p((el rrl peroypotprl orov nuprlvo. Otov ou1-r- -.:.,ogBovowor Kor to eforvro 15 r<or 16, r<drt nou - - ,r0cog ouprBo(ver oroug veupoveg, q r<rvdon Koreu- i, , i:or orrlv Kurroprrq pepBpdvq dnou enqped(er er6r' .:-c Sro0Loug r6wrov. Ie d).i.eg neptnriooerg, ro npoi6wo evoMoxrrr<r1g :-:cog(q 6,1ouv ow(Oereg ).etroupy[eg. Aur6 toX[er :-:.omtrd yro 6).o ro yov(6ro nou epnlSrowor orn :-:-Lot1 rqg on6mooqg. Kdoe yov(6ro er<<ppd(et tou- ' l,..croV 60o roopropg6q, pto nou npodyer rrlv on6- *ol.l Kor. pto dMq nou npoorore6er ro rdrropo ond ''-::roJorl. Oeropefuor 6l 11 ovoLoy(o rrrtv toopop<pritv -, . rv rov puopumo.rv qg on6mioorlg xooopi(er ov 0o . -.3r:rroeL to r<Urropo. -'J evoMorcrrxrl ouppogn prnope( en(oqg vo enrlped- ::. Srdcpopeg rdL6-rrlreg rou mRNA ouprnepil.opBdvov- -:: q nopoLe(nowog optopdvo puOprmrrd orotle[o -:- RNA ro ono(o prnope( vo pretoBd).).ouv no).U ro , ::vo q;-u(orqg rou mRNA. Ie noM6g neptmoroerg, xri- prog oron6g qg evoMor,rx{g ouppogrlq (ocog e(vor vo e[ovoyrdoer 6vo ouyrerprp6vo noooor6 rov nporoye- vriv preroyp6gov vo gdpouv 6vo np6opo xro6u<6vro 11 rcodrr6vro tepgortoprori Sror rilore ourd ro gerdypogo v' ono6oprlOoriv yprlyopo. Aw6 nr0qv6v owrnpooo- neUer pro evotrl,or,rrrrl orporqyu<r1 preroypogrrqq p6e- Utonq yro rov 6).eyXo qg orpOov(og erdrrciv mRNA oro rUrropo. O ouyxexprl.uivog prqlovrog6g Xpqorpronoe(- rqt yto vo enneulOe( oporoororu<r1 5r<gpoorl yro nol- ----*-":iffirg:-T' 6A ryryffi + rrdxsuon orn ueuopdvn dB*M@,r 6c wffi Evotrtrorlrrl ouppotpn rou yovrdlou CalVlKll: 0Lo- qopertro evotrl,orlro e[tlvto roreu0uvouv Trlv rLvool oe 6rorpo- penro rurroprro Dropep(oporo. ----+- IT6xeuorl orov nupqvo + Evr6nro4 oro xunopdn),oogro ?1.J4 H Evotrtrorrtrl Iuppoqn E(vot Kovovog,'OXr E(o(peo1 oroug [lotruxuncOC-a !-,::,--.: 603 17- Agorpo(o onoxtreuigevo e[6vro Evotrtrorlrdg 5' 06oerg ouppoqrlg Iuv6uoolrrl eniloyn eqoviov Evotrtrorlxoq unouvnrnq/ouppoQn Evotrtroxrrxrl noluoDevutr(oon/oupporprl
Lot3g pu0pror6q qq ouppoqrlq oe erdtxorig Kurropr- rotig dnoug rj tororig. Ie pto t6toto priO;uoq, 6vog er- 6tr6g 0eru<6q pu0prmqq rnq ouppoqrig ;rnope( vo enrlped(er rr1 6u<r1 rou evoMorrxrl oupporp{, o6qyc0v- roq o' 6wo[q ev6g e[trlv(ou nou nepr6Xet 6vo np6opo ro6x6vro rcpprorroprori. Aur6 droXere6er 6vo x).dopo rou mRNA rou npoq ono66gr1or1, per6vowog Sror 14 ouyx6wpoorl rcov nporeivc0v. Iuvenrig, 6tov n ouy- rSwpcoorl ev6g r6rotou Oenro0 puO;uord rrlq ouppo- rp4g ou(operdrveror;16oo oro rfrropo, q ouyr<6wptrloq rou ow(ororlou mRNA 0o orpo<pe( npoq rrlv ow(Oer1 roteU0uvoq. Motrov6l SXouv Xoporrrlprme( noAAd ouprBdwo evoMor<ru<r1g ouppocpr]g rr dXouv xo0oprore( or BtoLo- yrxo( p6l,or rov npoi6wcov evo.trl,ornr<r1q ouppo<p4q, ro nro Kqrovonr6 nopdderypro nopopr6ver 11 od6g roOopt- ol.rori tou <prJLou orn D. melanogaster, nou neprlopiBd- ver oMqLenrdpdoerg peto[ti yovrd(cov 6nou ouprBdwo evoMorcrxrlg ouppocprlg 6rorp[vouv r' opoevxd on6 rq 0ql,ur<d. H o669 no(pver rn Uopgn nou nqpouord(eror m4v EIKONA 21.23, 6nou q ovooy(o rcov X Xproprooolprd- rov oq npoq ro ouroocopxd Xprr:l.roodrpoto roOop(er qv 6r<<ppooq rou yovr6(ou sex lethal(sx/) r<or or oMo- ydg orqv 6r<gpoorl nepvoriv 6ro6oXrd g6oco rrov dM<ov yovr6(tov mo yov(6ro doublesex (dsx), to retreuro(o orqv o66. H o669 [er<rvd ;re gu].o-er6xr] oupporp{ rou sx/. To e[cbvto 3 rou yovr6(ou sx/ nepr6ler 6vo rco6rx6vro rep- prottoprof nou onorp6ner rrl o(v0eor1 l,erroupyrxrlq Avotroy(o X:A: 2:2 Ot- I * Irafira-Z Apvlrrdg pug;ttonlq rov yovrdltov n1q opoevtxflg 6torpoponoqoqg Apvlrr6g pu0Urornq rov yovrd[tov qg 01truxr1g 6rorpoponoirloqq / n( I Sxl off I Tra (xotroBo) ftKSl;P.f1.?3 0ro0opro;rogrouqUtrouorrlD.melanogasternepiloppdverprooddorrlvono(odrotpopeurdouppdvro0uppoQrc :- -:- ronorouvror oro 0qtruro. Kotrupporo oe onoloDqnore oroDro orrlg o6ou odrlyouv oe ovonru(r1 opoevuou. Errov((eror | 0upp0.- -- -, pre-mRNA nou puOp((eror ono qv nptorelvl Sxl, 11 onolo onortre(er Trl Xplol qg evotrtrontrlg 3' 0eo1q ouppqqlq, Korl 0uppc:- -- - : sx pre-mRNA nou puOpl(eror ono ng nporelveg Tra ror Tra2 oe ouv6uooiro pe dtrtreg npotrelveg SR, or onoreg enlped(ouv 0elL.: , - ro[1 rou evo],trorlrou e[ovrou. 604 KEOAAAI0 21 luppoqrl rt Ene(epyoo[o rou RNA rj ) Y ? Dsx npioteiVqg. Aut6 ro e(trrvto ouprnepil.oprBdvercL -: - RNA nou nopdyerot or' opoevrKd, evc,r nopo.).i.-:-- oro 0r1trur<d. 'Eror, p6vo ro Oqlur<d nopdyouv .*, -: -- refur1 Sx/. H nporetvq neprdler d<pOovo Boou<d c* . : l. : rot on6 ouq qv dnoqtrrl protd(er pre dMeg r:c* 1,:: nou npoo65vowot oro RNA. H nopouo(o qg npore[vqg Sxl oMo(er m ;-:: :: - rou yovr6(ou transformer (tra). N Etr6vo 21 2:3 1. , . 6tr our6 neprl.oprBdvet ouppo<pr1 prog oroOecr: : i= onq o' evoMorcnr<69 3' O6oetg (orlpreuoore o:L :---: : rp6nog e<poppr6(eror orn oupporprl t6oo ro- s, : r- Korrou tra).To (6ro np6runo oupporpqg roXJeL:::- - t' opoevrxd 6oo roL yro ro Or1).uxd r<or ono6i53. ..: : NA pr' 5vo np6orpo r<ro6u<dvro tepporrol.rori, H n::: - - : rqg npo.rre'lvr1g Sxl ovoord].,trer rrl Xpnorponorrl- -: ovodu<qg 3'06oqg oupporp4g pr6oor rqq noo:Ii- : rnq orn ouororl(o nuprl-udrvtov or1 05oq 6Lor).:l * - : tqg. Av nopo).eL<p0e[ouq q 05oq, 0o XpqoLl-ron: -:: - en6prevq 3' 0do1. Aur6 nopdyer 6vo mRNA e.1.,: : ro 0qLurd nou xro6rxonoe( ;lto npu:re'lvq. Iuvencog, 11 Sxl puOp((er r1v (6ro rrl oUQCCi- -. yto vo 6toorpoL[oer rqv 5xrppooq rnq oro 0r1],u < r . : - - tra nopdyet pto npo:te[vr1 pr6vo mo 01].uxc 3--: - Sxl, 11 nportelVq Tra elvot 6vog puOprorqg rrQ ]-::: enq. To tra2 6,Xer nop6proto Lenoupy(o fio 8.'.-. : - ).d er<<ppd(eror en(or1g Kor or'opoevrxd) , OL .:- . .: Tra rot Tra2 e(vor nopdyoweg ouppogqq SF -:- .-. vepyotiv dpreoo mo prerdyporpo-mdloug O. -'= .- Tra2 ouvepyd(owor (mo O1Lur<o) enrlpeolc,::: -
r-opoeq rou dsx. Ito yov(6to dsx, to Olluxd ouppd- -3.rv rrlv 5'05or1 tou twpov(ou 3 pe qv 3' 05or1 ou- -:- rou rwpov(ou. 'Etot, 11 pretdtppoorl teppror((etot : o rel.oq tou e(orv(ou 4. T' opoevtxd ouppdmouv qv : -:eoq rou rwpov(ou 3 r<oteuOe(ov m4v 3' Odoq tou tv- --:,,'iou 4, nopotre(nowog dtot to e[iovto 4 on6 to mR- ',: Kr 0nrrp6novrog vq ouvexrote( 11 pietogpootl I :leoou rou eforv[ou 6. To onot6].eopo rlq evoMo- - .(ng ouppocp4g e(vor 6tt 6totpopettr6g npolre[veg Dsx - : <iOe cp0lo nopdyowor.: ro npoidv ttov opoevtrtr.lv - : : e uno6 [(er rr1 0r1].u xrl <pu].etu<r1 dtotpopono(4 or1, evot -: :oor6v rc'rv 0rlLuro-lv xotoordMet qv dr<rppoorl eL6t- . * . yro ro opoevtx6 yovt6[ulv. 2 t.l 3 H lupporprl Pu0pl(eror on6 EvtoxuTdq ror AnooronnTCq nou BpioKovlot T60o oTo E[6vro Ooo ror oTo lwp6vro . - svotrtrorrtrut oupporpn ouxvo oxer((etoL pe ooOevefg 0eoetg : - cporprlq , l. otrtrntrouxteg nou neptpotrtrouv evotrtrorrlro e[(tvto ouXvo ..,,0r nro ouvrqprlpeveg e[etrtrrtro ono rtg otrtr1trouX(eg nou .e cLpotrtrouv ro rdroouotottro e[titvto. . l,SLxdq otrtrntroux(eq e[trlv[tov rot tvrpov[ov evtoX0ouv r1 ro- -:retr),ouv t1v enLtroyq qg Oeolg ouppotpnq. . ' en[Spoorl rov evroXur6v Kor Tov oTlootonrlrdtv rrlq ouppo- :'q 6LoUeootrope(rot ono et6tr6g yto qv otrtrr1trouX[o nporel- ,:q npoodeonq oro RNA, notrtrdq on6 rtg ono[eg pnopel vo :,3p[(ovior ovonru[roro ro/q vo exqpo(ovrot toro-et0xo. ' - rcxui1ro geroypoq1g eqpeo(eL dpeoo rrlv 6rpoo1 rnq . , ctr,ortrrlq ouppotpflq. - i,,oMorrrxq ouppo<prl yevtr<rilg olet((etot pe ooOe- ,.1 0eoerg ouppogrlg, rdtt nou orlpoivet 6rt to or1- / Uoro ouppoqrig nou ewon[(owot Kot oro 66o 6xpo rrrlv twpov[tov onor.l.(vouv on6 ro ouvowerxd orl;re ro ouppo<prlg. Aw6 entrp6net q pU0gtorl ttrlv oo0e- vc0v orlprdrov ouppog4g on6 6td<popoug trans- Epomrxorig nop6yoweq nou yevtrd e(vot yvooro[ og pu0lttot6g eva&axtur1q ouppaEfig. Aw(Oetq on' 6,tt 0o un60ere xove[q, oo16oo, ot ooOeve(g O5oetq oup- pognq yevxd e(vst nto ouwrlprlprdveq oro yovr6tripo- ro rtrrv OqLoontrciv oe oX6oq pe rtg t6toouorottxdg 06oerq ouppoqrlg. Auq q nopotiprloq owtt(Oetqt rrrlv owil.r1r[r1 6tr 11 evoMot<ctr( ouppoqrl 0o pno- poUoe vo nporUmer on6 Ld0q r;g Unxovnq ouppo- gr]g xor unootrlp((er 6tt noLLd ouptBdvto evoi.- Lorcnr<qg oupporprjg nt0qv6v e(vot e(efurcrtx6 ouwrl- prlpr6vo ciore vo dtoqpqOe( 11 pU0ptoq qg 6rrppo- onq roJV yovr6(trlv mo en(ne6o rqg ene(epyooiog tou RNA. H priOptoq qg evoi.l,orcru<r1g ouppotprlq e(vor nep(- nl,or4 6repyoo(o 11 ono(o nepil.opBdvet preydLo opr0;r6 frans-6poonrcirv puOpumcbv qg ouppogqg nou npoo- 66vowot oro RNA. 'Onrog rpo(vetot firlv EIKONA 21'24, ouro( or pu0puor6g nt0ov6v ovoyvtop((ouv orotXe(o R- NA r5oo oro e[cilvro 6oo rot oto twp6vto rowd or4v evollorcnrf 06o11 ouppoqrlg r<ot qoxo6v Oettrti rot opvqrurl en(6pooq orrlv entl,oyrltrlg evoLLorctrrlq 06- onq ouppognq. Ot pu0gtor6g nou npoodSvowot oe e[r6vro yro vo evtoXtioouv qv enil.oyrl e(vqt Oettro( pu0pumSg rnq ouppoerlq r<ot t' ow(orotlo cis-6poort- rd motXe(o ovoq6powor rog e(ovtro( evtoXutdg qg ouppogqg (exonic splicing enhancers, ESE). Ot npcrt- reiveg SR ovqrouv orouq nto r<oLd lopor,itrlptoprdvouq puOptordg nou npoo66vowot oe ESE. Aw(0eto, dMeg npcorefveg, 6nog or hnRNP A t<ot B, npoo6Svowot oe oMqlouX(eq e[cov(tov yto vo Kffioore^ouv qv enAoyti rcov 06oeolv ouppogrjq. T' ow(motXo cis-dpomtxd morXe(o e(vor yvoud rrlg e[ovtxo( onootonqrSg qg ouppo<priq (exonic splicing silencers, ESS). flopopo(- og, notrLdg np<oreiVeg nou npoodSvowot mo RNA enqped(ouv r4v enr).oyq ttov O5oe<ov ouppoEfq pSoto Anoouonnrnq qq ouppqQrig rov twpov(tov (tss) EvLoyurrlq Anootonlrrlg EvtoXur1g rnq oupp0Qrlq Tns ouppoQnq rnc ouppoqrlq rov e[ovirrrv rov e[ulvttrtv lov tvTpovtov (ESE) (ESS) (lSE) 0L otrtrntrouxLeq rov e[ovlrov xor rotv rvrpov[ov prnopel vo enlpeooouv rrlv eniloyrl qq 06olq ouppoQnq tretroupytirvroq - . .,.qureg 1 onoouonqr6g r1q ouppoQrlq. levrro, or npolrelveg SR npood6vovrot o' evtoXur6g rt1q ouppoQnq rov e(ov[tlv Kot ot npu]- ', ..: rnRNP (n.X. or otroyevereg A xot B rorv nptureivdrv nou npooDevovTor 0T0 RNA) 0e onootonlrdg e[trlvttrtv. Atrtreg nporeiveg nou - - - levovror oro RNA (RBP) treuoupyouv oq puOproreq rnq ouppoQqg peool qg npoobeolq Touq 0' evtoxuT6g 1 onootonlr6q rr1-q ouo- ,--: Toiv rvtpovltov. / ?1.'13 H lupporp1 Pu0p(etor onrl EvroXur6g ror Anooronrltdg nou Bp(orovtot Tooo oro E[6vto'0oo xot mo lvrptivto 605 eworeg
rov rwpovrKc0v oLLrlLouXrc6v. T' ow(ororXo 0errrd ror opvqrrrd cis-6pomrxd ororle[o ro].orlwor twpovtxo[ evtoyutdq rl anooon4r6g qq ouppaEryg @tronic sptic- ing enhancers, lSElintronic splicing sl/encers, /SS), ov- rroro(Xcoq. Ir4v nolunLor6qro outrj dplowor vo npoore- 0otiv or enr6pdoerg noMriv puOpuoroiv rqq ouppogrlg. To nro yvcoord nopo6etyproro e(vor or pu0pum6g qg ouppoqrlg rov orKoyevercriv Nova ror Fox, nou npoo- 66vowor mo RNA rr evrolriouv 11 r<oroordlAouv rnv enAoy4 rrlg 05o49 ouppocprlg, ovdLoyo pre q 06oq np6o6eorlg oe o16oq pe ro evol.Lorirrr6 e(civro. I-ro nopdderypro, 6nog go(veror orrlv E|KONA 21.25,1np6o- 6eor1 rov Nova xor Fox oe oMql,oul(eg rwpov(<ov nou Bp(or<owot ovo6rrd rou evoMorcru<ori e[ov(ou yevrrd odqyei oe roromolq rou elcov(ou, evdr 4 np6o6eori rouq or oMq).ou1(eg rwpov[ov nou Bp(or<owor Kq0o- 6u<d rou evotrl,orcnroU e[rov(ou ouXyd evroXriel rrlv enr- l,oyri rou e[ov(ou. T6oo q Nova 6oo ror 11 Fox ergpd(owor 6rogoperrrd oe 6rogoperrroUg rmoUg, t6(tog orov eyr<5goLo. Enopdvrog, 4 roro-er6u<rl pU0pr- on rnq evoMorrtrqg ouppogrig enmuyldvetor pr6oo qg rmo-erdrrqg drgpoorlq rov trans puOptorcbv rqg ouppog4g. Aev e(vor nLripcrlg rorovo4r6 nrig 5vo erdrrd oupr- Bdv evoMor,mu<qg ouppogqg pu0pr((eror pr6oo 6rdEo- p<ov Oetrxcilv xor opwlru<riv puOpumtiv qg oupporprig. Oeop4rrxd, or pu0ptar6g Lemoupyoriv evrolrlowog q rqtomdLLowoq rnv ovoyvcilproq ardrrrirv oq prdrcov ouppogrig on6 oproprdvo Boorrd ouororu<d rnq UnXo- vqg ouppognq. Or nro Korqvorldg neprmcboerg e(vor or nporeiveg SR xor or nporeiVeg hnRNA A r<or B yto ro 0enx6 xor opvrltrxd p5tro toug or4v ev(o1uon ri xo- toorolli u1g ovoyvrilprorlq rov O5,oeov ouppogqg, ov- norofX<og. H np6obeo4 rov nporeivriv SR oroug ESE npodyer 4 aroOeponore( qv np6odeoq rou U1 mqv 5' 05oq oupporpqg r<ot qg U2AF m4v 3' 06oq ouppo- rprjq. Iuvenrirg, 4 ouvop;roL6yrloq rou oopoud(ou ouppog{q y(vetot nro onore},eoprolxri nopouo(o rcov npoteivcbv SR. Aurdg o p6Log-r<ov npcoreivdrv SR ei- vor orlpowrK6g 16oo yio rnv r6rooumorrxrl 6oo xor yro rrlv evoMorcu<f ouppogri, xo0rmriwog nq npo- reiveg SR ovoyro(oug nopdyovreq ouppogrlg xcr puOprmdg evoMorcrxrlg ouppogrlg. Aw(Oero, or npo- reiveg hnRNP A/B go(veror 6n npoo66vowor o..ro RNA Kor. owoyov(owor qv np6odeorl rov nporeivri:v SR ror dLl.rov Boou<cilv oumotrrc0v rou oorpond(ou oup- pogriq xord -rqv ovoyvc0ptorl ).etrou pyrrcbv orlpdtcov ouppogqg. Or npotetveg SR 61ouv trlv rxov6trlro vo 6e- 4reriouv 6vo pre-mRNA mqv o66 ouppogqg, evri or nporretveq hnRNP owoyrov((owor our4 14 6repyoo[o. Ae6oprdvou 6rr or nporciveg hnRNP e(vor e(orpetrrd dgOoveg orov nup4vo, ncbg ot npotetveg SR owoycovi (owor onoreleopotu<d ng hnRNP orov nupqvo yro vo 6reuroLtivouv rn ouppogrj; I-'lporpovcbg, our6 enmuyXd- veror pr6,o<o ouyxcouoUoti r.1g preroypoqfq Ue rrl oup porp{ ;16oo orov nuprlvo rou rurrdpou (BMne Ev|trlro 21.7). Zuvenrilq, q dtepyoo(o qg ;retoypogrlg nr0qv6v eqped(et r1v evotrl,or,cu<4 ouppocprl, rdrr nou 6wcog rqXUer. H evqMoKru(4 ouppoqri go(veror 6l enqped(e- ror on6 et6roUg unor<rvqr6q nou Xprlorponororiwor yto vo npoo0rloouv rnv dr<rppoorl rrov yovr6(rov 6nog en[- onq Kor on6 trlv roXUr4ro rnq peroypqgrlg ro16 rr1 rpdoq enrprqr<uvonq. Arogopenro( unorrvqrdg prnope[ vo npooe],rUouv 6rogopertrdq opd6eq preroyporpu<civ nopoy6wrov, or ono(or pe n1 oerpd roug gnope( vo enqped(ouv rrlv eTrr- p{r<uvoq rnq Ueroypog4g. EnoprSvog, o (6ro9 Unxovr- opr6g prnope( vo euOriverqr yro rrlv en(6poon rqq enil.oyrig unorrvrpriv Kor rnq roXriqrog enrpuixuvolg rrlq Ueroypoqnq ornv evoMorcrxrl oupporp4. Or 6ro06- orgeg ev6e([erg unoderrv[ouv 6vo xrv4ru<6 prow6]ro oU;rqcr;vo pe ro ono[o n XoUnIf toXUqro ent;rrlxuvoqg rnq peroypoqnq eo noperXe oe Uro oo0evrl 06o11 oup poqrlq ovo6u6pev4 on6 ro oriprnlor<o r4g Pol ll nou enrprlr6veror opxer6 1p5vo yro vo (euyoprioer Ue rrlv ovo6rx4 06oq ouppogqg nporo6 etrrgovrore( pro r<o0o. U' .,r:r' j'.. '- - 0r orroydvereg npotreivdrv npoodl- qq oT0 RNA Nova ror Fox npooyouv 1 roroorer- trouv qv eniloyrl rrlg Oeor1q ouppotprlq ple rpo : ue[oprdrUevo ono ro ouprppo(opevo,,. H npdode:- qg Nova o' e(6vro ror neprpotrtrovro ovodrro .,- rp6vro ovooretrtret rqv evro[q rou evotrtrorrLx:- e[tov[ou, ev6 1 npoodeol qg oe neprpotrtrouc:: xo0odtr6g Lvrpovtreg otrtrqtrouXreq eXel ro ovrlOe : onoretreopo, H ddopeuol rrlg Fox orqv ovodtxn ,. rpovLrrl otrtr1trouX(o ovoorrltrtrer 11v evro(1 r: - evotr),orrrroI e(orviou evto 1 npoodeorl qq o:- , ro0odrrq tvrpovrxrl otrtr1trouX(o 6Xer ro ovr[0E.: onorel,eopo. 606 KEOA Al0 21 Iupporpq rr Ene[epyoo(o rou RNA ry
21.14 Ot Avrr6Pooetq Xpnotponoro0v MrrPd frons-ouPPoq6q RNA 6rq owoytrlvtortrrl 06o11 oupporprlq' Aw6 to pow6Lo unoVpouUi(tt pto tremoupyxrl en(mooq qq o0(eu(qq ue.qtO rnq LrerqYpqqriq r<ot rnq ouppqqfq tou RNA mov nup{vo, ore-mRNA ;retoypd<ptov tou 6tou yovt6(ou'-p6)rlov e(- ;; "q;p"ild *aut, enet6( ov {tov. ouxurl to e(druo ruii vo*otou 0o prnopo6oe vo oupntrrlpdrvouv to 6vo ;dII; owi v' ovtit<ouv oe pto provo6x( op66o oup- ntrrlpopronx6rrlrog. bp,opduot letptopro( unortvo0v trans-ouppqqn' >.o nopdday;ro nou nopouotd(erol c*3. r<dtt': pr6pog ;n; ;ft;;s ft.zo, ouunr.npgqoJx:s oMqloux(es et- or11Oqr<ov oro twpovtq aUb nun' To (euydptouo 99- o"rr' UetoqU rov o u pnr1 pul prot rrdrv o)r'r1l'o uXtdrv anu,ouovti'6vo p6pro oX4go-oq H' Aut6 to pt6pto il;pti'vo ouppoQe( e(te in cls, on6're ouppdmowot !g,6;. nou ,fuo, o1-totonotr6 ouv6e6eprSvo pr' 6vo tv- ijouo, eke in trans, on6te oupp6ntowot e(cbvto rov oG,^pio-,Ou prop(tov RNA' Kot ot 60o owtdpdoetg ou1't- Bo(vouv in vitro. '- - M,o 6trr1 nep(moon ornv ono(o pnope( v9 npoygo- ronotrlOe( trans-ouppoqn in vitro ouptBolvet orov nope- x;;;,;i; unomprir;roto RNA on6 t9.ono(o to 6vo f,;;;6x", u. 5' 06on ouppotpriq rot ro 6Lo gto 3' 05o'n "rbpaqiiq pro(( ;re rotdLt'r1Leq r<oOodrrdg oMql'ou{eq i",-"*its'p"ope(vo e(vot efue 11 en6prevrl 5'.06on oup- poqtis eke 6vog evrq4ut(q qq ouppoq(S) :-1'T1I; 1roiu.6qto, out6 prtpeftot rn ouppoqri.p6oto optouou I(*fru'(Bi5ne 6e[id na'eupd qq Erx6voq 21'11) t<ot oitxrt, oit, in vitro, ot 06oetq ouppoqqq cno 6e[td xot t' "p,*"pa bev e(vot onopokrlto vo Bp(ot<owor cno (6to p6pto RNA. --'nu-at' onotetr6optoto 6eftvouv 6tt 6ev undplet ptq- lovtotu<6 r<cbLupro nou v' onoyopeUet rnv trans-ouppo- lp,i inrons, onortre[ouv povr6[o ouppoErig nou npounoO6rouv rnv npoodewtr{ Uttgl.tygJ ev6g o<r;- liIioi"t ,roooqriq rot6 ;llroq tou RNA' 'Evq oogoil- 5. ouppo,pnq np6ner vo pnope( v' ovoyvop(et rtg 5' r<or 3' b6oetg oupporprlq 6torpopettxdrv RNA 6tov Bp(- oKowot row6, l-1op6tt rl trans-ouppoqq e(vot.qndvto oroug notr'u- norroi*s europuciteg, ewqt o Ku!t9.s prrlxovtop6q yto .nu ""tetiVoo(o ttov np66po;rov RNA oe 6pq'ro mRNA oe optolt6vouq opYovtouo6g 6nog ro rpunovooril;toto *o, -L uhUo.cilbq or<ou)'.r1xtq' :to tpunovootirl-toro' 6l'o i" v*to. ,^,pp&go*o, rog notruototpovxd ;retdypotpo nop6proto Ue rov porcrnp(u:v' Oo16oo' to RNA nou Ue- toyp6gqxe 6e gnope(,o pt-otppoct'ei ltop(g npooOrirrl rrili -ol,inLo uxlqq-bonvo6 37 vou rtr'eort6(ov nou <p6pe- tot 1.t6oto irans-ouppotpnq ytq rrl Uerqrponri ev69 noLy- oi*pou,^o0 RNA oe telcoptord Uovootorpovr6 RNA' H ol"r[rouX(o-o6nV6q 6ev xo6trorrorefuot ovo6xd rolv qeXroptorOv getoypotprxdlv pov66tov' Aw(Oeto' geto- Vp&,pt-o, o' 5vo ove(dprqto RNA, nou qdpet entnp6- t'g"itq otrtrr1lou{eq oro 3' drpo r.ou' on.o Uto enqvo- in;"h pou66o'no, ewon((etot rdnou.otrLot cno yo- ,,ioud n EIK0NA 21.27 deftvet 6tt our6^ro RNA E6pet tn, .ilULorX(o-o6qy6 rot oe-ro0o6rn 06on pto otr't1- LouX(o s' egirhq ouppoQris' Ot oMrl)rou{eq nou rodt- r.onoto0v rq mnNh'q6pouv pto 3' 06o11 ouppo<P{q oujorq nptv on6 rrlv ol'Lr1lou1(o nou undplet mo rilpqro mRNA. u 0r ovl[pooeLq oupporpnq ouvqOtog npoyporonototivrot Uovo in cls peto[u ouppohdrv ouppoQnq oro i6Lo ;r6pto RNA * irrr-orppoQn ouuBo(vet oro tpunovootituoTo Kot.T0 oroutrfl- rto onou Uro Bpoxeto otrtrr1trouXio (SL RNA) oupponretot oro 5' orPo notrtrdrv npodPoptlv mRNA' u To SL RNA exouv 6oun nou poto(eL ;re rn 06on npooDeorlq Sm ttrlv U snRNA. Tdoo on6 prqlovtottr<r1 6oo xot on6 e!,etrcrtxr1 or<ont6' r-orppo,pn'6eopr10qre og ev6o;roprorrl ow(6pooq' "o, luo,o*trd ouv(orotoi oe e}'eyx6pevq onoLotg( rr,.rv oLLqLouXttov tcov twpov(<ov oto en(ne6o tou R- ',tn n ,O yeveitxrl oxontd, q ouppo<pr1 ouprBo(vet pr6vo n "i.. nriO orlpo(vet 6tt prnope(vo ouppoQo0v pr6vo cL oMqtrouX(eg nou Bp(orowot oro (6to pr6pto RNA' To ndv<o trrdpoq rnq , r'' r'' ''': : : [s(1vs1 rnv Kovovt- <l rotdmoorl. To twp6vto pnopef v' o<potpe0oUv on6 <cOe pr6pro RNA, enttp6nowoq oro e(<r'rvtd rou vo oup- .o,poir, oMd 6e ouprBo(vet 6nptopnxrl oupporpq e[tr;- ..lcv preto(ti 6totpopettrov prop[tov RNA' Mo]'ov6tt ,urpiiouut 6tt ouprBo(vet trans-ouppo<pr1 preto(0 rov Kovovu6, ouppoqrl ougpoivet gdvo in cis EEtitvto'1 lvrpdvto E[titvto 2 E[titvto 1 t-*= Iuppoqn Unope( vo npoYgoronotrl0ef ln trans 0v {0 tvrpovto neptdxouv ou gntrqpolpolr6q olJtrltrouXteq --::-'r-.jji::::: .i.' H oupporpl ouvrlOoq npoyporonote(rot povo in cis -::o[u e(ov(tr:v nou pp(orovrot oro i5to pdpto RNA 0mooo' trans- - -:ootpfi'pnopei vo ouppet orov 61proupyq0o0v etbtreq xor0- - - e;dg nou unoonlpi(ouv ro (euyoptogo pooeov pero(u tvrpov(utv' 21.14 0t Avttdpdoetg trans-ouppoqnq Xpnstponoto0v Mxpd RNA flf'
Aro6oXu6q enovotrnqrerq qg povoDog odqyoU ffi I Y O0nVdq 100 Bdoorg 37 Booetov m:*wGUw Aptorep6 rvtpdvto? :eltoptm6g petoypoqrrdq govo0eq ffi I Y AtrLntrouxio mRNA ;g]mx{Qryse Ae(td rwprSvro? I 4t *w odrlYrls G Mtipto oXrlpotoq Y -.4. AG:g I 06r1yoq Atrir1trouX(o mRNA 37 pooeov . To SL RNA n0p6xer evo e[6vro flou ouvdeeTor Us T0 npdTo sEdvr0 evoq mRNA p6orrr lrans-oupporprlg. H ovropoorl nepiloU0ovtr rtq iorsq ovTrdp0oerq Us Tnv nuplvrKl crs-ouppotprl, otrtro nopoyer 6vo RNA 0nUoToq Y ovr[ yro trooo. SL 5' odoq oupporlfg Trans-ouppog( yo qv nopoXrl ev6g odnyo[ yro q gerdqpoorl x 5' 06o11 oupporprlg pA pA pA t_ I Trans-ouppo91 ou(euypevn pe notruobevutrioon yro rnv ene[epyooio J Tou no^uororpovrKou ueT0YpoQou IIII n IWreW IXKTo SL RNA rp00e6Tst evov o6rlyo yro v0 drsuKotruver i1 peroqpoon. Mo(i Ue nq ovTrdpooerq ndqnq Kor noluoDsvui.*- onq, 11 ilpooelKl Tou SL RNA Xprlotponots[Tot er[orlq 0Trl UeToTponrl rotruoroTpovrKd)v UeToypoQov or UovooroTpovtreg povodeg. 608 KEOAAAIO 21 Iuppoqrl Kr Ene[epyoo(o rou RNA Orov o odny6q Kor ro mRNA ouv66owor p6oo *.r: ow(6pooqg frans-ouppoenq, n 3' neproXq rou 06r'. : - RNA r<or n 5' neproxn rou mRNA oqv npoyporrKc:--: onoteloUv ro 5' Kor. 3' puod ev6g rwpov(ou. Orov :--- Bei ouppogq, oXqprot[(eror Svog 2'-5' 6eop6g p5oo. :-: ouvrl0topr6vqg ow(6pooqg preto[ti rcov GU rou 5 r,,:::- v(ou Kor rlq oI nlouX(oq dtox).d6olor1g xowd mc - ; tou 3' rwpov[ou. To 6[o rgrlporo-rou rwpov(ou a.,: o1-rotono).rrd ouv6e6epevo, oMd nopdyouv 5vo ;,c: : oXrlpotog Y ow( yro Ldoo. To RNA nou ouvrroe6per ro 5' e[ovro yro lre-j- ouppo<pr1 ro).ekoL SL RNA (spliced leader RNA). -: SL RNA, prp<oug 100 vour<l,eort6(ov, drntrovowc. :. pro r<orvr1 6eweporoyrl 6oprq nou dXer tpetg pr(o.;c,:- 0qLt6g Kot pto pov6r<trovr1 neproX4 nou prord(er r-: - 06oq np6o6eoqg Sm. Iuvencbg, ue(orovrot cr,c :, nRNP nou onoreLo0v p6q rrlg rorqyop(oq S- . nRNP. Kord rqv ow(6poor1 frans-ouppogqq -rc ! - RNA y(vetor Udpoq rou npoi6wog ouppo<pr1q c -: nopouoLd(eror oro ndvor prSpog rrlq ' . Cno.lg r<t dl,).o snRNP nou egnLdrowor orl oupoc-- (ercr6g rou U6), ro SL RNA <p6peL pLo rprge0u).rrr;i:,- r<o).[nrpo 11 ono(o ovoyvop((erot on6 6vov noc:, ).oyprdvo nopdyovro elF4E ome vo dreuKoluv8:,. - perd<ppooq. Iro C. elegans, nep(nou 70o/orav yovt6[r,-lv rne::-.
yd(owot trr5ool tou prlXovtoprori trans-ouppqqqq' Autd ro yov(6to rorotdooowot oe 60o roqyop(eg' H nptirq nepil,oprBdvet yov(6to nou ono6i6ouv Uovootorpovtrd pet6ypogo to ono(o ene[epyd(owot t6oo trre crs- 6oo ^o, pJ trars-ouppoQn. Ie ow6g rg neptmciroetg, evtir q crs-ouppogrl Xpnotponoekot yto rrlv oqo(peor1 eoore- prrtilv twpov(tov, evcit rl trans-ouppo<p{ nopSXet gto otr- irlLoulio-oO r1yo 22 vo u xLeottd(ov no u n podpletot on6 to St irun ciore vo 6euxotruv0e( 11 pretdrppoorl' H dlLn r<oqyop(o nepil'oprB6vet yovf6to nou ono6(6ouv no]'u- ormpovrrd getdypocpo. Ie ourdg ttg neptmtiroetg, 11 trans-ouppoqrl Xp4otpronotefuot Yto rn petotpon{ tcov noLuotmpovrrrilv getoypdqcov oe povootmpovtrd rt en(or1g nop6ler rrlv oMll'oufo-odqy6 SL yto q pretd- qpqon toug, 6nrog go(vetot oro rdtro p5pog qq Etr6- vog 21.28. Ito C. elegans undpXouv 66o trjnot SL RNA' To SL1 RNA (ro npcino nou ovorqtrf<p0r1r<e) Xp4otponoe(- 'ror ;t6vo yto rl'lv oqo(peoq tov 5' dxptov rov pre-mR- NA nou pretoypd<powot on6 Uovool'orpovtrd yov(6to' flrig Bp(oret to SL RNA rrlv 3' 06o4 ouppo<Prtq Yto rrlv 5vop(r1 rrlq trans-ouppoq{S, rot ndrg onoqe6yet 11 trans-ouppqqrl rov owoytovtop6 11 rrlv nope;rBolq pe qv cis-ouppoqn; H txov6rqto yto m6leuoq ptog tret- toupyu<rlg 3' 05or1q ouppo<p4g nopdXerot on6 rtg npto- tetveg tou SL snRNP. Fto nopddetypct, ro ro0op6 SL snRNP on6 to Ascaris, Uto flopootrtrq vqprot6l4tw0o, nepr5Xet 6uo er6tr5g nproretveq, pto on6 rrq ono(eq (SL - 30 kD) oMql,ent6pd 6peoo ps rrlv npcoreivrl np6ode- oqq orq 06o11 6roxi.d6ooq qg 3' 06oqg ouppoqqg' To SL1 RNA trans-oupp6metot p6vo orqv nprilq 5' ope- rdgpoorq neptoXq, oMd 6ev nopeprBo(ver oe ro0o6trd oulrB6wo cis-ouppoqriq. Aut6 ouprBo[vet enetdrl pr6vo 4 5' otrrerdrppqorn neptoX4 (xotre(ror ourp6vtolutron, B).5ne Err<6vo 21.28) nepfxet pto l,etroupyrxrl 3' 06o11 ouppo<prig, oMd dev SXet rqv ovo6u<r1 5' 05or1 ouppo- qiq ytq vo (euyoprioet Ue rnv xo0o6u<r1 3' 05oq oup- poaiq. To SL2 RNA ttg neptoo6tepeq qopdq Xploq-ronoe(- ror yro qv ene(epyoo(o noluotmpovtxdlv pretoypd<ptlv nou 6toXrop((owot p5oro ;rtog dtoorqprottx4g otrtrqLou- X(og 100 vouxtreottd(cov peto[ri 6Uo yettovtrcilv yovt- 6(tov. I' 6vo prtr<p6 r<l,dopro yovt6(olv 6nou to 6Uo yemovrrd yov(6to e(vot ouvdedeprdvo lcopfg vo nopep- BdMowot 6tomqponr<59 oMrl)rouX(eq, Yto ro dtoXcopt- opr6 roug Xprlotponotefuot to SL1 RNA. Kotd t4v ene[epyoo(o ttov notruotcrrpovtrd:v peto- ypdqorv on' onotod{note on6 outdtq SL snRNA, 4 ov- tidpooq trans-ouppogqq e(vor orevd ou(euypr5vrl pe ng owrdpdoerg n6$nq rot noluo6evul'(ooqq mo t6trog xdOe yovtdtor<rjg povddoq. Mto t6toto o[(eu[q <po(vetot 6rt euvoe(rqt on5 dpreoeg npoteivxdg oMrl}'ent6pdoetq pero[ri rou SL2 snRNP Kot rou nop6yowo 6t6yeporlq qg n6r.pqg CstF nou npood6verot orrlv nLorjoto oe U otrtrr1Lou1(o roOodtrd tou orlpotoq AAUAAA (BI6ne Ev6rrpa 21.15). Awo( ot pqXovroproi enttp5nouv oe oXerr(5pevo yov(6to vo pu0p((owot ouYxpovtopdvo oro en(nedo rnq UeroYpqqrlg (enet6r1 pretoypdEowot o:g notruotmpovtrd perdypoEo) xot vo puOgf(owot [e- lcoprord petd 11 ;retoypoqq (ener6q oL (eXopL::z: ':- vr6tor<6g provo6eq 6toltlp[(owot oq ouventto ric '-:t- (epyoo(oq tou RNA). 2l.l 5 To 3' Arpo To)v mRNA Anproupyo0wot pe n6{,n Kol noAuo6svulioon e H otrtrntroux(o AAUAAA e[vot 6vo ortUo neQnq yto rr1 6lptoup- yio ev6q 3'no),uodevutrto;ttlvou orpou mRNA. * H ovr[Dpoon onotre( 6vo nptlretvtro oOpn],oro nou neptexet evov nopoyovro et6trorrlrog, pto evdovourtreoorl rot noLu-A notrupepooq. * 0 nopoYovrog etdtroqrog rot 1 evdovourtreoorl n6nrouv ro RNA ro0odLxo qq AAUAAA. " 0 nopoyovrog etbtxoqtog rot 1 nou-A notrupepoorl npo- o06rouv - 200 rorotrorno A oro 3' oxPo. * H oupo notru-A etrSyXet rr1 oroOeporqro rou mRNA Kt srrlpe- o(er q perdqpool. + H notruodevutrurlorl oro rurrop6ntroo;to noi(et potro oTnv 0U- Bpurrrl ovonru[r1 rou XenoPus. To 3' dr<po tov mRNA nou UeroYpdgowot on6 qv Pol ll nopdyowot pe n6Qq nou ot<oou0e(rot on6 no- Luo6evutr(ooq. H npoo0rp<r1 notru-A oe nuprlvLr6 D- NA onotpdneror on6 to ovd).oyo 3'-6eo[uo6evoo(vq, 11 ono(o e(vot en(oqg yvoorq o.lg rop6uoen(vr1 @ordy' cepin). l-lop6rt 11 ropduoen(vq 6e mopotd rrl pero- ypoAn rou nuprlvtroU RNA, 11 npooO4r<n rnq nopeuno6[(et rqv suQdvton rou mHNA oro KUTrop6- nlool-to. Aur6 6e(Xvet 6tt 11 noluo6evutr(tr-loq e(vor onopoitqtrl yto rrlv otp(provorl tou mRNA on6 nupqvt- x6 RNA. H oupd no).u-A npooroteUet to mRNA on6 ono66prrloq pr6oto 3'-5' e[rrlvour]'eooov. Irrl ([pn, n oupd notru-A en[or1g prdMov eprnLdr<etot orrlv e[oyco- yr1 rou riprprou mRNA on6 rov nuprlvo Kot orn oro0e- p6qro qg r<oLUmpog. H nopoyotyrl tou 3' dr<pou nopouotd(etot or1v .t:':::.::t. :.: .'1r,. N RNA notruprepdOrl ;retOypdqet pretd rq 06o11 nou owtororXe( mo 3' drpo r<ot oMq).ou1(eg tou RNA ovoyvrr:p((owor otg m6Xot evdovourtreotrurLr<r1g ndqrnq 11 ono(o npoqye(tot qq noLuo6evu).iooqg. H RNA notrupepdorl ouveX((et q preroypo<pr1 prerd qv n6- rl.rr1, oMd to 5' drpo nou nopdyerot on6 qv ndqq 6ev npoorotetietot, yeyov6g nou orlporodote( tov repport- opr6 qg peroypo<pr1q (B).5ne Evorqra 21 .16). Iro neptoo6tepo pre-mRNA, 4 05oq ndqrlg/noluo- devutr(coorlg neptBdMetot on6 6Uo cls-6poortr<d orlpo- to: 6vo ovodtr6 prot[Bo AAUAAA, ro ono[o ouvrlOoq on6Xer 11-30 vour<l.eor(61o, xt tivo t<o0odtx6 motXe(o nlorjoto oe U 11 oe GU. To AAUAAA onotte(toL 16oo yto qv n6Qr1 6oo t<ot yto rnv noluo6evul'(toor1, 6nolg cpo- vepc0vet to yeyov6g 6tt onotrottpq r1 pero).LoEn rou eEoUepo0q AAUAAA ovom6Met rlv nopoYoYl rou no- l,uodevul.urlllevou 3' dt<pou. l; 1:{ To 3'Axpo rotv mRNA Arlgtoupyouvrot Us ileqrl xor [lo],uoDevuhltrlorl 609 Buorx6g ewoeg
Ev6ovouxtredorl AAUAAA opxereq evepy6rrlreg. 0t CPSF ror CstF onoretro0vror ono opre- reg unopovoDeg. To unotrorno ouorolro e(voL lLovopepl, H ouvo- trrrq po(o eLvor >900 kD. T0 mRNA or00ep0n0re[r0r U6oo rnq notrugodevul(oolg tlojvouK^eoorl * _.--"9 ,4il*orulcit flouodevutr (oor1 I S'Kotrunrpo AAUAAA An . : '.:: ', rr ' H otrtrr1trouXio AAUAAA efvor onopoiqrn yro Trlv n6gq rqq 5'rotrtinrpog tlore vo 6lproupyn0e( 6vo 3'orpo yro no),uodevuL(tooq, H ovdntu[4 ev6g ouorqproroq yro in vitro no].uo- 6evuL[coor1 dvor[e ro 6p6pro yro rlv ovd].uor1 rov ov- 116pdoeov. H 6qprroupy(o rot or Letroupy(eg tou oupn).6xou nou ovoloprBdvet qv 3' ene[epyoo(o no- pouotd(ovtor orlv :.' . ,:i ," . H 6qprroupy(o rrlg ooorqg 3' reLrrrlq 6oUnq e[oprdror on6 tov eL6txo nopdyovto yro rnv n6,Qq xu uyv no).uaevulfuo4 (cleavage and polyadenylation specific factor, CPSFl o ono(og neprdler no}.LonL5g unoprovd6eg. Mto ono rrq unoUovd6eg npoo66veror dpreoo oro proriBo AAUAAA Kor orov ndpoyovra 66yepor7q rnq nEqnq (cleavage stimulatory factor, CstF), o ono[og e[vor eni- orlg 5vo oripnLoxo pe noL).ontrd ouororrxd. 'Evo ono outd ro oumorrrd npoo6Sveror dpeoo oe pro ro0o- du<r1 o).qlouX(o nlorioro oe GU. Or CPSF r<or CstF evtoljouv o 5vog tov d.tro xord qv ovoyvtr;pLoq ror', o4ptdttrrv no).uo6evu).(oroqq. To er6rxd 5v(upro nou e!- nl6t<ovtoL e(vor 1-rro evdovour<Ledon (n unogovd-c tov 73 kD rou CPSF) nou n6nrer ro RNA Kor Uro no- Au(A) notrupep6o4 (PAP) nou ouv0SreL qv oupd nc- Lu-A. H nou-A noLupepdorl (poly-A polymerase) exg -- er6rrrl rorolutu<r1 evepy6rrlto. Otov ouvdud(erol "i .: un6lorno ouorortrd, q ouv0elrq ow(6pooq y(ver:. "- 6tr4 yto RNA nou neprdlet qv oMq).oux[o MU;-:-- - ow(6poor1 noLuo6evul.iuroqg e[e].(ooeror 6Lc-::: - 60o mo6[ov. I-lproro npoor(Oetor 1-uo oXerrro 3c:,i: oMrl).ouX(o o)lyo-A (-10 r<ordl.orno) oro 3' oi<oc :-- 4 ow(6poor1 e[oprdroL n].4ptoq on6 tqv oMr,.:,, : AAUAAA r<or 11 noLu(A) noLupepdorl rrlv 6LeEc,-. ,-: rrlv xo0o6qyrlon rou nopdyowo erdrxdrlrog I:: I"-- tepo md6ro, 11 nupqvrrq nporrelvr1 PABP I .: ., : binding protein) npoo6Sver rrlv oupo otrryo-A 3:.:::- nowoq tqv endmooq qg Sorq ro n).qpeg !rK3i - -200 xoto).o(n<rrv. H (6ro q notru-A noupepor -::- o06tet xotd).orno A mrlv 3' 05oq. O eyyevrc . --:: dpdoqg r1q e(vor vo 6lfororor rd0e cpopo nou n::,-i: ter 5vo voux..eot(6to. 0o16oo, nopouo[o rov CpS= . : PABP ll, Lenoupye( oStdLeLmo, enemefvowoq *.: ::- X<r-)pto,q oLuo(6o nou-A. Merd qv ow(6poor ,-'-:- 6evu).(coor1g, q PABP ll npoo65vetor fiorxao-.r: . : mrlv npo5rcrooq notru-A. Awd ;-re dyvooro lrit.r, --: neprop((er tq 6pdor1 rrlg noLupepdoqq no)..-: :: : -200 npoo0rlreg rorolo(nolv A. Kotd tqv e[oytrryq ttr-lv tr-lpL1-uov mRNA or:c ::. -- pnvo, n rurroponLooprortxrl nporelvq PABP .:-::: veror otrlv oupd nol,u-A. H PABP I d1r uc.: -:,- more[er ro mRNA on6 rr1 6poor1 tcov 3' r<oL 5 e!- , : - xLeooorv, oL}d en[or1q npoo6dveror orov ncc]. : ,- l 6vop(49 qg perdgpooqq elF4G, 6LeuxoAu,,'c.:r: - ;rerdgpoorl tou mRNA. 'Eror oro rurropon).::-: -- mRNA oplpror((er prro xLetmrl Or1LLo mqv c:3 : : - nporeivrx6 otipnoxo nept5ler tdoo ro 5' doc ,.:. : : dr<po rou mRNA (B).5ne Erx6vo 24.22 orqlt E,: --: 24.9). Zuvenrirg, 11 no).uo6evu).(oor1 enqped(e. :::: - oro0ep6r4ro 6oo ror qv 5vop[r1qg prerdgpc:*: :-: rurtop6nlool-ro. Kord qv epBpuirrl ovdntu(r1 rou Xenoprs - -:' ).uo6evuL[tooq 6re(dyetor oro Kurrop6ntr oc - : : i : o<poL((ovroq ro Unrprr<6 6l.eyXo rlq r:. - -- : epBpuoySveoqg. OpLopr5vo ono0qxeupdvo pqrc., : - : NA pnope( efre vo notruo6evu],t<l0oUv ond q', -:'- - noLutr-repdorl oro Kurrop6n),oo1.ro oore vo 6re ,"::i - Enrprlxuvorl rrrir> $sP$o ..- ., -net|trl ,,-.fi; , & 610 KEOAAAI0 21 Iuppoqrl rr Ene[epyoofo rou RNA To o0pntroro 6r[sr0T0r [er0 rnv npoo0{xr1' 200 xoro]o[nov A
Uerdgpoon e(re v' onoo6evulloeoUv, reppqr(owoq rn petdgpooq. 'Evo er6tr5 cis-6poonr6 qrorxe(o nLorioto oe AU (CPE) orrlv 3' oupd roreu0[ver rqv erdxr] yro 11 peuorxq top(provo4 notruo6evutr(rooq oro rur.op6nlo- opo, evepyonor<ilwog 6rot tq prerdrppooq oprog6vov er- drxdrv p4tpu<rriv mRNA. l-to q pU0puorl qg ono66- Unonq rou mRNA, undplouv roudlrorov drio rUnor cis-6pooru<rirv otrrlLouXrc0v orrlv 3' oupd nou nupo6o- ro6v rrlv onoo6evul(<oorl rou mRNA: 11 EDEN (embry- onic deadenylation element, orotle(o eprBpuir<tiq ono- o6evu),(oor1g), gro otri.r1Lou1(o 1 7 vour<.treorr6(rov, ror ro ororXe(o ARE, otri.qLouX(eg ntrotioteg oe AU, nou ou- vnOoq nepdXouv 6ro6o1u<69 enovolrlQerg qg AUUUA. Mto RNAdoq erdrr4 yro r4v notru-A oupd (poly(A)-spe- cific RNAase, PARN) epnMrerm orrlv ono66prrlon rou mRNA oro rurrop6nlool.ro. Aorpofuirg, 4 onoodevuL[ro- on rou mRNA Bpioreror ndwo oe owoycovrop6 pre 14 oroOeponofiorl rou mRNA: ouMoyu<d xo0op((ouv ro 1p6vo rlpr(ror1g [eloprorrirv mRNA oro xfrropo (BI5ne KegdLoro 22). 21.16 H EneEepyooio Tou 3' Arpou rou mRNA Eivor Kpioryn yro Tov TepUoTtoU6 Tnq MeToypoQfiq e Ynopxouv drotpopor rponor reppolopou qg peroyporprlq ono 6rorpopelxeg RNA notrupepooeq. * 0 oxlUolouog rou 3' orpou rou mRNA o1;roro6ore[ rov rep- polopd nlq Ueroyporprlg ond qv Pol Il. OL nlqpogopieg nou 6ro0Sroupe oXertrd Ue rnv ov- r(6poor1 teppotroporj yro nq europurrrrrr5g RNA no- uprepdoeg e(voL ).ry6repo Lenropepe(g ouyrprrrr<d lre rrq yvoloerg ;rog oXerrr<d Ue rnv dvop[r1. To 3' or<po rcov RNA nopdyowor Ue 6Uo rp6nouq. Opr- op5veg RNA nouprepdoerg teppor((ouv rrl UEro- ypoen oe Uro r<o0opro;r6vr1 oLtrqAouX(o tou DNA irepporrorqg, terminator), 6nrog <po(veror orrlv ., :i. H RNA no).upepdoq lll <po(veror 6l XpqoL- uonoteI ourrl tqv orporqyrxq 6ro06rovrog 1.rro 5ror<prrr1 o).).r1).ou1(o oAryo-dT nou onUoro6ore( rrlv cneLeu0dprr-ron rnq Pol lll rrtore vo repporrcrre( 11 pe- :oypo<pr1. l-to rlv RNA no).upepdoq l, ro provo6rr6 npoi6v qq leroypo<prlq e(vor 6,vo preydLo npdbpopro p6pro nou ne- cr51er ttg oLi.r1).ou1(eg rou rUprou rRNA. O repUorr- ;pdq npoyporonore(ror oe 6uo 6ror<prr6g 06oeLg (T1 <or T2) ro0o6u<d tou roptprou 3' dr<pou. Awo( ot reppo- :rordg ovoyvrop((owor on6 pro et6rr<q npcorefvr1 npdo- 5eoqq oro DNA (Reb1p orrl (Upq r1 TTF1 orov :owu<6). O repgotrol.r6q qq Pol I oler((eror en(oqq pr' ivo ou;rBdv n6Qnq nou dropeooLoBe(tor on6 rqv ev6o- W EIKONA 21.31 H UeroypoQn p6otrl rtrlv Pol I ror Pol ll Xpqotponor- e( erdx6q otrtrqhouXieq (repyolm6g) yro Tov repp0Trop6 qg pe- royporpflg. vouxi.edoq Rnt1p, 11 ono[o n6mer ro RNA nou opX((er vo 6qproupye(ror nep(nou 15-50 Bdoerq roOobrd rou 3'drpou rou ene[epyoop5vou 28S rRNA (BI5ne Ev6t4- ro 21.20). An6 ouri rnv dnoqrrl, o repporropr6g qg Pol I oXefl(etor l.r4Xovronr<d Ue rov repporropr6 qq Pol ll ener6rl ror or 6uo 6repyoo[eg (oog nepil.oprBdvouv 5vo ouprBdv ndqrqg RNA. Aw(Oero Ue rov tep;ronopr6 rov Pol I ror Pol ll, q RNA noLuprepdon ll ouvq0rog 6ev npoyproronote[ 6ro- rpn5 reptrlolopr6, ol)d ouvel((et vo pretoypd<per yro nepfuou 1.5 kb n6po on6 rq 06o11 nou owrororXe( oro 3' dxpo. To outrrBdv n6qnq ot4 05oq noluodevutr(oroqg nop6Xer ro 5vouo1.ro yro rov repprouopr6 on6 qv RNA noLuprepdo4 ll, 6nrrlg go[veror orrlv EtK0ilA 21.32, Auo prow6),o dXouv nporoOef yro rov repporropr6 rqg Pol ll.I0pgtrrvo prero oMoorepx6ltowi)to, n ndqn rou RNA orq 06o11 notruodevuL(ooqg (oog nupodore( optopdveg oMoy6g drop6prpoorlg 16oo oro oUpmloro Pol Il 6oo Kor ornv ronxq 6opu1 rrlg Xpcopror(vqg. Aur6 prnope( vo endyetot on6 owoMoy6g nopoy6wtrtv Kqrd t4v ow(6 poorl nolu o6evutr(oor1 q, n porol,c0wog nori orl qg Pol ll rr 6nerq oneLeuO6ptoon Uq qn6 ro DNA er<- goye[o. 'Evo evoMorcrx6 gow6tro, yvoor6 aq powilo top- ntAr1 ftorpedo model), npore(ver 6u ;uo er6u<{ e[<ovou- rledorl npoo66veror oto 5' drpo rou RNA nou ouvel((er vo geroypdrperor perd rqv n6q4. Ano6opre( to RNA nol,ri yprlyopo, tolUrepo on' 6rr ouw(0eror, on6te npogro(ver ulv RNA noLuprepdoq. 'Enemo, o.Lq- awarcffiffirymw P til Pol lll Poll affi* J xffiyffimffiw B.{ f ?i't* H Ene{epyooio rou 3'Arpou rou mRNA E(vorKproL;r1yto rov Tepgolopo rnc l'.le .:,:::-: 611 Boorreg fvuoreg
Pol ll gerd ro orlpo IIotru-A H rorq er06ret ro onpomdreuro 5'6rpo Anoddpqo4 qn6 eqovouKle6oq Erdyer Inv ofl oorooeponofi or1 n1g Pol ll Aneleu0dpooq qg Pol ll qnd ro eruoyeb EIKONA 21.32 0 oXqpolop6g Tou 3' oKpou rtov Pol ll peroypd- Qov 6reuKotrriver Tov TepUoTroUd Tnq UeTqypognq. Lenr6pd U' enrKouprK6g npoteiveq npoo6edepdveq orqv ropBo[ure].xr1 neptolq rqg no].upepdoqg. H o].- Lrllen(dpooq nupo6ore[ tqv one],eu06proo4 rqg RNA nol,upepdor;g on6 ro DNA, nporoLcirwoq repprorolr6 rnq Ueroypogrjg. Aw6 ro prowStro e(qye( ytor( or 06- oerq repporropro( qg RNA nolu;repdonq ll 6ev e(vor rold roOoprotrr5veg, oMd prnope( vo epgovroroUv oe drdgopeq 06o'erg pr6oo oe puo peydLq neproXll r<o0o6r- rd qq 06o49 nou owrororle( oro 3' dr<po tou RNA. H rUpro nepopronrrl dv6er[4 yro ro pow6].o ropnrtr4 efuor o p6troq r4g nuprlvrxrig 5'-3' e[ovoux].edoqg Ratl orq ([Un n trlg Xrn2 oro Oqtroonrd. H ono].ot<pq rou yovr- 6(ou ouyyd nporole( prerdBooq rnq Ueroypoenq oro en6pevo yovi6ro. Ie oproprdvo netpogoru<d ouorrjpoto, oqr6oo, peroMdlerg rou orlporog AAUAAA nou el,o- oOev((ouv qv n6q4 m4v xovovrrq 05oq noLuo6evu- Liroo4g 6ev nupo6ororiv ovoyroonrd oneLeu0Sptoo4 rnq Pol ll ofte ror perdBoorl oro en6prevo yov[6ro. Xu- venrig, ouui q 6v6er[4, po(i pe oproprdveq tonu<5q otr- Loy6g orn 6opn rnq Xpoprot(vr;9, euvoeI to oMoorepx6 pow6l.o. 'E1er nL6ov r<oroore( oog5g 6r ro 6vo prow6tro 6ev onor.).e(et ovoyroortrd ro dMo. Kor to 611o (otrrg owr- roromp((ouv oprop6veg xp(otpreg muX6g nou oXe{(ov- ror Ue rov rep;ronop6 rnq Ueroypocprlg p5oco qq Pol ll. Me onorov6rlnore 11 Kor pe rouq 6Uo ;rqXovtopo0g, e(vor ooq6q 6n o repporropr6g rqg preroypoqrjg pr5oor rrlg Pol ll e[vor orevd ou(euypr6vog Ue ro oxrlportop6 rou 3' drpou yro ro neproo6repo mRNA rov europur.rrttrri:v ruudprov. 21.17 O Ixnporop6q rou 3'Arpou rou mRNA Ttltv loTovd)v Anorrei To U7 snRNA " H 6rtppoorl rov mRNA rorv rorov6v e[oproror ono rr1 pero- ypotpl ror pu0p((eror roro q dtopxeto rou rurroprxou xu- xtrou. u To mRNA rov rorovrirv dev elvor notruodevutrropevo: ro 3 orpo rouq drlproupyelrot peoor lrrog ovridpoolg ndQrlg nou e[oproror ono q doprl rou mRNA. * H ovrOpoorl n5g1q nporino06reL npoodeorl tou SLBP oe gLo 6oU! Uroxou-0ntrrog ror (euyoprogo rou U7 snRNA Ue Uro y0r- rovrrrl povcirtro-rvrl fl sp roxn * H ovr[dpool neQqg rorotruerot ono 6vov nopoyovro Kotvo l.1t ro o0pntroro notruodevul[r,lor1g. H Broydveorl rov rmovurv etr5y1e-rot r<up(cog pdoo priOpuo4q qg o<pOov(og rrr.rv ow[orotlrov mRNA xoro rq drdpr<eto rou KurroprroU rUr<Lou. Irq prerdBooq G1/S, n ouyr<dwpcrroq rov mRNA trov rorovrrrv oulo- ver >30 gop6q L6yro ou(qpdvr;q pretoypogrlg. Auq q drepyoo(o puOpr((etor on6 ro o[gn].or<o Kux].[vqg El3dk2. H oU[r;o4 rov mRNA tov torovci.lv ot<o].ou- Oe(tor on6 toXe(o ono66prqorl rouq fio rSLog rqg gdor1g S. To mRNA rrov rorovc6v 6ev e[voL notruo6evu].uo;.-rdvo (ercr6g rou S. cerevisrae). Iuvenc6g, ro 3' dr<po roug 6q- puoupyotlwot pe 6to<popertx6 tp6no on6 q ouwovr- oprdvrl ovr(6pooq n6qtrqq/notruo6evu).(orotlg. O oXqprolopr6g rov 3' dr<prr-lv e(oprdror on6 pro e[orperL- xd ouwrlprlprdvq 6opq pr[o1ou-0qArdg nou ewon((eror 14-50 Bdoerg r<o0o6rr<d rou xrridtxov(ou tepprorroptou ror on6 6vo r<o0o6u<6 ororXe(o rorovcbv (histone down- stream element, HDE) -15 vour.treot(6to roOodrxd qg 6oUnq pr(cryou-0ql,rdg. H ndqurl npoyporonorekor preto- (U p(o1ou-0q).rdg r<oL HDE, o<prlvowog ndwe Bdoerg r<o0odLr<d rqq dopulg pr(o1ou-0r1Lrdg. MetoMd(erq nou nopepno6((ouv ro oxrlUorrop6 rou 6(x).covou pr(oXou ovomdMouv tr1 6qpuoupy(o rou drpou rou RNA. Aeu- teporoye(g 6o;rdq nou onoro0Lmo[v q 6[r].tovq 6opq (proi.ov6rr 61r onoporrrltc,:q rrlv opXLxrl o).Lq).ouX(o) onoxo0tmoUv Kor rrl 6qproupy(o rou 3' dr<pou, Auro uno6r1orver 6l o crlrlprortopdg qg 6eureporoyo0q 6o- prrlg e(vor nro orlpowrK6g on6 tqv oxprBrl oMll"ouX(o. H 6qproupy[o rou 3' dxpou rtov rmovtrrv <po(veroL orlv , . Auo nopdyoweg ononorjwor yto rov roooptopd rqg ow(6poor1q ndqUqg: 4 np<oreivq nou npoo6Svetor oro ;i(oXo-OqLrd (stem-loop binding pro- tein, SLBP) ovoyvop((er q 6opu1 1-riolou-0r1).tdq KoL ro 5' dr<po tou U7 snRNA (euyopover Ue Uro noOoro oe noup(veg oMqtroul(o prSoo oro HDE. To U7 snRNA e[- vor 6vo puxp6 snRNP nou onoretreftor on6 ro U7 snRNA (63 vour<Leor(6ro) r<or pro oprddo oprerorv npoteivorv or ono(eg olerf(owot pe lg snRNP nou etrrnL6xowor orrl -e__j-_reG Xrn2 I (A)n 612 KEOAAAI0 21 Iupporprl rr Ene[epyooio rou RNA Boorxeg 6worcg
A G SLBP UU ,'U jjc ffi,ffi ffic mRNA Lmovtirv Iugntrer(vr1 oupporprl tou mRNA (BLene Ev6t4ta 27.6). Movo6tr6g yro ro U7 snRNP e(vor d6o npcoretveg nop6proeg pe rtg Sm, or LSM10 ror LSM11, or ono(eg owroOtororiv ttg Sm D1 rot D2 qrn ouppoerl tov snRNP. Av ovoorqlei ro (euydpropro prero[U U7 snRNA xot HDE, q 3' ene[ep- yoo(o tcov mRNA r<ov totovtilv 0o 6toropol0e[. An5 qv d.tr.q nLeupd, owroro0prorx6g pretoMd(etg oro U7 s- nRNA nou onoro0roroUv q ouprnqpoptonr6tpo ono- xo0roro0v en(or1q Kot rnv 3' ene[epyoo(o. Aur6 unodqtrdrver 6n ro U7 snRNA Leroupye( pr6oo (euyo- pdrgorog Bdoeo:v pe to mRNA ttov tqrovdlv. l-16r[q yro rq 5r1p[oupy(o ev6q 3' drpou ouprBofuet oe moOeprl on6oroorl on6 11 06o11 nou ovoyvop[(erot on6 ro U7 snRNA, yeyov6g nou unodrltrcbvet 6tt to U7 snRNA etrrnMretoL orov xo0optopr6 qq 05ong ndq.trlq. O nopdyowoq nou euO0verot yto rrlv n5Qr1 e(vot q npone[vr1 CPSF73 (cleavage and polyadenylaflon speci- ficity factor 73). Iuven6q, ou'r6 to prdLog qg oroySvet- oq rov peroMo-B-Aor<ropoo<ilv no((et Bootx6 p6Lo orq 64puoupy(o rou 3' dt<pou t6oo yto to nol,uodevulttopr6- vq RNA 6oo r<or yro rq Un notruo6evuluo;.tdvo RNA ttov rqrovdlv. 'EXouv ovoyvoprore( opr<et6g nporetveg nou no[(ouv oqprowtr6 p5],o orrl 6qpuoupy(o tou 3' drpou rrov rorovdlv, pero[6 dMarv 4 CPSF100 rot q lugn].er(- vq (symplekin), oMd or erdu<5q 6pdoerg toug 6ev 61ouv xo0optore(. Aut6g or enrnp6oOereq npolteiveg nt0qv6v nop6louv trenoupy(eg rxptc0ptotog nou oro0eponototiv ro oripntroro ene[epyoo(og rou 3' dxpou. E(vot ev6ro<p5pov 6tr 11 dtotopoXrl rou (euyopdrtrro- roq rou U7 pre trg oLlrlLoul(eg-m6Xoug oro yov(6to rc,:v torov6v 6nog en(or1g ror q dropeool,oBotipevrl on6 siRNA e(dLetqrl dlLrov ouorotx6v nou egnl6t<owot orq 6qploupy(o rou 3' dr<pou rov torovdlv o6qyoUv oe CA-Py7-N2-u -AAAGAGCUGU-3, ttttttllll uuucuccAcn %e5' U7 snRNA nopoB(oor1 tou nSpotog rnq peroypqg{g rot oe TIo- l,uodevuL(cooq Xprlotpronotdrwog 5vo orlpro nol,u-A t<o- 0odrxd rou DHE. Iuvenc(lg, rot' owtorot{o pre to p6}'o qg n6gqg/notruo6evutr(oonq rou mRNA orov repport- opr6 qg preroypogrlq p6oco rqq Pol ll sro neptoo6tepo yov[6ro nou ro6rxonorofv yov(6to, 11 n6Q4 tou RNA nou peootroBe[tot qn6 ro U7 t<otd q 64;rtoupy(o tou 3' drpou Eofveror 6rr e(vot r<pfotprr; yro rov teppottopt6 rnq Ueroypoqnq oro yov(6to ttov torovrilv. neun U7 snRNP : H [rlproupyro rou 3' orpou rou H3 mRNA rotv torovtirv e[opToTor ono puo ouvrr1p1p6v1 qoupr6ro roL ;tto o),trr1LouX(o nou .e-yoptover ue ro U7 snRNA. 2 I .l I H luppogri Tou IRNA lleprAopBdvet Konh rr Enovoo0v6eon ot lextoprordq Avrr6pdoerq a H RNA notrugepoorl lll repuon(er rt,l UeroypoQn orqv 0trtrr1- trouXio notru(U)4 nou Bp(oKsTot ptloo oe Uto otrtrltrouxto ntro0- oro oe GC. * H ouppoQn rou IRNA npoyporonote(rot pe dto6oxtr6g ovrt- 6pooetg ronrlg ror orlvDeolg. e Mro evdovourtreooq roper ro npodpopo IRNA rot oto duo orpo rou rvrpoviou. * H onetreu05poorl Tou lvrpovlou dqptoupyei 000 utootRNA ue oouvrl0toro orpo nou nepr6Xouv 5'-0H rot 2'-3' rurtrtrd qo- oqoprro. * To 5'-0H oxpo rptootpoputrttlverot on6 pto notruvouxtreondtrl ruvoorl, 11 opo6o rou rurtrtro0 Qtrtotpoptrou ovo[yet ono pto rprootpodteorepoorl onodldovrog 2'-tpoorpoptro retrxo orpo rot 3'-0H oUo6o, ro orpo rolv e[ov[ov ouv66ovrot on6 ;tto RNA trtyroorl ror ro 2'-tptrrotpopuo otpopelrot ono gto Qo0Q0T0or1, :: 'i8 H Iupporpn rou tRNA l-lepilopBover Konfl rt Enovoorjvdeol oe :eXoprore_Q Avl6pooet-c 613 Lsm10 Lsm11 Boorreg 6woreg
Or neptoo6repeg owrdpdoerg ouppocp(g e(oprcbwor on6 BpoXe(eq ouvorverrK6g oMr.1).ou1(eg xor 6re[dyov- ror p6oto ovrr6pdoecov frans-eorepono[r1oqg mrg ono(eg 4 OpoUorl efvor ouwovropdvq pre q 64ploup- y(o 6eoprciv. H ouppogri rcov yovrS(ov IRNA enmuyld- veror p6oro ev6g drogoperrroU prqXovropro[ nou Boo[(eror oe [eXopror6q owtdpdoerg xonrlg rqr o[v- 6eo4g. [1epftou 59 on6 ra 272 nupqvrrd yov(6ro IRNA orq (UUn S. cerevisiae e(vor 6roreroprprdvo. Ko06vo riler 6vo ptovo6rx6 rwp6vro nou ewon((eror g6Lrg 6vo voux.l.eo- rbro prerd rrlv 3' nleupd rou owxul6u<ov(ou. To rwp6- vto r<ugo(vowor oe ;r5ye0og on6 14 6rrlg 60 Bdoerq. Ooo Bp(orowor oe oXerr(6prevo yov(6to IRNA SXouv nop6trloro oM4l,ouX(o, evdl ro rwp6vto oro yov(6ro tR- NA nou owrTrpoooneriouv drogopertrd oprrvo[6o 6e oler(owor. Aev undpyu ouvaweuxrj aAA4louyta nou 0a pnopoloe v' avayv@porc( an6 ru 6v(upa ouppa- g{9. Aur6 en(oqg roXUer oro 6rorerop;r6vo nuprlvrrd yov6ro IRNA rcov rpur6v, r<ov opuprB(cov ror rrov 0r1Lo- orrcbv. 'OLo to rwp6vro nepil.oprBdvouv pro oLl,rlLouX(o oupnL4pcol.rorxrj pe ro owrr<odm5vro rou IRNA, Aur6 6qproupye( pro evoMorcnxrl dropr6pgoorl yro to Bpo- 1(ovo rou owrxrrl6rrov[ou offlv ono[o ro owu<rrldrr6vro e(vor (euyopropr6vo, q44poil(owog 6ror puo npo6r,roorl rou ouvrlOoug BpoX[ovo. 'Evo nopd6erypo nopouord(e- ror orrlv EIK0NA 21.34. M5vo o Bpo(ovog rou owro6r- rov(ou enrlped(eror - ro un6lorno pr6pro 6roqpe( u1 ouvqOropdvq 6oprr1 rou. H orprBr1q otrLrlLouX(o r<or ro pr6ye0oq rou rwpovi ou 6ev 6Xouv o41.roo(o. Or neproo6tepeg preroMd[erg oro rwp6vro 6ev nopepno6[(ouv q ouppogrl. H oup- paEfi rcu IRNA elaptdtat xup[aq an6 u1v avoyvthpto4 ploq xo:fiq eutepotayolq 6op4q oro IRNA, aw[ yn pn xolr1 aMr1Aouy[a rcu twpov[ou. YndpXouv onpqwr- r<6q neptoX6g oe drdtpopo pr6p4 rou pop(ou, prero[0 d].- ).ov oLLrll,ouX(eg peto[O rou BpoX(ovo-66rcrr1 Kor rou ZeuydpoUo To twprlvro (euyoptirver ge q 0r[trd tou owtrordrrov(ou To rvrpovlo oT0 tRNAPrre r1q (uUlq (euyoprr-rver pe ro ovlrorDrrovro yro v' otrtro[er q dogrl rou ppoX[ovo TOU ovTrKo]- drrovlou. To (euyoptoUo pero(f pLog onortreropevlg poolg oro ploXo ror qq 0qtrrog rou rvrpov(ou rou npodpopou poprou rotrtg onoue [ror yro q ouppotpil. BpoX[ovo D, oMrll,ouX(eg oro Bpol(ovo T/C ror rd(cog oro BpoX(ovo rou owrKo6u<ov(ou. Aur6 Oug((er ng 6o- pm6g npo0no0Soerg nou r[Oewor oro IRNA yro rn pe- rdgpoorl (BMne KegdLoro 24). MoLorqUro, ro rwp6vro 6ev s(vor eweL6g doXero. To (euydpropro prero[ri puog Bdorlg orq 0r1].td rou rv- tpov(ou Kor Uroq o(euydpoqq Bdonq oro p(oXo e(vor onopofuqro yro rrl oupporpq. Mero).1,d[erg oe dtr].eg 06oerg nou enrlped(ouv out6 ro (euydprrtpto (n.X. 6r;pu- o u pyriwog evoMorcrxd np6runo (euyopcil poroq) enrl - ped(ouv 14 ouppogrl. Or rov6veg nou 6r6nouv rr1 6toOeorpr6qro rov npo6p6prov tRNA yro ouppo<pr1 ptord(ouv pe rouq ow(orotloug xov6veg yro rrlv ovo- yvciprorl on6 rrg opuvoor<ulo-tRNA-ouv0erdoeg (BLdne Ev6trya 25.9). I' dvo Oepproeuo(oOqro perd).troypro (0prqg nou onoruyldver v' ogotpSoer ro rwp6vto, droreroppdvo np6Spopo prdpro ouooopeUowor mov nuprlvo. To np66popo pr6pro lprlorponororiwor <og unoorp6proro yto rivo oUmqpro el,eU0epo rurrdpov nou er<Xu),((e- rqr on6 r0rropo dyptou rUnou. H oupporprl tou np6- dpoprou 1.rop(ou prnope( vo noporrlprl0e( L6yo rrlg enor6).ou0r1g pe[ooqg rou prey60oug. Aur6 onnKo- norefuot pSoo qg ol},oyr1g mr1 06oq qg (rrivrlg r<ord qv qLerrpoe6pnon oe nr1rr4, 6nog <porveror orrlv EIN0NA 21.35. H eLdrrcoorl rou pey60oug epprrlverieror on6 rqv eprgdvroq ;uog (cilvrlg nou owrnpoooneUer to rwp6vro. To eLe(0epo rurrdpov erlULrotrro prnope( vo xLo- ol.rotono rr1 0e[ el,5yXowo g rqv rxov6rrlro ou p poE4 g rou IRNA. H in vitro ow(6pooq onone( ATP. O lopo- rcrqprop6g rotv owr6pdoeov flou npoypororroroUwor pe { 1op(g ATP deiXver 6r ro 6uo leyuptord otd6n qq owbpaoqq xataiowat on6 lnEopetxa 6v(upa. Erd)ool ['erXUtrrogo llp6dpogo tRNA NG G -,ffi,# Ufl; :d,6nl ffiffi ./ ffi'8rry, TP ffiIvrprivto Oprgo tRNA CG GA AU GCm AV CG m AU cA UG wAvflK(l)6rK6vro tl ffi noo*,, ffi ry1 turpdvro ffi -r n*o [:'] cH"YdhW+ %, H),exrpoqopqol oe nrlrrrl ' H ouppoqrg rou IRNA Tnq (uUnS in vitro noparo- ),ouOe(ror p6oo qg ovotruolg rou npodpopou RNA xor Tov npo- iovtrov pe qtrerrpoqoploq oe n1rrr1. 614 KEOAAAI0 21 lupporprl rr Ene(epyoo(o rou RNA
o To npdno gnUo 6ev onore( ATP. l-lepiloprBdver n6- rlq <pootpo6ooreproU 6eogoti p6oto gtog dtunrlg ow(dpooqq vouxtredoqq. Kototr0etot on6 pto ev6o- vouxl.edoq. o To 6eUtepo FnUo onore( ATP xot nepfioprB6vet ro oXrlprotropr6 6eopro0. Eivot 1-tto owfdpooq o6v6eoqg xor 11 une60uvq ev(u;lxri evepy6qto neptYpd<petqt coq RNA lryrdo4 (RNA ligase). H ouppoqrl rou pre-tRNA Yto rrlv orpo[peor1 twpo- vfov e(vot onopo(tr1tq oe 6].oug rouq opYovtoproUg, otrtrd 6to<popettro[ opyovtopro( Xprlor;ronotoUv 6torpo- perrrotig prrlXovropro0g Yto rrlv en(teu[4 qq. Ito Bo- rcr{pro, to twp6vto ttov pre-tRNA ouro-ouppdntowot oq ouroKqrotrutrxd twp6vto opr66og I tl ogdSoq ll' Iro Aplo(o rot to Eurdpuo, n ouppo<pr1 tou pre{RNA nepil.o;rBdvet q 6pdoq tptc0v ev(0prcov: (1) gtoq ev6o- vouxtredorlg nou ovoYvopf(er rot nSmet ro np6Spopto Kor oro 66o drpo tou twpov(ou, ;rto fuyxdon nou ouv- 65er to e(drvto tou IRNA t<ot t1 2' 9t:ogotpovoqepd- on nou ogotpe( ro 2'-9rr:o9optr6 mo ouppog6vo tRNA. H ev6ovourledo'q rnq EUUnq e[vot 1-tto srepore- rpopepnq nproteivrl nou qnotel'e(tot on6 6uo xoto- l,uttr6g unoprovddeg, rtq Sen34 rot Sen2, rot 5uo doprrSg unoprovd6eg, trg Sen54 rot Sen15. Ot 5pd- oerq rrlq nopouotd(owot ornv Ell(0NA 21.36. Ot oXett- (6preveq unoprovddeq Sen34 rot Sen2 n5mouv ttg 3' ^oi s' e6oetq oupporprig, owtmo(1roq. H unoprovddo Sen54 (oroq roOop((er ttg 06oetg ronriq "petpri:wog" t4v on6moorl on6 6vo oqpre[o m1 6opr{ rou IRNA' AutS ro orlpre(o Bp[or<etot mov oyrdrvo qq (cbptpng) 6oUnS opi;rotog L. O p6Log qg unogovddog Sen15 6ev efuoiyvt:m6q, ol'Ld ro yov(6t6 r1g e(vot onopo(- rnro yto rn EUUn. To (e(yog Bdoeov nou oplprot[(e- ror ;rero[ti rqg nptirtqs B6onq orq 0r1].td tou owu<trrdtrov(ou xot qq Bdoqq ilou nponYe(tot qg 3' tlt(0NA 21.3? H evdovouxtredon rnq ouppoQnq rov IRNA rolv Apxq(ov r6pet xdOe rtr6vo o' 6vo tpouoxtol-to evdg ;roripou tpotl- orol po-6)rwo-tpouoro p0. 06oqg ouppoqriq e(vot ovoyxo[o yto ulv ron( qq 3' 05oqq ouppoqdq. Mto ev6roq5pouoo 5vdet[r1 Yto rrlv e[6].t[r1 rnq oup- pqgns tou IRNA npoqL0e on6 rg ev6ovouxirsdoeq rov Aplouov. flp6xemot yto opro6q.repri ri ogotxpopepi, oro ono(o rd0e unopovd6o 6Xet 5vo evepy6 r6wpo (nop6tt oro rerpopep6g treroupyotiv pr6vo d0o r<5wpo) nou 16- Ber puo on6 rtg O5oetq ouppoqriq. H unoprovd6o 61et oL- i4l,oux(eg nou oxet((ovrot Us ttq otrI'rlLouX(eq tov evepycirv rSwptrlv orq unoprovd6eq Sen34 xot Sen2 tou ev(riUou rnq EUUq. Om6oo, ro 6v(u;ro tcov oplo(tov ovo- yvrrlp(ouv ro unoorp6ptotd roug pe 6totpopunr<6 tp6no' Awlvq gerpo0vrqv on6oroc4 on6 ouyrexprydveg oMrl- LouX(eq, ovoyvop((ouv 6vo 6optr6 XqpoKrnptorx6 yvro' o16 coq e[6yr<o: go-6ltr<o-e(6yr<o go (bu I ge-hel ix-bu I ge)' H H(ONA 21.37 6e(yyu 6t q r<onq ouprBo(vet oro 60o e[oy- rrigoro. Euvenc0g, 11 ouppocprl rcu IRNA npoqyekot rou 6roXroproproU ttov Aplo(rov Kot rov europutottitv' Av npo- 5ruQe pr6o<o rqg erooyroyrlg tou twpov(ou oto IRNA, ou- t6 np6na vo {tov 6vo no)rU opXo(o ouprBdv. H ouvol,txrl ow(6pooq ouppo<pr1g tou IRNA ouvorf(- (eror orrlv Ell(0NA 21.38. To npoi6wo qg n6tl'r4q e(vot 5uo ypo1rg,n6 rwp6vto ror 6uo ptod g6pto IRNA' Autd ro ev6rdpeoo 6Xouv provodtt<d drpo. Kd0e 5' drpo ro- roLrlyet oe Uto udpo[uLopd6o, ev6 r<d0e 3' drpo oe puo 2',3'-rurtrxri rprrlo<popxrl ogd6o. ' To 6uo prtod IRNA (euyoprirvouv yto to olqpottopt6 ptog 6o;rig nop6protog Ue rou IRNA. 'Orov npoore0e[ ATP, npoyl.rotonotefuot 11 de6tepq ow[6poo4, 11 ono(o r<orol0etoi on6 6vq povodtx6 6v(upro pre noMontr6g ev- (upux5q evepy6qteg. 1. Evepy6tlta xuxLtxfiq EuoEodeorcpdo4q' Kot to 66o oouvriOtoro 6t<po nou nopdyowot on6 tlv evEovourtredorl np6net vo rpononotq0oOv nptv qv ow(6pooq ofv6eoqg. H r<uxl'txrl goo<poptxrl o;rd6o np6ro ovofuet onoS(dowog 6vo 2'-rpoorpo- pu<6 tetrtr6 dxPo. 2. Evepyotqta x:do7q. To npoi6v 6Xet pto 2'-qor- 5',3', FS pS fllS gI'{ hH Phi! h*ffi F$ryh/ H,{ l/- EqoYKouo mm-"-f ;+4-- E[6Yxtouo pd gt /&Ffif,* PS. lr/XopoKog 5' Kontl ll5', Senl 5 - O : Avrtrodtxdvto-tvrpovto (Al) (euyoq pooetlv ..,,r' . ' ::r 0r 3' rot 5' nerfetq oro pretRNA rou S, cerevisiae roroluovrot ono dtorpopertrSq unopovodeg qg ev6ovourtreoorlg' N4ro otrtrn unogovoDo ioolq ro0opi(et qv evrontol rotv Oeoeotv nrlfiq Uerpdrviog rrlv onooroorl ono qv 6ptUtt OoUn. En(onq, ol- povrtr6 e(vot ro (e0Yoq pooeotv Al. 3' Konrl Sen34 ?1."i.* H Iupporprt routRNA flepiloppover Konrlrt Enovoouvdeorloe:eyot0Lc :l - 615
3', 3', i,l i,n ry"pHry'#kry l?l qt,,, z',-3', P H rproogodteotepdorl ovo(yet ro grrrorpoprxd Oorrrjtrro AtruoLDq A),uo(60 tRNA Boon IRNA Boort oft-r'-r',+ 2'-3' xuxtrrxd groogoprr6 3',-0H, 2'rproogoprxri oqoprrrl oprd6o Kor Uro 3'-OH oprd6o. H 5'-OH oprddo nou nopdyerot on6 tr1v evSovour<).edor1 npSna vo eooeopulroOe( yro v' ono6c0oer S'-rptrl- o<popu<6. Aut6 nopdyet pto 05or1 orqv ono(o q 3'- OH Bp(oreror 6(nlo oro 5'-<proogoprx6. 3. Evepy6tryo ttyxdoryq. H oproronollrrl orepot6qro tr1 g noluvo u ri.eol6rruj g oLu o(6o9 qno ro0(ororor ;r6oco qg evepy6qrog l,tyxdoqg. To pr6pro nou 51er ouppo<pe[ efvot ouveX5g, pr' Svov 5'-3' groo<po- prr6 6eopr6 orrl 06o11rnq ouppoqris, q)ild eil(onq 6Xer gro 2'-<p<oogopxrl oprddo nou enrorlpo(ver ro ou;rBdv ndvo oro ouppoprpr6vo IRNA. Iro reLeu- to(o B{pro, outtj q enrnp6o0eq o;rd6o orporpeitor on6 pro g<r:orpordoq, q ono(o ;rero<pdper ro 2'-Ero- ogoprx6 oro NDP g4rlgor((owog ADP 1,2-xuxLu<rl qrooEopxrl prp6(n. H o669 oupporprlg rou IRNA nou neprypdgrlKe no- pondvo e(vor el,ogprilg 6rogopertrrl on6 qv owforor- Xq o66 rrov onov6u),orriv. l-1poto[ dpdoouv or RNA l,ryxdoeg, puo r<urLdoq nopdyer 5vo r<ux.l.rr6 2,3 rux.l.t- 16 dr<po ond ro opXx5 3'-groogoovoeoreprx6 drpo pr5- oo ev6g 3'-odevuLr<o1ldvou ev6topr6oou. H RNA Lryxdo4 en(oqg 6rogripet on6 ro 5v(utrro rnq (6pnq enetdrl pno- pe( vo ouv66ou 6vo 2',3'-xuxLu<6 EroogoSeor6po ror gto 5'-OH yro vo olqtrror[oer 5vo ouprBoru<6 3',5'-rpro- ogo6eoreprr5 6eopr6, oMd ourdg ot owrdpdoerg 6ev o<ptlvouv nLeovd(ov 2'-<p<oogopu<6. Ktrrovog K),ovoq IRNA IRNA 21.19 H Andvrnon oTnv Ilopouoio proq !e6rnAtrlpdvnq flpolrei'vnq Ix€Ti(srot U€ Tn Euppogfi Tou IRNA * H lrel e[vor pro eooreprrfl nupqvLrrl pepppovLq nporeivl pe qv N-retrrxq neproXl Tlq oTov outro rou ER ror q C-retrrx1 neproxn qq 0T0v nuprlvo. H C-retrrq neproX1 5Xer evepyoq- reg rtvoorlg rr ev6ovourtreoorlg. * H npdodeol ptog [edrntrop6vqg nporeivlg oqv N-retrtri1 neproxrl evepyonorel qv C-retrtxq evdovourl,eool Us 0uT0- tproorpop utr[too1. * H evepyonorlpevl evdovourtreooq rripet ro H,4C7 mRNA (Xbpl oto onovdutroro) onetreuOep6vovrog rlvo rvrpdvro ror dqpLoupytirvrog e[6vro nou ouvdeovror perol[ roug on6 pLo tRNA trryrooq. * M6vo ro ouppoptlvo H/C/ mRNA pnope[ vo perotppoore[ o 6vov peroypotprxo nopoyovro o onoiog evepyonote[ yov[6ro yro ouvo6o0g nporelveg nou po10o0v oro 6[n],topo rtllv [edr- ntro.t ptivrriv np toreiv6v. e H evepyonotlpdvq lrel enoyel ononrool rirov ro rurropo nepr6Xer nopo notrtr6q [edrntropeveg nporretveg. 'Evo oouvrlOroro otiorrlpro ouppoqrlg nou olet((etor Ue rrl ouppogrl rou IRNA e(vor 11 o66q ondwqoqg oe H ouppotptl rou tRNA onotre( (eXtoptoreq evepyorrireg vouxtreoolg u e[ovourtreoorlg. To 6pro e[otvfou-Lvrpoviou xopor- rot ono rr1 vourtreoorl nopoyovtog 2'-3'rurtrrxrj rporotpoprrd ror 5'-0H orpo.To rurtrlro qoorpoprrrl ovoryeL onodrbovrog 3'0H xor 2 q,;- oqoptr6g opo6eq To 5 -0H e[vot tptrtotpopu],urt1uivo. Mero qv one],eu0epoorl TOU rvrpovlou, ro pLoo popro IRNA dLntrovovror oe pLo dol* nopol-roro Us Tou tRNA nou r6po 5Xet ;rLo 3'-0H, 5-P 0po0o1 nou orppoyi(eror ono pro trryroo1. 0.16' KE0AAAI0 21 lupporp4 rr Ene[epyoo(o rou RNA H xrvdoq rpoorpoputrloiv€r To retrrK6 5'-0H 6xpo Boorxeg eworeg
(e6rn).oprdveg nporetveg (unfolded protein re- sponse, UPR) nou e(vor ouw4pnpdvn on6 q (tipq 5r,-lg ro OqLoortr<d. 'Onog ouvorp((etot orqv EIK0NA 21.39, 11 ouooril peu oq [e6rn],otrrdvr.ov npulreivcilv qrov qu^.6 rou evdonl.oopolxo0 6rrr[ou (EA) nupoSore( qv o66 UPR. Aur6 odrlye( oe ou(4prdvq ;reroypoqr; yovr6(crlv yro ouvo6orig nporeiVeg nou Bo4Ootiv oro dft).opro rolv nporetvri:v oro EA. Euvenrig, 6vo orjpo npSner vo perodoOe( on6 rov ouL6 rou EA orov nu- pnvo. O oro04rrlpoq Trou evepyonore( trlv o66 e[vor r1 nporetVq lre1, 4 ono(o ewon[(eror mo EA Ko/n rnv eororeprr4 nupqvu<r1 pepBpdvq. H N-reLu<rl neproX4 rqg lrel Bp(or<etor orov ou6 rou EA 6nou ovrXyeUer r1v nopouo(o (e6rnlcr:pr6vov npolreivriv, prdMov g6oro qg np6o6eonq rnq o' erre0erp6vo pror(Bo. To C-te).u<5 9ro6 r4g lrel ewon(erot e(re oro xurrop6nl.oopo e(te orov nuprlvo (L6yco qq ouveloUg pepBpdvqg rou EA Kor rou nuprlvo) xot 6ro06rer 16oo evepy6rrlro rrvdorlg Ser/Thr 6oo ror erdu<r1 evepy6ulro ev6ovour<tredo4g. H n p6o'6eor1 (e6rn).to prSvov npcoreiv6v n po roLe( ou oori- peuon Uovopeprirv lrel orrl gepBpdvq rou EA, odqydrv- roq qrrlv evepyono(qo4 qg C-relu<r1g neproXlq orqv H neproXrl orov quld rou EA npoo66vet qv [edrnAogdvl nporelvrl H neproX4 mov nupqvo / xunoprintroopo ropet ro H,4C7 mRNA +rc dtrl.q nLeupd qg pepBpdvrlg pSoro owogargopuXrrr- onq. H evepyonor4prdvrl C-relrrrj ev6ovour<Ledon 6xe1 npoq ro nop6v, pr6vo 5vo (oqpowu<6) un6mporpo: np6xeLror yro ro mRNA nou rro6rxonore( rov etdtr6 yro rnv UPR nopdyowo Ueroypo<pfq Hacl orn ([Un (Xbpl mo onovdulrord). Yn6 xovovrr6g ouvOqr<eg, 6rov 4 o669 UPR 6ev e(vot evepyonorrlpdv4, ro mRNA rou HAC| nepr6Xer 5vo rwp6vro 252 voux.treorr6(rov (To Xbpl nept6ler 6vo rwp6vro 26 vourd.eortS(rov). Irn (0prf , ro rwp6vro tou HAC| nopeprno6((er rn Ue- tdgpooq tou mRNA oe l,etroupyu<4 nporeivq, evdl oro 0r1Loonx6 ro rwp6vro rou Xbpl ennp6ner Uev rrl petd<ppooq oLl,d 11 npotetvrl ypnyopo ono6ope(ror on6 ro npcoreooopdrro. :rrlv ene[epyoo(o ouro[ rou rwpov(ou epnL6rowor oouvrlOrmo oumolrd ouppo- qnq. H evepyororrlp6wl ev6ovourl.edoq lrel 6po dpre- oo oro mRNA tou HACI (mRNA rou XbpT oro onov6uLrord) rou<6Ber lg 6uo ou;rBol.rig oupporprlg, ogrlvowog 5vo 2',3'-r<ur<Lu<6 googoprr6 mo 3' dxpo rou 5'e(olviou Kor pto 5'-OH mo 5' dr<po rou 3' e(o:v(- ou. 'Enetto, or 6uo oupBoLdg ouvdSowor on6 r1v tR- NA Atyr<dor1 nou 6po orrlv o66 ouppogrig rou IRNA. 'Eror, ol,6x).r1pq r; o669 ene[epyoo(og rou mRNA rou HAC1 (Xbp1) prord(er Ue rnv o66 rou pretRNA. Om6oo, undplouv oqprowx6g 6roqop6g prero[ri rtov 6uo o6drv. H lrel xor 11 ev6ovouxedoq IRNA 6ev SXouv nop6;roro oprol,oy(o ri orimooq oe unopovd- 6eg. H evepy6rqto ev6ovoux..edonq rnq lrel pu0pri- (etot oe peydLo Bo0p6 oro EA ru 61et pr5vo 5vo un6mpopro (pre-mRNA tou HACI). Aw(0ero, q ev6o- vour),edoq IRNA 5Xet no).),d unomptirporo, 6Lo pre xorv6 6(nLopo rou IRNA, pre ;lrprl npor(p4o4 yr' otr- l,4LouX(eg nou neprBdll,ouv rrg ouprBol6g ouppo- ens. Xprlorponortirvrog 1.rro tritoro o66 nop6proro pre rou IRNA yro rnv ogo(peor1 tou rwpov[ou on6'ro m- RNA rou HAC1 (Xbpl), ro riprpro mRNA prerogpd(e- ror Kor. nopdyer 6vov roXup5 preroypoprr6 nopdyowo pe por(Bo Boorrof-<peppoudp Leur[vr1g (bZip) nou npoo65veror o' 5vo r<orv6 prot(Bo (UPRE) orov unoKr- vr1r1 noLlcilv r<o0o6rrd:v yovr6(rrlv. To yovrbrord npo- i6vro npootore(ouv ro x0nopo ou(dvovtog rqv 5r<gpoo4 nporeivrilv nou Borl0otiv mo 6(nLtopo rov nptoreivrrSv. Av ro oUorrlpo UPR roror.truore( on6 [e6rn].opr6veg npoteiveg, n evepyoilornp6v4 neproXrl rrvdorlg r;g lrel npoo6Svetor oro pr5pro-npoooppromrl TRAF2 oro KUr- top6n).oopro yro vo evepyonorrloer rqv o66 qg on6- nroorlq Kor vo Oovorcboer to r0rtopo. 'Erot, to r0rropo Xprlo$onoe( pto oouvqOroq qrpqrrlyxq ene- (epyoo[og rou IRNA yro v' onowrloer oe (e6rn]lr:pr6veg nprrrreiVeg. 0o16oo, 6ev undpXer epqovnq oX6or; ;rero- E0 qq evSovoux.,edor1g lrel Kqr rnq ev6ovour<Ledoqg rnq ouppogr1q rou IRNA, on6re 6ev e(vor npogovSg n6g 0o pnoporioe vo SXet e[e].tX0e( ow6 ro efet6rreu- pSvo orior4pro. | rulr.ooooon I nnooounon I tnr,rn i,v*oon MeroQpoon i i .] HAC! '! j9 . '.;6P I H onovqol oTrlv nopouo(o prog [edrntropevlq rporrelvlg npoyporonorelror Us illv evepyono[101 erbrxqg oup- roqflg rou HACl mRNA yro Tr'1v nopoytrty1 evoq UeToypoQrKou ropoyovro nou ovoyvop[(er rqv UPRE, UPRE ;i.i* HAnovqorloqvllopouo[oprog:e0rntropevqgl-lporeivrlglXen(erorpeqlupoc:-,,-::',- 617
21.2O H Ilopoyrrrvfi rRNA [lpoUno0drer IugBdvro Koniq ror fleprAopBdver Mrrpd RNA * H RNA notrupepool I reppori(er rn Ueroyporprl o' evov reppo- norl 18 pooeolv. * To Ueydtro rot ro prrpo RNA onetreu0eptirvovrol ge ronrl on6 evo rotv6 npo6po;ro rRNA. To 55 rRNA peroypoqerot [eXrrrpr- oro. * H oUodo C/D rtov snoRNA e[vor onopolqrq yro qv rpononol- loq Tnq 2' Oeor1g qq pgo(qg pe npoo0rirl prog ;re0utropo- doq. * H ouo0o H/ACA rov snoRNA elvor onopo(rr1T! yro q UeTo- rpoq rnq ouptdivrlg oe qreudooupr6[vn. * Ie roOe neplnroor1, ro snoRNA (euyoprlver pe gro otrtrqtrou- Xro RNA nou nepr6Xer q pooq-oroXo riore vo dqproupyrloer 1.uo tunrxfl Dop1 nou elvot ro unoorpo;ro yro rrlv rponono(- ton. To r<rlpro rRNA ouw(Oewor oq U6poq ev6g povo6txorl npo-lroyevoUg preroypdgou nou unoBdMeror o' ene- (epyoo(o yro v' onodriloer ro roptpro npoidwo. To np66popro pr6pro neprdxer nq oM4).ouXieg rov 18S, 5.8S, and 28S rRNA. (H ovoproro).oy(o tov drogoperr- r<cbv prBoorr;prrcbv RNA Boo((eroL oe npot;reg prei.dreg ro0((r1or1g nou 6re[4X0r]Kov oe 6roBo0pr[oerq ooKxo- p6(qg ro 1970.) Iroug no).urrirropoug europucbreg, to np66popro pr6pro noipver ro 6voprd rou on6 rnv ro- XUqro xoO((qoqg rou oq 455 RNA.Iroug govorfr- ropou g/otrtyovr<Unopo u g eu xopu c6reg e(vor puxp6re- po (orn (0prl, 35S). To cbprpo rRNA onel.euOeptr:vowor on6 ro np66po- po ;16pro pr6oo evdg ouv6uool-roti ouprBdwrrrv r<onr1g rot owt6pdoeov {,ro}.t6(ouoroq nou ogoLpo0v t6oo ro e[olreptrd 6oo xor ro eooreprKd preroypo<pdpevo dro- orqprotu<d rpqproto (external/internal transcribed spac- 5,ETS 18S ers, ETS ror lTS, owroroiXtog). H Ell(0t{A 21.40 6efi3rer rr1 yevrxrl o66 orrl (0;rq. YndpXouv 6rogop6g orq oerp6 rrrrv ouprBdwrrlv, oMd nop6proreg owrdpdoerg eprnM- Kowor oe 6Loug rouq euKopu6teg. To neproo6repo 5' drpo nopdyowor dpeoo ;16oo ev6g oupBdwog ronrig. To neproo6repo 3' drpo nopdyowor prSoro ron{g nou oroLouOe(ror on5 puo 3'-5' ow(6poon qrofu6(oporoq. Aurdg or 6repyoo(eg r<o0op((owor on6 noMd crs-6po- orrrd prot(Bo mo ETS ror ITS ror dre[dyowor Ue rrl 6pdon > 150 nopoy6wcov ene[epyoo(og. l-lotrLdg prBovouxLedoeg epmL5xowqr ornv ene(ep yoo(o rou rRNA, pero[U d],],rov opropr6vo oumorxd rou e(roool.rqr(ou, ro ono(o e(vqr pro ouvd0poropo op rerc0v efrovouxtreoocbv nou enloqg outrrprerSler mrlv ono66;rqoq rou mRNA (FL6ne Ev6-r4ta 22.5). MeroMd- [erg oe (eXoprord 6v(upro ouvr1O<og 6ev nopepnodi(ow qv ene[epyoo(o, yeyov6g nou uno6r1],civer 6l or evep y6r4t6g roug e(vor nl,eovd(ouoeg ror 6n yro rnv nopo- yoyf rov drprprov prop(rrlv prnope( vo Xcrlorponorrl0o0v 6rogoperxoI ouv6uool.ro[ ndryeov. Yndplouv ndwo nollonl,d owrypogo rnq Uero- ypocpxrlg provddog yro ro rRNA. To owiyporpo opyo- vrivowor rog ruXo(eg enovoA4Qerg (BA5ne Ev6qta 7.8). To yov(6ro nou ro6rr<onoro0v rRNA preroypdrpowot on6 tqv RNA noLuprepdoq I orov nupqv(or<o. Aw[Oero, ro 55 RNA peroypdrperor on6 [eloprord yov(6ro pre rrl 6pdoq qg RNA no).uprepdorlg lll. l-evrd, ro 55 yovi6ro e(vot ouyrewptrlpr6vo, oMd 6roXrop((owor on6 ro yovi- 6ro yto ro r0pto rRNA. Yndpler puo drorpopd orqv opydvroon rou np66po 1.rou 1.rop(ou oro Borcrripro. H oMrlLoul(o nou owrorot- 1e( oro 5.8 S rRNA oXqtr-ror[(er ro 5' dxpo rou preydou (23S) RNA, dpo 6e ouprBo(ver ene(epyoo(o ;rero[ri ou- rdrv rtrlv oLLq).ouXrcirv. H EIKONA 21.41 de(per 6n ro np66popo prdpro nepdXer en(or1g ro 55 rRNA r<r 6vo 4 60o IRNA. Zto E. coli, ro 7 onep6vrq rrn e(vor drdonog ro oe 6l.o ro yovr6[tr;1-ro. T6ooeprg yevetxo( r6not rrn nepr6louv 6vo yov(6ro IRNA peto(ri rrr:v oMrll,ouXrdrv 165 ror 23S rRNA r<or or dMor yevenxo( r6nor rrn ne- E OC lTSl r'eu lTS2 28S 3, ETS Evdovouxtredorl i t Evdovourtredorl I EvdovouxLedoetg t * EErovourtredon E[rovourtredoetq ++ Ev5ovoux.edor1 |re 185 E[tovoudte6oerg ++ 585 : ' To topt;to europuortro rRNA nopoyovrot ono ro nporoyev6g peroyporpo peoto oupBovrov rondg ror rl.ro),rdrogorog. 618 KE0AAAI0 21 Iuppotprl rr Ene[epyooio rou RNA Boorxeg eworeg taa
yov[dro yro rRNA roL IRNA. To orpLBeq plrog rov pe- royporprrrv e[oproror ono roug unorrvlr6g (P) xot rep;loloreq (t) nou XprtorUonorouvTor. Ko0e npor6v RNA np6ner v' onetreuOepoOer ono ro geruyporpo 1r' evrop6g oe roOe n),eupo. pr6louv 6uo yov[6ro IRNA oe ouqv qv neptoXrl. Mept- r<69 qop6g, peto[ri trlg o].].qLouX(oq 55 rot rou 3' drpou ;rnope( vo undplouv enrnp6o0eto yov(6ro IRNA. 'Eror, ot owr6pdoetg ene(epyoo(oq nou onorroriwor yro rrlv oneLeu05poorl rov npoi6wrov e(oprrirwor on6 ro nepeX6prevo rou ouyrerpql6vou yevenroU r6nou r- rn. T5oo oroug nporopurrireg 6oo ror oroug europuri- reg, rord qv ene[epyoo(o tou rRNA nopdyoweq ene- [epyoo[og ror prBoorol.ur<69 nporeiVeg (iocog xor dMeg nproretveg) npoo66vowor oro np6dpo;ro ;r6pro, on6re un6orpopo yro rnv ene[epyoofo 6ev e(vor ro e],e0Oepo RNA oLLd 5vo prBovouxleonproreivrr6 o(;rnl.oro. 'Onog q ene[epyoo(o rou pre-mRNA, q ene[epyoo(o rou rRNA npoyporonoekor oe ouylpovtog6 Ue rn Ue- roypo<pr1. 'Eror, or nopdyoweg ene[epyoo(oq 6ron].6- Kowor pe ptBoorol.ux6g nporeiVeg oX41"rot((owoq ro prBooroprdrro, ow[ npdlro vo ene[epyd(owor r<r 6nemo vo ouvoplloLoyoriwor oro6roxd ndvo o' ene(epyoopS- vq rRNA. H ene(epyoo[o ror rpononofio4 rou rRNA npouno- 06rer pro ror4yop(o ptr<priv RNA nou rqloUwor snoR- NA (small nucleolar RNA, pmpd RNA rou nupqv[orou). Iro yovr6(opro rou S. cerevisiae Kor rov onovduLorcbv undplouv exorowd6eg snoRNA. Opropr6vo rrodrxonor- o0wor on6 [eXoprord yov(6ro, dMo erEpd(owor on6 nol,uormp6vto r<or nolLd npodpXowor on6 twp6vro trov yovrS(tov-[evrmciv. Aurd to snoRNA ug[mowor no).6nLorq ene[epyoo(o ror orp(povo4. Opro;r5vo s- noRNA onomo6wot yro rnv xon{ rou np66poprou prop(- ou oe rRNA, 'Evo nopd6erygo e(vor ro U3 snoRNA, ro ono(o e(vor onopofurpo oro npri:ro ouprBdv ronrlg. To oUpntror<o nou nepdlet U3 owrororle( oroug urellKoUq r6pnoug, mo 5' dt<po ttov veooXqprott(6prevov rRNA peroyp6tprov, ro ono[o efuor opord oro rltrempovu<5 pu- rpoor6nro. Ae yvop((oupre n p6l,o no(er ro snoRNA orqv n6q4. Oo prnoporloe vo e(vor onopokrpo yro ro (euydpopo pe erdxSg oLL4Lou{eq rRNA tirore vo q1n- gorrme( gro 6eweporoy{q 6oprf nou ovoyvop((eror on6 puo ev6ovour).edo4. Auo ogddeg snoRNA onono6wot yro rrq rpononor- rloetg nou npoygoronoroUwor oe Bdoerg rou rRNA. To p6Lq rdOe o;rd6og ovoyvrop[(owqr on6 no].ti pur<p6q ouw4pqpr6veg oMqLoul(eq Kor rotvd loporcrrlpronrd 6eureporoyo0s 6o[nq. H opd6o snoRNA C/D onotrefuqr yro rrlv npooOrp<r1 tRNA tRNA Pl P2 165 rRNA 23S rRNA 23S rRNA ilt2 165 rRNA 23S rRNA I3OS RNA llpotrivro **Y' 5S rRNA Y tRNA proq preOuLoprd6og orrl 2' 06o111119 prB6(49. Ie ouwq- pqp6veg O5oerg rov rRNA r<ov onov6ul<otrilv un6plouv >100 2'-O-pre0u),opddeg. Aur{ q oprddo snoRNA no(p- ver t' 6voprd t4g on6 6rio prxpd ouwqpr;pr6vo pror(Bo otrL4LouXrc0v nou rotro[wor rou'nd C xor D. Kd0e s- noRNA nepdler pto oLi.r'1Lou/o rowd oro rour( D nou e[vot oupn].r1po1.rotrq Ue pro neprolq tou 18S 11 rou 28S rRNA nou e(vqt pre0ui,uopr6v4. H onri:trero ev6g ouy- rerpql6vou snoRNA nopeprnod(er q preOuL(roon ornv neproXri tou rRNA Ue rrlv ono(q e(vor ouprntr4pco1.rorr6. H EIK0NA 21.42 6e(yyer 6n ro snoRNA (euyoprirver pre ro rRNA yro vo 6qgroupyrloer q 6(xLovrl neproXrl nou ovoyvrop((eror og un5orpopo yro rq peOuL(rrloq. H Ue- 0ul(ooq ouprBofuu prSoo orqv neptoXrl ouprnLqprrlpron- r<6111o9 oe Uro 06oq nou on61er oxprBcirg ndwe Bdoerq 5'rou rowto0 D. l-lt0ov6v, xdOe oupBdv pre0ul,(too4g ro0op(eror ond 5vo 6torpopenx6 snoRNA. Ie owrlrrlv rponono(qo4 epnl,Srowor toutrdltmov 40 snoRNA. Kd0e C+D rout( snoRNA ouv66eru Ue rperq nproret- veg Noplp (oro onovduLrord, gq.rnprMopwn - fibrillar- in), Nop56p r<or Nop58p. Or peOuldoeg dev SXouv XopoKrrlproref nArlpog, nopdrr q rUpro snoRNP npo- re(v4 Noplp/grpnprMop(vq e(vor doprxd nop6proro pre ng pe0utrorpovogepdoeg. Mro dLLq opd6o snoRNA epntr5reror srrlv rpono- RNA-m6Xog 3, RNA-ortlXog EIKONi{ 2'!.42 'Evo snoRNA (euyoprirver Ue Uro neptoxn rRNA nou npdretrot vo peOutrrtll0e[, BoxD' ctD snoRNA box D 21.20 H llopoyroyrl rRNA tlpoUno06rer luppdvro Komjq ror llepiloppdver Mrxpd RNA 619
00H P/ EIK0NA 21,43 H ouprdtr4 gerorp6neror oe tpeudoouprd[vq owr- ro0rortrlvrog rov N1-oorXopurr6 deopo p' 6vov C5-oorXopLnr6 0eopd roL neprorpdgowoq rn Boq oe oX6oq pe ro ooxXopo. no(r1o4 Bdoeov prerorp6nowoq rnv oupr6(vq oe r,Ueu6o- oupr6(vq. Yn6plouv -50 xotdlorno oro rRNA rnq (6UnS r<ot -100 oro rRNA rov onovduLrotcbv nou rpononor- oUwor ;r5oo rpeu6oouprdw(aroqg (pseudouridination). It4 ouyrerprprdvq ow[6poo4 {EIKONA Z1.4tl, o N1 6e- opr6g prero[ri tou oupr6u],rro0 o(5og Kor rnq prB6EnS ondet, 11 Bdo4 neprorpdrpetor r<or o C5 enovoouvd6erot oro odrlopo. O oXrlponopr6g qUeu6ooupr6(vr1g oro rRNA onome( qv oprdSo rrov H/ACA snoRNA. l'lo(pvouv t, 6voprd toug on6 rrlv nqpouo(o truog tprnAsrog ACA oe on6- oroorl rprcbv vourLeor6(rrlv qn6 to 3' drpo Kol. proq Ue- prrdrg ouvt4prlpr6v4g olLr1trou1(og (r<our( H) nou Bp(oreror prero[ri 6uo 6o;rcbv goupr<6rog pr(o1ou-0q- l,rdg. Ko06vo on6 owd to snoRNA 61er pro oM4Lou(o oupn).4pto1.rolrq pre ro rRNA prd,oo mo trrioXo rdOe goupr6rog. H EIKONA 21.44 6e(yyer q 6opri nou 0o no- poy6rov trrdoro rou (euyopcbproroq pe ro rRNA. Mdoo oe rdOe neproXrl (euyopd:prorog, undpXouv duo o(eu- ydpcoreg Bdoetg, on6 lg ono(eg q pro e(vor oupr6(vr1 nou perorprlneror oe rfeudoouprd(vq. snoRNA 5' dr l-1 : .. . To H/ACASnoRNA eXouv duo Bpoxereq ouvTnpl- g6veq otrtr1trouy[eg ror duo 6opeq qouprerog, ro0epro on6 rrg onoleg eXer reploxeq oro pfoXo nou e[vor oupntrqptopolxeg pe ro rRNA. H Qeu0ooupLdivq oXrlpol(erot UeToTpenovToq pLo o(eu- yopotrrl ouprDLvrl luioo orrl oupnLqpotporLrl neptoXrl rou rRNA. 620 KEQAAAI0 21 Iupporprl xt Ene(epyooro rou RNA To H/ACA snoRNA ouv66owor pe r6ooeprg er6u<dg npcoreiveg rou nup4v(orou: Cbf5p (oro onov6uLrr:rd, duorepfuq - dyskerin), Nhp2p, Nop10p r<or Garlp. E(vor orlpowu<6 6l 11 Cbf5p/6uorep(vr1 e[vor 6opurd nop6- Uoro Ue yvroordg ouv0doeg rfeu6ooupr6(vr1g ru 6ror nr- 0ov6v nopSler r4v ev(u;rrrr1 evepy6rrlro orrlv roOodr;yoriprevq on5 snoRNA ow6pooq qUeu6ooupr6r- v(too'r1q. I-loMd snoRNA Xpnorponororiwor en[or1g yro vo xoOo6rlyrloouv rponoTrot{oerg Bdoeov oro IFINA Kor sro snRNA nou epnl6rowor orrl oupporp4 rou pre-m- RNA. Aurdg or rpononot4oerg e[vor r<p(orpreg yro ng Aer- roupy(eg rouq olq owfmorleg owr6pdoerg. 0o16oo, noMd snoRNA 6ev 51ouv e;rgove(g or61oug. Awd ro snoRNA r<olorivror opEava RNA (orphan RNA). H Unop(r1 roug uno64Lcbver 6rL notrtr5g BroLoyrx6g 6rep- yooleg (oog 1p4or;ronoroUv prrllovroproUg r<o0o6qyo6- Uevouq on6 ro RNA yro vo rpononorr;oouv Leroupyrxd dtri.o er<<ppo(6prevo RNA pe nto nou<[],oug tp6noug on' 6,tr yvop(ou;re oripepo. 21.21 IlepiAn{n H ouppogq enmuyXdver qv ogo(peoq rtov rwpov(rrrv xor rq oUvdeon rov e[covirrtv orrlv c0pq-rq oM4].ou1io rou RNA. Yndplouv roui.dlrorov r6ooeprg r6nor ow[- 6pooqg, nou 6ror<p(vowor on6 rg onomqoerg roug rn vitro xot ro ev6rdpeoo nou nopdyouv. To ouorrlgcrro nepil.ogBdvouv ro euKopuortKd nuprlvrxd rwp6vto, ro rwp6vro o;rd6og I r<or II Kor ro rwp6vro rou IRNA. Kd0e ow(6poor1 ouv(ororot oe oMoy4 opydvoorlg, pr6oo o' 5vo (eXo:pror6 trl6pro RNA, dpo e(vor dvo cis- 6pooru<6 ougBdv. H ouppog4 rou pre-mRNA qKo).ou0e( npol;rcirpre- veg oMd 6Xr unoXpeonr5g o6orjg. Anopo(rrlteg e(vor p6vo noLU pu<p6g ouvorvettx6g oLl,rlLouX(eg, evc6 or un6Loneg oMr;l,ouX(eg rcov rwpov[cov e(vor odrdqopeg. Oqr6oo, oLl4Aoul(eg 16oo rrov e[otv(rov 6oo ror r<ov rwpov(ov prnope( vo enqpedoouv 0etxd 4 opvrlnr<d rqv enil.oyrl rnq yerrovrKrlg 05orlg oupporp4g. O).eg or 5, O5oerg ouppogrlq pdMov e(vor roodtjvopeg, 6ncog ror 6Leg or 3' O5oerg ouppocpqg. Or onopo[rrlreg oM4],ou- X(eg 6(vowor qn5 rov xov6vo GU-AG, o ono(og nepr- ypdger ro drpo rou twpov(ou. Anopofu4rrl e(vor enioqq n e6on droxlddrrrorlg (UACUAAC orn (UUn, ri pro Lty6- repo xoAd ouwrlpqprdvq ouvorverrrrl oMrll.ouX(o oro rwp6vro rrov 0qLooru<rrlv). H owidpoorl pe rrlv 5, 06o11 ouppogrlg nepil,opBdver ro oxnpqrropr6 ev6g Adoou nou evdlver ro GU dxpo tou rwpov(ou ;r6ocrl 2'-5' 6e- opo0 trre rrlv A orrl 06o11 6 qg 05or1g 6rorLddolorlg. 'Eneno, ro 3'-OH drpo rou e{ov(ou enn(Oerot orrlv 3' 06oq ouppo<pr1g, 6ror cilore vo ouv6e0ofv ro e(c6vro ror ro rwp6vro v' oneleu0epo0e( un6 propE4 Ldoou. O oXrlgotropr6g Ldoou e(vor unerj0uvog yro rqv enr).oy{ t4g 3' 06oqg oupporprlg. Kor ot 6uo owr6pdoerg e(vor trans-emeponor{oerg ortg ono[eg ouw4porjwot gro- oqodreoreprxo( beotrrof. Apr<erd or6bro qg ow(dpoorlg ononoriv 11 u6p6luoq rou ATP, nr0ov6v yro vo npo. 0 P s /!-'-' I e(rr ")-o.i i( )r" w00H P/ 0 ']* /'*, nspmpogn 180 oC -)> Iuv0dorl qeu0ooupt6tuqg P I o -'- I I RNA-orrlXog
oenoouv olloydg dtopr6prpotoqg oro RNA rot/n oe nproreivtrd ouorortrd. Evotrtrorrtrd np6tuno ouppo- griq nporotrotiwot on6 npcoteivtrorig nopdyoweq nou efue dteye(pouv q Xprlorl puog v6og 05or1q efue TIopeU- no6[(ouv rn Xpnon qg npoenil.eyp6v4g 06or1q. H ouppogq tou pre-mRNA npouno0Stet ro oxrlUo- roprS evdq oorptou6(ou ouppoqqg - 6vo peydlo oopo- t(6ro nou ouvopptoloye( ng ouvoweru<69 otrtr1ouX(eg oe dropr6p<pcooq r<or6Mql,q Ytq vo ou;rBe( ow[6pooq. To oropoilEto ouppoqrlq 6rlprtoupye(tot on6 tr; 6tepyo- o[o opropori twpov(cov, 4 ono(o nept],oprBdvet ovoyvtil- pron rnq 5' 06o119 ouppogriq, qg 05or1q 6toxLd6toorlq Kor rrlq 3' 06o119 ouppoqrlg. Aut6 toltiet yto 1-ttrp6 w- rp6vro, n.X.rnq (riUnq. Av, toor6oo, to twp6vto efuot preydl,o, 6nrog to twp6vto tov onovduLrotdlv, n ovo- yvdpron tov O5oerov oupporprig orotrouOe( q 6epyo- o(o rou optoproU efov[ov, nou nepil.ol.tBdvet oMq]'e- nrdpdoerg 6ro;r5oou tou e(o:v(ou ovdpeoo orqv 3' 05oq ouppogrlg Kot rrlv roOodx4 5' 06o11 ouppoqrig. Aw6 6nemo peron(met oe ou(euypdveg owtdpdoetg 6topr5- oou rou rwpov(ou yto ro petoyevdmepo mddto rqg ouvopprotr6ynorlq rou otr:1.ton6(ou ouppo<pt'1q. E(te pe rov oprop6 rwpov(<ov e(te pe tov optop6 e[ov[rov, q op- Xrr4 depyoo[o ovoyvd:ptoqq rqg 06oqg oupporprlg 6e- opre[er to pre-mRNA un6orpo;to orqv o66 ouppotp4g. To orj;rntroxo pre-mRNA nept6Xet U1 snRNP Kot opt- qr5voug rp(orproug npcoteivu<otig nopdyoweg ouppo- <pr]q, pero[ti ttov ono(ov o U2AF-rot o nopdyowog nou npooddveror orrl 06oq 6toxl.d6cooqg. Ito nolut<frropo euxopurr:nrd x0rropo, o oxrlportou5g tou outrtntr6rou 65opeusnq onotte( r1 ouprtrreroXri rov npoteivdlv SR. To oo;rot[dto oupporpqg nept5Xet to U1, U2, U4lU6 r<ot U5 snRNP, 6nolg enioqg rot ;reydtro optOpr5 nopo- y6wrrlv ouppoqrjg. To U1, U2 t<ot U5 snRNP nepdXouv 6vo provodtr6 snRNA rot opretdq nptotefveg. To U4lU6 snRNP nept6Xet 6[o snRNA t<ot opretdg nprrlteiveg. Oprou5veq npotefveg e(vot t<owdg yto 5tro to orrrpo{- 6ro snRNP.To U1 snRNA (euyopdrvet pe rnv 5' 06o11 ouppogrlg, to U2 snRNA Us rnv oMrllouX(o 6ror<Ld6o- onq Kor q U5 snRNP ouyrpote(rq 5' Kot 3' 06oetg oup- poqns ;16oo prtog oLLrlLoul[og Oq]'tdq pSoo oro orrlpor[dro ouppoqrjg. M6]lg to U4 qne],eu0ep6oet to U6, ro U6 snRNA (euyoptilvet pe rnv 5' 06o4 ouppocprlg xor to U2, nou nopogSvet (euyopcop6vo, ge rqv oMrl- l,ouX(o 6toxtrd6trroqg. Aut6 nt0ov6v drlprtoupye( ro Ko- rol,utrr6 rdwpo yto rn oupporprl. Mto evollorcrtr<r1 oprd6o snRNP nop6Xet ovdLoyeq Lettoupy[eg Yto rrl ouppoEq t4g U 1 2-e(opttirprevrlg unor<oqyop(oq twpo- v(tov. O xotol.uttr6g nuprlvog Oup((et tov owlorotlo nuprlvo rov ouroKoroluttrriv twpov(ov opddog ll. Iu- venri:q, to otol.tot(dto ouppo<p4g prnope[ vo e[vot Uto Yt- ydwro Unxovn RNA (6ncog to ptBoocoprdtto) orrlv ono(o to Booxd ororle(o RNA Bp(orowot oro rSwpo rnq ov- il6pooqg. H ouppotp4 ouvrlOog e(vot evSoptoptorq. 0o16oo, oro rpunovootilporo Kot ro vrlporr664 orouLqrto oup- Bo(ver trans-ouppoqn (dtoproprorrl ouppotp4). l-lept}'o;t- Bdver ;uo ow(6pooq prero[6 ev6g prtr<poti SL RNA rot rou pre-mRNA. Ito oxoul.rlrto undplouv 6uo tfnot SL RNA: o 6voq Xprlot;ronoteftqt Yto'rn oupporpr; mo 5' drpo ev6q mRNA xt o d).l.og Ytq'rrl ouppogn oe Pto eoorepwi 05or1 cirore vo dtoonoore[ ro notruotorpovt- 16 np66popo RNA. H erooyoyri"rou SL RNA o' ene(ep- yoopSvo mRNA nop6Xet onopo[tqro o41-toto Yto rr'l prerd<ppoo'q. H xov6tlto tepprottoprori r1g RNA nolupepdoqg ll e(vor mevd ouvde6ep6vn Ue ro olrlpottopr6 tou 3' 6rpou tou mRNA. H oM4Loul(o AAUAAA, nou ewon[- (erot 11-30 Bdoetq ovo6md rnq eSonq ronrlg, nopdXet ro orlpro yto xon4 on6 pto ev6ovout<Ledon Kil nol'uo- 6evu(trloq on6 r4v notru-A nol,u;rep6oq. Aw6 evtoXrie- tor ptSoto tou oupnL6Kou nou e(vot deotr;eupr5vo mo nLoUoto oe G/U orotXelo to ono(o Bp(oxerot rq0o6Kd rrlg 06oqg ronrlg. H ;retoypogrl tepproil(etot 6tov 1.tto e[covoux].edoq nou npoo66vetqt oro 5' dr<po tou RNA to ono(o opXi(et vo dr1;rtoupyefuot on6 trlv r<onri npo- <ptdoer qv RNA nol,uprepdorl. Otro to ;ret6ypotpo Pol ll e(vot notruo6evul,tcrlpdvo, er<r6g on6 ro mRNA tcov torovdlv, nou orjte nepr6louv rwp6vro o6re onomoUv oup6 nol.u-A. O oXrlpottopr6q rou 3' 6r<pou rou mRNA ttov torovr.riv e[optdtot on6 ;to 6opq pr(oXou-Or1Ltdg t<ot on6 to (euydpotrro ev6g xo0o6rxori o.rotXe(ou pre to U7 snRNA drme vo npoerot- proore( puo ron{. H 6opri pr(oXou-0q}.tdg nt0ov6v npo- sroteUer to 6rpo, 6nog oro Botctipto. H ouppo<pri rou IRNA neprloprBdvet [elroptmSg ov- ndpdoetg evdovouxl,edonq Kot llyrdoqg. H ev6ovou- rledorl ovoyvtrrpi(et 11 6eweporoYn (n tpttotoyrl) 6oprl rou np66poprou pop(ou rot ndntet rot ro 60o drpo rou rwpovfou. To 6uo prtod IRNA nou oneleuOe- pri:vowor pr6oo qg on6tretog tou twpov(ou ouv65ov- tor on5 qv IRNA Ltyrdoq nopouo[o ATP. Autl 11 o669 u:pf;rovorlg tou IRNA oftonoe(tot on6 qv o66 ondw4- onq oe (edrn).ctprSveg npote(veg oro EA. H ene[epyooio tou rRNA npoypotonotefuot orov nupqv[oro, 6nou to U3 snRNA nupodore( pto oetpd dpdoetov ev6ovoux.treoori:v rqt e(ovoux.Leood:v nou 16- Bouv r<ot qrqfud[(ouv entnp6oOereq o].trr1]'ou1(eg mo np66popro rRNA, nopdyowoq felcoptm6 p$ootogort- r<d RNA. Er<orow66eg 6o9 ltltddeg ptrl r<codxonoqtxd RNA exgpd(owot oro europuconrd xrirropo. Itov nu- pqv(oro, 6uo oprddeg t6totcov pr1 ro6tronotrlttxtilv R- NA, nou ovoprd(owot snoRNA, e(vot une0Ouveq Yto ro (euydpopo ;re rRNA oe O5oetg TIou rpononoto0wot. To snoRNA t4g oprddoq C/D ovoYvtop((ouv 05oerg-m6- Xoug ytq preOuL(ooq, evc0 ro snoRNA qg optddog H/A- CA r<o0op[(ouv ttg 05oetg 6nou 4 oupt6(vr1 perorpSnerot oe tPeu6oou Ptdtvn. BrBAroyporpio :i i rir-:t,ti ttrrl I Lewin, B. (1975). Units of transcription and translation: sequence components of hnRNA and mRNA. Cell 4,77-93. BrB),.:.::: : 621
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Ero0ep6rnTo Kor Xropo0drnon Tou mRNA ffiHS ErooVoyn W To Ayyetrrorpdpo RNA E(vorAoro0rl Mdpro * H ooroOeto rou mRNA oqeiLeror oq 6poo1 prpovou- rtreoo6v. a 0r prBovour),eooeq droQ6pouv oq rpoq rrlv npor[pr1orl yro ro un6orptop0 Kor Tov rp6no eni0eorlg. * To mRNA nopouoro(ouv eupe(o drorupovorl ro:v Xpo- vtrrv 11p(0r1q. * H dLorpopun oro0epdrr[o rtov lroplrov mRNA enr;peo- (eL rr1 ouyx6vrptoo4 roug, ouven6q rot ro qoogo rov nporeivdrv nou nopdyovrot oro rUnopo. ffiffi To Europuonro mRNA YndpXouv un6 Moptprl mRNP ono q l-evvr1or1 6o9 ror ro 0ovor6 roug s To mRNA otrtrqtrentdpo g' 6vo peropotrtr6pevo ntrq0u- opd nporreivdv rord rrlv orpilrovor'1 Tou 0T0v nuprlvo Kor Tnv nopoUovr1 rou oro rurropontroo;ro. * Meptr6q nporetveq rov nupqvxtitv mRNP 6Xouv ptitro rot oro rurropontroopo. <e Ynopxouv nopo notrtr6q npulreiveg nou npoodSvouv RNA. 0r neptooorepeg dev SXouv Xoporrqprorel ordgo. * Atotpopelro mRNA oXeri(ovror pe [eXoprordq, otrtro otrtrr1troenxotrunr6peveg, oU0deq pu0Urolx6v nporei- vtilv, DqproupyrirvToq T0 RNA peyroutrdvro. 626 KEOAAAI0 22 lro0ep6rnro rot Xropo06rnon rou mRNA o Equinox Graphics/Photo Researchers H Anoddprlorl rclv flporopurolrdrv mRNA l-lepilop$oveL l-lotrtro'Ev(upo e H onodounq rov porqprordv mRNA opxr(er ue oQoL- peol ev6g rupoQooqoprKou ond ro 5'drpo. * To Uovotprrrorpoputrulgevo mRNA ono6o;to[vrot xoro rr UeroQpool o' 6vo rurtro Duo prlporu:v nou nepiloppo- ver evdovourtreotrurrr6g ewopdg, ttg onofeg orotrou- 0e( n5(l4 rolv rpqporov nou npoxunrouv oe roreuOuv- ot 3'-+5'. e H 3'notruodevutritool gnopel vo dreurotruver Trlv 0n0- 6opr1o1 rtrooporov mRNA nou nepr6Xouv deureporoy'; doun. * To rOplo ev(uuo Uq onoddunolg Leuoupyo0v oq ouu- ntroro nou ovo go(eror orr.rpor(6to ono6o gqorl g. To [lepLooorepo Europuorrro mRNA Anodopouvror Mrloto Auo Movonou6v E{oprtirpevtov ono flo},uo6evu- trioo1 * 0r rpononorrloerq Kor oro 6uo orpo rou mRNA ro npo- ororeuouv ono q 6poor1 rov e[ovour],eootirv. * To 0uo rupro lrovonorLo onoouvOeorlg rou mRNA opXr- (ouv pe onoodevutrltrlo1 11 onoio rorotrueror ono no- tru(A) vourtreooeq. * Mero rqv onoodevutr[oor1 pnopei v' orotrou0rloet e[re oqorpeol qq rotrunrpog ror 5'-+3' n6(rr1 ono ev6o-
vouKtrooon, siTe rdqJn ord eqovouKtreoorl oe K0TeU- 0uvo1 3'-+5'. o To 6v(uuo orpo(peonq rqg rotrunrpoq ovroyov[(etot pe ro oupntroxo 6vop[qg rnq UerdQpoonq yro npdodeon oqv 5'rotrunrpo. * To e[ulotoudlo, ro ono(o rorqluer qv 3'-+5'n6Un rou mRNA, e(voL 6vo peydtro, e[etrrrlrd ouwqplp6vo o[p- ntroro. e H ono66utlotl ouppofvet o' euOtorpuo rutopontrooport- ro oopor[6ro nou ovopo(ovror orrrpor6ro ene[epyoo(og. @ nffo Movonolo Ano6riprlorlg lroXe0ouv Iuyrerprpdvo Mopro mRNA e T6ooepo enrnpdo0ero povondlo ono66pr1or1g enue- trouv q pu0;u(opevq onodoglorl erdLrtlv mRNA. o Atpo[peorl qq rotrUnrpoq ove[dprnrrl on6 qv onoode- vutrltoor1 e[etr[ooeror rlrov unopXer pro lroxpro notru(A) oupd. * H qno66unol rov un notruodevutrur:1.uivtr:v mRNA ro:v rorovdrv nupodorelror ge qv npoo0rlKrl Uroq notru(U) oupoq oro 3'orpo. c H ono66uno1 oprop6vrov mRNA pnoper v' opX[oet ge n6fi ond evdovourtreool erdrq yro qv otrtrr1trouXio 11 q doPr1. * 'EvoS oyvtrlorog opt0gdg uop(tr:v mRNA oroXeuovrot yt' onoddgnon n Heroqpoolrn roroorotrrl ond microRNA. ffi O Erdrrdg Xpovoq HU(urnq Ko0e mRNA Etr6yXeroL ono AtrtrqtrouX(eg q Aopeg M6oo oro mRNA * Elduo crs-ororXe[o evrig mBNA en4ped(ouv rnv roxri- Tnro Trlq onoddp4o4g rou. * frorxe[o onooro0eponoi4o4g gnope( vo enuoXtlvouv rr;v onoouvOeorl rou mRNA, ev6 ororXe(o oro0eponof- lonq vo qv neptop[oouv. * To n),orjoto oe AU ororXe(o e(vor rorvd motXelo ono- oro0epono(4or1g oro 0qloolro oro ono(o npoodevov_ ror notx[],eg npo:reiveg. * Optopeveq npoTe(veq nou npoodevovTor or ororXe(o onooro0eponoirloqg otrtrr.1trentdporjv ge ouoronrd qg Uqxovng onoouv0eoqg roL potrtrov ro enrorporeUouv yr'ono6opr;0r1. e lrorxe[o olo0eponoqolg undpXouv oe oprop6vo notr( oroOepo mRNA. * H roxut1ro ono66p1or1g rou mRNA pnopet v, otrtro[er oe onovrqol npog norr[tr0 oriporo. ffi fo Neoouvre0erpevo Mrlpro RNA EtreyXovroryro Etroruo- poro MSoto Evog llup4vuorj Iuorlgorog Enurlpqorlg * To etrorrotpolrti nupqvrrd RNA ovoyvropr(ovTor Kor K0_ roorpdqovrot ond 6vo ouorrlpo enunplonq e To nuprlvtr6 elooouorto tretroupyeI rooo <o:: .*, ene[epyoo(o rpuorotroytrdv RNA unoorpoigtoTurv,:: ror xoro TIV KoTooTpoqq rov etrorrtrlltoiu6v RNA. s Iu (uul, ro oUpnl,oxo TRAMP entorporeuet ro e[o- orol-rono oe ovtopotro popto RNA xot 6teuro),uver qv 3'-+5' e[trlvourtreotrunrf 6poot1 tou. a Iro unootpriruoro yr' onoDdurlon 96oo rou oupnLrkou TRAMP-e[ooropotlou nepilopBovovrot ptrl ouppoppevo 1 ovrirpotro ouppoppevo npo-mRNA xot tro0og reppo- ropevo perdypoqo qq RNA notrupepoonq ll Xtopiq no- Iu(A) oupo. e To neprooorepo geroyporpo rrlg RNA notruuepoonq ll [oog e[vor rpuqo ooroOq peroyporpo nou roroorp5- govror yprlyopo orov nuprlvo. ffi O'EtreyXoq llororrpog qg Meroqpooqq rou mRNA l-lpoy- goronore(ror ono Kurropontrooportrd luor4poro Enurl- pnortq * 0 unxuvtoudg onoouv0eoqg NMD oroxeuet mRNA pe npoorpo rtodrrovro reppouogou. * H oroxeuorl Tov unooTpouororv rou NMD onouei 6vo 6torqpqp6vo ouvotro nporeivtirv UPF ror SMG. " Ie notrtrofq opyovrolroug, urtrerd0, yro rr;v ovoyvdrprorl npooptrtv rodrrov[trrv reppologou e(vor 11 oouvrl0rorrl doUl n ro oouvrlOroro pqrog qq 3' UTR. Iro 0r1trool- ro, o 1r4Xovto1r6g ovoyvtirprorlg entoqg o[totroye( ror qv nopouo[o roOodutirv oupntrdrtov oupBotrqg e[o- v(rov. * 0 Unxovroltdg onoouv0eo4g NSD oroXerier mRNA Xu.r- p(g evrdg nloto(ou ro:drrdvro repgouogorj ror poot(e- T0r 0' rvo ouvr4p4grivo orjvotro nporeiV6v SKl. " 0 priXovropog onooUv0eonq NGD oroyeuet mRNA pe ordorpo prpooropriuo or4v xrodrxonor4rtrrl nepto_ xn. ffi UeprxO Europuolro mRNA Evronr(ovTot 0r Erdudg l.1e- proXdg rou Kurropou + 0 evrontoprjq rov mRNA e(unrlperer drorpopenr6g let_ roupy(eg oe ;lepovtrt;rdvo rUrropo ror ovonruooripevo eUgpuo. * 'Exouv roroyporpe( Tperq Unxovropo( yro rov evronropo rtov mRNA. * 0 evronropog onorre[ crs-orotXero oro mRNA_oroXo roL lrans-nopoyovreg. * 0 ruproq p4Xovropog evepyou perorpopoq nepilogpo_ ver qv roreuOuvripev4 x[v404 rtov mRNp rord p(rog odtiv rou rurropooxetrerou. @ YrffiS; IleprtrnUn 22.1 Erooyayh ]9 nrun 61er rewpu<6 p6),o mr.1v 6r<gpoorl rorv yovr_ 6icov. ApXu<d XoporcrqpiorrlKe oq 6vo ev6rdpeoo npo_ i6v rrlg npotelvooriv0eo4g. Irjpepo,'rrlor6oo, yvcop[(ou;re no].].d enrnp6o0ero RNA nou no((ouv dogrxorjg q Letroupytroriq pdLouq oe dLtro ord6ro qg Srgpooqg rcov yovr6(tov. H ou1-rprero14 rou RNA oe noM6g i.etroupy(eg nou ogoporiv offlv 6r<gpoo4 rcr.rv yovrdfurv unomnpf(et rq yevtxn dno{q dtr 4 6L4 dtepyoo(o nr0ov6v e[e].rX04re o, 6vqv "r6opo RNA,, orov onoio to RNA oplu<d rjrov ro evepy6 ouororu<d yro rn drorfpqo4 Kqr rrlv 6r<gpooq rov yeverrxc0v nAqpogopuirv. Meroyevdfiepo, or nporetveg unoBofl_ Oqoov ri ovdAoBov onox,,ermtxd noMdg on6 our6g rrg Lenoupy(eg, pre ouvdnero qv orl[4or1 r1g eue]l_ [(oq r<or rqg onote].eopotrx6r4rog. To'regd].oto our6 !io:',:,* 627
eond(er oro oyyefuoq6po RNA (messenger RNA, m- RNA). Or Lefioupy(eg r<ov d].l,rr:v xurroprrri:v RNA onoretroUv owxe(trlevo dMcov reqotro(rov: snRNA xor snoRNA oro KegdLoro 21, IRNA rqr rRNA oro Kerpdloto 24, miRNA xot siRNA oro Keqdl.oro 30. To unooUvotro lov RNA nou 51ouv droqprloet qv opX6- yovn evepy6rlro r<ordLuorlg e(vor owtre(pevo rou Keqol,o(ou 23. To oyyel,tog6po RNA no[(er rov r<Upro p6].o mrlv Srrppooq rov yovrd(rov nou ro6rronoro(v nptoreiveg. Kd0e ;r6pro mRNA g6pet ro yeverrr6 r66xq yro rrl orivOeoq ev6g er6xo0 notrunemr6(ou rord q prerdrppo- on. En(onq, 6vo mRNA g6per enmp6o0ereg n).qpoqo- p(eg, 6ncog n6oo ouprd 0o prerorppd(etor, n6oo 1p6vo 0o enrBuiver rot oe nor6 orlpe(o rou rurdpou 0o pero- rppd(erot. Awdg or nLqporpop[eq prerog6powqr un6 Uo pen cls-oro r1e(tov r<or oXer(5 prevov n pcoreiVcirv. l-l ol.- Mg ewon((owor oe rpqporo rqg oll,r1l.ou1(oq rou mR- NA ro ono(o dev epnl,Sr<owor oneu0e(oq orrnv rro6u<o- no(4or1 npolrefvrlg. H EIKONA 22.1 6eiyyer oprotrrSvo qn6 rq tunxd 6opr- xd loporcrqptonrd rov mRNA orouq ilpoKopucbreg ror rouq euKopurilreg. To drpo ttov Borcrrlprordrv mRNA 6ev rpononoro6wor pretd q pretoyporp{, on6re op1(- (ouv pre ro 5'tprgroorpopx6 voux.leorbro nou Xprlotpo- nore(ror orrlv 6vop[q rnq Ueroypogrlg r<or re],euivouv pe ro refur6 voux).eor(6ro nou npoo0Srer q RNA notru- Uepdon nprv rov reppotropr6. To 3' drpo noMciv mR- NA rou E. coli opl;ror[(et pro 6oprf goupr<6rog nou epnl,Sreror orov evdoyevrl (rho-ove[6prrlro) repport- o96 qq peroypoqqg (BI5ne KeEdLoro 19). To euro- purortxd mRNA onorroUv rol.0nrpo rot oupd noluo6evulrxoU ev6oro preroypdgowor (B).5ne Keqd- Loro 21). Or neproo6rspeq Un xtrldrxonorqnr<5q, pu0pu- mrr6g nlr.qporpop(eq rp5powor mtg 5' ror 3' oper6- epooreq neprolSg (untranslated regions, UTR) rou mRNA, oproprSveg 6prrlq ewon((owor Kor or4v ro6xo- noqnr<f neproXri. Evri 6Lo ro pr6pto mRNA e(vor ypopr- prrrdg otrL4Lou1(eg vour<Leot16(rov, pnopeI vo oXqponoro0v 6eweporoye(g xor rpmoroye(q 6op6g ge ev6opropror<6 (euydpopro B6oeov. Aut6g or 6opr6g pno- pe( vo eivor onl,5g, 6ntog or 6op6g piolou-Oqtrldg (stem-loop) nou Tropouotd(owot orrrlv Eu<6vo 22.1, i nro oriv0ercg, pre 6roxi,o6roprdveg (branched) 5oU6S { (A) 5'UTR s'UTR 3'UTR Xoporrrlprolro Tolv npoKopuoTu6v ror rov euKopuoTrrorv mRNA. (A) 'Evo runtro porqptor6 mRNA. Aurd e(vot 6vo povc- olorpovtro mRNA, oporq ro porqptoro mRNA pnoper en(o1g vo e(vot ror notruororpovtro. l-lotrtro porrrlptoro mRNA retretdrvouv Ue p,c 6opfl p(oXou-0qfuoq. (B) '0tr0 ro europuo-rnro mRNA opx((ouv pe puo rotrunrpo (m?G) roL oXe66v 6tr0 retrerorvouv pe gto notru(A) 0up0 t notru(A) oupo entrotrunreror ond nptorelveg [poly(A)-binding proteins, PABP]. To europuolro mRNA pnopeivo tlXouv 1-uo fl xot rept00.- repeg neproX6g Deureporoyoug dogflg, runLro otrg 5'ror 3'UTR. (f) Iro 0r1troonro, ro mRNA nou rtodrronolouv lg rupLeg toroveq ov, qq notru(A) oupoq eXouv evo p(oXo-01trro. (B) (r) M(oXog-011trro KroDrxonoulrtrl neptolrl (0RF) 628 KE0AAAI0 22 lro0epoqro ror Xopo06rnoq rou mRNA 3' UTR
(euydpopo vourtreotr6(ov preto(0 onoporpuotrrdvov neprolti:v rou pop(ou. H 6repe0vrloq tov prqlovto;rtirv Ue rouq ono[oug onoKpurnoyporpoUwot ot puOptoru<dg nLqpo<pop(eg rou mRNA ror ror6ntv Xprlotponorofwot on6 ro pqlovrop6 nou e(vqr une60uvog yto rnv onod6- Unon, rn ;retdgpoorl Kqt rov ewontopt6 tou mRNA e(- vor onUowr.K6 ne6(o qg oriylpovqg poptorfg Bto],o- Y(oq. 22.2 To AyyeArog6po RNA Eivor Aoro0ri M6pro r H oord0ero rou mRNA orpeitreror oq Opdon p$ovouxtreoodrv. o 0r pt9ovouxtreooeq drotpdpouv oq rpoq rrlv npoqrloq yto ro undmpopo rot rov rpdno en[0eor1g. o To mRNA nopouord(ouv eupe[o drorupovon Tov Xpdvtov nut- (tons. o H drorpoptxrl oro0epdrrlro rov ;rop(ov mRNA ennped(et rn ouyxdvrptooq roug, ouven6g ror ro rpool-ro rov nptoreivtitv nou nopdyovror o' 6vo rUrropo. T' oyyelrog6po RNA e[vot q4enrd ooro0rl p5pto, oe ow(0eoq pre to DNA Kor, oe plxp6repo Bo096, to r- RNA r<or IRNA. E(vor o).r1Oero 6l or <pootpodteoteptro( 6eopro( nou ouvdSouv to prBovour<treot[6to e[vqt t<6- nog oo0ev6orepor oe o16or; Ue rouq 6eopo0g nou ouv6Souv ro 6eo(uprBovoux.l.eor(6ro L6yro rnq no- pouo(og rnq 2' udpo[u].opr66og orr'1 ptp6En. H oord- 0ero rou mRNA 6ev oqefteror 6prog rup(tog oe outq rq drogopd oMd oro yeyov6g 6rr ro rfrropo nept6- Xouv 1luprd6o 6v(uprov ono66;rqoqg rou RNA, to ono(o ovoprd(owor pr povouxtredoeg (ri bonucleases), prepx6g on6 rrg ono(eg oroleUouv er6xd"ro mRNA. Or prBovouxtredoeg e(vor 6v(upro nou r6Bouv toug groogodemepxorig 6eo;ro(g prero[ri rrov ptBovouxtre- on6(olv oro RNA. Eprgov((ouv erepoydvao enedrlnd- l,5g 6rogoperx6g nporeivtr6g neptoX6g (domains) od- Krnoov evepy6qto ptBovouxtredorlg r<otd rnv eqffuqrl. Tq ondvto nopodefu;roro yvoordlv prBoev(fpttov (xcro- Luru<d RNA) nepil.ogBdvouv noLL6g ptBovouxhedoeg, Svder(r1 yro rrlv opXdyovr'1 npoSLeuoq oufiq rqg or1- prowrrqq 6poorqpr6rrlrog (pL6ne KegdLoto 23). Ot pt- Bovouxl,edoeg 61ouv nollo0g p6l,oug oro x0r.opo, roOdrq ouprper6louv srnv owtypo<p4 rot qv enr6r6p- Ocoor1 rou DNA, mqv enefepyoo(o v5tov geroypdgtov (pre-mRNA, IRNA, rRNA, snRNA t<ot miRNA) Kqt ornv onod6gqoq tou mRNA. Or ptBovouxl.edoeg efuot efue evEoprpovouxtre6oeg efue e[oprBovouxtredoeg, 6nog rpo(veror or4v EIK0NA 22.2 (Blene eil(onq Ev6tqta 3.2). Or ev6ovouxtredoeg r6Bouv 6vo g6pto RNA oe pto eoo- repxq 05oq ror prnope( vo npouno06rouv r1 vo nport- profv ouyrexprprSvrl 6opn n ol.L4l,ou1(o. Ot e[tr:- vour<l,e6oeg orpoporiv vouxleot(dto on6 to 5vo 6xpo rou RNA ror 61ouv ro0optopr6vq roteriOuvoq (notrrr6- rqro) en(Oeoqg - efue 5'-+3'e(te 3'-+5'. Optol-tdveq e{co- vourtredoeg nopo;r6vouv npooxotrl4p6veg oto un6mptotr-ro evc0 orpotpo6v 6to6oXtxd vourtreot(6to, evrir 6Meg rorotrriouv t4v ogo(peoq ev6q { },fuov vou- xtreorr6[rov ror 6riorowor on6 to un6mpol;ro. To neproo6repo mRNA qnoouvt(0evtot rulo(o (6noq q onooUv0eoll rov po6revepyriv toot6ncov), on6te 11 oro0ep6rlro rou mRNA ouvrlOcog sr<gpd(etot rog 1p6vog nyr(oliq (half-time, t1z). O 6pog onooriv- 0eoq rou mRNA (mRNA decay) Xpnorpoilo€fuor ouvrl- 0og evoL..ortrrd rou 6pou ono66prlorl rou mRNA (mRNA degradation). l-ltrr1pogop(eq yro rrl oroOep&r1- ro ev6g ouyrexprprdvou mRNA e(vor r<to6txonotqpr6veg orrq crs-oM4l,ou{eg (BI6ne Ev6qra 22.7) ru dpo 1o- porcrqprorr<6q tou r<dOe mRNA. Arogopertrd mRNA epr- Eov((ouv o(roorlpre[oto 6rogopetu<4 oro0ep6tnro, U' erjpog dtor<riprovorlg 100 eop6q 11 rot nopondvco. Eto E. coli o runtx6q 1p6vog ng(oiq ev6g mRNA e(vot nep[- nou 3 tremd, 6pr<oq optoprdvo mRNA prnope( vo 61ouv Xp6vo qpu(toriq U6fuq 20 6eutep6tremo ri or6trro r<ot 90 Lemd. Irrl (0pr4, xotd tq rpdoq erBldornonq, o Xp6- vog rlpu(<or1g rov mRNA xupto[vetot on6 3 6tog 100 tre- 'e^* e^4 3 4ro4a s4+/a Prpovourtreooeg. 0r e(tovourtreooeg treuoupyouv npog ;lro rore0Ouvorl (unidirectional). flenrouv ro RNA e[re ono ro 5' oxpo e[re ono ro 3'orpo, onetreu0ep6vovroq Eextoproro vourtreor(dro. 0r evdovourtreooeg ropouv ro RNA o' eotoreprro0g rptrrotpodteore- prrouq deopoUq. Mro evdovourtreool ouvrlOolq oroXeuer etdrreg otrtrr1trouXieq ro/l deureporoyelg 6o;t69. Ev6ovouxtredorl tt.? To AyyetrLorpopo RNA Ervor Aoro0l Mopro 629 Boorreg dworeg E' Q' EItrlvouxtredoq EIovourtredorl
m6, ev6 cno petd(too, on6lemd 6crlg cirpeq xot, cmd- vto, or6po xot ;r6peg. T' ovritrrotro mRNA oroXeUowot yro notr0 roXe(o xoroorpogrl @6ne Ev&ryeq 22.8 rot 22.9). fevv<o, o Xp6vog nUt(onq npoo6top(etqt Ue na- poMoySg rrlg pe066ou nou rpo(vetot or4v EIKONA 22.3. H ogOov[o ouyr<er<prpdvov mRNA mo r<Urropo e(- vor ouvroropdvq qg rol0qroq o6v0eo4g (petoypo- <pr1 rr ene(epyoo[o) Kot rrlq roXrjqtoq ono66p4or1g. 'Orov out6g or nopdpetpor nopop6vouv oroOepdg, ro en(ne6o rou mRNA grdvouv oe oro0epri xordoroo4 (steady state). To <p6opro tov nproreivc0v nou ouw(- Oewor on5 ro rfuopo xup(og owtKoroTlrp((er qv oE0ov(o trov ow(ororlov mRNA (prolov6tt rdnoto p5- tro no((ouv en[or1g ror ot 6totpop6q qrnv onoreeopo- rrr6rqro 14g prer6qpoonq). H oqproo(o rnq ono- oUv0eoqq rou mRNA ovo6etrvUetot on6 pretr5teg pe- VdInS rL(goxog mrg ono(eg eletdmqr<e 11 oXettrrl ou- verorpopd rrlg rolrirqrog onoofvOeor1g Kot rnq rolrlrrlrog getoypocpqg orq 6rorpoptxti og0ov(o tou mRNA r<ot Bp50r1xe 6rr xupropXe( 11 np6q. To preydLo n).eov6rcr1po trlv ooroOc0v mRNA e(vot 4 trov6tqto roug v' oLLd(ouv ypiyopo qv on66ooq qg trretd- qpoonq ;16oo oLl,oyrilv orrl o0v0eor1 tou mRNA. Ilpogovrrrg, our6 ro nleov5xtqpo e(vot opretd or1- powrr6 r6ore vo toooro0p(oet rrl qorvopevtrrl onord- trn qq 6rlpuoupy(og Kqt 16oo ypliyopnq r<otoarpoEtlg rov mRNA. Avc0polog 6LeyXog tqg oro0ep6qroq rou mRNA lopor<rr1p((er no0oloyrxSg t<otomdoetg, 6ntrlg o xopK(vog, or Xp6vreq <pl,eypov5g Kot n oreQo- vro(o v6oog. 22.3 To Europuorrtrd mRNA Yndpxouv un6 Mopcpr'r mRNP on6 Tn [dvvnon dog rot ro @dvor6 roug e To mRNA otrtrr1trenrdpo g' 6vo peropotrtropevo ntrn0uouo nporeivdrv rord qv trlpipovon Tou orov nuptlvo Kot TIlv tt0p0- govrl rou oro rurropdnl,oolto. o Meprrdq nporetveg tov nupr;vlxtitv mRNP 6xouv pdtro rot oro rurropdntroo;to. * Yndpxouv nopo notrtreq nporeiveg nou npoodevouv RNA. 0t neprooorepeg 6ev 6Xouv Xoporqptorei o16po. E Arorpoperuo mRNA oxett(ovrot ge [eXoptot6q, otrtro 0trtr1tr0- enrxotrunr6peveq, oUd6eq pu0gtonr6v nporeivdtv, 6lUtoup- yrirvrog ro RNA peyroutrtivto. An6 q rn peroypogq ru.lv npo-mRNA orov nup{vo ;r5Xpr ror rrlv Korqorpog4 roug oro rutop6nLo- opq, ro euKopuorrKd mRNA o).).rllent6potiv pr' 5vo pretoBoM6 prevo penept6 p to n porteiv6v. To oti prn).o- ro RNA-nporelvc0v ovoprd(owot ptBovouxtreoTrpo- reivrrd oopori6ro (ribonucleoprotein particles, RNP). I-lo).i.69 on6lg npcuretveg nou npooSSvowot oe npo-mRNA ouprprer6louv oE owt6pdoetg ouppo- eriq Kr ene[epyoo(oq (B].6ne Kegdtroto 21), evcir d].- Leg e;lnSxowor orov 5Ley1o not6trlrog (ou(r1te(tot otrlv Evdtrlra 22.8). H op(;rovorl ev6g mRNA mov nuprlvo nepil.otrrBdveL notrtrd Bqpoto ovodtopr6pgco- oqg (remodeling) nou enqpped(ouv t6oo qv oMq- LouX[o rou RNA 6oo rot rtg oletr(6Ueveq npo- refveg, To drprpo mRNA e(vot xotdtrXq].o yto e(oyco- yn U6vo 6rov 6Xet otroxl.qpo0ef 11 ene[epyoo(o xot 11 oL).qLen(6poo4 rou pe to orr:md npcoteivtxd oUpr- nloro, preto(ri dtrtrcov ro TREX (transcription export, e(oyoyrl petoypo<pqq), ro ono(o preooloBe( ottl oriv6eorl pe rov uno6o15o e(oytoyrlg on6 tov nupn- vrr6 n5po, To cirprpro mRNA 6totqporiv nollonL6g 06oerg np6odeo4g (cls-orotXe(o) yto dtd<popeg puO- ;rrmrrdg nporeiveg, nto ouXvd prSoo ortg 5'n 3' UTR rouq. Evcb noLLdg nupqvu<5q npotetveg onoBdtrLowor nptv 11 rord tr1 6tdpxeto rqq e(oytoyrlg rou mRNA mo ruttop6nLoopro, d}'),eg ouvo6e[ouv to mRNA xt 6Xouv p6louq oro Kurrop6n).oopro. l-to nopd6etypto, pr6).tg Bpe0e( mo rurrop6nLoo;ro, to nuprlvtr6 o0pt- nloxo nou npoodSverot orrl rotr0nrpo ou;rper6Xet oro npoto oupBdv qg pretd<ppoorlq rou veou mRNA, tov rotroril.revo (nporon6po r0rl.o" (pioneering round) trlq prerdqpqolq. Autn q npurtrl Svop(r; qq pretd<ppoorlg e(vor r<p(otpr1 yto to v6o mRNA: ov ro eK- proye(o Bpe0e( eLorropotu<6, 0o xotompo<pe( dpeoo qn6 6vo oforrlpo entr;prloqg (B].5ne Ev6r4to 22.9). KdOe mRNA nou 6Xer nepdoet rov 5ley1o qg prerd- epqonq 0o (o65r,Uer tr1v un6troLn4 (ot1 tou ouvdede- p6vo pre norr<(treg npotetveg nou eL5ylouv rt'l t-- --------::- I trz=zumtn I 75 z. E. E >= 35 >E .oo b* o o E a a a a0 /10 ,/ Aroxonrl ueroYpoQrlq RNA Polll 20 40 60 B0 Aenro pero Tn 0roKonn 100 Ell(0NA 22.3 Mro U60o6oq yro Tov npoodtoptop6 tou Xpdvou nu(onq rtrlv mRNA. H petoypotptl on6 qv RNA notruuepoon ll dror6meror, e[re ge xdnoro rpopporeutLrrl ouo[o e(re otrtrd(ovroq q Oepporpoolo oe oretr6xrl Ue 0eppoeuo(001r4 peto]rtro[1 oro yov(6ro qg Pol ll. l-lpoodrop[(ovrot to en[ne6o ouyxerptpt6vov mRNA g' erruntrlo4 Northern Tl RT-PCR oe dLotpopetxd Xpovtxd olyproruno pero rr1 6Loxonq. H onoddpnon rou RNA, otpou [ert- vrloer, ouvrl0org e[voL rrioo ypnyopn 6me to evdtogeoo npoi6vro rnq 0repyoo(uq dev ovrXve0ovrot. H Xpdvoq lut(onq e(vot o Xpti- voq nou onque(rqr tiore ro mRNA vo n6oet oro Uto6 rr1q opXxfg nunq. 630 KEOAAAIO 22 Iro0ep6r4ro rot Xtopo06r4oq rou mRNA Boorxeg eworeg
UerdQpoon, rn cnoeep6rnro Kot, UeptK6q Qop6q' rnv ;;,i;n i;, oro r0rropo' H "nuprlut''{ torop(o" ev6g *nNrn eivot rp(otprrl rot r<oOop((et ro nenpou6vo tou crro rurroP6ntrooPto. 'Hdn yvop((ou;re nol).6q 6to<popettx6g nproreiVeg npOoieons'-b nnn (RNA'binding. proteins' RBP) r.rt, t ; Bdil qv ovdl'uor1 tou rlovtdttiruoto3',ovog6ve- io,' *- .i"apxouv nol0 neptoo6tepeg' l'1 po BL6nerot 6tt io VourOfouo rou S. cerevisiae rodtronote( nep(nou 600'6togopettr6g nporetveq np6o6eoqg oro RNA' ne- pino, .o 6vo 6riroto tou ouvol'tt<o[ opt0pori rov Yovt- bi,ru -or. Me B6o11 nop6proteg petpqoetq' 0o.nep(pmve xove(g 6tt to ovOpc0ntvo yovtd(topo vo Tleptexet Tlqvo) on6 zboo t5toreg npote"weg' AwSq ot errrprioetg Boo(- io*o, *nu nopouo(o xopoKrnptop6vtov neptoxtiv ipOoJtons mo RNA roi qo(verot 6tt peLl'owrxd 0o Bpegofv xot dltreg neptoX5q' To RNA-or61ot xot ot tr'et- i5;;Gs;;" neptoo6tepcov RBP efvot 6Yvooro' coor6- oo' n,ott,it-oi 6., nolL6g o)'l'r1l'ent6poriv Ue npo-mRNA n mRNA. Aut6 to e(5og ov6}'uor1g 6ev nept- LoprBdvet to peydtro opt0p6 npcote'twilv nou 6ev npoo- 6rivowot oneuOe(oq mo RNA, otrLd oup;ret6Xouv oe o6pnl,oro nP6o6eorlq oro RNA' - -'inuo*,^gg ev6e(ietq yto to tr6yo [-nop(r1g t6oo pe- VoLou opt0poti 6toEopetrr<6v npoteivdlv np6o6eo4q *o ,nrtfn'nporj).0ov on6 to eUpq;ro orl ro mRNA ouvd6owot pe (eXo ptor6q, o)}'6 otr'rlLoenu<otrum6 ge- veq, oUaOeS nee.- Vtel,etag ortg ono(eg 6ywe owtoro(- Xrol etdtr<rirv RBP pe ro mRNA-or61ouq onordLuqUov 6tr owd to mRNA rtodrxonoro(v nptoretvsg pe xowd XqpoKrrlptfixd, n.x' ouprpretopi oe nop6Uoteq Kurro- ii'lgi o"pvoo(eq 11' nop6ptoto ew6ntorl' ..ruvendrq' to penept6pto trov npoodedepr5vtov npolteivrilv KornYo- bil"J,tito mRNA. l-to nopd6etygo, erotowd6eq mR- NA tqg (ri;rqq e(vot 'or6xot" yto np6o6eo- ::: * : " "tptLbitptg on6 ttg 6 ouyyeve(g npcltarvac = -- : Puft xot Puf2 npoodevowot rup(o:g oe mRNA :3- '-- 6rxonototiv trreprBpovtr<6q npultetveg' evrrl 11 Puf3 oo -:- NA yto 1-tttolov6ptot<6g npotetveq K'o'K IUptcptrlvo * ero iiJ*,p"io pow6tro o ouwovtopevoq 6eyXoq rov Ueroperoypotprxov dtepyoottilv tou mR- r.rn ir"ooLoBe(toL on6 rn ouYxpovtog6vq 6pdoq noMo- niiu nap rot ;-rotd(et notr0 pre ro ouwovtou5vo eleyxo -ni ur-ovpoqrlq trov yovt6(trrv nou geoo)'oBekot on6 -ouq or-o,ig ouv6uool.toriq tov ;ietoypocptr<dlv nopo- V6wrov (B).6ne Keqdtroro 28 H opd6o trov mRNA nou uotoo(owot dvo ouyxerptp5vo tUno RBP ovo<p5petot Kor'coq' RNA peyxoulovro (RNA regulon)' 22.4 H Ano66pnon T(DV l'lpoKclpuoTlKdv mRNA I'lePrAoUBdv€l noAAo'Ev(uPo * H onodourlorl rolv porrrlptor(rv mRNA opx((et pe oqoipeorl evog nuporpolotpopLro0 on6 ro 5'orpo' * To Uoro,pro,poputrttrl;t6vo mRNA onodogo0vrot xoro rn 6top- Keto Tlq UeroQpoonq o' tlvo xurtro 6uo Bnportov nou nept- ioUpdr" ev6ovourtreotrurLr6g evio;t6g, rtg onoieg orolou0el neqtn rov rprlgdtirlv nou nporunrouv oe roteu0uvon 3'npoq tr a H 3'notruodevutr(toot1 pnope( vo Dteuro)'0vet rrlv onoOoprlorl rtrooporulv mRNA nou nepteXouv deureporoYrl 6oUI' * Tu x0pto rlv(upo rrlg onodounonq )'etroupyouv toq ouptntroro nou ovoprd(erot otr:1tot[Dto ono6oUnorlq' , H yevtrl rorio svoq RNA peyroutroviou (RNA regulon). rro europu0rtro mRNA npooorlvovtot notrteq npoTeiveq 0t oflottq .i.eyxouv rr1 perorppoorl, ,nu r*in,on roLq oro'eporrro rouq. To unoouvo),0 rov mRNA oro ono[o npoob.veror 11 rbto nptrlrelvq 'eo- : eiror rplpo rou lbtou peyroutroviou. Iro DLoYpoUUo, ro mRNA o rot 0 elvot Uepr] rou peyrou]'ov(ou 1 ' ro mRNA 0' Y K.t 0 rou peyroutrovi- - r 2 r.o.r. 2?.4 H Anooounon ttrlv l-lpoxopucorrxtilv mRNA t]eprtrouBoveL [lo},6.Ev(uuo 631
Or yvdroerg Uqq yro qv ono66prnorlrov npoKopuon- rc0v mRNA npodplowor r<up(rog on6 preA6reg oro E. coli. MSXpt onygriq, or yevtrdg opldg roXriouv Kor fi' dLLo Borrrlproxd e(64 nou 61ouv ;relerq0e[. Iroug nporopuri:teg, 11 onoS6gnon rou mRNA oupBo(vet rord r1 drdpr<ero qg prerd<ppoonq. To nporopurotrxd prBooro;rdrro ferwofv q pretd<ppoorl nporo0 oLox).q- ptoOe( 11 preroypo<pri: npooSSvowor oro mRNA oe pro 05o4 5vopfig xowd oro 5'drpo ror ouveX[(ouv npoq to 3' drpo. lloMonld ptBooroprdtro pnope( vo [,exwq- oouv rn perdgpoorl rou r6(ou mRNA, 6ro6oXrrd, q44- pori(owog 6vo notruprBootopdno (polyribosome) q notruoop6rro (polysome), 6qlo6r1 5vo mRNA pre noMd ptBoocopdrto. To mRNA rou E coli onodoprofwor on6 5vov ouv- 6uoopr6 evepyor{rolv evdovoux},edorlq Kor 3'-+5' elto- voudiedo4g. To rUpro 1rovondn onod6prqorlg mo E. coll efuor puo drepyoo(o noA.trrirv mq6(ov nou rpo(veror orqv EIKONA 22.5. To opXrr6 Brlpo e(vot q ogo(peor1 rou nupogooqoprroU on6 to 5'drpo, pe rordLorno puo provodtrfl grrlogopu<rl pi(o. H Uovogooqopu].uo;r5v4 popgn dteye(per rqv rorol.unrrl 6pdoq prog ev6ovou- xledoqq (RNAorl E), 11 ono(o npqyporonoo( 6vo opXr- 16 r<6rprpo rovrd mo 5'drpo rou mRNA. Aur6 ro r<6t!rpo oqrlver 6vo 3'-OH oro ovodrr6 ri.dogo r<ot 5' Uovog@oqopr6 oro rq0odtK6 xLdopro ror xoroorpS- rper Lemoupyrxd 5vo povoororpovlx6 mRNA, eq6oov to prBoocopdrro 6e pnopo0v nL5ov vo (erwrloouv rrl prerdrppooq. To ovodrx6 xopprdn ono6opefror on6 pro 3'-+5' e(ovouxtredorl (PNPdoq = gooeoputr6o4 tov nol,uvoux.A,eott6(o:v, PNPase = polynucleotide phos- phorylase). KoOdlq neptoo6repo RNA err(Oeror perd q Suileuorl pgootopro{<ov nou erlov [exw4oer rn Ue- tdtppoo4, our6g o r<Ur<Log 6ui: Brltrrdrtov qg prBovou- xLedorlg enovoLoprBdveror rord pr4r<og rou mRNA oe roteriOuvorl 3'-+5'. H 6repyoo(o efeL(ooeror ro10roro, 6nog cpovepcbvet to yeyov6g 6tr BpoX6o rLdoproro nou nopdyowqr on6 qv RNAoq E ovrlveUowor pr6vo oe perotrtroypSvo r<rirropo nou epqov((ouv e).orrcopro- nrq evepy6rrpo e[olvoux],edo4g. H PNPdoq, 6ncog r<or df,l,eg yvom6g 3'-+5' e[ro- vour<Ledoeg rou E. coli, o6uvote[ vo npolrrlprloer 6ro- prSoou 6(rlovtov neprolrriv. Iuvenci:9, n 6oUn ;r(oXou-0qLrdg mo 3'drpo nolLriv Bor.rqprordlv mR- NA ro npomore0er on6 r4v dpeoq 3'en(Oeor1. En(- oqg, optopr5vo eooreprKd x}do;roto nou nopdyowor pe ndQ4 on6 r4v RNAoq E 61ouv neproX6g 6ewepo- royorig 6oUiq nou 0o pnopo[oe vo noporco],0oouv q 6pdoq trlg e[olvouxLedoqg. Om6oo, r1 PNPdorl ei- vor rrovrl vo n6nrer 6(rloveg neproldg ov undpXer 5vo rprrlpro pov6x.Lrovou RNA prrlroug rouLdlrorov 7- 10 voux.treorr6(ov r<o0o6rrd (3') rou pr(o1ou-04l,rdg. H ptov6xLcr;vr1 ol,l,r1l,ouX(o qo(veror vo Lenoupye( oov rxp(<o;ro yro ro 6v(u;ro. O Rho-ove[dprnroq repUqn- op6g o<prlvet pto ;rov6x.,ovq neptoXrj q ono(o e[vor noLti ptrp4 yro vo l,enoupyrloer oov u<p(ropro. l-to r4v enil.uoq quroU rou npoBl,ri;rorog, puo Borcrrlpror<ri no- Iu(A)- notrupep6o4 (PAP) npoo0Srer oupSg noAu(A) Merdqpooq + I v 5'pp p Y- r.,e rY 'r- 3' 3' - 5'E[tovourtredorl - Ell(0NA 22.5 Anodoglorl rolv porqpLoxtirv mRNA. H onodoplorl (rlv porqpror6v mRNA opX((er ge ro xdtlllro tou rprqooqopxou 5'orpou, ono6f6ovrog Uro UovoqooQopxq opd6o.'Eneuo, ro m- RNA onoDogorivror o' rivov rfrtro 6uo prlpdrrov: ev6ovour],eo)u- rur1 drdonooq, qv ono(o orotrouOe[ 3'-+5'n6Qq tou r],doporog nou qnetreu0epdlveror. 0r evdovourtreotrulr6g dLoondoerg oulr- po[vouv oe rote00uvorl 5'-+3' novo oro mRNA, oro],ou0rirwoq rr1 d r6treu or1 ro u te,Jeuro(o u p rpootrrpor(o u. 10-40 vour<i.eotr6(rrrv oro 3' drpo, rdvowdg ro euo(- o0qro oe 3'-5' ono66trlqoq. To rldo;roro RNA nou xorol,qyouv oe 16rokepo moOep6g 6eureporoye(g 6o;t5g (otog ononoUv enovoloprBov6;revo Brlproro no- i.uqdevu^,[oo4q Kor n5qnq on6 efcovour],edor1. Aev e(vor yvom6 ov 11 noAuo6evul(oor1 e(vor ndwq ro evoprcr{pro B{po yro qv ono65pr4orl rou mRNA 11 ov Xpqorpoilore(ror ;r6vo yrq vo Borl0rloer qv nopoyoyr] t<Loogdrrrlv, prero[rl dtrtrtov Kclr. ro 3'rs}.rr<6 r<].do;ro. Meprrd nerpdproro unodqLrilvouv 611 4 n6tlq evdg m- RNA qn6 qv RNAoq E [oroq e(vor onopo(rqt4 npo- un60eoq yrq rnv evepyono(r1on rnq poly(A) nolu- pepdo4q. Aur6 nr0ov6v eppqve[er yror( r' ordporo mRNA 6ev ono6oprorjwor on6 ro 3'dr<po. H RNAo4 E ror q PNPdo4, po(i pe pro eLrrdoq rr I k$d&' l^N ---&---&---# 632 KEOAIIAiO 22 lro0epdqro ror Xropo06qorl rou mRNA
5vo entnp6o0eto entrouptr<6 6v(upo, oXqpor((ouv evo rroir"pr-tlvr16 oti prnl'o ro yvoord cog -ot'l porf6 lo """iOirq"nq (degradosome)' H RNAorl E 6Xet 6trr6 ;4il il otunlo'<o. H N-te)'tr<( neptoxq qg nop5xet i.lu euepyO-qro ev6ovour<Ledor1g, evtir 11 C-reLtrq ne- propj 6vo txp(opo nou ouYKpore( to un6l'otno ouoro- ,,^&. ru,i, n RNAon r<ot 11 PNPdorl e(vot ot nipreg 6v6o- xot eqovo'uxLedoeq otqv onod6prlon rou mRNA' undplouv en(onq rot 6trtre9, p6LLov pe nto nepropt- ;uJvous p6Loui. o p6Loq tov dtr'I'tov vourLeoocbv mrlv ono66prnon rou mRNA drepeuvrl0qr<e o(toLoytirv- roq rouq qotvot6noug pretoMoylrdvov rurtdptov yto nodSuo onb ro 6v(ugo. l-to nopddetygo, 11 o6povono(- non rnq RNAoqq E enrBpo60vet tqv.ono66pnon rou mniln'Xop(q vo rrl moptot6 eweAdrq' Metol'Ld[etg "ou qapouonototiv ttlv PNPdorl q onotodrlnote on6 rq 66o yvtlordq 3'-+5'e[covourtredoeq.6ev 51ouv ou- oromrr6' rog(o en(ntoon crrn ouvotrrx{ mo0ep6qto rou mRNA. Aur6 onorotrUmet 6tt 6Uo onoteo6{note e[c,lvour],e6oeg pnope( vo Sternepotdloouv prdtrtrov *ouou,oO t4v ono66prlorl rou mRNA' Motrororho' pr6- vo 6uo on6 t,g tpe(q e(ovour<)re6oeg (PNP6or1 rot RNAoq R) e(vot tt<ov5g vo nSntouv t<)'1o"y1to pe otq- 0epdq bewepotoye(q 6opdg' Aut6 ono6effinKe oe pe- L6ieg 6tnl,d perol'troypr5vov rundporv, oto ono(o e(Xov o6povonotrl0e( t6oo 11 PNPdorl 6oo t<ot 11 RNAon R. Ie outd to xtittopo, 11 ouyr<6wptoorl rov x).oop&tov mRNA nou nept6louv Seutepotoye(q 6o- p6g ou(dvet. ' 'noila epcorriproto nopop6vouv ovondwrlro oxett- rd pre qv ono66prr1or1 tou mRNA mo E' coli' O yp6- vog rlpr(oriq tcov 6to<p6pov mRNA eprgov((et eupe(o 6toxtil-rovor1, 100 rpop6q rotnl'6ov' H Bdoq outcirvttov orpo(tov 6to<popdrv 5ev SXet rotovoqOe( ntrr1pu:g' oL- trd <po(vetot 6tt orpefietot r<up(<og oe 6uo nopdyoweg' i euotoeqo(o ttov 6toEopettr6v mRNA oe n6qr1 on6 evdovour<Iedoeg notxftet: Svov 9q0p6 npomoo(og npoo<p6pet 11 nopouo(o 6ewepotoyo0q 6oUnS oro 5' a^po.' Op,o1r6vo mRNA ;retocppd(owot qno6ottK6te- Jo o"O 6trLo, o6rlydrwqq oe nurv6tepq ouoxeuoo(o icov ptBootopdrtov nou rcl npomore[ouv' Aev e(vot yrtoorA ov undplouv entnp6o0eto ptovondrto ono66- Uron, tou mnttlA. Ito E. coll 6ev 6Xet Bpe0e( 3'+5' lqt:vouxLedo4, nop6tt rivo t6toto 5v(upro 5xet touto- notrlOe( oto Baciltus subtl/is. l-lt0ov6v or 6totpopettr<69 evdovour)e6oeq Kt e(ovouxLedoeg 61ouv 6torpno0g p6l,ouq. Mto trreL6tr1 UtKpoouorotxtriv o' en(ne6o Yovt- Sui,lroioq npoodr6ptoe to en(ne6o 4000 xot nLtiov mnfln oe o0.ropo pretoLl'oyprdvo Yto rllv RNAoq E' qv PNPdorl n d).Lo oumottrd tou otol'tott6(ou ono- O3prlorlq. To en(ne6o noMrirv mRNA ou[r1Or1rov mo freioLLovpdvo r6rropo, rot6 to ov.opev6pevo (pre(- oon rrlq' ono66prr1or1q). AI'l'o, om6oo, nopSpetvov sro'(6tq'enine6q 11 ox6pro xot getrilOlrov' O Xp6vog rlptEonq erdtrriv mRNA prnope( v' oAl'd(et ond 6totpo- petir6g' rotomdoetg qg rpuototroy(og rou rurrdpou' n.1. ootr(o ri dL}'o e(6r1 stress. Ot uneU0uvot Urlxovt- opro( yt' ow6g rtg oMoydq nopog6vouv dyvtlorot' 22,5 To lleproo6tepo EuKopuoTlK6 mRNA Ano6opo0vTol M6oo Auo MovonoTl6v EEoprdlUevov on6 tloAuo6evuAitrlon r 0t tpononotfoetq xot mo 660 6rpo tou mRNA ro rpoorq- reuouv ond rn 6p6on rtov e[ovour],eoodtv' r To 6uo rupto l.tovondtto onoouvOeoqg tou mRNA opxi(ouv pe onoodevu)(trtorl 11 onoio rorotrUerot on6 nolu(A) vourtred- oeq. " MeiO rqv onoodevul(tlon Unopei v' orotrou0rloer e[te oqo(- peon nlq xoh6ntpog rot 5'-+3'n6r'[Jrl ond evoovourledol' e(- ie n6qn ono e[otvouxtreooqq o roreuOuvorl 3'-+5'' r to dviuUo orpo(peonq Uq rohrjntpoq ovroyrovi(etot ge ro orjpntroro evopEnq Uq UerdQpoonq yto np6o0eor1 oqv 5' rol6mPo. r To e[ootogono, ro ono(o rorotrrjet tr1v 3'-+5' n6$1 rou mR- NA, eivot 6vo UeYoho, e[drtxttxd ouvqprlp6vo otjpdroro' I H ono66prlon ouupo(vet O' eudtoxptto xurroponhooportxd otopor(Oto nou ovopd(ovrot otlltotfdto eneEepYooioq' To euxoputottrd mRNA npoorote0owot on6 ttg e(co- vour<ledoeg gSoto ttov rpononotnp6vtov drpov toug (Err6vo 22.1). H xol'rimpo 7-geOul'oyouovoofuqq no- p61", t po*qo(o on6 5' en(0eoq, evri: q oup6 poly(A)' lroil p," rtg npoo6e6e;r5veq npcoteiveq, nop6Xet npo- *.ioio on6 3' en(Oeor1. E[o(peor1 onote]'oUv to mR- NA tcov torov6v oro 0r1Loonr6, to ono(o rototrrlyouv oe Uro 6opri ;r(o1ou-0q}'r6q ow( trlq ou.pdq poly(A)' Irou g eu r<opudlreg, en(0eoq evdovou rtreootirv ove- (dpqq on5 tr1v oMqAoul(o, evoprcrrlptoq Pnxovt- o;r6q nou XpnotUonote(rot on6 to Bort(pto' ou;rBo(vet ondrto 11 not6. Molov6tt 11 onoo0vOeorl tou mRNA (mRNA decay) loporcrqp(orqre tremope- pSorepo orov (uproprfKnro, ro neptoo6tepo euprlpo- to tcrxtiouv Kol, Yto to r6rtopo ttov 0qtromtrtirv' H' ono66;rqorl rrlq ouwptmtrrlg netov6trltog ttov mRNA e[,opt6tot on6 onoo6evu]'(coo4, 611o6{ opX((et ond(owoq rnv npoororeurxrl oupd poy.(A) H veooxrl- Uoi,oUgun'ou'pd poly(A) (r1 ono(o o-rn.(0Un e(vot nep(- Iou io-so vouxleoidto o6ev(vqg, evci oro Oflr'oortr<d nepinou 200) entroLtirrerot on6 nporeiVeq np6o6e' onq oe polyin) [poly(A) binding proteins, PABPI' H orpa pofv(A)-u,p[oro.o, oro6toxrl BpdXuvorl ;r6Ltq ne- paot,' *o' rurrop6ntr'oopro, dtepyoo(o no-u t<ototr0erot on6 erarx6q vouxtredoeg poly(A) [ovogd(owor enioqq xor onoo6evuldoeq (deadenylases)l' T6o'o orn E[Un 6oo ror oro OqLoonr<d, 11 oupd poly(A) opltxd 9poX6- verot on6 to oUtrtntrot<o PAN2/3' Arol'ou0e( tolUrepq n6rlrlrotv 60-80 r<orol'ofnov A on6 ro orlprntror<o CCR4- ruOf, to ono(o nept6xet rrlv e[rovour]'edoq Ccr4 rot routrdXtorov 8 enrnp6o0eteg unogovddeg- A(tooqpre(- ,.o e'fro, to yeyov6g 6tt nop6;roto otiprnloxo CCR4- NOT eprnl'6r<ovrqt oe norr<(treg 6tepyoo(eg trlg 6r- ti.I To l]eptooorepo Euxopuorrro mRNA AnoDououvrot M6oto AuO Movonort6v E[opttirgevov on6 tloluodevutritool 633
epoonq rov yovrd(ov, Uero(ri dtrLrrtv 11 evepyono(r1oq rnq peroypqgnq. To oripnLoro Oeo:pekor orroutrlevtr6g puOprorrlg qg 6r<gpoonq rov yovrS(tov, svonot6wog rn Ueroypogri ;re qv ono66prqo4 rou mRNA. T6oo orrl ([pq 6oo Kor. oro 0q],ooru<d undpXouv r<or dMeg vou- xi.edoeg poly(A). O L6yog our4g rqq norrd.6rqrog 6ev eivor or6go oqgnq. A0o Srogoperrxd piovondrro ono66gqorlg tou mR- NA nupodotoUvtor on6 rrlv ogo(peor1 tr.1g oupdg poly(A), 6ncog go(veror orrlv EIKONA 22.6. Zto np6ro pro- vondrr (ErrSvo 22.6A), n n6{n rqg oupdg poty(A) oe ollyo(A) prrlroug *10-12 A nupodore( rrlv orpo[peoq qg xolfmpog on6 ro 5' drpo tou mRNA nou rqto].Uerqr on6 ro ev(uprx6 oripnloxo ogoipeoqq rrlg xoAri- rrrpoq (decapping enzyme complex). Irrl ((9r1, ro oUpnl,or<o onoretre(ror on6 5uo npcorelveg (Dcpl r<or Dcp2), evc0 oro O4l,oonrd nepil,opBdver rrg opr6l,oyeg nporeiveg 6ncrrg en(oqg xor oprotrlSveg enrnp6o0eteg nporeiVeg. H o<po(peo4 qg xoL0nrpog ono6(6er 6vo RNA pe 5' povogoogopuLuopr6vo drpo, un6orprrr;ro yro trlv 5'-+3' e[<ovoux].edoq XRN 1, 4 ono(o n6met ro- lriroro ro mRNA. Ir4v npoygorx6t4ro, ouq 4 n5rfrl efuot 16oo ypnyopn ciore ro ev6tdpreoo nopdy<oyo qg frov odtivoro v' ovoyvoproro6v ;r51pr nou or epeuvrl- tdg ovor6Luquov 6rr ;uo oerpd vour).eol6(cov youov(- vqg (poly-G) pnoporjoe vo moprorloer r4v npoS).oor1 qg Xrnl orn EUUq. 'Onog qo(veror qrrlv ElK0llA 22.7 ot epeuvrlrSg roroore0ooov ;r6pro mRNA c6me vo nepr6- Xouv eooroprr6g neproXdg poly-G ror Bprjr<ov 6rt ouo- oopeu6rov ro ollyoodevu)lo1.ulvo 3' drpo rov mRNA. Aw6 66er[e 5tr (1) q 5'-+3'n6r,Uq on6 r4v ev6ovouxAed- oq qrov to rfpto 1-rovondtr qg onooUv0eo4g r<or (2) 4 ogo(peorlqg roLrimpog ouv6Bowe npw onS qv nlri- pr1 orpo[peorlrrlg oupdg poly(A). Kovovu<d, q r<oLimpo e[vor ovOercru<rj orqv ocpo(pe- orl tnq rotd r4v evepy6 perd<ppooq enerdq oLtqLenr- 6pd pe rrlv Kurropontrooponx{ nporefrv4 np6o6eo4g orq xotr0nrpo (cytoplasmic cap-binding protein), 5vo ouororx6 tou ougnl,6r<ou rou euKopucolKo0 nopdyov- ro 6vop[r1g 4F (elF4F) nou e(vot onopo(rrlrog yro rrl petd<ppooq (BL5ne KeqdLoro 24).'Eror, or prlXovropro( qg prerdqpoonq Kor qg ogo(peorlg rqg r<oi,rimpoq ov- royov((ovror. l-lrig 6poq n onoodevul.[coor1 rou 3' dxpou rou mRNA roOrord qv xoLtjnrpo euno0rl oe ogo(peor1; H prerdqpoo4 nepil.oprBdver gro rpuorxrl oL- ),qtren(6pooq pero[ri qg PABP nou e(vqt npoo6ede;r6- vrl oro 3' dr<po Kqr rou oupnl6r<ou elF4F-xoLfnrpog mo 5' dr<po. H one).eu06poon rnq PABP prSo<rl rqg qnoodevu^,fooqg nroreUeror 6rr onomo0eponore( qv oMqLenbpoo4 el F4F-r<ol,rimpog, orprjvowog ouyy6re- po erre0erpSvrlrrlv roi.rimpo. O ;r4Xovropr6g 6ev e(vor 16oo onL6g, xo0rig e(vor yvcom6 6rr orqv ogo(peoq qg roLUmpog epn).dxowot enrnpdoOereg nptorelVeg. 'Evo o0prnLoro emd oXetr(6prevcov np<oreivcilv, Lsm 1-7, npoo66veror orrlv neproXri oligo(A) ;rerd rr;v on6l.ero qg PABP xor e(vor onopo(rqro yrq rnv orpo(peoq rrlg rol[rnpoq. Enrn]*iov, 5Xouv ovor<o],urp0e( Srdrpopeq npcoteiveg nou onou[dvouv rnv ogo(peor1 qg r<o],ri- rnpoq. Or pullovropro( pre roug ono(oug ourSg ot npro- tetveg dreye(pouv rqv oqo(peo4 5ev e(vor nl.rlpog r<otovoqto(, nop6tr golveror 6n 6pouv eire enrmpo- te0owog ro Unxovrop6 ogo[peorlg qq r<otrUmpog e(re ovoor6Mowog q prerdqpoo4. Iro deUrepo trrovondrr (Err6vo 22.68), q onoo6evu- Nourtredorl poly(A) nno,oFl I [m?c F ". /r'--: it it -, A10 (A) 3'-5'Arrcanvooy/ - r'onooortro, (B) EIrootogolo ^,rS 4+/a t4+4a : , .. . To r0pto povonorto onoouvOeolg nou e[oprtivrol ono qv onoodevu],iroor1 otoug euxoputr.rreg, Auo povonolo opXi(ouv ge onoodevutr(too1. Kot oro duo, t'l oupo poly(A) rovro(ver ono ;.rro vour),eoo1 poly(A) eotg orou qrooeL oe plrog 10 A,'Eneuo 5vo mRNA pnope( v' onodoprl0eI p6oto rou 5'-+3'11 rou 3'-'>5'ptovonolou, To 5-+3' ;rovonon nepiloppover orpoLpeol qg rotrunrpog onrl ro ev(u;ro Dcp rot ndQl on6 r1v e[ovour],eooq Xrn1, To 3'-+5' povonon nepiloppover n5fi ono ro oupn],oxo rou e[trlotrlpor[ou. 634 KEOAAAI0 22 Iro0epoqro ror Xtopo0erlol rou mRNA
Xprlot;ronoilorl ptoq otrtrr1trouXiog no- Iu(G) yto vo roOoptore[ 11 roreu0uvol rrlg onoofv- 0eonq. Irl (0U1, ov ltto ohtrrltrouXio nou(G) erooX0ei p6oo o' 6vo mRNA, 0' ovoorefieL Trlv npo- titrooq rov e[tlvour],eoodtv. To 5'11 ro 3' rtroopo rou mRNA nou ov0(ororot oqv onodoprlorl 0o ouootrl- peuOet oro rutopo rot pnopei v' ovtXveu0e[ p' erru- nool Nofthern. 3' -5' Anootxty/ 5'- 3'Anooljveton L(oor1 npoq o)rtyo(A) orohouOe(tot on6 3'-+5'ndqr1 on6 pro e(ovour<tredor1 rou r6ptou otirprotog tou mRNA' Au- iJto'Briuo ono56pr1or1q t<ototrUetot on6 to e[tootog6' rro lexoiome), 6vo 6orruhtoet66q orjgnl'or<o on6 5vqv ewe.opepTi nuprlvo, ndvo oto ono(o npoo6Svowot 1'tto ri neptoo5tepeq entnp6o0eteg npotelveq' Mto np6- o,poin ovogopd 56et[e 6tt to e[,<oocrlpot(6to 61et en[- onq'apdo'n ev6ovout<tre6o11g: 11 or1;roo(o trlq ouy'rerptpevrlq ev(u gtr<r1 g evepy6qtoq orrlv-onooUv0e- on'rou tnnrun nopog6vet dyvomrl' To e(ulotoptdtto undpXet oe nop6proro Uopqn oro Aplo(o r<ot en(oqq e[- ,o. oudtroyo rou otopor(ou ono66ptqonq rov Borrrl- p(cov on6 t4v dno$r1 6tr or unogovd6eg tou nuprivo oXeil(owot 6oprtr<d Ue rnv PNPdon' Iuvenrilg, to e(o- or,:1.tot(6to e(vot 6vo opX5yovo ouorottr6 Utoq popto- xnq UnXovnq. To e(ooopdtto no((et en(or1g orlprowtr6 p6Lo crrov nuprivo (BL6ne Ev6rrya 22'2)' H olettrrl orlpooio tou t<dOe pr4lovtopro0 6ev e[vot yvoorn;nop6rt orn E6pn <pofuetot vo uneptolUet to 1rouor,6-, otpo(peorlg qq rotrfntpqq il9u e[optdtot on6 onoo6evulrdron. To govondno ev ;r6pet roLOmov- rot. Erotowd6eg mRNA rnq (6pnq ;retr'eu10r6ov pe gt- Kpoouorotx(eg oe rrirtopo otq ono(q e(Xe onevepYonotrlOe( efue to 5'+3' e(te to 3'-+5' provondl' >e xd6e nep(molor1, pr6vo 5vo prtxp6 noooor6 preroypd- rprov ouEri0nre oe cr16or1 pe ro quotoloyxd r0nopo' Aur6 to erlprlpro uno64lrivet 6tt nol,0 L(yo pr6pto mR- NA trlq (riUnq eEopttirwot on6 to 6vo ( to 6Mo ;tovo- ndtt. "Ltot, npotdOrlr<e 6tt qutd to ptovondtto nou e(opttirwot on6 onoodevul'(oor1 e(vot ro npoenrLeypS- vo lrovondtto ono66prqonq Yto 6Lo to notruo6svutr'to- pr6vo mRNA, ptotrov6tt unoo0votro mRNA pnopef vo e(vot or61ot dMtov et6trrilv l.tovonotttilv nou neprypd- eowot orqv Ev6tqto 22.6. Ar6pro Kot ro. mRNA nou xpqotp,onoiotiv to npoenil,eypr6vo povondtto, ono6o- Uo.i*o, pre 6togoperxrl toX0t4to et6trci yto r60e mR- NA. Ndeg 5peuveg npore(vouv 6t 11 onoS6ptrloq rou m- RNA oulrBo(vet o' eudtdrptro ooltrro{6to oe 6o ro KUr- top6nLoopro, yvomd og oopori6ro enefepyooiog (piocessing bodies, PB). Aw6q or 6op6q, opot6q ge io ,pr-or,*6 pttt<poor6nto, e(vot ouvo0po(oporo Un Ue- rogpo(6prevov mRNA Kqt notK(Iov nproteivcbv nou oxe- t((ovtor UE rotootoLq rnq pretdqpootlq Kor onoo0v0eorltou mRNA, preto(0 dltrtov 11 prr11ovr1 oqo(- peonq rnq rotr0mpog rot q e(tovourledorl Xrnl' l-evt- rd, orq pg Oev undplouv npotetveg np6o6eorlg oe poly(A), yeyov5q nou uno6qL6vet 6tt npw on6 rnv eY- no**oon oe our5g ttg dopr6q oupBofver onoo6evu- iiron. To oto;ror(6to ene(epyoo[og.e(vot 6uvo1-ttt<d' ou[operdlvowot oe g6ye0og ror opt0;r6. ri or6pro ru e[o- qov((owor un6 6roqoperr5g rur-optx6q rot nepo;tott- r<6q ouv0rlreq TIou enrlped(ouv tr1 petoqp-ooq Kot rnv onoo0v0eoq. l-to nopd6elYUo, n onetreuO6ptoorl rov mRNA on6 to noLuoto;-tdtto pr' 6vo qdpptqKo nou ovo- or6Uer rqv 6vop(r1 qg perdqpoor'1q nporol'e( peyolq o[[r1or1 tou opr0ptoti Kot rou prey60ouq tov PB To 6to onot6f,eqto 5Xet rot n Ueptr<( o6povonofioq ouorort- r<c0v t1q onooriv0eorlg. Om6oo, 6tro to mRNA nou e6pe0ouv oro PB 6ev efvot xotodroopt6vo yt' onoofv- 0eor1: optol.t6vo prnope( v' onetreu0epo0oUv yto pet6- epoorl,'Xop(q 6pog vo 6Xet qnoooqnvtore( otte ndrq enil6yowot npoq onel,eu06ptooq ofre Kot ytor(' E(vot 6yv<omo rotd n6oo 11 ono66pr1or1 6}'crlv ttov mRNA ou;rBo(vet oe owd to ocopot(6to 11 o161lo rot tt tret- roupy(eq entteLo(v. Mto npoEovrlq t66o e(vot 6tt 11 ory^3*pror1 rolu ptiv Koroorpenrtr6v ev(tipttov oe onopovcoptdveq neptol6g ro0tord rrlv ono66prlon rou mRNA nto oo<Pohi rot ono6ottt<ti. !e et6xo0q runoprorig ttlnoug undplouv otopot(- 6ro nou nepr6louv mRNA t<ot gotd(ouv pe to PB' ro- 0cirg nept5Xouv rtq neptoo6tepeq on6 ttg nptoteiVeq nou'eptnl6t<owot qrnv r<otoorotrrl qq ;retdqpoorlq Kot rrlv onooriv0eo4. M4rptx6 xoxx[q mRNA (maternal ,hrun granulesl undpXouv mo coor6rtopo noL)'riv opyor,o-1rt'i,u. Aurd to xoxr(o onopt((owor on6 pr6pto niirrrn nou 6roulpo6wot oe pretoqpocrnrrl r<otoorotrq 6cog 6tou evepyonotrl0ofv rffid tqv enox6tr'ou0q ov6- rnuEn. H rotooroLrl ennuyl6vetot pr' ercretopr6vrl qTIo- odevutr(tooq Kot n peroYpoqn Ue notruodevutr(coot1' En(or1g, outd to rorr(o (oog pretoqdpouv mRNA oe ouyxer<prg6veq neptol5g rou oorurdpou (pl'5ne Evd- qia 22.70). To veuptovtx6 xoxxio (neuronal gran' uiesl ovoyvtop[mrlxov oe veuprilveg qq Drosophila' 'Onoq xot to l.rqtptrd rorx(o mRNA, q Lemoupy(o ou- ttilv ttov rorr(rov epntrdretot ornv Koroorol'ri qq petd- qpoonq Kor rn peroQopd er6trrirv mRNA' Mto t6roptrl ^o-nVopio otol-tott6(ov nep0.oprBdvet to xoxxio stress (stress granuiesl. To rorr[o stress 61ouv opretd 6to- ,poptt,o{ orioroorl on6 toug npoqyoigevoug tpe(q li- noug. Oot6oo, en(or1g nept6Xouv peto<ppootxdrg ovevepyd mRNA to ono(o ouvoOpo(owot oe ondvrn- on npoq gto yevrxrl ovoorol,ti r1q 5vop(r1g trlg petd- Qpoonq. *t"S To tleptooorepo Europuolro mRNA AnodouoUvror M6otrl Auo Movonorttilv E[oprdrpevov ono [lc]'Lorle' -r ^l- 635
22.6 AAAI Movondrro Ano66pnonq IroxeUouv tuyrexprpdvo M6pro mRNA * Teooepo enrnprlo0ero povonono ono6o;u1or1g enuetrofv rrl pu0pr(opevq onoddprlol er6trov mRNA. * Arpo[peon qg r<otrunrpog ove(oprrlq ono rrlv onoodevu],uo_ or1 e[etrlooeror orov unopXeL pro porpto notru(A) oupo. * H onoboplol Tov Un notruoDevutrur.rlrevov mRNA Tov r0To_ vtlv nupodore[rot pe npoo0rlrrl ptoq notru(U) oupog oro 3. orpo. qn H onooounorl optopevov mRNA prnopeL v, opXioet Ue neurl ono evdovourtreooq erdrrd yro qv otrtrr1trouX(o r.1 U 6oprl. e 'EvoS oyvtoorog opt0p6q popur;v mRNA oroXeuovrot yr, 0n0_ 6opr1o1 1 gerorppoonrf roroorotrrl on6 m i cro R NA. 'E1ouv neprypoge( r6.ooepo enrnp6o0ero provondrto ono66pqoqg rou mRNA to ono(o ouvotf((owor 6ro_ ypopl.rorrxd ouq :;iljiil:i :j,:..ii r<or ,til :i pio(( pe ro 6rjo r<ripro l.rovondrLo. E(vqr er6u<d yro unoo0vo).o pop(cov mRNA ror nepil.o1_rBdvouv ouprBdwo pu0pr(6pev4g ono66;rr1o4g. 'Evo provondtt nepiloprBover o<po(peo4 u1g xoLrj- mpog ove(dptrlq ond t4v onoo6evu).luloq, 64],o64 11 oqo(peor1 e[e].(ooeror or<6tr1r1 r<or drov undpXer Uro Ue- yd).r1 oupd poly(A). H orpo(peo4 rrlg xo).Unrpoq oKo- Lou0e(ror on6 n5q4 pe Xrn1. H nopdxopqrn rou Brlporog rqg onoo6evul,(rrroqg npouno06rer 6vov ;r41o- vrop6 yro qv enrorpdreuon rnq Unxovnq o<po(peoqg rqg rotrUmpoq Kor u1v ovooroL4 qq np6o6eorlg tou elF4F Xropig q Boq0ero rou ougn}6xou Lsm 1-9. 'Evo on6 ro mRNA nou ono6oproUwor qn6 our6 ro govon6- u e(vqt ro mRNA RPS28B, to ono(o xto6rxonoref rr1 p- Booroporrx4 npcoreivq S28 ror e(vor 6vog ev6tog6pov 1.r41ovropr6g ouro-pU0puoqg. Mro 6oprq p(o1ou-0r1Ar6g m4v neproX4 3'UTR eprnLSKeror orrlv enrmpdreuorl evdg evrq4u'r4 ogoipeoqg trlg r<oi,0mpog. Enrorpfrerr oq ouprBofuer pr6vo 6tov q nporetvq S2g npooddvsror orrl 6opr4 pr(oXou-0q).rdg. Iuvenrilg, 11 nep(ooero eleri Oepqg S28 oro xfrropo 0o nporoAdoer enrdluvoq r4g onooUv0eoqg rou ow(ororlou mRNA. 'Evo derirepo er6trd povondrt lp4orpronorehor oro 0qLooru<d yro rrlv ono66grlor; rou mRNA rrrlv rqrovdnr. Awd ro mRNA e(vor une00uvo yro rr1 ofvOeoq rou rE- pdonou opr0pro0 rov rorovcilv nou lperd(owor xmd rnv owrypoe{ rou DNA. IuoocopeUowor pr6vo rcnd 14 qdon S xor onodo;rotiwot rolfroro oro t6l,og rnq. To ;tr1 noluo6evuAur:1.rdvo mRNA rcov rmovd:v reppcrri(or ror Ue pro 6opr1 pr(oXou-0r11,(og nop6proto ge r4v oupri noMri:v Borcrqproxrirv mRNA. H ono66pr1o4 roug $1a o(rooqpre(coreg opror6r4req pe rnv onooUv0eo4 rou ln- RNA oro Borcrrlpro. Mto no).upep6o4 6ogu<d nop6poo trre q Borcrrlpror4 poly(A) nol,uprepdoq npoo06rer +rlo prxprl oupd noLu(U) owf qg oupdg poty(A). H pxpfl oupd ),er-roupye( oov nl,otg6ppro yrq ro oripdoxo Lsml-7 xqr/n ro e[<oocoprdrto, evepyonouilwoq ro Kt> 0repopr6vo ;rovondrro onoorivOeoqg. Aut6q o rp6nog ono66pr1or1g nopSXer Uro onUowrKri e(e).rrcnx4 ow6e- AqoLpeorl (A) rrlg xo),[nrpog oveIriptrlrrl onrl r4v onoobevutr(roor1 AAAAAMAA Poly(U) notrupepdorlnpooodKn olryo (B) (U) mo mRNA rov totovov, poly(A)notrupepdorl ,rrrr1-) Ev.ovourtreotrurLxn _ Ev0ovouxtreoorl (f) *nrn' pz6'*----------[ft'.-'.'|'iJ*- ev6ovouxtreoorl -Anoouiqq peootropotjpevrl ond miRNA ii{*l*il l? t Atrtro povonorto onoouvOeolg or0 tuKopuoTtKo rurropo. Anerrovr(eror ro opxrxo oulrpov oe roge povonol. (A) Ie oc :-i.vo mRNA 11 rotrunipo otpope(roL nprv vo ouppei onoodevu),irrlo1. (B) To mRNA rov rorovdrv onorrouv pro notru(U) oupo 11 ono(o 0c , ,. ro unoorpogo rrlg onoouv0eols (r) H onodoploq optoprlvtrtv mRNA ;rnopet v' opXLoet pe pro evdovourleotrurLrfl xonrl e(oprtirpev- --,(1v oqou1h. (A) 0progwu mRNA oroler.tovrotlt unobolrqoq roruororqq peroqpuoqq un6 oupnrlpoporyo mRNA-i:-l - 636 KEOAAAI0 22 Iro0epor1ro rorXorpo0eqorl rou mRNA (a) Buorx6g eworeg AMAAAAAA
Movondrr Evoprurlprooup$fv MeroyevdmepopnUq(m) YnompriIots 5 - 3'ndqrn Anoo6evutr[ooq e{optogevrl on6 npoq otriYo(A) qv onoodevutr(trlorl 3'- 5'ndt}ln Anoodevutr[trlorl e[oprdgevl ond npoq oItYo(A) rrlv onoodevutr[oor1 Aqoipeorl rrlq Aqo(peoq rqq xotrunrpogove(dpqrrl xotrunrpog ond onoo0evutritrlorl Ev6ovourtreolulxo Ev6ovourleotrurtxil govonol xonrl lMovondl rorv Otrtyoouptduvttr[oor1 mRNA rov totovcitv Movonol rolv Zeuyoptrlgo Pooeov miRNA ue to miRNA oro o[Untroro RISC on qvdpeoo oro ouorrluorq onooUveeonq rov npoKo- puordv Kot Tov euKopuor6v. 'Evo tpko Uovondfl opxi(et Ut pto ev6ovouKleolu- nKn Konn e(optri:prevq on6 rrlv oMqlouX(o n q 6oUti. H xonrl oKolouee[ror on6 5'+3'4 3'-+5'n6rprltov r]'o- opdrov. 'Eneno, dvo erro0optoru<6 (scavenging) 6v(u- Uo ogo[peonq rnq Ko^Unrpoq, 6togopertr<6 on6 ro o[pnloKo Dcp, prnope( v' ogotpSoet rnv xo].Untpo. 'E1ouv ovoyvoptore( 6rd<popeq ev6ovouKledoeq nou droonoUv er6K5q 06oetq-or6Xouq oro mRNA. Mto ev- 6toq6pouoo ilep(nroon e(vot 11 oroXeupr6,v4 dtdonoorl rou mRNA CLB2 orrl(6pn, n ono(o ou;tBo(vet g6vo oro 16l,o9 r4g pr(roo4g. H evdovoux).edoq nou rotoLUet rqv ronri, yvtoorq cog RNAo4 MRP, r<otd to preyoLfrc- po ;rdpog rou KurroptKoU r0xi.ou neptop((etot orov nu- prlv(oro Kor ro ptroX6vdpto, 6nou egntrdretot m4v ene[epyoo(o tou RNA, oMd pretotp6perot oro rurrop6- nloollo oro t6Log tqg prfucootlg. To r6ropro trrovondrt e(vot to govondtt ttov microR' NA (miRNA), nou odrlye( r<oteu0e[ov o' evdovoux],eo- Lutrxrl ronrl tou mRNA r1 oe rotootoLq tqg per6gpoorlg tou. Itq ouyrerpqrdvq nepfmtrloq, to m- RNA oroXefetot on6 to (euydptopro pru<pd.tv (19-21 bp) ouprnl4poprortrriv RNA (miRNA-o64yo(, guide miR- NAs) oro ntro(oto ev6g nporeivtro[ oupn].6rou nou ovoprd(eror RISC. To miRNA-o6qyo( npo6plowot on6 preroyporp6prevo yov(6to miRNA ror 64ploupyotiwot on6 q 6r6onoor1 preyol.Uteprov np66popttov RNA. Iuve- ilci)q, n onooroOepono(qoq tov mRNA-ot61ov el.6yXe- ror on6 q pu0pl(Sgevrl Ueroypo<pr1 tov yovtdi<ov ttov miRNA. Ot l,emoprdpeteq rou prrllovtoprori 0o neptypo- qoriv oro Ketpdtroto 30. H oqpoo(o ouro[ tou vSou ;to- vonqrroU otrl ouvotrtr<q onooUv0eon rou mRNA nopop6ver odreurp(vtorrl, oMd 0o ;rnoporioe vo efuot Ueydln. 'EXet 6totun<o0e( 11 np6Bler.frl 5n orov dvOptrt- no Lemoupyoriv routrdXtorov 1 000 miRNA. Iiprgcovo trr' 5vo olor<trqpolpr6vo pow5tro Ytq rnv ono66pqoq tou mRNA, to l.tovondtto onooUv0eoqg nou e[oprrilwor on6 rrlv onoo6evu],(toorl owmpooconeUouv ro npoenil,eyp6vo ouorr]prorq Yto rrlv onoo0vOeoq 6}'tov rcov notruodevutrtcop5vtov mRNA. O Fo0p6q rrlg onoo6e- vuL(o:oqg ro/n dMo Brlporo r4g ono66prqo4g pt6oto ou- rdlv rrov l.tovonotuilv etr6yXovrot on6 cls-dpoottrd ororle(o rou rd0e mRNA t<ot trans-6pooru<otig nopdyov- req nou undpXouv oro r(nropo. I-lopdMql,o Ue ro npo- enAey;r6vo ptovondtto, trettoupyotiv Kot ro 1-tovondno onootivOeorlg nou otoXeUouv etdrd mRNA. npooOson tou oupntrdxou Lsm oro otrryo(A). Arpo(peol rnq rotrdnrpoq 5' - 3' eEorvourtreotruttrn ndQn ond rnv XRNl 3' - 5' e[tovouxtreotruTtKn ndq]r'l onri ro e(oroopdrto 5' - 3' elovouxtreotrulrrl n6Qq 5 -3 xor3 -5 e[ovourtreotrurtxn ndqln on6 ro e[cootogorto lpdobeon rou oupntrdrou Lsm oro 0kyo(U). AQoipeon trlg xotrunrpoq roL 5' - 3' e[orvouxtreotrurtrrl ndQrl ond tqv XRNl 3'- 5'n6Qr1 ond to e[tooogdrto Ev0ovourtreotrurtxrl 0toonqon n roroorotrq qq petogpoolq ftOovriv to neproootepo notruodevutruog6vo mRNA nr0ovrlv ro neprooorepo notruodevutrurr;ievo mRNA 0prog6vo erdtro mRNA 0propdvo erdlKo mRNA To mRNA rolv Lorovtitv oro 0qtroortro l-lotrtro mRNA (dyvooroq opr0pog) EIK0NA 22.9 Kprotpo ouoronro rtrlv l.rovonoldrv onoouv0eorlq rou mRNA oT0 euKopuoTtKd r0topo. 22.7 O Er6rrdq Xp6voq HUt(tufiq Kd0e mRNA EAdyxeror ond AAAnAouxieq rl AoUdq MCsq OTO MRNA * Erdrro cls-ororye(o ev6q mRNA enlpeo(ouv rrlv roXurrlro qg ono6opqoqg rou. s Irorxe(o 0n00T0etpon0tnonq Unopei vo entroXuvouv rriv onoo0veeorl Tou mRNA, sv6 oTotxe(o oroOeponofr1or1q vo qv nepropfoouv. * T0 nLo00r0 0e AU oTotxslo elvot rotvo 0T0tXe[0 onooro0epo- noinonq oro 0r1troortxo, oro ono[o npoodtlvovrot notr[treg nporeiveg. e Opoudveq npoTelveg nou npoo66vowot oe moryelo onomoOeponoi rlqq ol.ltrenLDporiv Ue oumorxo qq $Xovnq onoouv0eorlg rot gol.ov ro enorporeuouv yo qv onodoUnol. * Irorxe[o oroOeponol1or1g unopXouv oe optogtlvo notrf oro- Oepo mRNA. s H ToX0TrlTo onodounolq rou mRNA pnope( v' otrtroEer oe onovrqorl npoq notxftr0 or]Uoro. :] ;r 0 Erdrroq Xpovoq Hpt(orrlg Ko0e mRNA Etr6yXerotono Atrtrrltrouxreq rl AoUeq Meoo oro mRiA 637 Boorx6g eworeg
Ie rr oqeil.erot q preydA4 6roxriprovoq ro:v lp6vcov npr(ofq 6rogoperxc0v mRNA rou r6(ou xurrdpou; Er- 6u<d crs-morXe(o puloo mo mRNA enqped(ouv 14 oro0ep6rqrd rou. Iuvrl0og ewon(owor ornv 3' UTR, nop6rr undplouv ror oMori. Meldreg o'eninedo o.1,6- xLqpou rou yovr6rcbprotog onord),urpov noMd t6tofue- po ouwnpnpdvo pror(Bo 3'UTR, trov ono[rov 6porg o p6).og nopopSver dyvororog. Optotruivo e(vot 06oerg- o161or yro np6o6eo4 miRNA. A).Lo e[vor 06oerg np6odeorlg yro rrq RBP, r<dnoreg on6 rrg ono(eg 61ouv yvcoorSg Lenoupy(eg orov 6Ley1o rqg oroOe- p6qroq. H roXriqro onoo6evul,(roorlg drogoperu<cbv mRNA pnope[ vo noxfi.er noAU rr 6Xouv neptypo<pe[ oMqloul(eg nou rrlv enqped(ouv. KoLUtepo pel,errltrrdvo e(vor rq oror;1eio qnooroOe. ponoirloqg (destabilizing elements, DE). To rprrlpro yro vo oprore( pro oM4LouX(o og ororle(o onooroOepo- nofiorlg e(vor vo nporole( enrdXuvoq rrlq ono66prqoqg ov eroolOe( o' 6vo oro0ep6 mRNA. H orpo[peor1 ev6g rtltorou orotle(ou on6 ro mRNA 6ev ou(dva ovoyKclorr- rd rq oroOep6rrlrd tou, 6v6er[4 6rt prnope( vo nepdler neproo6tepo on6 6vo orotXe(o onomoOepono(qor1g, 416;ro noLunLox6repo, n nopouo(o ev6g DE 6ev roo6u- vope( pe o6wopro Xp6vo qpl(oqg un6 6Leg trg ouvOr1- req, ener64 dMeg oMql,oul(eg rou mRNA pnope( vo tpononor{oouv r4v onore}eoproru<6rr1td rou. O r<ol,Urepo preLerrlprSvog r0nog DE e(vor ro ntroU- oro oe AU ororyeio (AU-rich element, ARE) nou Bp(- oKeror orrlv 3' UTR nep(nou tou 8% rcov mRNA r<ov 0ql.ooru<rirv. To ARE e(vot erepoyewi r<or 6roxp(vowot oe oprerorjg unotUnoug. 'Evog ttinog onoreLefuot on6 rrlv newqpep4 oL).r1Lou1io AUUUA, o' rivo 4 enovo- LoprBov6gevo ow(yporpo. 'Evog dtrLog rUnog 6ev ne- pr6Xer qv oMr1trou1(o AUUUA oMd e(vor nlo0orog oe U. 'EXouv rouronor4Oe( noM6g nporeiveg np6o6eoqq oe ARE pe erdrr6rrlro yto erbrrofg trinoug RRE r<o/n rUnoug rurrdpov. l-lc0g 6prog l,enoupyo0v ro orotle(o ARE yro vo 6reye(pouv 14 yp4yoprl ono65pr4o.n; l-lol,- l,6q nporeiveg np6o6eorlg oe ARE oLLq).enrdporjv pr' 6vo r1 neptoo6tepo ouororrrd rou trrqlovro;ro0 ono66- Unonq, prero(0 dMov ro e[o:oroprdrro, or onoo6evu].d- oeq Kor ro 6v(upro ogo(peoqg tqg r<oLrimpog, yeyov6g nou uno6qL6ver 611 6pouv enrorporetiowoq ro Unxovr- opr6 ono66gqo4g. To e(cooolprdno pnope( vo npoo6d- veror oileuOe(og oe oproprdvo ororle(o ARE. To morXe(o ARE oproprivcov mRNA enrol0vouv ro Brjpo onoodevuL(oonq rnq onooUv0eoqg. En(or1g (oog 6pouv deurol0vowoq rnv onoteLeogoru<r1 np6o6eor1 rou mRNA oro ool.rori6ro ene[epyoo(og. l-loMd morXe[o DE nAoUoro oe AU ror dMo e(6r1 onomoOepono r4rrr<cilv mo r1e(tov rowono 14 0r1 Kclv oro mRNA rnq (0UnS xor dMov np6runov opyovtopr6v. t-ro nopdderypro, oTn ([pn or npcoretveg Puf npoo66vowot oe erdmd ororXe(o ntrorjoro oe UG ror enmoltivouv rqv ono66prr1or1r<ov mRNA-or6Xov. Ie ouq qv nep(mo- on, o Unxovropr6g onooroOepono(r1or1g enmoX0vet rrlv onoodevuL(<oon eTtorporstiowog r4v onoodevuLdorl CCR4-NOT. Mro ovdl,uorlrov 3'UTR qg (Uprrlg touro- no(r1oe 53 morXefo oMrll,ouX[og ro ono(o oXer(owor pe ro 1p6vo nprEons ttov mRNA nou ro nepdlouv, eU- pnUo nou unodrlLrirver 6n o opr0pr6g rrov 6rogoperu<cirv ororle(ov onomoOepono(qoq g (oog e(vor ;reyol,fte- pog. It4v EIKONA 22.10 ouvorp((owor or yvcrror6g 6pd- oerq rov ororle(orv onooro0epono(4or1 g, Irorleio oroOeponoi4o4g (stabitizing etements, SE) touronorq0rlKqv oe Liyo oouv4Ororo moOepd mRNA. Tp(o mRNA nou peLerrl0nKov oe r[rropo Oq].ooru<oiv 5Xouv orqv 3' UTR otri.4LouX(eg oroOepono(4o4q nLori- oeq oe nupqlr6fuq. Ot nporefveg nou npoo6Svowqr oro ouyrexpq.dvo ororle(o rou mRNA qg oqopfuqg o.lr.qle- nr6porjv Ue rnv PABP, 5v6er(q 6tt 0o pnopo[oe vo ler- toupyoriv npomoteriovrqq rrlv poly(A) oupd on6 ono66prqor1. Ie opropr5veg neprmc0oerg, 6vo mRNA prno- pe( vo oroOeponoq0e( ovootdMowoq to DE tou. Flo no- pd6erygo, ouyr<erpq.r6veg nptotelveg np6odeoqg oe ARE epno6(ouv t4v onooro0epono(qoq rou mRNA on6 ro ARE, prdMov ovoordMowoq rr1 06oq np5o6eorlg ARE. 'Evo nopd6ery;ro pu0pr(6;revqg oroOep6rrpoq rou mRNA nop6ler to mRNA rrlq rpovogepiv4g (transferrin) rcov 04- l,oorrxrilv. Iro0eponore(rqt 6tov puo erdu<r; nporetvq EnrorpdTeusn rou grllovrogorj oroddUnonq Iuvopgo)dyqorl rou PB; 06oq m6Xog yro evdovoudeosn Avumotrrl npdodeorlg rnq npoteivnq onooro0eponofioqg PABP-poly(A) Dcp ffi Nouxtreoorl poly(A) _ ")-> I l<_-___)ur M'.&/ MqlovrolroL pe rouq ono[- oug Leuoupyouv ro orotXe[o onooroOepo- norqolq (DE) xor oroOepono[qorlq (SE). 0r enlnroroerg rov DE rot SE oq oro0epoqro rou mRNA ;:eootrogouvroL rup[og ono lq nprrlrefveg pe rrq onofeg otrtrrltrentdpouv E[orpeol onore],e[ 6vo DE ro ono(o ],er- roupyer olg ordXog yro evdovourtreooq. 638 KE0MA|O 22 Iro0epdqro ror Xtopo06rqorl rou mRNA
H pti0Uton rrlg oroOepoqtog rou mRNA *c rpovogeptvlq on6 ro en[nedo rou otOflpou To IRE :rlv 3' UTR etvot 0eor1 npoodeorlq yto rrlv nporetvl nou :roOeponotel ro mRNA. H nporreivq nou npoo66verot :ro IRE eivut euo(oOr1rr1 oro en[ne6o otdnpou (Fe) tou ..rropou: npoo6everot oto IRE g6vo 6rov ro enlnedo ot- lnpou e[vot Xopntro. En(neDq otonpou XoUrltrd To mRNA tns tpqvoQep(vnq e(vot mo0ep6 llpotreil'1 :: - npoodevero,:.: :i AAAAMAAA @ AAAAAAAAA npoo66vetot oro orolleio qn6wnonq oro o6rlpo (iron' response element, tnel nou nept61et, 6vo orotXe(o to ono(o ewon((etot qrnv g UTR rot onotelsftot on6 notr'- Ionl6q 6op6q ;r(cn1ou-0qNdq, 6nog qofuerot ornv EIK0NA nfi. H ouyy6veto rrlq npcoteivrlq yu to IRE otrl'6(et pe rqv np6o6eor1 otdrlpou: e(vot 1opr1trrl 6tov 11 06oq np6o- deorlq tou ot6rlpou e(vot roteilrlprpr6vq r<ot ro ow[orpo- go. 'ci.ou q ouyr<5wpcoor1 otdripou mo xirropo e(vot Xo1.rnNl, Xpetd(etot neptoo6tepq tpovorpep(vq Yto etoq' vrjv,r oianpou'on6 rrlv r<urlotpop(o rou o(poroq, on5te ro m RNA rqq tpovorpep(vr1 g oro0eponoekot' H nporchnl np6odecrrlq'oro IRE oro0eponoe( to mRNA ovoor6Mov- rqq rn leroupy(o ttov yerovxrilv oMqlouXttilv onooro- 0epono(qo"r1q. E(uor o[toorlpre(ulrg 61] (6to npotetvrl npoo66vetoi en(or1g o' 6vo otorye(o IRE tou mRNA rrlg eeppmvnq rot to puOg((et ;re no}'0 6togopenr<6 rp6no' H qapptt(vn e(vot pto npoltetvrl np6o6eoqg o't6ripou q ono(o eVr<ttopt(errnv nep(ooeto otSripou oro xfrropo' H npcrnetvq np6o6ecrqg oe IRE npooddverot oe g(q4ouq-Oq- tu5q IRE oqu 5 UTR rrlq Eeppr(vqq 6rov 19 ertue6o ot- dqpou e(voi Xoprl}'6 r<ot ovoor5trtret n'1v oMqMn(6poor1 rou ouprnl6r<ou np6odeoqq onlv ro}'rimpo Ue ro mRNA rnq qeppmvng. 'Erot, 4 pretdgpoorl tou-mRNA rnq Qep- pnfis nopelinodi(etot 6tov ro en(ne6o ot6tipou oro L*r"p" e(vot loprqLd, ouvOrir<eq un6 tg ono(eg oro0epo- noeftot rot prerorppd(etot to mRNA tqq tpovorpep(vr1q' I-1oM6g'npr.iiueq np6o6eorlq oe cis-orotle(o utpi orcrwcrt tpononoqoerq ot ono(eg go(verot 6tt enqped- (ouv u1 trettoupy(o rouq, n'X. tptoogopul'(toor1, peOu- )t(oorl, oMoydg Stop6ptpoorlq e[otr(oq qq np6o6eorlg rel,emov Kqt tooUepe(oor1. T6roteg tpononot(oetg (ocrlg euOrjvowot Ytq nq oLLoydq rrlq tolUqtoq qno- 66Unonq tou mRNA nou endyowor on6 runoptrd of- Uoiq. H onoo6v0eorl rou mRNA oMd(et oe ondwrlorl npoq eupU qdopo neptBotrl'owtr6v rot ev6oyevdrv oq- g6ttov, 1.reto[ti dM<ov 11 np6odog rou KurrclptKori 16- x.ou, 4 rurroptrri 6to<popono(r1or1, oppr6veq, n rpo- rpo6oo(o oe 0pemr6g ouo(eg xot toyeve(g Lortrldr[etq' Mdr.fteq Ue UtKpoouototX(eg Sdet[ov 6rt to 50% nep(- nou rov oLloyriv oro en(ne6o tou mRNA nou npoKo- Lo[wot on6 xurroptr<d oqproto oqeilowot oe ouU- fdwo oroeeponohorlq q onooroOepono(r1or1q r<ot 6xt oe oMoydq rnq UeroYpoqnq. To ncirg enmuyXdvowot our6g ot oMoydq oe prey0to 9o0U6 nopog5vet 6yvt'l- oTo. En(fledo otonPou uUntrd 22.8 Io Neoouvre0elPdvo U6Pto RNA EA6yxovrot Vlo EAoTTdLtoTo Mdoo Evoq tlupnvrro0 Euorhporoq Entripnonq @L9--l , o To etrorroUortro nuprlvtro RNA ovoyvtrlpi(ovrot rot r0r0- orpeqovrot ono 6vo ouorrlpo entrnplolq * To nuprlvtxo e[ulotrll.totto tretroupyet rooo roro rnv one(ep- yooio tpuototroyLrtirv RNA unoorptlgorolv ooo rot roto u1v roroorporpl rctlv etrorroportrtirv RNA. * Iu (0un, ro oupntroro TRAMP entotpore0et ro e[ootr-tltotto oe oviilpotro popLo RNA rol dLeurotr0vet qv 3'-+5' e[ovou- rtreotrurtxfl 6Poo1 tou. * IT0 ufl0oTptlgoro yt' ono66p1or1 p6ool rou oupntrorou TRAMP-e[oroopor[ou nepfioptpovovrot Un ouppogU6vo rl ovtilgoho ouppopp6vo npo-mRNA rot tro0oq repportoptlvo gerdypotpo rnq RNA notrupepoorlg ll Xtrlpiq notu(A) oupo e To neptooorepo peroypotpo uq RNA notruuepoorlq ll totr:q ei- vot rpuqo ooroOn UeroYpoQo llou roroorpdqovroL yprlyopo orov nupflvo, Otro to veoouweOet;r6vo RNA unoBdMowot oe nol- ond Bqproto ene[epyoo(oq petd tr1 peroypoqrl rouq (Bi6ne Ke<pdoro 21). Ze r<dOe Brlpo prope( vo y(vouv ).d0r1. Evto to d0q oro DNA dtopOtr'lvowot on6 notrfto ouorqpoto ent6t6p0cooqg (B6ne Ke<pd- Loro 16), ro Ld0r1 nou ovtXveUowot oro RNA owtge- rconf(owor Ue Kqroorpocprl rov e]'orr<optottt<tov RNA' Iuorriporo enlr{pqoqg rou RNA (RNA survellance systems) undpXouv t6oo orov nuprlvo 6oo rot oto r<urrop6noopo Yto vo 6toXetp[(owot 6tocp6pov et- 6ov npoBlrlpoto, H entrrlprloq nepAoprBdvet 6po- orrlpr6tqteq 66o etdtov: Uto Yto v' ovoyvtlp((et Kol' vo or11lo(vet ro etrorro;trtqrLr6 RNA un6otptol-to Kot Uto dM4 yto vo to rotoorP6<Pet' d *o-o*po.p5oq e(vot to nupqvtr6 e(olotop6rro' O nupqvoq (core) tou nuprlvLt<oti e(cuoopot(ou e(vot oXe- 66v 6protoq Ue ro Kurroponoo1.totLx6 e(u'lotr:pdtto' no- p6tt otrtrq).ent6pd pe 6togopettr<o0g npo:teivtr<oUq ouprnopdyoweg. Arpotpei voux..eot(6to on6 tq sroxeu- pSvo mRNA trre 6p6o'r1 3'-+5'e(rovour<)'edoqg To nupq- To mRNA qq tpovorpepivrlq elvot omo05q 21.* To Neoouvreoetptevo pdpto RNA Etr6Yxovrot yto Etrorrdrporo Mtlotrl Evoq l-luprlvtxou Iuorqpotoc EnL:-c';'c 639
vrK5 e[oooUdrro 6Xer noMonLsq Leroupy(eg, peto[rl dMov 11 ene[epyoo(o oprqrdvtrtv 1.u1 r<to6u<onor4urc0v RNA peroypdcpcov (snRNA, snoRNA xor rRNA) ror q nlnpnq ono66pqorl rtov el,orroportxc0v RNA. To e(to- orrrpdno entorporeUeror orcl unompcitl.roro nou np6- Kerror vo ene[epyoore( tri5oo nproreivrrc6v oupn].6r<ov nou ovoyvop(ouv er6u<69 oMqLoul(eg RNA n do;rdg RNA/RNP. l-ro nopd6eryuo, ro Nrdl-Nab3 e(vor rivo nproreivu<6 6rpep6g pe erdu<6r4ro yto ouyKeKprgdv4 oL- tr4LouX(o ro ono(o enrorporcUer ro e[rrloroprdrro oe qu- orol,oyrxd unootpcbgoro snRNA/snoRNA. Aur6 ro (euydpt nporeivrirv npoo66veror oe ororle(o GUATA/GI Kqr UCUU, owrororXcog. O ouprnopdyowog Nrdi-Nab3 epnA6xeror en(o4g orov reppono;r6 qg peroypoqqg rov pn noLuo6evuLurt;r5votv RNA nou preroypdgowor on6 qv RNA noLuprepdo4 ll. Aur6 uno6q).dtver 6rr ro e(ootopdrro foog enrorporeUeror oneuOe(og mq 06o11 qg oUv0eo4q rouq. AvripoLo ene(epyoo;r6vo, rpononorqpuivo q roxo6r- n).copr6vo pr6pto RNA onomo[v d].troug nproreivrxorig oupnopdyoweq yro qv ovoyvcbpron Kr entorpdreuo4 rou e(oorol.rorfou. Ir4 (rjprq, ro rripro nuprlvrr6 o6pr- n).oxo nou errele( ourq q l,enoupy(o ovoprd(eror TRAMP (or<pcovripuo tov nporeivc0v nou ro onoreLoriv) ror undpler rouldXrorov oe 6Uo Uopqdq nou 6rogd- pouv oq npoq rov rtno qg poly(A) no).uprepdoqg nou nepr5louv. To oriprnAoro TRAMP enrlped(er qv ono66- Unon Ue 6rdEopoug rp6noug: 1. AM4),enr6pd d;reoo pe ro e(cooto;rdrro, 6eye(pov- roq rnv evepySqro e(ovour<Ledoqg nou 6ro0drer 2. l-lepil.oprBdvet ;.uo eAu<do4, q ono(o prdL),ov e(vor onopo(rr'1q yro ro {erri}.rypro qg 6eweporoyo0g 6oUnq ro/r1q preror<[vqoq npo:reivriv nou npoo6d- vowor oe RNA on6 RNP unompd:goro xord rrlv onod6prloq. 3. llpoo0Ster puo ;lrpri 3'otrryo(A) oup6 oro uno- orp6poro-or6Xoug. I1toreUerot 5l 11 oLryo(A)oupd rdver ro oroXeuprdvo RNP xolrirepo un6orpopro yro ro pqxouogd ono65;r4or1g, 6no4 orpgdrg Kor oro Borcrrlpro. H EIK0NA 22.12 ouvotp((er rouq p6Loug tou TRAMp Kor rou efootol.rot(ou. E(vor nfuiov oog6g 6tr 4 ono66- Unorl rou RNA oro Borcrrlpro ror r' ApXo(o rot r1 ono66- Unorl rou nupqvrro0 RNA oroug euxopuolreg oXer((owor ovomu(ror<d. H opor5qrd toug uno6r1).ril- ver 6tr opXdyovog p6).og r4g noluo6evul.(oo4q r.1rov vo 6teurotrUver rrlv ono66prr1o4 rou RNA r<or 6tr q poly(A) npoooppr6or4re opy6repq fiouq euxopucbreg yro rrlv nopd6o(o ow(Oerr'1 Lenoupy(o r4g oroOepo- nofiorlg rov mRNA oro rurropdnlool.ro. floro efuot ro unoorpripqro rrlq ono66g4oqg prdoro TRAM P-e(cooopo{ou ; To oti pn}.or<o TRAM p ovoyvcop(- (er peyd,l.q noxil.fo e].ortrr:;.rotxcbv RNA nou peroypd- gowqr xor qn6 nq rperq RNA noluprepdoeg. Aev e(vor yvom6 ncbg enmuyXdveror our6 roOrig ro RNA-or61or 6ev 51ouv rorvd Xoporrrlprorxd. OproprSvor epeuqtdg unomq p((ouv d,vo prow5.tr o r<rv4ru<ori owoyrovrollori, uno06rowog 6l unoorpci:1loro ono66pnonq on6 ro e(cootrtpduo 0o y(vouv RNA ro ono(o dev ug(mowor ene[epyoo(o oUre xot ouvopltoA.oyolwat 1yxatpa oe puo oprorxrl [opAn RNP. Aur6g o;rrlXovro;rdg onoge0- yer rrlv npoun60eo4 er6u<r1g ovoyvroprorlg rrov ovop(S p4tov nt0ovcbv elorrolprdtcov. Tr e[6oug ovopro].(eg xorodu<d(ouv ro npo-mRNA oe Korqsrpotprl mov nup4vo; 'EXouv ovoyvroptme( 6uo e(6q unoorpoprdrcov. O 6vog r0nog neprloprBdva Un ouppotrU6vo 4 ovcbpoLo ouppotrrprdvo npo-mRNA. Tq ouoronrd rou ocol.rot(ou oupporpqg ouyxpororiv rriroro pretdyporpo Scog 6rou e(re ono6oprqOoUv on5 ro e(oocopdno e(re oLor<i.r1pco0e( 4 xordM4).r1 ouppo- An (qv our6 e(vor erpucr6). l1tmeUeror 6rr ro pow6]to rou KrvrlnKo0 owoyrovropro6 prdl,trov tolrjer Kor orrl ouyrerprp6,vrl nep(nroon. 'Evo npo-mRNA ro ono(o 6ev 6Xer ouppo<pe( Kor ouoKeuoore( onoteLeoporxd ep<pov((er ou[r1g5vo rfv6uvo vo y(ver or6Xog rou pn- Xovrol.rori ono66pr;o4g on6'ro elroou:prdrto. O rp6noq ovoyvrilpro4g rov KoKoouppo;r6votv npo-mRNA 6ev ei- vor yvcoor6g. O deUrepog ttinog unoorpoprdrov nepr- LoprBdver npo-mRNA Xop(q oupd poty(A) e[orr(og LdOog repponollori. Evti 4 noLuo6evul(oo4 npooro- EnrmporeUer to nuplvtxri e[ootopdrto vovrot ono npotreivtroug ougnopoyoweg ot ono[ol entorpore0ouv ro nuplvrro e[ootr;polo. Iq (uUl, o oupnopoyovrog ervor ro ourr-. ,. xo TRAIVIP. H poly(A) notrugepool (PAP I Td4) rou TRAMP npoo0erer pro prrpfl oupo poty(A) oro 3'orpo rorv oroxeupevorv RNA. Etronroporrrd RNP . KoKodrntroUdvo RNA . Ad0og ene{epyoo(o 040 KEQA Al0 82 Iro0epdqro xor Xropo06qorl rou mRNA
re6er ro yvnoro mRNA, orrlv npqyporrK6rrlro Unope( v' onomoOeponote( ro xpuqd ooro0{ per6ypogo (cruptic unstable transcripts, CUT). Aurd ro RNA nou 6ev ru:drronororiv nporelveg (B).6ne en(oqg Evri- rrya 30.3) peroypdgowot on6 r4v RNA noAupepdorl II r<or 6ev rodrronoro0v ovoyvtop(orpro yov(6ro. 0o16- oo oulvd enrroLUnrowor (xor (otog puOpri(ouv) yov(- 6ro nou xtodrronorotiv npcoreiveg. To ouyrerpt;r6vo prerdyporpo noluodevuluilvowot on6 5vo ouoronr<6 rou oupnl6rou TRAMP (Trf4). Aror<p(vowor on6 dtrl.o gerdypogo Ue dyvtoon'l Lenoupy(o on6 u1v unrip;re- rprl omd0erd roug: r<ovovrx6 onodoporiwor on6 ro ofprnLoro TRAMP-e[oocopor(ou opr6ocog perd tr1 o6v- Oeoq roug, nt0ov6v ener64 moXe[owor on5 notruqde- vuL[oo4 e[oprriprev4 on6 rov Trf4. H rjnop[r1 r6rorov peroypdrprrlv onodeffiqre nermrrd yro npd:rr1 <popd oe metr61q (UUnS U' e).orrcoproru<q ono66pqoq RNA orov nuprlvo. Fldvtrl on6 ro rp(o t6ropro rov Uero- ypdgo:v qq RNA noLu;repdoqg ll gnope( v' oTrore- Loriv pr1 r<odrronorqru<d RNA Kor vo un6xerwot oe ypnyopn ono66prr1or1 on6 ro e[ooro;rdrro! Oprotrrdvo CUT go(veror 6l npo6plowor on6 r.!eu6ri 5vop[r1rqg peroypoqrlq Kor ro (6rq to BpoX[Bro RNA npoi6wo tunrrd 6ev epgov((ouv rdnoro Lertoupy(o (n.1 6e dpouv in frans). Om6oo, npoorp6pouv 5vo nopd6ery- po yro ro p6Lo ourrlg r<oOournq rnq petoypocprlg orrl priOproq yeuovxriv r1 enrrol,um6pevrov xrrl6rronorrln- xc6v yovr6(rov (B).5ne Ev1tqra 30.3). 22.9 O'EAeyxoq llor6rnroq Tnq Merdgpoonq Tou mRNA npoyporonolsiTol ond KurroponAoopcrrlKd Iuorfrporo Enrrfipnonq * 0 Ulx0vroU6g onoo0v0eorlg NMD oroXeuer mRNA pe npo- opo rodtrdvto reppono;.rou. * H oToxsuon Tov unooTptlUorov rou NMD onorter 6vo Drorri- prlpevo o0votro nporeiv6v UPF ror SMG. * Ie notrtrouq opyovrolroug, urtrer60, yro qv ovoyvtirproq npo- opov rordrxovlrov reppolopou e[vor 11 oouvq0roq 6opq r; ro oouvr'10Loro g1rog qg 3'UTR. Iro 0r1troolro, o U1xovropoq ovoyvrirptolg eniorlg o(totroyel ror rqv nopouo[o ro0odtrtov oupntrorov ouU0otrnq e[tovltrlv. e 0 Unxovropog onoouv0eorlg NSD oroXeuer mRNA Xtoprg ev- rog ntrotolou rordrxovlo repporropou ror poo((eroL o' evo ouvqplpevo ouvotro npotrelVdrv SKl. * 0 plXovLo;rog onoouvOeolg NGD oroXeuer mRNA pe 0T0orU0 ptpooopolo orqv rodrronotlrtrl neproXrl. Optoprdvo e(6q BoBc6v tou mRNA o[to].oyoriwor p6vo r<ord rr1 perdqpoorl. To ouorqproro ennqprlo4g e[e- LfiOrp<ov yro vo ewonf(ouv rp(o e(6q eontoprotu<rirv mRNA nou enoneil.oriv rqv ntor6rrpo rqg prerdgpo- onq Kor vo ro oroxeUouv yro ypnyopn ono66prqor1. H ': .r,i' i r.: i,,nopouord(et ro unofipoproto yt' ourd ro AAAAAM AAMAAA Ynoorp6poro rov rurropon),ooportr6v ouoqpororv enunplolq 0 gqXovro;rog NMD onodoper mRNA nou 6Xouv npotrrpo '-SLxovlo repgortopo0, nptv ono ro rovovtro. H onoouv0eol NSD onodopei mRNA Xtop(g evrog nLoroiou xo6rrovro reppolo;tou. H ono- , - J0001 NGD onodogei mRNA nou 6Xouv evo oroorlro ptBootrrgolo orrlv rodrxonorrlnxrl neproxrl. Jf .i] 0 'EtreYxoq llotorqrog rrlg Meroqpoorlg rou mRNA llpoyporonoreirot ono Kurropontroopolro Iuoqporo Enrqprlol_c 641 Buolxeg dworcg
rp(o ouor{Uoro. Kor ro rp(o nepfioppdvouv ouppdwo ovr.i: 1.roLo u repporropori n1 g ;rerd<ppoorl g : ouvenrig, e[- vor or6nrprl pro enrox6nqon rnq depyoo(og rou qu- orotroyu<o0 rcpporrolloU (BMne Ev6qto 24.15). 'Orov 6vo ptBooro;rdno nou prerogpd(er rprdoer oro ro6116- vro repUorrol.ro0, 6vo (eUyog nopoy6wtov qnetreu06- poonq (release factors) (aroug europucbreg, eRFl xor eRF2) ero6pXeror orrl 06o11 A rou prBoocol-rot(ou, 11 ono(o rord rlv enqlfruvoq xovovxd e[vor efvor xoret- InUU6vn on6 eroepl6prevo IRNA. To oripnLoro rov no- poy6vtc,lv onel.eu0Sporo4q peoo).oBe( orrlv onetrsuO6proon rou oLoxLrlpcopSvou notrunemt6(ou rr Sneno rou mRNA, rov IRNA nou on6pewqv Kor roJV unopov66rov rou prBooroproilou. H onoo6v0eorl nou peoo)roBehor on6 perotrtr6lerg lr,rpig v64po (nonsense-mediated decay, NMD) oro- Xe0er mRNA nou nepr6louv 6vo np6opo r<rodu<6vro rspporropori (premature temination codon, PTC). Om6oo, np6copo r<ro6rr5vto repporrol.ro0 pnope( vo npox0rlouv ror pe d..trouq Unxovrogorig. l-ro nopd6ety- 1.ro, yov(6ro Xtop(q ;retoMd(erg pnope( vo 6qproupyq- oouv ovrilprol,o perdypogo nou nepdXouv PTC L6yro (1) ).dOoug qg RNA noLupepdoqq n (2) prrl olorl,rlpo- U6vnq, eogol4r6vqg 11 evoMor,ctxrlg oupporpqg. Erctprd- ror 6n qXe66v ro 50% rov evolloKrxd ouppoprprdvrov npo-mRNA 6qproupyo0v roul.dlrmov Uro Uopqn Ue PTC. l-1ep(nou ro 30% rov yvoordlv voooy6vrrrv oMq- Lo;r6prp<ov pdl),ov r<o6rronoro6v 6vo mRNA ge PTC. 'Evq r6roro mRNA 0o nopdyer nolunembro pe roLo- Bcop6vo C-rcllrd dxpo, ro ono(o 0eorporiwor r6rofuepo ro[trd yo ro rUnopo e(on[og qg rdorlg rouq vo Troyr- 6e6ouv noMd dMo 1.r6pro oe pr1 Lemoupyrrd oUpnl,o- Kq. To govondrt NMD Lerroupye( oe 6Loug roug euxopurilteg. H or6Xeuoq mRNAnou nepdlouv PTC npouno06ret vo ouprBe( perd<ppoorl xr efoprdror on6 6vo ouwqpqpt6- vo orlvoLo nporeivxc0v nopoy6w<ov, nou nepil.optBdvet nq rpe(g nptoreiveg Upf (Upf1, Upf2 ror Upf3) r<ot t6o- oeprg enmp5oOeteg nporeiveg (SMG1, 5, 6 xor 7). An' 6leq trg nporeiveg NMD, npc0rrl 6po 4 Upf1, 11 ono(o npoo6Sveror oro p$oorol.rdrto nou Bp(oreror oro rrodt- r6vro repporrolloU, er6x6repo oro oriprnLot<o rou TIo- pdyowo onetreuO6proorlq. H np6o6eo4 UPF orlproSe0et ro mRNA yto yprlyoprl onooUv0eorl. l-1op5rr or er6u<o( p6Lot rrov nopoy6wrov NMD nopop5vouv o6teuxp(vt- oror, roor6oo tdto(tep4 oqpoo(o 61er q googopul(ooq qg Upf nou e(vot npoo6e6ep6vq oro p$oo<rl;rdtto on6 rrlv Smg1. Or ouvduooll6veg dpdoerq rouq roto6tx6- (ouv ro mRNA yro ono66pr4or1 on6 ro yevrr6 prllovtopr6 onooriv0eo4q xot 6eye(pouv rn ypiyopn onoo6evuA(to- on. To mRNA or6Xot onodo;roriwor 16oo on6 to 5'-+:r" 6oo rqr on6 ro 3'-+5' ;rovondrr. l-ldrg 6prog 6roxp(vowor ro PTC on6 ro rovovu<6 t<t+' 6x6vro tepproloproU nou Bp[oxeror opretd nto rqOodr- r<d; O gqXovropr6g elet pretrerrl0e[ errevcbg 16oo mn ([pq 6oo Kor oro 0ql,oonxd, 6nou brorpdper rdn<rrg {El K0NA 22.14). Iro 0fli.oonxd, to r0pro o(pro nou rouro- nore( 6vo PTC e(vor q nopouo(o grog ouprBolqg oupporp{q, nou orlgo(vmot on6 ro oripnloxo ougpo- tr{g e[ovitov (exon junction complex, EJC) ro0o6x6 rou np6ropou ro6rrov(ou repporro;.ro0. Enerdrl ro ne- ptoo6tepo yov(6ro rrov ovri:repov suKopuordrv 6ev SXouv rwp6vro rou vo 6tox6mer qv 3' UTR, r' ou0ewt- xd xo6rx6vro repproropro6 6ev orotrou0o0wor on6 06- on ouppqqrlg. Kord rov rrporon6po yfpo rqg yer6gpooqq (pioneer round of translation) ev6q xovo- vtxoU mRNA, 6l,o to EJC Bp(orowor p6oo orqv xodrxo norqnrf neptoXr] rr ercron(owot on6 to 6repX6pevo , '',:'' , Auo plxovrolroi yto qv ovoyvri-rproq evog rodrrovlou repponogou og npourpou. (A) Iro O1trootuo, 1 nopouolo evog oulL- ntrorou ou;rpotrnq eltovur.rv roOodrro rou rtrldxov[ou repplonogou oroXeuer ro mRNA yro NMD. (B) Moltrov oe otroug roug europudlrac pro orpuotro Ueyotrn 3'UTR ovoyvrrrpi(eror ono qv onoorool ovopeoo oro roldrrovro repporropou ror ro oupntroro poly(A)-PABP. Ie xoOe nep(nrooq, rl npo.rrelvrl Upfl npoodSveror oro ptpoorrrpolo repgolopou yro vo nupodorloer rrlv onoo0vOeo1. 642 KE0AM|0 22 lro0epoqro rorXtopo0er4oq rou mRNA Avoyvtirproq PTC p6oro ro0odxori EJC Avoyvtilprorl PTC ond r1v no),rl gurprd 3'UTR
" t-l -'- u[m,oFTffi- . ,r' ,: .,: To onoretreopo rrlq npoodeolg ev6q PABP t<ovto o' evo npoopo rtodrrovro reppouolrou. 'Evo yovidro PABP gerorponlxe (rl01t v0 erqpo(et gto neptoXrl npoodeolg oe RNA evoq qoyou H neproXrl npoodeonq evoutpor60lre o' evo doupoorLxo yov[6to-uno- oTpopo yto NMD. To npoodedepevo PABP epnoOroe rr1 ouv101 yplyopl onodoprlorl ourou rou mRNA ono ro prqXovro;.ro NMD. Aurrl q p6- 0oDog eXer notrtreg eqopgoy6q orn goprorrl protroyro. OXr NIVID t,'t . prBoorotrrdrro. Kord rov nporon5po yUpo qg prerdqpo- onq yro 6vo un6orptrrpo NMD, or nprore"[veg Upfl xor Upf3 npoodSvowor oro(o) xo0o6x6 rordlorno(o) EJC, oroXerlowog ro yro ono66prqor1. To neproo6repo yov(6ro rou S. cereyr.srae 6ev 6ro16- firowor ro06],ou on6 twp6vro, on6re o prqXovtotrr6g ev- ronroprori rov PTC np6ner vq e(vor 6rorpoperrr6g. Ie ourn rrlv nep(moor1, orlpro rrv6rivou e(vor ;uo orpriotro peyoln 3' UTR. Aur6 ono6etX0rlre on6 to erjp4pro 6tr r1 entg{ruvo4 rnq 3' UTR ev6g quoroloyrKorl mRNA 0o pnopo0oe vo ro Uerorp5r{.rer oe un6orpo;ro yro NMD. 'Evo rp61ov ;rowSlo npore(ver 6rt o ooor6q repUorr- op6g rqg prerdgpoo4g o' 5vo ro6rx6vro reppronoproU npo0no06rer 6vo orlpo on6 6vo r<oww6 PABP. MoLov6rr or 3'UTR 61ouv noLri 6rorpoperx6 pnKoq, q <puou<r1 on6- oroon peto[ri rou ro6rrov(ou reppotroprorj ror qg poly(A) oupdq 6ev e[oprdror onoxlerorxd on6 ro Un- xog roOc0g 6eureporoye(g 6op6g xor oMqtrsnr6pdoerg pero[ri RBP prnope( vo rrl oupnr6oouv. H ovdyr<4 yro PABP ono6etl0qr<e oe noMorig opyovrogo0g npoo66- vowoq pro PABP r<owd oro PTC (B).6ne EIK0NA 22.15), on6re ro ow(ororlo mRNA 6e oroXeu6rov n).6ov yto NMD. En(orlg, 4 ovoyvrirproq rou PTC ouprBo(ver ove[dp- rqro on6 q oupporpq or4 Drosophila, o"ro C.elegans, oro rputd ror oe optopr6vo mRNA rrov 0qLoonxri:v, yeyo- v5g nou uno6rlLrilver 6rr ro ptixog ror q 6opq rnq 3' UTR e(vor r<p(oq.ro yto rnv quoroLoyrrl 6ro6troo(o repporr- ogoU qg perdqpoorlg oe 6Loug roug opyovroporig. Opropr6vo guoroLoymd mRNA orolefowor on6 ro g4Xovropr6 NMD. Aurd ro mRNA routonor{Oqxov on6 nerpdproro mo ono(o elorc60rlxov ro en(ne6o rqg Upfl, pe onor6l,eopo peydlrl oU(4oq ev6g unoouv6- Aou geroypdg<ov. O xotdLoyoq rov euor.ol,oyxriv unoorpoprdrov yl.o NMD nept}opBdver pr6pro mRNA pe r6rokepo peydLeg 3'UTR, ro mRNA nou ro6rronoroUv oeArlvonporeiveg (or ono(eg Xpnoqonoroilv ro ro6u<6- vto repUorrol.rorl UGA rog xrodx6vro oelqvoxuoretvqg) xt Svov dyvocro opr0pr6 evqMoKrmd ouppogpr6vov m- RNA. Me Bdorl 6oo yvrop((oupre pr61pr oriprepo, 6s pno- po0pre vo npoBL6goupre 6Lo to unomptilUoro nou onorelorlv m6Xoug NMD. To NMD (oog ono6erX0e[ 6vo oqpowrr6 povondrr yptiyopns onooUvOeoqg yro notrfi.o BpoX0Bro mRNA. Teprlrl 06o11 nprjodeonq H onoorjv0eorl non stop (non stop decay, NSD) oroleUeL pr6pro mRNA lrop(g ew6g nLoro[ou xrodtr6vro teppono;.rori (peoo(o oxnpo orqv Eu<6vo 22.13). E[or- r(oq qq onorulfog reppotrol.rori, 6vo prBoootgd-no ou- veX((er vo preroqpd(er ornv oupd no).u(A) r<oL nL0ov6v orol.rord mo 3' drpo. Ynoorpor;roro rqg onoo6v0eorlg non stop 611;;oupyotiwou<up(cog on6 np6copo repUol.- op6 qg peroyporpqg ror noluo6evul(roo4 orov nuprl- vq. T{itoro np6opo no}.uo6evu},rcop6vo prerdyporpo e(vor nopd6o{o rotvd. H ovd}.uor1 tulo(otv n}.r10uo;rd;v oDNA or ono(or npodpxowov on6 mRNA qg (Upr1g r<or rou ov0pc6nou uno6rlLrriver 6n ro 5-10% rrov ouprBdv- rolv no).uo6evu).(roo4g (orog ouprBo(vouv oe ovo6tr<d "KPUtPd o4pe(o" to ono(o prord(ouv pe to ou0ewu<6 or1- po nol.uo6evuL(oor1g. H or6Xeuoq rov unoorprr;prdrrov non stop nepil.o;rBdver 6vo oUvoLo nopoy6wurv nou ovogd(owor nporei'vsg SKl. To prBooroprdrto oneleu- Oepcbveror on6 to mRNA pe q 6pdor1 qg Ski7. H SkiT 6,1e1 pro neptoXrl GTPdo1g nop6;roto Ue rou eEF3 ror nr0ov6v npoo66veror m4 05oq A rou prBoocol-rot(ou yro vo deye(per rrlv onetreu0Spcooq. H enor<6Lou0r1 enr- mpdreuo4 trov un6).orn<ov npoteivriv SKI xor tou e(o- orrrpor(ou 6Xer og onor6Leopo rqv onoo[voeoq rou mRNA pe xoterjOuvorl 3'+5'. H onooJv0eorl rcov uno- otprrlpdtov non stop prnope( en(or1g vo ouprBe( onouo(o r4g Ski7, on6te xor npoyporonore(tor pe ogo(peo4 r4g r<oLrjmpog ror n5r1.rr1 oe rqreUOuvoq 5'-+3'. H euoroOq- o(o mr;v o<po(peor1 qg r<o).rinrpog 0o pnopo0oe vo o<peftetor o' ercr6nroq rorv PABP xo0rig to npcoron6po prBooroprdno 6rorp5Xet qv oupd poly(A). H VpnVopn onoo6v0eoq rov unoorpto;.rdtrov non-stop o6qyei 61r p6vo oe npooroo[o on6 ro[u<d no],unem(6rq oMd r<or oe oneleu 06pooq noyr6eu p6vcov prBooro prot[ov. Ev6ro- g6pov efvqt ro yeyov6g 6rr ro E. coll lprlotponore( 6vo e[er6rxeugSvo pr; r<o6rxonorqtrr6 RNA (tmRNA) ro ono(o 6po 16oo ctg IRNA 6oo Kor oq mRNA yro vo 6to- ocioeL ptBoorrlprdrro moprorqprSvo o' 5vo nonstop mR- NA. To tmRNA xqreu06ver qv npooOrp<q ev6g pu<pori nenl6(ou nou moleUer ro eLorrol;-torLr6 npoteivtx6 npoi6v yr' ono66pu1or1, nop5ler 6vo xro6rx6vro reppol- o1-roU nou ennp6ner r4v ovor<tix),ooq rou prBooropor(- ou Kor oroxeUer ro e),ortco1lolr<6 mRNA yr' ono66pu1orl on6 r1v RNAo4. TeXvrltd PABP ge v6o neproXrl npdoDeolg :I.-q 0 'EtreYXoq l-lotoqrog rrlg Meroqpoolg rou mRNA l-1poy;loronoLeuor ono Kurropon),oogolro Iuorluoro ErLrc-:-l 643 l-lpoo0ebepdvo PABP
H onoofvOeol no-go (no-go decay, NGD) mo- Xeier pr6pto mRNA nou 61ouv ordoqlo ptBoorrlpdno oe ro:6u<6vrq u1g rro6rronorrlru<r1g neproXrlg toug (rdro oXnpo oqv Ex6vo 22.13).1-1opo6u<r1 r1 noporcropr6vrl oroorg6rqro prnope( vo npor<iqOe( on6 <puou<6 Xopo- rcqprolx6 opropr6vov mRNA, 6no4 roXupSg 6eutepo- toye(g 6opr6g xor ondvro r<rrl6rxdvto (rrov ono(<ov r' owimorXo IRNA Bp(or<owor or XopqLdg ouyr<ewpdr- oerg). Aur6 ro v6o povondrr enrrnpnonq 6Xer pe),eqOef pr6vo orq (r1;r4 r<or e(vor ro trry6repo xorovoqr6 on6 ro rp(o. H m6leuoq rou mRNA nepd.oprBdver rrlv enr- orpdreuoq rov nporeivriv Dom34 rot Hbsl, or ono(eg e(vor o;r6l,oyeq rov eRFl r<or eRF3, owrmo(Xoq. H ono66prqoq rou mRNA op{(er p' rivo ev6ovoux.,eol.ul- 16 r6rptpo r<or ro 5' r<or 3' xldoproro ndmowor on6 ro e(coorrlprdrro Kot rIV Xrn1. H Dom34 0o pnopoUoe vo e[vor 11 ev6ovoux.l,edoq, erp6oov truo on6 ng neproXSg ng ;rord(er pre vour<Ledo4. l-ror( 5vo rovovu<6 mRNA vo riXer oMql,ou{eg nou e(vor 60oxo},o vo Uerqgpooro6v ror pnope( vq ro rorodxdoouv oe yprlyopq ono66prr1- o4; Trftoeg oMqLouX(eg 0o gnoporioe vo 0eopq0oUv rrlg 6vo dLLo e(dog onomoOeponor4-nro[ morle(ou. E[eLurtrd, n 6roqpnon epnoS(tov yt' ono6orrrrl perd- gpoon uno64Lrilver 6tr e[unrlpetoUv rdnoto o41.lowrxr1 l,enoupy[o mov 5trey1o rou 1p6vou npr(ofq owdrv rtov mRNA. 22.1O Meprrd Evronl(ovror oe Kurrdpou Europuorrrd mRNA Er6rrdq lleproxdq rou " 0 evronrogog rov mRNA e[unlperet drorpopelreg treuoup- yieg oe pegovtripevo rurropo rol ovonruooolrevo 6pppuo. * 'Exouv roroyporpeI Tpsrq Urlxovropoi ylo rov evronropo rov mRNA, + 0 evronlop6g onotreI cls-ororXe(o oro mRNA-oroXo ror Irans-nopoyovreg. * 0 rupLoq plXovropoq evepyou perorpopog neprtroppover qv roreu0uvopevl r[v1or1 rcrlv mRNP roro plroq oddrv Tou KUT- ropooretrerou. To rurrop6ntroopo e(vor rivog noLuo[lvooroq 16pog nou roto).oprBdveror on6 uQq).q ouyr<6wpolon npo- reivov. E(vor dyvr.ooro nog prnope( vo droXSowor to aLerjOepo noluoopdrto Kor ro neproo6repo mRNA prdMov prerogpd(owor oe tulo(eg 06oerg or ono(eg xoOop[(owor on6 ro oqpreio ero66ou rouq oro Kurro- p6ni.oo;ro Kor rrlv on6oroo4 r;v onofo SXouv 6rovri- oeL on6 ow6. Opropr6vo mRNA piero<ppd(owor pr6vo oe ouyKeKprtr-i6vo oqpre(o, xo06g 11 pretdrppoo4 roug r<oromSLLetor ;rdXpt vo rprdoouv mov npoopLo;.r6 toug. Pu0pr(6pievog ewonroprdg dXer nepLypoge( yto ndvol on6 100 erdrrd mRNA, opr0p6g nou oorpo).rog owrnpoooileUer 6vo pirrp6 trr6vo r<),do;ro tou ouv6- 644 KEOAAAI0 22 Iro0eporrlro rot Xtrrpo0eqol rou mRNA Aou. O ewonrql6g tou mRNA e(unrlpere[ drdgopeg oqprowu<5q l,eroupy(eg o' euKoputorrr<orig opyqvr- o1-rorig 6l,rov rrov r0nrav. Tperg xp(orpreg Lenoupy[eg nopouord(owor ornv ElK0tlA 22.16 r<or ou(qrofwor oq ouvSXero. 1. O ewonropr6g erdu<cOv mRNA oro ooxUfiqpo notr- Lrirv (tirrov e(un4pere( t4v eyro0(6puor'1 peMowu<rilv nporUnov oro 6prBpuo (6nrog o d[ovog no].rr6rq- tog) ror op(er ro ovomuftor6 nenpop6vo rurr6- pcov nou edpe(ouv oe drorpoperrr<d o4pe[o. To ewonrotrrdvo prqrprrd mRNA rro6ronoro[v peroypo- gu<o0g nopdyoweg 4 d),Leg nproreiveg or ono[eg puOpr((ouv rqv 6r<gpoo4 rrov yovr6i<ov. Ito rrroxUrro- po rnq Droshophila, ro mRNA rrov yovrd[rov bicoid Kor. nanos ewon((owor mov np6o0ro ror on(o0ro n6lo, owroro(1oq, xor 4 prerdgpoorj rouq nou oKo- l,ou0e( rq yovtporro(r1or1 SXet cog onordl,eopo q 64- l.uoupy(o 6roBo0p(oeov rcov npo:teivrx6v npoi6wrov roug. Or 6roBo0p(oerq Xpnorponorodwor on6 ro xrirropo oe opXrxd md6ro rrlg ovdmu(r1q yro rov npoodroprol.r6 qg npoo0ron(oOrog 05oqg oro dp- Bpuo. To bicoid r<ro6rronoe( 6vov petoypogrr6 no- pdyovro, evcil to nanos Svov xorooroL6o tqg perdgpooqg. Opropr6vo mRNA nou ewonf(owor oE er6u<dg O5oerg rro6tronoroUv roOoprmdq rou ne- npoprdvou rou rurrdpou. l-to nopdderypo, ro mRf.lA rou yovr6(ou oskar ewon((eror oro on(oOro pr5poq rtov rooxurrdpov Kor nupodore( 14 6repyoo(o nou o64ye( m4v ovdnruEn rcov opX6yovrov BLommdrv xurrdpov cro 6prBpuo. 2. O ewonrop6g rou mRNA en(o4g no((et pSLo mrg ooril-rperpeg 6rorp6oerg tou rundpou, 6r1Lo6r1 cng trrm<olr5g 6rorp6oerg ot ono(eg ono6(6ouv 0uyorpud rUrropo nou 6rocp6pouv ro 6vq on5 ro dMo. Atn6 enmuyldveror pe oofprpetpo droXropropr6 rcov xo0e prorrilv rou nenpopSvou rrrlv xurrdprov, ot ono(or pnopef vo efvot nptoteiveg xo/ri ro mRNA nou nq r<co6rronororjv. Iro dprBpuo qg Droshophila, rc mR- NA rou yovt6(ou prospero ror ro ow[ororXo npoi6v (5vog geroypo<prr6g nopdyowog) ewon((owor oe pro neproXrj rou neprqeperqroU rplotori rou ep- Bpriou. Ie preroyevSorepo ord6ro r4g ovdmulnq, n npooovoroltop6vrl xurropx4 6to(peoq rov veupe BLoorrilv 6rooqo).((er 6n pr6vo to e(c(lroro Ouycnpr- 16 rUrropo 0o nepr5ler prospero,6eopreriowdg to vo yfuer rivo pr4rpu<6 rUrropo yoyyl(otv. H oo0p4re- rpq rurropxri 6ro[peoq Xcnoqonore(ror en(oqg on6 rn (UUn yro q 6rlpuoupy(o ev6g Ouyorpxotl rurr6- pou dro<poperxo6 ou(eurcnrotj rUnou on6 to grppr- 16 rrjrropo. 3. O ewonropr6g rou mRNA orouq 6rorpoponorr;pr6- vouq Kurroprxorig t6noug tcov eql,(r<o:v e(vor Svog prlXovropr6g yro rn 6ropeproprorono[qor1 tou rurr6- pou. O ewonrog6g [orog Xpqorpronore(ror rilore vo 6toogol.rme( 5tr ro oumonxd trrrv nol,unpcureivr- rc0v ouprnL6rtov 0o ouw(Oewor xowd ro 5vo oro dMo ror 6rr or npotetveq nou oroXeUowot oe opyo- v(6ro ri e[erdu<euprSveg neprol6g rou rurrdpou eni orlg 0o ouvr(0evror or Kovrrv6g O5oerg. O Boorxeg evvoreg
EIK0NA 22.18 Tpetg r0pteq tretroupy(eg Tou evTontopou rou mRNA. IXnUottoUriq nporunov r<ot npodtoypoqrl rou nenpogdvou toorur-roPulv ror egBpriov AnUroupy[o 6totpoPettrtlv 0uyorprxtilv runopotv roro rrlv oouggerpq 6ro[peoq rou runoPou Arogeproporono(qor1 rou rurropou o' e[erdrxeuPdveq neproydq ./ @ @ ewonroU6q rou mRNA e[vot tdto(tepo orluowtK6q yro r0rropo pe noltK6rnro, 6noq ot veup6veq. Evd) ro neproo6repo mRNA Ueroqpd(owol' oro Kurrqpt- x6 ocirpro rou veup6vo, flo^.Id mRNA ewon(owot qro veupdqovo Kot rouq 6ev6pkeq. >e ourd flept- LoprBdvetor xot to mRNA rnq p-oKrivnq, rou ono(ou ro npoi6v ouprpret6Xet orrlv ovdmu[q tolv 6ev6ptttirv Kor rov veupo(5vttv. To mRNA rnq p-oKrtvnq ewo- n((erot oe O5oerg evepyo6 petor<ffioqg oe UeY6l'n norxil(o rwqrdlv run6pov. O ewontopt6g tou mR- NA oe veupu<5g getoouvomtr6g O5oetq qo[vetot 6tt e(vot onopo(trlroq Yto rtq rpononoqoetg nou oXetL (owot pre q prd04or1. Iro rfrropo qq ytro(og, ro mRNA rrlg Booxriq nporetvnq qq pruel'ivr1g (myelin basic protein, MBP), ro ono(o t<todtronop( dvo ou- orotrr6 rou el[rpou qq pueL(vr1q' ewon((etot o' 6vo ouyxerptpr6vo 6topr5ptopo 6nou ouw(Oerot pue- L(vr;. Ito <purd, to mRNA ewon((owot orq 9).otdr6r1 nepropj rou rurrdpou. Meptxdq eopdq, o ewont4t6g tou mRNA nepilopt- Bdvet pretoqopd on6 6vo rUrropo o' 6vo 6I'o. Er1 Drosophila, to pqtptxd mRNP ouw(Oewot Kot ouYKev- rpd:vowot oe neptB6trowo tporpodorxd r0nopo rot preroq6powot oro ovonruoo6 gevo olor0nopo pr6oul xurroponlool.tottrc0v dto[],ov. To tputd e[dyouv RNA droprSoou ttov nl.ool.todeopdttov Kot ro Uoroq6pouv oe proxpw6g onoordoetg pt6oto tou oyyetororj ouoni;rotog rou gLouiprotog, Meptr6q qopdq, ro mRNA ;reto96- powot pro(rd oe xoxxio mRNP. H oUorooq outtiv rov xorr(ov nopopdvet o6teurp(vtorq. 'EXouv r<otoypocpe( rpetq Unxovtqro( yto rov ewoTlL- opr6 tov mRNA: o To mRNA t<otov6getot o;rot6prop<po oMd ono6oge(- ror oe 6treg ng 06oetg erc6g on6 rn e6oq qg pretd- Qpoonq. o To mRNA gnope(vo dtolSetot etre0Oepo otrtrdnoyt- 6e0eror orn 06on trlg pretdrppootlg. o To mRNA petoq6perot evepYd mrq 06oetg 6nou 0o geroEpome(. H evepy6q pretorpopd e(vot o r<riptog prrllovtqr6g yto rov ewontop6. H trreroqopd enmuyXdverot Ue rrl pero- r6ntorl rrvqrrlpttov npoteivr6v xotd prrlxog o6tilv rou xunopooretrerori. Itrg olettr5g drepyoofeg, o(tonot- o6wot Kot ot rpetq t0not ptoptot<drv r<wryriprov: 6uvetveg ror rwrlo(veg, ot ono(eg ro[t6etiouv rord ptirog rtov Ut- r<poocotrqv(oxov npoq ow(0eteq roteu0Uvoetg, Kot UUo- o(veg ot ono(eg to[t6eUouv r<otd prir<oq ttov wt6(rov rqg qlctvnq. Aut6q o rp6nog ewontopoti qnore( routrdlt- mov 4 ouorotu<6: (1) crs-motXe(o cno mRNA-or61o, (2) trans-nopdyoweq nou dpeoo ri 6prpreoo npooddvouv ro mRNA orrl oolorri r<wqtrlpto npoteifi, (3) trans-no- pdyoweq nou xotoot6trLouv tq ;retdqpoorl rot (4) 5vo oU arrl pro oy ru po B6Lr1 onq ornv ent0 u prrl( tonoOeo(o' M6Xpr oqgepo 61ouv XopoKnlptfie( pr6vo Liyo cis- orotle(o nou peptr6g Qop6q nopoprotd(owot Ue roXU' 6polirxorig xdrdlxeg (zipcodes)' To motXe(o outd no,r<ilouv rot, nepil,opBdvouv t6oo tdto(tepeq 5op6q 6oo t<ot tdto(tepeg ollrll'ouX(eg RNA nou pnope( vo Bp(orowot onou6rlnote oro mRNA, nop6tt ouvriOcoq ewon((owot orrlv 3' UTR. H ovoyvtilproq toug r;rov 5U- or<o1,4, prdMov enetdrl noMd onotel'o0wot on6 nept- nLordg dewepotoye(g rot rpnotoye(g 6op6g. l-lotr'71'o( frans-nopdyoweg 6Xouv ouoletore( pe tonrrrl UerqQo- p6 mRNA xot xoroorol{ qg prer6qpoorlg. Optopr6vot e[vot ouwrlprlpr6vor oe dtotpopettxoUg opyovtoprofg' l-ro nop66etygo, 11 nporeiwl sfaufen nou npoo66verot oe 6(rtrovo RNA eprnL6retot orov ewontop6 mRNA sro coor6rropo rrlg Drosophila rot tou Xenopus, r<oOdrq en(- orlq Kor oro veuprr6 o0orrlgo qq Drosophlla, tov 0r1- tromtr6v ror (oog rov oxoulrlrttilv t<ot tou rfopto6- ii t?."1S Meprro Euxopuortxo mRNA Ewoni(ovTqt 0t Etdtxriq l-leptoyec rou Kuncoou 645 ^^iA
(5ppq.Aw6g o notrurdl,qwog nopdyowog 61er noMo- nldq neproldg or ono(eg ouv66ouv oritrlntror<o oe Uovo- ndtto petogopdg nou e[optrirwor 16oo on6 orr(vr1 6oo xor on6 pu<poorrrL4v(or<oug. IXe66v ilnoro 6ev eG vot yvroor6 yrcr ro r6topro onopo(rr1ro ouororx5, 611- Lo64 rouq prrllovroproUg oyrupoB6Lrloqg. A0o nopo- 6efyporo pr4lovrop6v ewonropro6 nopouord(owor orrl ouv6Xero. O ewonropr6g rou mRNA qg B-orcr(vqg 6Xer ;releq- 0e( oe tvoBldfieq Kor veupriveg oe xoMdpyero. O ,,rq- lu6poptr6g rc06rrog" e(vor 1.rro ol),qLou1(o 54 vour.treold(rov ornv 3' UTR. H np6odeoq qg npo:rei- v4g ZBPI oe ourl rrlv oMrl).ou1(o oe ouvepyoo(o pre rn Ueroypoqrl e(vot onopofurlreg yro rov ewonropr6, ye- yov6g nou unodrllc0vet 6n to mRNA r4g B-orrr(vr1g 6e- ope0eror oe 6repyoo(eg evtonroprori nporori rov unoBl,q0e( o' enel,epyoo(o r<t e[o10e( on6 rov nuprlvo. E(vor ev6rocpdpov 6t o ewontopr6g rou mRNA rnq B- omfuqg e[optdror on6 or6poro rv(6to orcr(vqg oflq rvo- Bl,doreg ror ox6porouq UrKpoooL4v(oxoug orouq veupc0veg. H yevenrf ovdLuorl rou ewonrogorj rou mRNA rou yovtS(ou Ash'l qg (0p49 npooSeepe rnv nro otrox).q- prop6vq dnotfq yro ro prrlXovropr6 rou ewonroproti xor nopouor6(eril ornv ElKOilA 22.1?. Kord rrlv exBLdorq- orl, ro mRNA rou yovtd(ou Ashl ewon((eror orov ovo- muoo6pevo Bloo16, on6re nporeivq Ashl ouw(0erot pr6vo mo veooxnporropr6vo 0uyorptr6 rUnopo. H Ashl e(vor rorooroL6og rnq Ueroypogrlg nou onoyo- peUer rrlv 5r<gpoo4 r1g evSovouxledoqg HO, puog r Evronroptog rou mRNA qg Ash1. To veoe[oyope- vo mRNA Ashl npoorotrtroror oqv uvlrlpro Uuoo[vn Myo4 Ueoto evog oupntr6r0u Ue Trq nporetveg She2 ror Sh3. H xrvqrlpro nporreivrl peroqeper To mRNA roro pdrog rov rvrdlrrlv orr(v1g rou ovonruooopevou ptroorou. 646 KEOMA|0 22 Iro0epoqro rorXtopo06rrlorl rou mRNA npordvqg nou onome(tor yro rrlv evoMoy4 rou ou(eu- rrrrof rUnou (B].6ne Ev6t4ro 15.20).'Eror, evoMoyrl ouprBo(ver p6vo mo p4rprr6 xUnopo. To mRNA rrlg Ashl tpdpet mqv rodu<onotrlur{ neptol( r6ooepo ororXe(o ewontotrrorj pe doprj p(oXou-0q),r69 oro ono(o npoo66veror q npotetvrl She2, prdMov orov nuprlvo. H npcoreivrl She3 Lerroupye( <og npooqppoorqg nou npoo65veror 16oo mq She2 6oo ror orrlv xrvqrrlpro npu:tefvq Myo4 (en(or1q yvoorri Kqt oq Shel). H npo- reftrr1 Puf6 npoo66verot oro mRNA, roroorSMowog rq perdqpoorl tou. H xrvrlrrlpro np<ote[vq perocp6per ro mRNP qg Ashl xord prp<og rov notrxriv wr6(cov orr[- vnq nou o6qyoUv on6 to ;r4rprr6 rfrropo orov ovo- nruoo6pevo Bl,oot6. Enrnp6o0ereg nporetveg onoro6wor yro ro otoqr6 ewontopr6 Kqr rnv Sxgpoorl tou mRNA qq Ash1. l-ldvrrr on6 20 mRNA rnq (6gnq Xprlorponororjv ro (6ro ;rovondn ewonrotrroti. MrlXovrol.ro[ ewonropo0 or ono(or dev nepd.oprBd- vouv evepy5 petogopd 61ouv onoderX0e( oogcig pr6vo yto L(yo ewonrop6vo mRNA mo roorUrropq Kqr rq npcbtpro SpBpuo. O pr4lovropr6g qq ronrrqq noy(6eu- o4g droxe6prevcov mRNA Boo[(erot or! oupperofn oy- rupc0v nou 51ouv ronoOerq0e( er tov npo-r5polv, or ono[eg 6puog 6ev 61ouv touronorrlOe(. Iro coorrirtopo qg Drosophlla, ro 6roXe6prevo mRNA rou nanos nqyr- 6eUovrot oro on(o0ro uBl,oorrrd ntrdotrro, (germ plasm), pto e[er6treupr6vq nepropi rou rurropon],6- oUoroq xdro on6 ro gLor6. Iro <r:orUrropo rou Xeno- pus, ro mRNA nou evron((ovrqr oro gurrr6 n6lo npriro noyrdeUowor oe xdntog pruor4prc66q, pregBpovo- Bpr0rl 6opn yvomt| org purolovdpror5 vdEoq (mito- chondrial cloud, MC), q ono(o opy6tepo perovoore6er oro qurtr6 nSLo ouprnopooripowog ro mRNA. O UnXo- vrop6q tonrrrlg oro0epono(r1onq rou mRNA 51er nept- ypoge( yro 6vo mRNA ro ono(o en(or1q ewon((eror orov on(o0ro n6lo tou epBpriou qg Drosophila. Zto nprilrpro orddro rqq ovdmu[r1q, ro mRNA rou yovr6(ou hsp38 r<otov6;reror opror6propgo oro e;rBpuir<6 rurro- p6ntroopro, otri.d opy6repo onodopefuot nowo0 err6g qn6 rov n6Lo. H nporeivrl smaug epnl5reror mqv onooroOeponofqor1 rov neproo6repov hsp38 mRNA, rup(og enrorporeUowoq ro otipntror<o CCR4/NOT. Aev e(vor yvoor6 nrig 6rorpe0youv ro mRNA nou ewon((ov- tor orov n6tro. 22.11 [IepiAn$n To rurropxd RNA e[vor q4errrd ooro0rl p6pro ],6y<o rnq nopouo(og runoprrc6v prBovoux).eoo6v. Or prBo- vouxl,edoeg 6to<p6pouv rrlq npoq rov rp6no en(0eor1g ror e(erdrreUowot yto 6rocpoperrr<d unompdrgoto. Aurd ro dv(u;ro ono6Sprqorlg rou RNA 61ouv nol- Lotiq p6loug oro rUrropo, prero(0 dl,l,cov Kor rnv onooUvOeoq tou mRNA. To yeyov6g 6rt ro mRNA e[- vor BpoldBro enrpdner rn ypnyopn npooopproyrl rou <pdoprorog r<,r:v nporeivcbv nou ouw(Oewor on6 ro rUrropo, puOprl(owog qv roXUrrlto rnq Ueroypoe]iq H nporetvrl She2 npood6verot mn doUn U(oxou-0ntudq rou mRNA H nporeivrl She3 ouvd6er qv She2 ge q puoo[vr1 H Myo4 npoo66veror o'5vo w(6to onfuqg lv(Dto orr[vqg
rov yovr6(ov. Ayyelroq6po RNA pe drorpopettr( oi,- LqlouX(o eprgov((ouv notrU 6rogoperrrri euoto0rlo(o or4 6pdoq rov vour.treqod)v, pre lp6voug nptEorliq nou norril.ouv xqrd 100 gopdg r<or neproo6repo. To mRNA ouv66etot ;r' 6vov petoBoLi.6pevo nLr10u- op6 npu:teivriv xord rqv cop(provo( rou mov nup{vo r<ot q (o( rou oro rurrop6nl,oopro. Yndplouv ndpo notr)69 nporeiveg np6o6eoqg oro RNA, or neproo6re- peq on6 trg ono(eg 6ev 6Xouv or6pro Xoporrqptme(. l-loM6g npcoretveg ;re p6Loug orov nuptlvo onoBdMov- ror nprv ri rord qv e(oyoyrl rou mRNA oro rurrop6- ntroo1.ro. AMeg ouvo6eUouv ro c0prpro mRNA t<ot 6pouv Kor oro rurrop6nloogo. To mRNA ouv66owor g'eu- 6r6rpro, oMd oM4lenu<ol.um5pevq, oUvo^.o np<otei- vdrv (RNA-binding-proteins, RBPs) nou trerroupyo0v orrl gerdqpoon, tn oro0ep6rqro Kor rov ewonropr6. H oprd6o tov mRNA nou protpd(owor 6vov r6ro(repo t6no RBP ovorpdperor Kor. cog RNA peyroutr6vto. H ono66prqoq rou mRNA opX(er Ue rnv orpo(peoq ev6g nupogroogoprrori on6 ro 5' dr<po. Aur6 to Brlgo nupodore( 6vqv xUKIo ewopuilv on6 ev6ovourtredoeg, nou orotrouOe(ror on6 3'+5' e[ovour<],eoAunr{ n6rpr1 r<ov el,e60epcov xtrooprdrcov. H 6o911 g(oXou-Oqhdq oro 3' dxpo nol,trrilv mRNA rq npoororeUet on6 3' ent06- oetg. H 3'-+5' evepy6rqro e[ovoux].edoqg 6reuro].Uve- rot qn6 rqv nouo6evul,(ooq rrov 3' 6xptov, q44;ror((owoq Uro nLorrp6ppro yro ro 6v(u;ro. Or r<0- preg npotetveq nou epnL6rowor mqv ono66ptrlorl rou mRNA trenoupyoriv rog o6prntroxo yvroor6 pe tov 6po oropdrro ono66prrlo'r19. H ono66grlon rov neptoo6te- prov mRNA orn (rjpn, nt0ov6roro Kor ficl Oqtrooru<d, onone( cog npri:to Brlpo illv qnoo6evutr(@o4. H ercre- vrlg Bpdpvorl qg oupdg poly(A) ennp5net vo e[e].t1- 0e( 6vo on6 ro 6rio govondrto ono66prqor1g. To povondrr 5'+3' onoorivOeorlg nepil.optBdvet oqo(peoq rrlg ro).Umpoq Kor 5+3'n6{q on6 e(ovour<}.edoq. To govondrr 3'-+5' onoo0v0eonq roro.l.Uerot on6 to e[co- orr:pdrro, 5vo peydLo orj;rnLor<o e(covouxLedonq. H pe- rdrppoorl Kor rl onooUv0eoq p6oco rou 5'-+3' povonorrori e(vor owoytovtmrr6g 6repyoo(eg ener6ri t6oo ro oritrrnLor<o 5vop[qg qg prerdgpooqg 600 Kor to 6v(utrro ogo(peoqg rrlg r<otrUnrpog npood6vowor cr4v ro).0mpo. To oroprdrro ene[epyoo(og neptSXouv mRNA r<or nporefveg nou eprn].6Kowor orqv onoo0v0e- orl Kor rrlv Koroorol{ qq perdqpooqg rot nrme0erot 6l e(vor oqpre[o onod6gqorlq rou mRNA. 'E1ouv neprypoge( dMo rdooepo provon6rro ono- dSUnonq nou oroXe6ouv er6xd mRNA. Ko0dvo Xpnor- ponore( rov (6to prrlXovropr6 ono66grloqq Ue ro povondrro nou e(oprdrwot on6 rqv onoodevuL[oor1 oMd opX((er drorpopertrd. Iuyrexprpr6vo, opffiouv pre (1) ogo(peor1 qg rol,0mpog ove[dpqrq on6 rrlv qTro- odevuL(u:oq, (2) npoo0rjrq puog oupdq noLu(U) (3) ev- 6ovour<Leol.uttx5 r<6Qr;ro er6tr<6 yro ouyxexpr;r6v4 olrlLouX(o/6oUn (4) (euydpropro Bdoeov pre pr6pro mi- croRNA. Or 6togop6q orouq Xp6voug npr(oiq ttrlv mRNA ogeil,owor oe er6xd cis-morXe(o pr6oo oro mRNA. 'E1ouv neptyporpe( ororle(o onomoOepono(r1or1 g ror oro0eponofionq. Tq neproo6repo ewoni(owot ornv g UTR xor Spo6v toq oqpe(o np6o6eoqg yto npoxefveq 4 microRNA. To ntroriotq oe AU motXe(o (AU-rich ele- ments, AREs) onooroOeponototiv peydlo opt0p6 po- p(ov mRNA oto xrltropo rov OqLoorrxc0v. Ot npu:teiveq nou npoo66vowor oro orotle(o onooroOe- ponoqorlg (ocog 6pouv rup(og entorpoteUowoq opt- opr6vo ouorolrd rou pnxovtoprorj ono66gnonq. H oro0ep6qto rou mRNA prnope( vo puOpr((etot onov- tc0woq oe KurrqprKd oqproro pe rponono(qoq tcov n poo6ev6 prevov n potetvcilv. YndpXouv ouorrlporo ennrjpqoqg rot etrdylou not- 614ro9 nou lemoupyo[v otov nuprlvo rot to xuttop6- nl.oo1.ro r<or oroXeUouv elonopottrd pr6pto RNA yto ono66prr1oq. Itov nuptlvo, ro ef,orotoprdtro 61et 6trr6 p6tro: ene[epyd(eror to <puototroyrxd RNA Ko] Koro- orp5qer to etrort<r:1-rorrxd. To elonopottxd RNA ovo- yvcop((ovror on6 notr(Loug oupnopdyovteg rou e(coorll1.ror(ou nou 6nero entorpote0ouv to e(cootogd- tro. Irq (UUn, o r<0prog oupnopdyowog e(vot ro oripr- nloro TRAMP, ro ono(o 61er opr6troycl Kcrr oe dMouq euxopucineg. An6 ro pretdypogo qg RNA Pol ll uno- crrpcir;roro yr' ono66prqon orov nuprlvo e[vot 6oo 6ev 61ouv ouppoge( oromd rj 6e q6pouv rpuototroytt<6q poly(A) oup6g. To neproo6repo prerdypo<po qg RNA Pol ll (orog e(vor r<pugd ooro0f prerdyporpo (cryptic un- stable transcripts, CUTs). l-lorx0.o mRNA molefowor on6 ro rurroponLo- ol.rorrd ouor{proro enrnpnonq. Kor oro tpto g4etu<d ouor{ goto enmq pqoqq epn},6xowor ou pBdwo ovcOpro- Lou tepporropro6 qg prerdgpoo4g. H onooUv0eor; N- MD oroXe0er g6pro mRNA pre np6opo rodtt<6vto tepporroproU.'Evo ouwrlpqprdvo oUvotro nopoy6wtov (npcoreiveq UPF ror SMG) eprnL6rerot qrnv ovoyvcirptoq Kor rrlv nopddooq ev6g unoorpc0ptotoq NMD oro pqlo- vropr6 yevrrrig onooUv0eoqg. 'Evo np6olpo t<to6tr6vto repporropoU ovoyvrrrp((erot xotd tov npoton6po ytipo qg prerdgpoorlg on6 rnv nqpouo(o ev6g xo0o6rroti oupntr6r<ou oupBoLqg e[ov(rov r1 ev6g oouvrlOtoro pe- yd).ou 3'dxpou. En(or1g, 4 NMD epntrSreror orrlv qno- 66prqoq optop6vrov guototroyu<drv ooroOc0v mRNA. H onoorlvOeol NSD oroXe[et mRNA Xcop(g ew6g ntrotol- ou xto6x6vro tepprortopori rot onore( 6vo ouw4pqpr6- vo orivoLo nporeivc0v SKI ciore vo e[co0rloet tqv oneleu0Spoorl rou noy6eupr6vou prBoorol.tot(ou rot vo enrorpore0oer ro pnxovropr6 ono66prnonq. H onoo6v- Oeo'q NGD orole0er pr6pro mRNA nou 51ouv ordotpto prBooroprdrro oe rro6rr6vto rrlg ror6xonotrpu<r1g nepto- Xrig xor npoxoLe( oneLeuO5ptoorl rou ptBootol.tot(ou rot ono66prqoq. Oproprdvo mRNA ewon((owor oe et6u<69 neptoX6g rou rundpou xor 6e prerogpd(owot pr6Xpr vo qtdoouv orov npoopropr6 roug 96oo oro rrlrropo. O ewontopt6g qnore( crs-orotXe[o ndvo qro mRNA-or6Xo Kot trans- nopdyoweg. E[unrl pere[ rperg rU preg ].enou py(eg: 1. Iro cooxrirrqpo ouvetog6per orrlv eyro0(6puor1 peMowu<cilv npor0nov rou eprBpriou Kot ornv npo- 6royporpri rou ovonruftor<oU nenpopr6vou r<urt6- pov nou e6peUouv oe 6rogoperx5g nepro16q. 3I 11 l]eoltrrun 647
2. Ie rUnopo nou 6torporiwor ooriprprerpo, e(vor rivog prrlXovro;r6 g r<orovo prrl g n p oreivt rciv n opoy6wrov p6vo oro 5vq on6 ro 6rio 0uyotpu<d r(rrropo. 3. Ie oproprdvo r0rropo, t6[rog oroug notropr6voug ri- noug rurrdpov, e(vor rivog pqXovrog5g yro rrlv Ko- 0r6pooq unoKurroprxrilv 6ropeptopdrov. E(vor yvooro( rperg prqlovropro( ewontoprori: o. ono66prqorlrou mRNA oe 5l.eq trg O5oerg er,r6q on6 q 06oq-or51o B. enilerctrf oyxupoB6Lrloq 6roXe6prevcov mRNA orq 06oq-or6Xo y. roreu0uv6pevq perqqopd rou mRNA ndvu: oe o6o09 rou KurropooreLerori. H teLeuro(o e(vor 4 nro rowri pr50o6og xor ofronoe( rouq UoproKotig r<rvrlr4peg nou Boo((owor orrlv oKr(vn Kor rouq trlrxpooro),qv(or<oug. BrBAroypoeio Ievrxd Houseley J., and Tollervey, D. (2009). The many pathways of RNA degradation. Cell 136, 7 63-77 6. 22.2 To Ayyetrtorpopo RNA Eivor Aoro0rl Mdpro 'Lgeuvo Dolken, 1., Ruzsics 2., Rddle B., Friedel C. C., Zimmer R., Mages J., Hoffmann R., Dickinson P., Forster T., Ghaza P., and Koszi- nowski, U. H. (2008). High-resolution gene expression profiling for simultaneous kinetic parameter analysis of RNA synthesis and decay. RNA 14, 1959-72. Foat, B. C., Houshmandi, S. S., 0livas, W. M., and Bussemaker, H. J. (2005). Profiling conditionspecific, genome-wide regulation of mRNA stability in yeast. Proc. Natl. Acad. Sci. (USA) 102P, 1 7675-80. 22.3 To Eur<opurrrrtxo mRNA YnopXouv uno Mopqn mRNP on6 q [ew1oq rltog ror T0 00v0r0 Touq 'Epeuvo Hogan, D. J. et al. (2008). Diverse RNA-binding proteins interact with functionally related sets of RNAs, suggesting an extensive regulatory system, PLoS Biology 6(10), e255. Avorxonnsrl Keene, J. D. (2007). RNA regulons: coordination of post-transcrip- tional events. Nature Reviewsl Genetics 8, 533-543. Moore, M. J. (2005). From bifth to death:the complex lives of eu- karyotic mRNAs. Science 309, 1514-1518. 22.4 H Ano06p1o1 rov Iporoputolrdrv mRNA ilepilogpovet l-lotrtro'Ev(upo Bernstein, J. A., Khodursky A. B., Lin P. H. Lin-Chao, S. and Co- hen, S. N. (2002) Global analysis of mRNA decay and abun- dance in Escherichia coll at single-gene resolution using 048 KEOAAAI0 22 lro0epoqro rorXtrlpo0rlqol rou mRNA two-color fluorescent DNA microarrays. Proc. Natl. Acad. Sci. (us/) 99, 9697-9702. Celesnik, H., Deana, A., and Belasco, J. G. (2007). lnitiation of R- NA decay in Escherichia coli by 5 pyrophosphate removal. MoL Cell27,79-90. Mohanty, B. K. and Kushner, S. R. (2006). The majority of Escherichia coli mRNAs undergo post-transcriptional modification in exponen- tially growing cells. Nucleic Acrds Bes. 34(19), 5695-5704. Avaoxdqoq Carpousis, A. J. (2007), The RNA degradosome of Escherichia coli: an mRNA-degrading machine assembled on RNase E. An- nu. Rev. MicrobioL 61 ,71-87 . Condon, C. (2007), Maturation and degradation of RNA in bacteria. Current 1pinion in Microbiology 10,271-278. Deana, A. and Belasco, J, G. (2005). Lost in translation: the influ- ence of ribosomes on bacterial mRNA decay. Genes Dev. 19, 2526-2533. 22.5 To ileproodrepo Europuolro mRNA Ano6opouvror Mriotrr Auo Movonolrov E[oprtipevov ono [1otruodevutrltoor1 Avorxonqoq Franks, T. M. and Lykke-Andersen, J. (2008). The control of mR- NA decapping and P-body formation. Mol. Cell32, 605-615. Parker, R. and Song, H. (2004). The enzymes and control of eu- karyotic mRNA turnover. Nat. Struct. Mol. Biol. 11,121-127 . Parker, R, and Sheth, U. (2007). P Bodies and the control of mRNA translation and degradation. Mol. Cell25, 635-646. 'Ipeuva Sheth, U. and Parker, R. (2003). Decapping and decay of messen- ger RNA occur in cytoplasmic processing bodies, Scrence 300, 805-808. Zheng, D., Ezzeddine, N., Chen, C. Y., Zhu, W., He, X., and Shyu, A. B. (2008). Deadenylation is prerequisite for P-body formation and mRNA decay in mammalian cells. J. Cell Biol. 182, 89-101 . 22.6 Atrtro Movonol0 Anod0Un0nq lroleuouv Iuyrerprp6vo Mdpro mRNA Avuordnqorl Filipowicz, W,, Bhattacharyya, S. N., and Sonenberg, N. (2008). Mechanisms of posttranscriptional regulation by microRNAs: are the answers in sight? Nature Rev. GeneL 9,102-114. Garneau, N. 1., Wilusz, J., and Wilusz, C. J. (2007). The highways and byways of mRNA decay. Nature Bev. Mol. Cell Biol. 8, 1 13-l 26, 'Epeuvo Mullen, T. E. and Marzluff, W. F. (2008). Degradation of histone m- RNA requires oligouridylation followed by decapping and simul- taneous degradation of the mRNA both 5 to 3 and 3 to 5. Genes Dev. 22,50-65. 22.7 0 Erdudg Xpdvoq HU(onS Kq0e mRNA EtrdyXeroL ond AtrtrntrouXieq n AoUeq M6oo oto mRNA irvlijll ilti:t,:I Chen, C Y. A and Shyu, A. B. (1995). AU-rich elements: charac- terization and importance in mRNA degradation. Irends Biochem. Scl. 20, 465-47 0.
von Roretz, C. and Galtouzi, l. E, (2008). Decoding ARE-mediated decay: is microRNA part o{ the equation? J. Ceil Biot. 181, 1g9- 1 94. 22.8 Tq Neosuwe0etyrivo Mripro RNA EtreyXovror yro Etrotrtoporo M6oto Evrl g lluprlvuou luorrlporo g Enrinpnonq 'Epeuva Arigo, J. T., Eyler, D. E,, Carroll, K. 1., and Corden, J. L. (2006). Termination of cryptic unstable transcripts is directed by yeast RNA-binding proteins Nrdl and Nab3. Mol. Cett24,7g5-746. Davis, C. A. and Ares, M. (2006). Accumulation of unstable pro- moter-associated transcripts upon loss of the nuclear exosome subunit Rrp6p in Saccharomyces cerevisiae. proc. Nail. Acad. Sci. (U SA) 103, 3262-3261 . Kadaba, S., Wang, X., and Anserson, J. T. (2006). Nuctear RNA surveil lance in Sacc h aro myce s c e revi s iae; Trf4p-dependent polyadenylation of nascent hypomethylated IRNA and an aber- rant form of 55 RNA. RNA 12.508-521 . Auco'xcnqaq Houseley, J., LaCava, J., and Tollervey, D. (2006). RNA-quatity control by the exosome. Nature Rev. Mol. Cett Biot.7, S2g- 539. Houseley, J. and Tollervey, D. (2008). The nuclear RNA surveil, lance machinery: the link between ncRNAs and genome struc- ture in budding yeast? Biochem. Biophys. Acta 1779, 239-246. Villa, T., Rougemaille, M., and Libri, D. (2008). Nuctear quatity control of RNA polymerase ll ribonucleoproteins in yeast: tilting the balance to shape the transcriptome. Biochem. Biophys. Acta 1779,524-531. 22.9 0'Etrey1og [lordrrirog rrlg Merorppoorlg rou mRNA llpoyporonore[ror ono Kurropontroopouro Iuorrlporo Enrnlprlolg - ,:iii.llliliiil lsken, 0, and Maquat, L. E. (2007). 0uatity controt of eukaryotic mRNA: safeguarding cells f rom abnormal mR- =,-:: :- Genes Dev. 2l , 1 833-1 856. McGlincy, N. J. and Smith, C. W. J, (200S). Alternative spticin_c .;- sulting in nonsensemediated mRNA decay: what is the meanrr; of nonsense? Trends Biochem. Scr. 33, 385-393. Shyu, A. B., Wilkinson, M. F., and van Hoof, A. (2008). Messenger RNA regulation: to translate or to degrade. EMB} J. 27,471- 481. Stalder, L. and Mohlemann, 0. (2008). The meaning of nonsense. Trends Cell Biol. 18(7),315-32i. 'Ipe uvtr Wilson, M. A., Meaux, S., and van Hoof, A. (2008). Diverse aber- rancies target yeast mRNAs to cytoplasmic mRNA surveillance pathways. Biochem. Biophys Acta 1770,5S0-5S7. 22iA Mepuo Europuoruo mRNA Evronl(ovror oe EL6uriq l-leproy6q rou Kuropou 'Epsuvo Blower, M. D., Feric, E., Weis, K., and Heatd, R. (2007). Genome- wlde analysis demonstrates conserved localization of messen- ger RNAs to mitotic microtubutes, J. Cell Biot.179, i 365-1373. Lecuyer, E., Yoshida, H., Parthasarathy, N., Atm, C., Babak, T., Cerovina, T., Hughes, T. R., Tomancak, p., and Krause, H. M. (2007). Global analysis.:of rnRNA localization reveals a promi- nent role in organizing .cellular architecture and function. Cel/ 131,174-187. Avooxtinrloq Bullock, S. L. (2007). Translocation of mRNAs by molecular mo- tors: think complex? Seminars Cell Devel. Biol. 18,194-201. Du, T. G., Schmid, M., and Jansen, R. P. (2007). Why ceils move messages: the biologicalfunctions of mRNA localization. Semrn. Cell Dev. Biol. 18,171-177. Giorgi, C. and Moore, M. J. (2007). The nuctear nurture and cyto- plasmic nature of localized mRNPs. Semin. Cett Devel. Biot. 1g, 1 86-1 93. Martin, K. C. and Ephrussi, A. (2009). mRNA tocalization: gene ex- pression in the spatial dimension. Cell 136,719-30. i.::.-,:,-: 649
@ Nucleus Medical Art, lnc./Phototake/Alamy lmages ffiffs ffi l(oroAunK6 RNA Erooyoyrl To lvrpovro rqg 0podog I AuroouppdnrovTor Ue trans- eoreponofiorl * 0t U6vor nopoyovreg nou xpeto(ovror ro rvrprivro rrlg ogo6og I yro Trlv ouToouppoqrl l/? vitro etvot 6uo perotr- trrxo r6vro rr 6vo vourtreor[dro youovoo(v1q. * H ouppoQrl npoyporonore(ror pe duo trans-eoTepoflor- loetq Xorpig ovoyq evepyerog. * To 3'-0H orpo rou oupnopoyovro qg youovoo[vr;g enu(0eror oro S'oxpo rou rvrpovlou orrlv nptirq trans- eoreponoqo4. * To 3 -0H oxpo nou dlgroupyrlOqxe oro r6trog rou nptir- rou e[oviou enu(Oeror oU ouUBotrrt pero(U rou rvrpo- v[ou xol rou de0repou e{ovrou roro rr1 detireprl frans- eoreponolrlorl, * To rvrpovro onetreuOeprirverot un6 poptprl eu00ypoUuou pop[ou nou oX4por((er rur],0 orov ro 3 -0H oxpo rou enueOe[ o' 6vov 6eogo oe pro on6 600 eooreprr6g 06oerg. * Zto Tetrahymena, 6vog eoo:repuog 6eop6g rou tvrpo- viou nou orpoLp60qre eniolg 06Xeror en(0eoq on6 dtr- tro vourtreor[dro oe Uro neproXrl trans-oupporpflq. ffim To lvrprlvto qq 0po6oq I lXlpori(ouv Mro Xopoxqprotr- ru1 Aeureporoyrl Aopq * To rvrpdvro rqg opodog I oXrlpori(ouv 1uo deureporoyl 6op1 U' evv6o d(rtroveg neptoX5g. * To rdwpo rov neptoXrirv P3, P4, PO ror P7 tlXouv ro- rotrunr1 dpoorl. * 0L neprox6q P4 ror P7 oXqgori(ovroL pe (euydpopo ouvtrlplg6vtrlv ouvorvrtrd:v otrtrntrouXLrov. * Mro otrtrrltrouX(o 6intro oqv neproXrl P7 oXqpori(er (eU- yn pdoeorv pe pto otrtrqtrouXlo nou nepr6Xer rq dpoort- Krl G. To Prpo5v(ugo'EXouv fl orr[treg Koro],ulr6g Apooerq * MeroBdtrtrovroq U 06on np6odeonq rou unoorp6po- rog o' 6vo rvrp6vro rqg opodoq l, elvor nr0ovrl 11 eroo- ytrrytl evotrtrorildrv otrtrr1trouXulv nou otrtrntrenrdpot.rv pe q dpoortrrl G. * 0r ovl6pdoerg oroLou0o0v rtroorul rrvlnrrl ev(upov Ue XoUntrrl rorotrurtr1 roXuqro. * Avrtdpooetq nou XprlorUonototioov 2 -0H deououq 0o pnopouoe vo e(vor 11 poorl yro rqv e(etrt[4 rtrrv ou0evl- rtlv xorotrunrdlv evepyorrlrtov rou RNA. ffi 650 KEOMAIO 23 Korotrurtxo RNA Enry€leta KeEaAa{oa: Douglas l. Briant
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rouronorleei otg prBo5v(upro (ribozymes), 6nou ro R- NA evepye[ og ro roto].utrr6 ouororx6 xor 4 nprotet- vr; cog rxp[copro. O yevu<69 5pog plpo6v(upo 1prlor;ronoefuor yro vo neptypd{.rer 6vo RNA pe roto},uflr<ri 6pdor1. H ev(utrru<rj evepy6r4ro prnope( vo neprypoge( 5nrog 11 ow(ororXq evdg nro oupBorrro0 ev(Uprou. Opro;r6veg xorol.urrr6g Dpdoerg tou RNA roreu0fvowor 5vowr [eXopror6v unoorpo;rdrolv (6topro p tor<69), evril dl.l.eg e(vot ev6opro- ptordg, yeyov6g nou neptop((er rrlv rotol,uur<r1 evepy6- rrlro o' 6vo pr6vo xrir<Lo. To 5v(upro RNAdoq P efvor pro prBovour].eonpco- refvq nou nepr6ler 5vo provodLr6 pr6pro RNA ouv6e6e- p6vo oe pto nporetv4. H RNAdoq P Aerroupye( 6roproproxd ror onorel.e( nop66etypo prBoev(Upou no u roroL(er owr6 pdoerg nolLon).rj g ovor<ri r.tr oor1 g (multiple turnover). Evc0 oplrrd ovoyvrop(orqre oro E col/, ori;repo yvcop((oupre 6rr 11 RNAdo4 p e(vor ovo- yxo(o yro rq Brootpr6qto 16oo rov npoKqpuor6v 6oo Kor rov europuror<i:v. To RNA 5Xer trlv u<ov6rqro vo r<oroLfer q 6rdonoorl ev5g IRNA unoorpc0trrorog, evcil ro npore'(vrr6 ouorotx6 qg RNAdoqq p no((er Sprpre- oo p6Lo, gdtrl.ov mq 6rorqpr1on rnq 6oUnq rou Kclro- ).uru<ori RNA. Or 6Uo r<oqyop(eg rov ourooupponr6pevov rv- rpov(rov, o;rd6eg I ror ll, e(vot Xoporcr4prmrrd nopo- 6e(yproto prBoev(rlpr<rlv nou Lerroupyoriv ev6o- l.ropror<d. To rwp6vto 16oo qg o;rddog I 6oo r<or r4g opddog ll 6Xouv n'1v txov6rlro v' oqorporiwor on6 pr6vo roug on6 r' ow(ororXo npo-mRNA. Yn6 rovovr- r<59 ouvOrp<eg, 4 ow(dpoorl ouroouppqgnq e(vot ev- 6o;roprorrl ror, ouvenc0g, Uovtiq ovoxUr<},oor1g, Om5- oo, ro rwp5vro rqg opd6og I pnope( vo ole6romo0v 6ror rime v' onoddloouv pr6pro RNA nou 61ouv opre- rdg dMeg roroLutrrdg evepy6qteg oXerr(6preveg pre qv ouOewrrrj. To rotv6 orlpre(o tov owrdpdoeov nou 6rexnepot- cbvowor on6 ro roroluru<6 RNA e(vot 6l ro RNA ovo- LopBdver pto ev6opropror4 11 6roproptorrl ow[6poor1 4 ono(o nepil.ogBdver 6rdonoo4 rl Sqproupy(o gooqo- 6eorepx6v 6eoprcirv in vitro. l-1op6r 11 er6rr6rrlro qg ow(6poo4g ror 4 roto).urrrrl evepy6rrlro nopdlowor on6 ro RNA, or npalreiveg nou oLLrllenrdpo6v pe ro RNA (oog e[vor ovoyro(eg yro rqv onore],eoprorrrrl npoypqrono(qo4 qg ow6poorlg in vivo. H ouppogri tou RNA 6ev eivor ro p6vo ;r5oo pre ro ono(o ero5plowor oMoy6g oro n),4pogopror<6 nepre- 16;revo rou RNA. Kord tqv ene(epyoo(o rou RNA (R- NA editing), oMoySg etodyowor oe (eXopror5q Bdoerg r1 Bdoerg npoor(Oewor oe ouyKeKprprdveg O5oerg o' 6vo mRNA. 'Ev0eoq Bdoeov (ouvrlOrrlg rordlotno oupr6(- u1g) npoypotonoefuor oe noMd yov(6ro oro pmoX6v- 6pro opropr6vov l.rovorUttopov/o}.tyoKrjrropov europucorcbv. 'Onrog q ouppo<prj, 6ror ru q ene(epyoo(o tou RNA nepiloprBdver dtdonoorl rr enovdvoorl 6eoprcbv pero(0 vouxleorrd(ov, coor6oo en(onq onore( 6vo ex- poye(o yro rnv rto6rrono(r1o4 tov nlqporpopuriv rrlq v6- og oMqLoul(og. 23.2 To lvrp6vlo rnq AuroouppdnTovrot l.Jr Opd6oq I frons-eorcponoinon * 0r Uovot nopoyovreg nou Xpero(ovror ro lvrpovro rrlg opodoq I yro qv ouroouppogrl in vitro e{uot drjo gerotrtrrro rovro rr rivo vourtreor[6to youovoo[v1 g. " H ouppoQn npoyporonote[ror pe 6uo trans-eoreponouloetg Xop(g ovoyrq evdpyerog. * To 3'-0H orpo rou ouprnopoyovro qg youovoo(vqg enul0eror oro S'orpo rou lvrpov[ou or4v npdrrrl trans-eoreponoqoq. e To 3'-0H orpo nou d1;rLoupyrl0rlre oro rdtrog rou np6rou e(ovrou enu(0eror oq ougpotrq pero[u rou rvrpov[ou ror rou 6eurepou e[ovrou orq 6eureprl trans-eoreponoqorl. e To rvrpovro onetreu0eptiveror uno poprprl euOuypoglrou lLopL- 0u nou oxrlpoTr(et xurtro 6rov ro 3 -0H orpo rou enlreOel o' 6vov 6eopo oe pto ono 6uo eoorepLr6g Oeoetg. * Zro Tetrahynena, iuoc, eooreprrog 6eop6q rou lvrpov[ou nou otpoLp60rlre en(or1g 0SXeror enr0eorl on6 otrtro vour),eo- r(6ro oe pro neproXq frans-ouppoqrlg. lwp6vro rrlg oprddog I Bpfor<owor oe notx[].eg 05,oerg xor oprOporlv ndvco on6 6Uo 1illddeg pr6Lr1. Awi0ero ond r1v RNAdorl P, 6av e[vor onopo(rrlro yto r; Burr- otp6qto. To rwp6vro qq o;rd6og I Bp(or<owor oro yov(6ro nou rtodrronoroUv rRNA orov nupqvo rov Uo- vo rUrropo-rv/oryoxUttopov e u Kop u an rhv T etr a h y - mena thermophila (Blecpopr6og6po) xor Physarum polycephalum (pu(oprUrrlrog). Efvor xorvd oro yov(6ro puxrlrcov xor nptor[ortllv, evo onov((ouv orouq npo- r<opud;reg ror ro (c6o. To rwp6vto r1g opddog I 6Xouv pto ev6oyevq rxov6rrlro vo npoyporonoroUv ouro- ouppog{ (sellf-splicing, autosplicing). (Tr;v (6to rdr6r4ro en(oqg 6ro05rouv Kor ro rwp6vto rrlg otrro- 6o9 ll, nou 0o nopouotooroUv orqv Evotrlru 25.6). H ouroouppog4 ovor<o).0g0qxe og t6r6qro rotv pieroypdrpcov rov yovr6(<ov rou rRNA oto T. thermophi- /a. To yov(6to ttov 6tjo r<[prov rRNA d,Xouv rq ouv1Orl opydvoroq r<oOrrtg Kor ro 60o er<<ppd(owor rr.rg pr6pog lrtog r<oLvrlg preroypoqrr<qg provd6og. To npoidv e(voL dvo np66popro 35S RNA Ue rnv oMrllouX(o tou 1-rLxpori (17S) rRNA oro S'dr<po Kor rnv oM4).ouX(o rou preyd- ).ou (265) RNA npog ro 3'drpo. Ie opropr6vo orel6pl T. Thermophlla, q oMq),ouX(o nou rodrxonore( ro 265 rRNA, 6ror6metot on6 dvo pro- vo6Lr<6 BpoXri Lwp6vto. Orov ro 35S npddpopro RNA snald(eror in vitro, q oupporprl ougBo(ver og our6votru1 ow(6pooq. To rwpdvro onox6meror on6 ro np66popo p6pro r<ot ouooope[erot iog eu0rjypol.lpo r<],do;ro 400 Bdoeorv, ro ono(o 6nero perorp6nerot oe xurLrr6 R- NA. Aurd ro oupBdwo ouvotU((owor crrlv ti,r,'ii,,,. ,_.. H ow(6pooq onorre( 6rjo prero).).rxd L6wo Kr evo vouxLeor(6Lo youovoo(v4g rog oupinopdyowo. Ko;rrd dM4 Bdon 6e pnope( vo unoKoroor(oer rr1 G, evc0 6ev 652 KEOA Al0 23 Kototrunro RNA Boorx6g eworeg
EE6vto 2 tr- :uPPoQn I I ow(6pqon, sfl1v oTlo(q rl veoouweeetp6vn 3'-OH ol.e ;;;" 6ripo tou eqov(ou 1 enn(oerot oro 6etnepo r(,,i,*o Or 'Orlo trretoqopdq. ouvddowot' xo0tilg 6ev no- A;n;;,i*", el'e0oepo'e[ti''to, q ouvdeoq touq trr6Mov lruBlttt, coq p6poq qg i6roq.ow(6p1on1lo.y onetreu- e"f,i,ut, to twp6vto' To twp6vto onetr'e-u0epoverot coq rr'eJvpoUUo p6pto, oL)'d 11 tp(tq ow(6poorl uoroqo- pdg to PetorP6net oe rrixi'o' ' kae, orddto t4g ow[dpoonq ourooupporpfg oup- Bo(vet ;re trans-ecnepono(nqn, rotd.qv onoio 6voq <po- L,oootr6c eor6poq petotp6netot 6;reoo o' 6vov 6ltro xipis t"i,outon uopOxuon' ot 6eopo( evotrdooowqt ad"o rot rl ev5pyeto 6totrlpe(tot, ouvenriq 11 ow(5po- oi-otu ono,.e( nopoxrl evepyetoq on6 qv u6p6uor1 iou nrp q tou GTP.'kdee enor<6ou0q ow(6poor1 t r.-r*rpono(qor1q 6ev neprLopBdvet r<oOopr1 peto- il;iti -ns ev5pyeioq. rto x0rropo, 11 ouyr<6wptoorlrou bip'tlro, uqnlrl ouyt<pttxd pe q ouyr<6wpoorl rorr RNA r<r 6tot npoo0e( qv ow(6poon npoq-rnv Kortu- gruon xotd tqv ono(o ;tto pretoB-oM trlg 6ewepoto- v"li a"uns tou RNA 0o nopepno6(oet rrlv ow(orporPn ow(Opoo'r1. nw5 enttp6net cnrlv ow(6po9n vlle:I1- pqoei p6xpt vo ol,ortrr1poOe( owivo nepooet oe Korct- L'"tn ,oippor,tos ou&p"oo oro ouppoupr6vo npoi6v Kot rq pn ouppopg5vo np66po;ro g6pto' Metoyporpn I HleKrpoQdPnon oe rtnmn 35S RNA --# +!€!# KuKlorornon | * t Kurtrtxo rvrPovto -,A U EuotlYpouuowrpovto- ' - - H ouppoQrt rou 35S np66popou rRNA rou Tetrahy- mena nopoKolou0e[rot pe qerrpotpoprlorl oe nrlrq H otpoipeorl iou ,uiporrou onoxol'unrerot ono rrlv epqovtorl ;rtoq ptrprlq (6- ;r* ".; rtve(rot yprlyopo, '0rov ro tvrpovto ylvet rurtrtro' peroxt- veirot ppo50repo, 6ntlq tpo(verot ono rrlv utllltroTepl (6W' onome(tot tptqroorpopu<{ op66o: Xpnot- trlonoto6wot ot GTP, GDP, GMP rot 11 i6,o ,1 yououoo(vq, ouventilg 6ev onot- te(toi ev5pyeto. To vourtreor(6to u1g youovoo(vqg npSnet vo nept6Xet pto op66o 3'-OH. To nenPoPdvo tou vourtreott6(ou youovoo(vqq tXvrltrote(rot pe podtoo(- Uqvon. H podevrlpyero oplrrd eto5ple- ror oro onoroppSvo eu06ypoppo twp6vto. To t<otd,tr'otno qq G ouv66etot ue to 5' drpo rou eu00ypo1-t1'tou twpovt- ou g' 6vov t<ovovtr6 <ptooqodteoreptx6 6eopr6. tf ftXOtU 23.2 6efyet 6tt ouprBofuouv rpetg ovrt6p6oerg petotpopdq' Etqv npOq getoqoP6, to voudreot[6to r1q youovoo(vr1g oupneptq5perot oq. ouU- nopdyowog nou nopSXet qv el'er3Oepr1 3'-OH oPrddo q ono[o enr[0etot mo 5 6rpo tou twPov(ou. Aw( q ow(6Pooq OnUrupVti 5vo 5eopr6 getofiti G-twpovl- ou'Kot 6r1;rtoupye( pro 3'-OH ogddo oT o 6t<po tou 5' e(ov(ou' H de6teprl peto- tpopd Boo(etoi oe tr to nop6proto xnptKn G-OH EEtJvto A V.- c '- -- EEovto B P,.<3 tttaa..rrt' I '@*P# + (Itl G- p J (c*Qll ; tta. tt' , !ijt :, l. i i:' .,:,1':.!l'r: To 3'-0H rrlq G entriOerot oro S'orPo rou tvtPov[ou 3.' PG-0H Y pNpNPNPNPNPN t:::.1:: ii. : ;: ;:'':: ;/'": i' I I Y PNPNPNPNPNPN-OH + PG i i ll r'.i:li i'r:'i "li i"r'':' 'r* AE[TEprl geroqoPd To 3'-0H tou e[olv(ou A entr(Oerot oro 5'6rPo tou e[olviou B Tpfu peroqoPd To 3 -0H rou tvtPov(ou entriOerot oTo 6sogo nou on61et 1 5 Pdoetq ono'ro S'orPo : H ouroouPPotP1 nPoYPoronotet- ror pte ovrt6po00tq trans-soTepono(qolg ortg onoieq oupBoiver opeorl ovrotrhoyrl 6eog6v 0t Oeopto( nou DrlpLoupyouvrot oe roOe orobto or1- polvovror pre ro oxtoopevo no(oto' I c-P 8-0H + ?* ? To lvrpovto rnq 0go6oq I AurooupponrovTot Ue Irans-eoreponoiqorl 653 . ,. .. ldffiit& Er.'[i
t Avostpoqn rurtronofqorl ,ro I 6"s" llpoidwo I"O eu0uypdpprorlq oG4140H AvrfDpoorl trans To onorop;.ulvo lvrpovro ;lnopel vo oXqgorloer r<u- rtroug Xplorponouovrog Uro ond 0uo eoorepirdq e60erq drore v' ovlDpooer pe ro 5'orpo ror [ovovolyer roug rur],ouq pe ovr[6po- o1 pe vepo 1 otrryovour),eor(6r0. H rxov6rrlto rnq ouppoqrlg e(vor ev6oyev4g yro ro RNA xor ro o6orrlgo e(vqr Kov6 vo npoxope( in vitro Xopfg npoo0rl<r'1 rovev5g npcoreivxori ouorfirKo0. To RNA oXqpor[(et pro er6rxrl deuteporoyrl/rpnoroyrj 6o- pn, ornv ono(o or oXenr<5q opddeg rpdpowor owKpu- ord 6ror riore 6vo vour<Leor(6ro youovoo(vqq vo npoo6e0e( oe Uro er6trrl 05oq ru 6neno vo pnope( vo ouprBoriv ot owrdpdoerg 6t6onoo4g xt enovdv<ooqg rou 6eopro0 nou ropouord(owor ar4v Er6vo 23.2. MoLo- v5tt onoreLe( r6r6qro rou (6rou rou RNA, 11 ow(6poor1 e[vot notrri Bpode(o in vitro. Aur5 e[qyehor enerdf rn ur'- vo n ouppoq4 rov rwpov(ov r1g oprddog I unoBorl0e[- ror on6 npcorer*veg, t<ov ono(cov 11 l,emoupy(o 6yrenor or4 oroOeponornon qg 6opr1g rou RNA oe Uro 6rog6p- qcooq euvoir<4 yro 11 ouppogrl. H rxov5rrlro ouggeropig oe t6roreg owrdpdoerg gerogopdg Syxenot orqv otr).qLou1(o rou twpov(ou, ro ono(o e[oro].ou0e( vo efvot 6poonr6 prerd qv onoKo- nrl rou un6 popgrl eu0Uypo;r;rou prop(ou. H Ell(0l'lA 23.3 ouvor.!((er rrg xoroLutrrdg 6pdoerg rou onoxol.rpdvou rwpov[ou tou Tetrahymena. To 3'-0H rnq G41 4 enufOeror onlv pAl 6 q qv pU20 S'exoG.. UUUpAI 6CCUpU20UG To rwp6vro rurLonore(ror 6rov n G oio 3' dr<po (OG) entte0e( oe puo eooreprr4 06o11 rovrd mo 5 'dr<po. O eooreprr6g 6eopr6g droondror r<or ro v6o 5' dxpo pe ro oplu<6 e[<oyevdg voux].eor(6ro youovoo(vr1g (exoG) oneLeu0ep6veror oq eu0Uypoprpo xldo;ro, To ruri.onorqpdvo twp6vto prnope( vo eu0uypopprore[ Ue erdu<ri u6p6).uon rou 6eo;ro6 ovdpeoo orrlv OG r<or ro eoorepx6 rordl,orno nou erxe rl.e(oet rov xrixl,o. Awrl 4 ow[6poo4 ovogd(etor avdotpoE4 xuxAonotqoq (e- verse cyclizafion). Avdloyo Ue rn 06oq qg npororo- yo0g r<uxLono(noqq, ro euOrjypoprpro g6pro nou npoS- ruqe on5 qv u6p6Luor1 nopopSver 6pooru<5 Kor Uno- pe( vo npoyporonotqoer puo Sewepoyev4 r<urLono(- non. To reLrr6 npoi6v rrov ou06ppr4rov owrdpdoeov nou oxoAou0orjv r4v oneleu06proon rou rwpov(ou ulg o;rd6og I ao Tetrahymena e(vot ro L-19 RNA, 6vo euOrj- ypogpo pr6pro ro ono(o nopdye'ror ovoorpdeowoq rn BpoX[rep4 xur].rxr1 6o;r{. Aur6 ro pr6pro 5Xer ev(uplxrl evepy6qro, Xdprl or4v ono(o rffioltier r4v en6moorl Bpo16c'lv oLryovou rLeolS(rov. H owr6poortr6qto rrrlv onorop;r6vrov rwpov(rov enerrefveror n6po on6 qv onLrl ovoorpoe{ rnq ow(6poor1g r<ux}.ono(qo4g. H npoo0fr<q rou oLryovoux).eon6(ou UUU (ovovoryer rov nporotoy4 rUx),o owrdpc0woq pre ro 6eopr6 gero[ri OG-eoorepmoU vouxl,eotrd(ou. To UUU (ro ono(o pror- d(er ge ro 3' dxpo rou 1Sprepo0g nou oneleu0epc00rl<e on6 rrlv npcoroyevrl rurlono[r1o4) y(veror ro 5' dxpo rou oxnporr(6prevou eu0riypo;rpro u prop[ou. l-'l p6remor yro drol.roproxri ow(6pooq nou ovo6exv0er r1v xov6- rqro orivdeorl g 6ti o drogopenr<iv ;ro p (cov R NA. Awrl4 oerpd owrdpdoeov <povepri:ver 6n q owoxo- roi.unrrl 6poorq pr6r4ro owrKoromp((er puo yevmeu ;16- v4 rrov6rrlto rou prop(ou rou RNA vo oX4pror[(er 6vo evepy6 rd,wpo rrov6 vo npoo66ver ouprnopdyoweg youovoo(v4q, v' ovoyvop((er ollyovouxleot(dro Kor vo g6pver rrg owr6pc6oeq oprd6eg oe pro 6topr6peoorl nou entrpdnet q 6rdonooq rr enqv{ivoorl 6e4rcirv. 14..trq rv- rp6vro qq ogdSog I 6ev 61ouv 6repeuvrl0e( 16oo tre- mopeprirg 6oo ro twp6vro tou Tetrahymena, uror6oo or r6r6qr5g roug yevu<d e[vor nop6poreg. H ow(5poo4 ouroouppoerig e(vor evdoyevrlg rdr6- rrlro rou RNA ln vrtro, oMd o p6loq rcov npolteivciv ln vivo 6ev SXer xoOoprcrre( nl;1pog. Oproprdveg evde([erg yro rn oupUeroXrl nporeivdrv nopSlowor on6 Urroxov- dproxd oumrlporo, 5nou 4 oupporpq tcov rwpov(trlv rrlg oprd6og I qnone( ro frans-6poorrrd npoiSwo dl,- l,rov yovr6(rov. Mtq ewunooroxrl nep(nroorl e(vor to cyf78 geroMoypr6vo m6l,e1og rou Neurospora crassa, ro ono(o eprqov((er el,orrogonrc] ouppoqrl noMdrv gr- rolov6prorcbv rwpov(<ov oprdSog l. To npoi5v our6 rou yovr6(ou Bpd0nxe 6rt e(vor q pmoXov6proxrl ouv0etdorl rou rupooulo-RNA. Aw6 e[4yefuor on6 ro yeyov5g 6rr ro rwp6vto prnope( vo npooLdBer Ulo rprroroVn SoUri ninou IRNA, 4 ono(o oroOeponotefuor qn6 q ouvOerd- oq npodyowog 6ror r4v roroluru<r1 owbpooq. Aurrl 11 oXSoq ovdpeoo orq ouv0erdorl Kor rn ouppoqrl e(vor o0prpovrl Uo rnv r55o 6rr 11 oupporprj [er(vqoe og ow(- UUUpAtSCCUpUzoUC UUU ,1 -> oG414oH j -/ ,S'exoG.. U UUpAI6CCUpU20UG 1.15 RNA L-1 9 RNA 654 KEOAAAIO 23 Korotrulro RNA Kurtronohon
6poor1 dtotr-reooloBo6prevrl on6 to RNA, q ono(o mq ouv6Xeto unoBor1O40qre on6 npo:teiveq nou npoo65' ,our-iNA, ot ono(eg opXtrd etXov dtrsg leroupy(eq' H xov6rlro ouroouppogrlq in vitro (otog owtnpoooneilet rn Pooirn BtoXrlpurl olJr4Len(6poon. H 6oUn tou RNA }qtrttoupyei rc evepy6 xdwpo, oMd treroupyef onore- r'.#Uoixa in vivo g6vo 6tov unoBoq0e(tot on6 6vo nptrrte[vtr6 o6Pn].oro. 23.3 To lvrp6vlo Inq Opd6oq I Ixnpori(ouv Mro XoporrnploTlKrl Aeureporoyr'r AoUf mena. Evrit ot 6oprtr6g ovolfoerg xcn6qepw vu 6a qort(oouv u1 6eutepotoVi 6opn rou twpov(ou 116 oprd6og l, 11 tpttotoynq 6oUn rou onorotuiggnKs Ps to, np6o,poro npoo6toptotrr6 qg xpumoMucqg 6e prdq. l-1oli.5q rpuoroMtrsg doprdg tcov twpov(cov rrlg opddog I 6youv 6to)teuxov0el xot enrBeBotr'tlvouv no- l,or6rejo prow6lo 6eurepotoyoriq 6ognq' A0o orn6 r,q.,ep,oX6q, tcov ono(cov or Bdoetg e(vot (euyopro;r6- ,ri p"-oti iouq, 6r1;rtoupyo0wot on6 to (euydpopo u"id,ir ouwqprll.rSvcov oMrlloulttilv nou e(vot row6g oro twpOvro ini oudOoq I' H P4 xotooreu6(etot on6 rrq onn*rlLouX(eq P rot Q, ev6 11 P7 on6 ng oMrltrou- X(eg R xqt S. Ot 6)ieq neptol6g notr<fi'ouv. oq npoq irlv'oMrlLouX(o oe 6togopetu<6 twp6vto' H ovdLuorl preroMd[eov ovoyvrop((er 6vov 'nuprivo' rou twpovi ou nou nepdXet trg P3, P4, P6 rot P7 xot nop6let tr1v eLdltorrl neptoXq 11 ono(o prnopef v' ovo]'dBet Uto Ko- roLutrrf ow(6poor1. To prrlrog rov twpov[ov rnq oprd6oq i notr<fiet notr'ri rot ot ouvotvertrSq oMql'ou- X(eg ewon(owot opretd proxprd on6 ttg npoyprotrr<69 ouprBoLSg ouPPo<Pfg''6prqr6veq owt6p6oerg ori(eu(r1q <p6pvouv nq ouU- BoLeq ouppoqnq ot gto 6rop6ptpoorlnou unoorqp((et inu euEuU,r<ri o*iOpoon. H Pl neprl'opBdvel to 3'dr<po rou S'e(ov(ou. H otrr1l'ouX(o oro eocotepu<6 rou tvrpo- v(ou, nou (euyoprirvet pe to e[,c0vto, t<otrefuot eotoreptrq o).trr1loufo-odnV6q (internal guide sequence, IGS)' H ovoproo(o IGS owovot<Ld to yeyov6q 61 rl neptoXq 3' npoq rnv oMqLou{o lGS, nou nopouotd(erot or4v Et- xovo 23.4, opltxd 0eopri0rp<e 6tt (euydptove Ue rrlv 3'ou;rBo).ri ouppoqrlq, rp5pvowog 6tot oe enorpd ttg 6tio'ou;rBotr5g. Autri 11 oL}'r1)ren(6poon Unope( 6wcog vo ou;rioiver, oMd 6e go(vetot onopo(tr1tq' Mto notrti BpoXeio olnqtrouX(o -rdnoteg qopdq, ;rriroug p6l'tg bOo'[dotrr- preto[ri tov P7 t<ot P9 (euyopdrva Ue gto otrql,ouX(o nou Bpioxetor opr6ocog nptv-rn Spoonr<{ G (OG; e6on 414 oro Tetrahymena) oro 3'dr<po rou twpo- v(ou. H oqproo(o tou (euyopri:poroq rov Bdoeov Yto rn aqgroupy(o tou onopoknrou (nupdvoD rou RNA rov[- qeiot on6 ttg t6t6rqteq rtov cis-dpooru<drv (cis-acting) ;retoMd[eolv nou nopepno6((ouv q ouppoqli roJV tv- tpov(tov tqq ogdSog l. T6toteg preroMd(etq 6Xouv ono- provo0e( oe ptttolovdptord tvtp6vto metre1<ilv nou o6uvoto6v v''ogotp5oouv 5vo twp6vo in vivo' En(oqq' 6,Xouv onopovo0e( oro twp6vto rou Tetrahymena Uero- ,p6po*oq trlv owbpooq ouppo<p{q 9- 9Yq Borcnlptor6 neprBdl,trov. H xotooxeurl rr'1q'EIK0NAI 23'5 enttpSnet t1v noporo)ro0Or1oq qg ow(6poor1q ouppoqrlq oro E' coli. To ouroouppom6gevo rwp6vto tono0etefuqt oe pto 06o11 nou 6tor6met to d6roto t<todu<6vto rrlg olr'r1- ioux(oq Yto rrl B-yoLorro(tddor1. Euventirg, 11 npoteffi gnoie( vo peroqpocne( enttuXtirq on6 to, RNA pt6vo o<p6rou ogopeOe( to twp6vto. H oUvOeoT l F-Votrot<co(t- O&onS on6 to E. colioe out6 to o0orqpo uno6rltrrirvet 6tt 4'ouppocpri prnope( vo npqYporonotrl0e( un6 ouvOtl- req opxetd Stocpopenr6g on6 ere(veg nou enu<poto6v arc Tetrahymena 4 or<6trro Kot in vitro' Evtil to twp6vto * To rvrpovro rnq ouoOoq I oXrlgor(ouv uto deuteporoYl 6oun p' evv6o 0ixtrtlveq nePtoX6g. a To x6vrpo rov neptoXtilv P3, P4, P6 rot P7 tlXouv rorotrurtrrl 6pdor1. e 0r neptox6q P4 xor P7 oxrlpor(ovrot ue (euYoptrlUo ouvupn- gdvtov ouvotverLrdrv otrtrltrouXttitv" * Mro otrtrrltroux(o dintro oqv neptoxrl P7 oxrlpori(et (euyn F0- oeov Ue pro otrtrr1trouX(o nou nept6Xet rrl dpoortr1 G' Otro to twp6vto qg opr66oq I opyovcovowot oe Uto XopoKrnptortr<r1 6ewepotoyrl doprrl.g' ewSo 5)'txeq tiri-pzl. H !:Kcl;& ;:.,i nqpouotd(et 5vo ;row6}'o qq 6eutepotoyotig 6og(q rou tvtpov(ou rou Tetrahy' rqovto r fo-oH P 5,CUCUCd flpdrrn 5'UGCGGG 3 i,eOenee UetuQopd 3'ACGCCC 5' _IGS O P8 s 5' UAGUC 3' 3'AUCAG 5' R 2bp oXrlgot((owot mo 3'drpo rou rwpovlou, , To tvrpovto Uq oUodoq I 6Xouv rotvrl deurepotoytl dogrl nou oXrlgori(etot ono evv6o (euyuptrrpeveq nepLoX6g 0t otrtrqtrouXieq rtov neptoXtitv P4 rot P7 eivot ouvqpqgtveq Kt 0v0- yvolp(ouv ro ororXe[o P, Q, R rot S. H Pl nponinret ono ro (eu- yopopo ovopeoo oro drpo rou optorepo0 e(oviou rot rrlv IGS rou rvrpov[ou: 11 neptoXrl Uero[u Uq P7 rot rrlg P9 (euYoptlvet Ue ro 3'orpo rou tvrPov[ou. ?*:.* To lvrpovLo rnq 0Uo6oq I lXrlpoti(ouv Mto Xoporqptortq Aeureporovl AoL'r 655 4 P6
Ynorrvrlnlq lwprivto evotoltottitvetot oro rtodtxdvto 10 Ktodrxdvro Ktrl6xovro Kol0x6vro 23.4 lo PrBodv(uuq 'Exouv llorriAeq KorqAurrrdq Apdoetq * l4eroBotrtrovrog rr1 06011 npoobeolq lou unoorptitltoroq o' evo lvrpovto rrlq oUo6oq l, elvot nt0ovtl 1 etooyoyq evotrtro- rlrtirv otrtr1trouXulv nou otrtrrltrentdpouv pe rr1 DpoorLrq G, i 0r ovndpooeLg orotrouOouv r).oorxi1 uvllrq ev(uptrtv pe Xo- UItr1 rorotrulrul roXu11ro. " Avldpooerq nou Xpnotuonotouoov 2'-0H deopoug 0o pno- pouoe vo e(vor 1 pooq yro Uv e[etrt€n rotv ou0evrtrdv roro- trurutlv evepyorrlrov tou RNA. Mol.ov6tt 11 rorolu-ru<q evepy6qrd roug ovot<otrri<p01- r<e Xdprl orrlv rxov6tlrd toug v' ouroouppdmowot, to rwp6vto rrlq oprd6og I e(vot Lt<ovd v' ovoloprBdvouv Kt dMeg xoro).utu<69 owr6pdoetg rn vitro. OLeg out6g ot owrdpdoeLg Boo((owot oe trans-eoreponotrloetg. Oo rrg ovoAriooupe on6 rr1 oxontd rqg oXSorlg rouq Ue qv [6ro rqv ow[6poor1 ouppo<pr1g. H r<oro).uru<r1 evepydrqro ev6g Lwpov(ou rqq opo- 6o9 I otpeiLetor otrlv u<ov6qtd rou vo nopdyet ouyKs- r<prpr6veg Eeureporoye(g r<or tpttotoye(g 6opr6g nou drlprLoupyo0v evepyd r<6wpo toodrjvopto pie to evepyd r<6wpo rcov ou;tBotu<trrv (npoteivtxt-lv) ev(0ptulv. H ': , ,;i, : '' nopouotd(er rrlv ow(6pooq ouppog4q oe o16 or1 pre ourd to r<6wpo (np6retrot yto rrlv (6to oetpd owtdpdoeov rrlg Etrdvog 23.2) To x6wpo np6odeoqg tou unoorpu.rgoroq oxnpo- r((erot on6 qv 5l.Lro P1, ornv ono(o ro 3'dr<po tol nporrou rwpov[ou (euyopc0veL Ue l'lv lGS. 'Evo x6wpo np6odeoqq youovoo(vr1g oXrlpot[(erot on6 oMrl].ou- Xieq orqv P7. Aur6 ro rdwpo nt0ov6v r<ototroptBdvetot eke on6 dvo ele60epo e(oyevdg vout<l.eot(6to youo- voo(vr1g (exoG) eke on6 to rord).otno OG (06oq 414 oro Tetrahymena). h'1v nptrrrrl ow(6poor1 petogopdq, ro rdwpo np6o6eorlq youovoo(v1g t<otolopBdvetor on6 ro etrerJ0epo vouxtreot(dLo youovoo(vr1q. Metd r1v one).eu06ptooq rou tvrpov(ou t<otoloptBdveroL on6 ro OG. H 6e0repr1 preto<popd onel.eu0eputveL ro evo:prdvo e(ovto. H tpitrl pretoqopd dqpLoupye( to xu- r<Lrr6 rwpdvto. H np6o6eor1 mo un6orpopro nepLl.oprBdvet ;rto oL- l.oyr1 6ropr6peoonq. l-1ptv on6 tqv np6odeorl rou uno- mpcoprorog, ro S'drpo qq IGS Bp(or<etot r<owd mtg P2 xor P8, evc,r prerd t1v np6o6eoq, 6tov oXrlpot((et rqv 6tru<o Pl, Bp[or<etoL xowd oe ouwlprlprdveq Bdoetg pe- ro[rj torv P4 r<ot P5. H owi6poon omtKonote(tot on6 enogdg nou ovrXve6owot fin deutepotoytl 6opt4 tqg :':r'.r:' : ' . :rrlv tpttoroyrl 6opr1, to 6Uo r<6,wpo pre to ono(o Spletot evoMorrLrd o' enogrl q P1 on6louv 37 A, yeyov6q nou unodqtrulvet onpowtKrl petox[vr1oq orrl 05or1 tqg P1. Opropr6veq neporr6por ev(uprtr6g owt6pdoetg nou I rr,teroypo,prl I ruppo,pn Mntre ntrdreg dqgroupyorjvrot pe Xptloq yto p-yolono(6dor1 I Mero,oooon V B-yotrorro(r0oon I I ,r'- ,-.a "' .: EIK0NA 23.5 H rono06qoq rou rvrpov(ou tot Tetrahymena oqv rtodrronoultxf dJrlloux(o rnq B-Ydroxto(tddonq drlptoup- ye[5vo npooDtoptolto yto TI'lv ouToouppoqn oro E. col1. H ouvOe- on tnq F-yqtrono(tdoonq eEerd(etot Ue npoo0nrn ptoq Xlgxrlg dvoorlg nou y[veror pn]re on6 to 6v(upo. H otrtr1trouXio peroq6- peror ono 6vo porrqpLorpdyo, ouven6g 1 nopouo[o gntre ntroxtitv (nou nepr6Xouv potruop6vo BoxrnpLo) unodl)rtlvet entruXlg6vrl ouppoQn. qq oprd6og I rou Tetrahymena o<potpeftot p6vo rou on6 ro mRNA rqg B-yoLorcro(tddoqg orc E. coli,6ev eL vot ooq6g ov 4 ow[6poor1 unoBoqOefuot on6 Bomqpto- 169 nporeiveg. Ie out6 tov npoo6toptopt6, petoL- l,d[erg orrq ouvorvetu<5q otrtrqLou1(eg qq oprd6og I nou droropdooouv ro (euydpoprd toug oropototiv 11 ouppo<pr'1 ror dpo nopeprnod[(ouv rqv 6rEpoorl rnq B- yoLorcro(t6dor1g. Ot petotrl.d[erg prnope( v' ovoorpo- cpo0v pre owrpponrorrx6g oMoydq nou onoxo0toroUv ro (euydpopo rov Bdoecov. Or preroMd[erq onq ow(orotleg ouvolettr5g oM4- LouX[eg rov prroxovdproxrilv twpov(tov rrlg optddog I 6Xouv nopSpoteg entntrioetq Ue ttg ow(orotleg nou nopor4poriwor oro Tetrahymena. Mto prerdtrLo[q oe Uro ouvorvermrl oll.rll.ouXio ;rnopef v' ovqorpoqe( on6 gro gerdLtro(q oe pto oupntrqprr:prottt<ti ouvowenKn oI- Lrll,ouX(o onoro0torc0wog 6tot ro (euydprr;pro: Yto no- pddety;ro, preroMd(etq orn ouvotvertrci oLLrlLouX(o R prnope( v' owrppoTlrl0o6v on6 petotrI.d[etg orrl ouvot- verrcj dtrqtrouX(o S. Iuvotrrxd, ourd t' onorctr6opoto uno6flrcirvouv 6tt q ow(6poo4 ouppog(g qq o;rd6og I efoprdtot on6 to q4r1;rotropr6 6ewepotoyo0q 6oUnS ovdpeoo oe (e[y4 rov ouvorvrrrrdlv oMqLoulrc0v oro eorotepm6 rou tv- tpov(ou. H opfn nou roOtepdrOqre on6 ouq rqv epeu- vryxr] epyoo(o e(vor 6tr aMqLouybq nou aniyouv an6 nq oupBoLiq ouppaertg anarcolwotyo va oyqpauorc( rc evepy6 xiwpo to ono[o xalotd eELxtr'1 rnv aurooup- paEi. 656 KE0AAAI0 23 KorotrurLrd RNA Boorr6g 6worcg
To rorotrulro RNA dxsi Uro 060n np006s0!g rnq youovooivnq Kor Uio gSon rpo06s0nq rou unootprirporoc 0don npoodeolq iilg youovoo[vr1g -3' d414 :-.'&a.'- ''- 0don nodoOeonc -ffiffi rou unomotiruotoq GGGAGG UUUACCU I fleprdXet Al 6 xor U20 iis:;;z:t:t:;:"t::+**i H G-OH rorotropBovet rn 06on nprlo0eolq G. To 5' e(6vro roro)'ogpoveL q 0do4 npoooeonq Tou un0oTp6U0T0q. -3' 8414 ,tw,6H ':' ,*J;A.GGGAGG#:. -3' 8414 ---> w G*l 5',-@cu . GGGAGG ..' :ii::1r;:.i ,.i;-',:t:.*.:: H G414 Bpioxeror rq 06o11 npoobeorlg G. To 5' e(civro pp[oreror q 0eon npoodeoqq rou unoorpoUoToq + : l-',.j(-ij:' i:i:.:'; ;$:ao;p: H G4l4 Bp[oKeroi oq 06on npoobeoqg G. T0 5' 0Kp0 rou rvrpov[ou pp[oreroi rr1 06011 nprlodeorlq Tou uttooTpopoTo Al: -t-1i"'- / + G,-,S[EffiffiCU.r, GGGAGG..,i; G _j,-i.,:a..:.;{:'.; H e rrop1 Tou ivTpov[ou qq opo0og I oro rRNA rou Tetrahymena 0uUpo[ver Us 0rodoxrKeq 0vTr0p00erg 0v0Ur00 oTrg 0v0l0srq nou KoTo),ouBovouv Trl 0eo1 npooDeor.lq Tnq y0u0v00i vlq Kor ul e60l rpooosqg Tou urooTpidUoToq. T0 oprorepo e[6vro eLrov[(eror p0( ev6 To 6eqrd Uop. 6Lexneportrlvovror on6 LVrpdvLo rlq oUd6oq I rou Tetrahymena ovoertpowor orrlv i ": . r . To prBo- dv(upro l.erroupye( o.rg ev6oprBovour<l.edoq er6rr<q yro ouyr<er<p r pdveq o^.InIo u X(eq Xp! or Uono Lovrog rrlv rKov6rnro rnq IGS vo npoo66ver ouprnLqptrrprotrx6g oLlrl).ouXieg. Ie our6 ro nopdderypo, ilpoo66ver 5vo e[rr;teprr<6 un6orporl.ro ilou nepr6xer rqv o]..qtrouX(o CUCU, ow( rnv ovd).oyq ol,l.r1trouX(o nou ouvneoq nepLSleror mo dr<po rou S'elov[ou. 'Evo vour<].eor[6ro nou neprdler youovoo(vq Bp(oxetor mo x5wpo np6o- 6eor1g youovoo[vlg xt enrr(0eror orrlv o].].r;].ouX(o CUCU r<ord rov (6Lo or<ptBtrlg rp6no Ue rov ono(o 66- leror en(Oeoq ro eEolvro mqv npcoq ow(6pooq ;rero- eopdg. Aurq q ovr(6poo4 6roond rlv oLLqLouX[o-or6Xo o' 6vo 5';-r6pro nou prord(er pe ro lr (f(l(lALTLr EIK0NA 23.7 H 06oq qg IGS otrlv Tprroroyn DoUrl otrtrd(er 6rov oxngonoTe[ n Pl p6oro rpdodeorlq Tou unoorp6U0T0q. S'e[c0vto rr rivo 3'pr6pro, nou <pdpet 5vo re].rx5 ro16- Lorno G. MeroMdooowoq ro ororxe(o lGS, oLLd(er Kor n er- 6rr6rrpo rou prpoev(fpou ro ono(o ovoyvop((er nl.6ov ol,t1).ouX(eq ouprntrr1popronr<5q pre q v5o otrtrr1lou{o orrlv neproX4 lGS. Autrl n rponono[non rqq IGS rirore v' o).d(er q er6tr6rr;ro rou x5wpou np6odeo4q rou unoorp6gorog ennpSner oe dl,i.q RNA-cr61oug vo ugiorowor ene[epyoo(o on6 ro prBo6v(u;ro, to onofo Unope( en(ong vo enrrelSoer owrdpdoerg RNA-tuyrd- o4g. 'Evo RNA nou Korolnyer oe 3'-OH npoo6Sveror oro raiwpo rou uTroorp6poroq rt 5vo RNA nou Koro- )rrlyer oe 5'-G, npoo65vrror fio K6wpo np6odeoqq G. H en(Oeor1rnq u6po(ulopddoq oro gooeopur6 6eopr6 ouvdSer ro 66o pr6pro RNA ;.re onrirLero rou Korolo(- nou G. H ow6poo4 rnq eooqqrdo4g 6e q(er(Eeror dpeoo pe ng owr6p6oerg ;rero<popdg rord rq ouppogrj. Mro ol,ty ovo u rl,eond u<r1 otrLqtro uX(o ou prnLrl p ro prot u<ri pre qv IGS nou roro).r1yer oe pro 3'-groocpopu<q opd6o 66- Xeror en(0eor1 on6 r4v QG. H rpoorpopu<r1 oprddo ;rero- qdpetor orrlv OG xr 6nerro oneLeuOeprirverot 5vo olryovouri.eot(6ro pr' 5vo el.efOepo 3'-OH. Iq ouvS- Xero, n gooEoprrui oprddo prerog6peror efue oe 5vo ofu- yovoux.treor(6to nou rorotrrlyer oe 3'-OH (ouorootrrd, ovoorp6qowoq rrlv ow(6pooq) efue oro vep6 onel.eu- Oepd:vowog ov6pyovo rpd:orpopo r<r ol,or<Lr1 privowoq puo ouOewrrrl ow[6poo4 groorpordoqq. Ot owt6pdoerq nou rorol,riowor on6 ro RNA nepr- ypdrpowor 5nog or r<i.oox6g ev(upru<69 owtdpdoerg ;re 6poug rrvrlrxqg Michaelis-Menten. H EIK0NA 23.9 ovo- tr(er rrg owr6pdoetg nou rorolfowor on6 ro RNA. Or np5q qg Kn,r Tu)v owt6pdoeov nou roroLtlowot on6 ro L: GGGAGG Enoqdg gero qv npdo0eorl rou unoorptiporog Iii.4 To Ptpo6v(upo'EXouv llorx[],eg Korotrunxec Apooerg 657 w Enog6g nprv on6 qv npdodeoq rou unoorpolcio( s', @@s,cu GGGAGG
'Ev(u;ro Yndmporyo KM (mM) Avur6xtrosn flmin) t i5'ffi*3' GGGAGG 5' * S,*CUCU-0H + loooet6Sq 1 9 pdoeov lwprlvLo L-19 RNA RNAdsnq P nlripnS RNAdon P RNAdoq T1 p yotrorro(tddorl RNA 24 pooetov 0.0006 0,5 cccccc 0.04 1 7 npotRNA 0.00003 0.4 pretRNA 0.00003 29 GpA 0 05 5,700 trorro(q 4,0 1 2,500 5'_Y@3' +u + 5'UCU-oH 3 ' EIK0NA 23.8 0r KoTotrurKdq qvTrdpdoerq rou p0oev(6[0u rspr- trogpdvouv trans-eoTeponoqoerg ovdgeoo oe Urq oUd60 Tnq ee- onq npdoDeonq Tou uflooTpouqroq Kor Urq oud6o TIq e6onq rpdooeonq rnq G. RNA efuor XoUnMq, yeyov6g nou uno6rlLri:ver 6rr to R- NA npoo66var ro un6orpo;r6 rou Ue uqrnln er6u<5r1ro. Qor6oo, or oprOpro( ovor<UrLoorlq yro nq owr6pdoerg nou roroLriowor on6 ro RNA e(vor 1.uxpo[, yeyov6g nou owtxorompi(er q Xoprllll rorol,uurrl roXriqro. ['lpo- mxd, ro pr6pro RNA ou;rnepup6powor Uetov 6ro yevr- 16 rp6no 6nog xor ro nopo6oorord 6v(upro, ov Kot e[vor oletrxd opyd ouyrpnrrd pe rouq nporeivrro(g rorol,Ureg (6nou ro runw6 e0pog tov oprOpcirv ovorU- rtroonq efuor 103 dcog 106 minl). Ot 6poor4pt6qreg r<ov prBoev((pulv enerctdO4rov Ue rnv ovor<dl,urlq 6n prnope( vo pu0;uoroUv on6 ouv- 66teg (BL5ne Ev6r4ta 30.3). Aw6g or pu0pronr6g cis- 6poorrx6g neprol6g xol,otjvtor prpo6ro16nreq (riboswitches). Ee 6l,oug q(e66v rouq prBo6ror6meg, puo geroBolrj dropr6pqooqq r<o0op(et rlv xotdoroorl on/otf. A[rooqpe(corq e[o(peor1 e(vor ro yov(6ro g/mS rtov Gram-0erx6v Borcrrlp(ov, ro ono(o g4rlpor(er 5vo ouro6roonrirpevo ppo6v(ugo nopouo(o 6-rproogopm{g yluro(opr[vqq (GlcN6P). H tlK0NA 23.10 ouvorf((er 11 p[0pror1tou prBodror6mq g/mS. (l-ro neproo6tepeg ].e- mop6peeg, Bl$ne Ev&1ra 30.2). Av ro evepy6 r<5wpo e(vor 1.uo enrgdvero nou er06- rer pro oerpd evepyolv opddrov oe ndyo prero[rl rouq ox6on, y(veror rorovoqr6 ndlg ro RNA e(vor rxov6 vo nopdoXer 6vo rorol,utrr6 r6wpo. Ee pro nporelw1, or 658 KEOA AI0 23 Korotrurrxd RNA EIK0NA 23.9 0t ovn6pdoerg nou rorofujovror on6 ro RNA 6Xouv ro Oro XoporrrlpLolxd pe lq ovrL0pdoerq fiou rorotruowor on6 nptoretveg, gotrov6r 4 roXUqrd roug e[vor BpodUrepn. H Krvr 6(ver q ouyx6vrptoorl rou unoorptirgqroq flou qnquefror yro vo enr- reuX0e(ro pro6 qg Udyromq roXUrrlrog: e(vor 6vo ovr(orpoqo p6- Tpo Tnq ouyy6verog rou ev(Ugou yro ro undorproUd rou. 0 optOpdg rov ovorurtrtiroeorv e[vor o optOpdg rtrlv lrop(trrv rou u[o- orp6gorog nou gerogoprpt0vowor oT1 povddo rou Xprlvou o' 6vo p6vo rordruilxri x6vrpo. 6poorr6g opddeg nop6lowor on6 ng nAeupu<69 otru- o6eg ru:v o1.uvo(6rov, or ono(eg epr<pov((ouv r6ro(repr1 nomil.io ror nepr).oprBdvouv Oenr5g r<ot opwlnx6q rowr- 169 oprddeq, r<o0rilg r<or u6p6<poBeg oprddag. Iro RNA, or 6ro06orpeg p((eg e(vor neptoproprdveg ror onotetrotiv- tot t<up(tog on6 rrg er,ee0eqdveg oprd6eg rcov Bdoecov. Bpole(eq neproX6g rou RNA ouyr<poro6wor oe ouyKe- rp rp6vr1 6eweporoyrl/tpnoroyrj 6ro 96 pgoorl, nop6- Xowoq 6ror puo evepy6 entgdveto r<ot 6roqpriwog 6vo nep$dMov oro ono[o ;rnope[ vo droonooroUv Kqr vq oXrlporroro0v 6eopo(. Oo(verqr ovon5rpeurcro 6n q o).- LrlLen(6poo4 pero(U rou xorol,uru<od RNA Kor rou unoorpdlprot6g tou Boo(etor oro (euydpropro tcov Bd- oeov npoKerp6vou vo 6qproupyq0e( 6vo evepy6 nepr- BdLl,ov. Arnotru<d Kqldwo (ouvriO<og Mg2*) no[(ouv orlpowrr5 p6Lo mr1 6oprl ror runxd Bpiorowor mo evepy6 r<6wpo, 6nou ouwov((ouv rtg 06oerg tov drd- eopov oprd6tov. To 6rno).txd gerotrtru<d Kffir6wo en(- oqg nof(ouv dpreoo p6].o mrlv evdovourl.eo],ulrr1 6pdoq tov prBoev(ripov tcov roet6cbv (BL6ne Ev6rrrya 23.9). Or e[efurcrx6q npoerrdoerg our6v rcov ovor<otrri{re- <ov e[vor o[roo4pre(orreg. H "6tXoo;r6v4 npooonu<6upo" rou yeverrKori prrlXovtoproU -orov ono(o ro RNA Bp(oxe- ror oe 6l,o ro ouoronrd, oLld ot nporeiVeg ovotroprBd- vouv r<oro.truru<69 owr6pdoetg- onoteloUoe ndwo 6vo yp(go. Oo[veror on(0ovo ro ev6eX6prevo ro npdrro ou- oniproro owrypoqqg vo nepre(1ov r<or voux.trsivurd o[6o xor nporeiVeq. Yno06ore, to01600, 6rr to npoto ou- orqporo nepe(Xov g6vo 6vo ouroqwrypog6gevo vou- rLetvu<6 o[U pe nprrlr6yoveg roro].wrr6g evepy6qreg -g6vo 6oeq Xperd(owor yto rq 6rlproupy(o r<or rq 6td- onoon rpoogo6teoreprrciv deoprdrv. Yno0riore en(or1g 6l q ouprpreroXrlrov 2'-OH 6eopc0v onq onpepvdq ov- ndpdoerg ouppogqg ovdyeror oe our6g rrg opXdyoveg r<oroLuttr6g evepy6rqrcg. Me ourrig rtg nopo6oXdg, 0tooqordorl RNA trtyxdorl
G-C P3 G -e) o-o o-o a- o- o- o- a- o- o o O P3,1 o o o a U .0r0xl rou mRNA To oIIo[0 KoorKonotel T0 ev(uuo nou nop0Yet :-:oorpoprril ytruro(opivn (GlcN6P). '0rov 1 GlcN6P npoodevetot ' , pLpo6v(upo, onoono ro 5'orpo rou mRNA, odpovonot6vroq : <L epno6[(ovT0q rrlv nepou6po nopoYoYrl rou ev(rjpou. 0t ne- : :Xeg, nou e(vot orlpovrLr6q Yto fll Dtorrlplon rrlg evepyou rptro- :','ouq doUnq, errov((erot pntre, evdt I0eol Dtoonooqg i .ol1polveror ;t' 6vo roruvo petroq. (Avorunoorl ono Curr. Opin. :.'uct. Biol., vol. 17 T. E. Edwards, D. J. Klein, and A. R. Ferr6-D' --rar6, Riboswitches: small-molecule recognition . . ., pp. 273- --9 Copyright 2007, pe qv odeto rou Elsevier [http://www.sci- . -.r e d irect. com/science/jou rnal/0959440X) . -:ope( vo 6rorunto0e( to entXe[prlpro 6tt ro ou0ewtr6 .cur<.treivrr5 o[U rlrov to RNA, r<o0cog ro DNA 6e 6to05- iL 2'-OH u6poluLopd6o xot ouvenc,.rg 6e 0o prnopo[oe . : drexneporooer tdtoteg owrdpdoetg. Ot npu:tefveg -:,,ov npoor50rp<ov Xdpn orrlv tt<ov6tqtd toug vo -oOeponotoriv q 6opr1 tou RNA. Xdprl orq preyotr0te- :- eue).r((o rouq, ot nptoteiveg orrl ouv6leto ov6LoBov trg r<ototrulx6q owLdpdoetg, odrlyo.lwog teAtxo c:: ' nolrjn).or<o oriyXpovo prllovtopr6 trlg 6xgpoorlQ rt"; yovt6(rr.lv. 23.5 AptapCvo tvrp6vro Tnq Opd6oq I Kor6rronoro0v Ev6ovourAedoeq nou tlpodyouv Tnv KrvnrrK6rnro * To rtvlro rvrpdvro e[vot trovo vo eloXtopouv oe v6eq Oeoetq. * To rtvlro tvrpovto Uq oUodos I rtodtronotouv gtu ev6ovou- rtreoo1, q ono[o npoyporonote( pto 0trtrtovr1 evro;lr1 oe pto 0doq-oroXo. * To tvrpdvto perorlOeror oq 06orl rr]q dLrtrovr]q evroplg p5- ou.r evoq ovlypoqtrof glXovtopou nou 6togeootrope(rot ono ro DNA. Optopdvo rwp6vto t6oo trlg opddog I 6oo r<ot ttlg oprd6og ll nepr5Xouv ovotXrd nLo(oto ovdyvtr;or1g nou prerogpd(owor oe npora(veg. H 6xgpooq rov npco- teivo-lv enrrp5net oro twp6vto (e(te mrlv ou0ewtr<q Uopen rou tl:q RNA eke cog 6vo DNA owtypogo rou RNA) vo r<Lve(rot, on6re e(vot txov6 vo eroXtope( oe Uto r<otvoripro yevoprxrl 06oq. To twp6vto t4g oprddog I ror ll e(vot 6ro6e6oprdvo t<ot undpXouv t6oo os npo- ropuorreg 6oo rot o' europutrtteq. To Lwp6vto trlg o;rd6og I petovooreUouv ;t5oto prrllovLopttrtv nou 6to- preootroBorjvrot on6 ro DNA, evot ro twp6vto tqg oprddoq ll petovooreUouv prSoo prqlovtol.ttov nou dto- ;reootroBotiwor on6 ro RNA. H r<rvqttr6rrlro rcov twpov(ov npotoovtlver-i01re on6 6tomouprrroetq ottg ono(eg t' oMrlL6propqo rou olelro6 yovr6(ou 616<pepov oq npoq qv r<otoXtl rou tv- tpov(ou. l-lotru;toptptopol ovo<poptrd pre qv nopouo(o 11 qnouo(o rwpov(rov e[vot t<otvo( oro pirol6v6pto rov pu- rr[cov. Aut6 e(vot ouprBot6 Ue rnv dnoQrl 6tt ro ouyre- r<prprdvo twp6vLo npoqL0ov on6 eLolopqon oro yov(6to. Mto ovdluoq ovoouv6uooproU ttDv 6tomquptooerr.rv mqv ono(o Xprlotponotrl0r;re to yov(6to tou peydtrou r- RNA tou puroXov6p(ou rnq (UUnq 6pt[e qcog oqv ntOo- v4 6tepyoo(o etolrrrpqorlg ttrtv Lwpovlo.lv. To yov(6to tou peydtrou rRNA tou prttoldv6ptou qg (Upr1q 6XeL 6vo twp6vro oprd6oq I nou nept6Xet Uto Ko- 6u<onotqrLr( oMrl).ouX[o. To twp6vto Bp(or<erot ot opt- opdvo oreLdxn (0UnS (xootiwot to*), evo onouotd(et on6 dMo (to ). Ot on6yovot t<ov yevettr<lv drooroupto- oeov petofri rou o+ Kor rou ro 6ev 66otoov rov ovope- v6pevo yovotuntr6 l.6yo, xoOrig ouvqOotg rltov co*. Av Oeoprlooupre ro ordLeXog ot oq 66q r<ot to ordLeloq to rog 65r<111, odrlyotiprome ornv dnoqrn 6tt oe 6to- oroup6oerg o+ x 0)- nopdyetot 5vq v5o owtypo<po tou rwpoviou oro o- yovt6(topro. 'EtoL, 6l.ot ot on6yovot e(- vor o+. MeroMdletg prnope( vo ouprBoUv oe onotodqno- I Ktodrronororjv Evdovourtreooeg nou llpooyouv qv KLvnrLxo:-.: 659 aa o-o o-o a-o G -C o-o o-o G -CDOO lJ -tJ c -c G G."y' o-o o-oP2 o-o o-o a-o orffiou' ox- O' UA o-o o-o o-o o-o o-o O-O A.A A.O AA Oa O'A t.o-o o-o o-o o-o o-o var a oA t -G - c P2,1 a U GA Go oo A- i :<-'t- 1oo.o-t?O-O-l 23.S 0prop6vo lvrpdvro rnq OUd0oq Boorxdg dworcg AC G
RNA I Ev6ovourtreoorl Fa A'*'qOeor1-oroyo I*u**u7. *R;{& To rvrpovro ovlypdtperot rr 6nerro evoorpor6veror > &6u{{ .{Xb{,& 'Evo rvrpovto xui6tronoLe( ;:ro evdovouxtreoorl nou npoyporonorel pro 6frtrtov1 eviop4 DNA. H otrtr1trouXio Tou rvTpo- vlou ovrtypoqeror rr eneuo evoopordtveror oqv evroprl. 660 KE0AAAI0 23 Korotrulro RNA re norprr6 ordl,eXog pe onor5l.eopo n Ul Mev6SLeto yovotunrrrl r<orovoprrl vo Xdvetor. OptoprSvo meL5X4 epgov((ouv rovovtr6 drolropropr6 pre ioo opr0;r6 o+ ror co- onoy6vrov. 'Otov Xoproypoeo[wor, ot preroMd(erg rou ore).5Xoug ro- Bp(orowor r<owd orqv neproXrl Snou 0o pnoporioe vo eroxoprloer ro rwp6vro. Or trreroMd- [erg ev6g ore].ri1oug co+ Bp(orowor oro ovoyvtoonr6 nLo(oro rou rwpov(ou xor eprno6((ouv tqv nopoyruyq npcoreivqg. To yeyov6g out6 unoorrlp((er ro prow6).o qg EIKONAI 23.11, orqv ono(o n npotretvrl nou ro6u<o- noefuor on6 ro rwp6vro o' 6vo or6Lexoq o*, ovoyvop(- (et 14 06o4 6nou 0o 6npene vo eroXcopqoer to rwp6vro o' 6vo mSl,eXog o- Kor euvoe( rqv rlrlpovoprrr6tqrd TOU. OproprSvo rwp6vro r4g oprddog I r<o6xonoroUv ev- 6ovouxledoeq nou to xq0rmoUv rrvrlrd. YndpXouv roul.dXrmov n6we oxoy6vereg yovrd(iov yro ev6ovou- r).edoeg enorrtoprori (homing endonuclease genes, HEGs). Ario ouvrl0ropr6veg ou<oy6veeg HEG eivor r1 ev- 6ovoux),edoq LAGL-IDADG ror q ev6ovoux.l.edon rou rourro6 His-Cys. To rwp6vro r4g opd6oq I nou nep6- Xouv HEG owrnpoooneUouv 6vo ;urp6 noooord rou ouvo).xo6 oprO;rof rov nuprlvKcbv rwpovirrrv qg opd- 6o9 l. Ev6 51ouv rouronot40e( nep(nou 1200 nup4vu<d twp6vro opddog l, Lry6repo on6 30 nepdXouv HEG. To rwp6vto rrr nepr6ler 6vo yov(6to HEG nou rodxo- noei "rr1v ev6ovouxl,edorl l-Scel. H l-Scel avoyvap((et rc yov(6Lo u-uq m6yo yo 6kAuv4 0pa0or7. H l-Scel ovo- yvrop(er pro oMqLou{o-or61o trrrlxoug 18 bp, nou ne- prSXer 14 05oq ero1c6pr1o4g tou rvrpov(ou. H otri.rlLouX(o-o,6Xog 6roondror oe rdOe xLdrvo rou DNA oe on6orooq 2 B6oerg on6 rrlv 3'neproXrlqg 06o119 er- ox6pnonq. 'Eror, ot 06oerg 6rdonoo4g onSlouv 4 bp t<or 64 puo u pyo6v enxpeprdgevouq provori g r<Lrirvo u g. Au- rn n Uopqn drdonooqg oXet((eror Lre ro XqpoKrqptonrd rov rpovoflo(ovfcov 6rov perovooreUouv oe vrleg 05- Eq6vro lvrpovro E[rrtvro &h..6kfir"/&&t& lrrrrrarrrr.rr.rrrra 060r1-oroXog *b#&ift{g-&ffi oerg (Btr5ne KeqdLoro 17). H 6(xi.rovq ewop4 pdLtrov nupodore[ pto 6epyoo(o yovtdroxrlg perotponrlg, oqv ono(o 4 oMqLouX(o rou yovr6(ou o+ owrypdeeror yro v' owrroroorqoet qv oMq).ouX(o rou yovr6(ou co-. H owbpooq ouv(orqrot oe prerd0eoq prSoot ev6g Unxovr- opori owryporprlg r<or npoyporonoekor ;r6vo oro en[ne- 6o tou DNA. H erq4ripqoq tou rwpov(ou dror6met rqv oMrll,ouX(o flou clvoyvopf(eror on6 rqv ev6ovoux.tre6- o4, 6roogo).(owog Stor rr1 oroeep6rnro rou 1lop(ou. l-1op6poro rwp6vro ouprd ;reroq6pouv opr<erd 6ro- <popenrSg ev6ovour.ledoeg. Yndplouv 6rorpop6g ong l,emopSpeeg qg erqXd:pnonq: yro nopd6erypo, q ev- 6ovouxl,edoq nou rco6tronore(tor qn6 ro rwp6vto td rou gdyou T4 droond puo 06or;-or6Xo 11 ono(o Bpiore- tot 24 bp ovo6rrd tqg 05oqg etoXdrpqoqg rou rwpov! ou. H drdoroon rnq otrL4),ou1(og rou twpovlou on6 qv oMrl).ouX(o r4g ev6ovoux.tredoqg rov((eror on6 ro yeyov6g 6rt or (dreg oMqloufeg ev6ovour<].eoorilv Bpi- oKowor ong rweiVeg (oMr1Lou1(eg nou rco6rronororjv ourooupponr6preveg npcotetveg - BL5ne Ev6r4ra 2s.12). H norril.(o rov ev6ovourl,eoocirv oqpo(ver 61 6ev undpXer opoloy(o prero(0 rcov oMlLouXuirv rrov ow! ororxov 05oeov-m6Xov. Or oXerrrdg 0rioerg-m6XoL 6Xouv ;reyd).o pqrog (oe r<L(proro 14-40 bp), dpo ror Ueyoln er6x6rr1to. H erdu<6rrpo e(oogoL((er 6tr ro w- tp6vto 0o noMonl,oord(etor pr6vo prloto eroXdrpqoqg oe pro 1.rovo6rxr1 05or1-m6Xo xor nou0evd oMoti oro yovr6(opro. Aur6 r<oLe[ror enolxrop6g rtov lwpovirov (intron homing). lwp6vto nou peroE6pouv ollqLouX(eg or ono(eg ro6tronoroUv ev6ovour<Ledoeg Bp[oxowor oe notx0.o Bomrl pro rot ;rovo rUrropo u g/ollyo x6rropou g eu xo- purilreg. Aurd t' onore).64roro evroXriouv rrlv dnor[r1 6n ro twp6vro nou prerorpSpouv rtobxonorqnrSg oMrl- Loul[eg [ex(vrloov trtg ove[dpqro ororle(o. 23,6 To lvrp6vro Tnq Opd6oq ll 1ooq Ko6rronoro0y l-loAuAelToupytKdq fl poretveq * To rvrpovro r1q ogodoq ll ourooupponrovror ln yltro, otrtro ou- vrl0trl g unopol0ouvrot on6 nptoretve g no u rtrld txono lo uvror ono ro rvrpdvro. * 'Evo U0v06rKo rodrronotllr6 n),0(oro rodrronore( Uro npul- relvrl p' evepyorqro ovlorpoql g peroyporpoorlg, ;uo evepyo- qro poroupooqg, 6vo gor(Bo npoodeolq DNA rot ;uo DNA ev6ovour),eoo1. s H evdovourLeooq droono ro DNA-oro1o enupenovroq rrlv evotrrportool rou lvrpovlou oe gLo veo 06o1. I H ovr(oTpoq1 peroypotpoorl d1;uoupyei evo DNA ovr[yporpo rrlg otrtr1trouX(og RNA rou evoogoropevou rvrpov(ou. To ro).rirepo XopoKrrlpro;-r6vo r<rvq-rd rwp6vro rrlg Buorxfg evvoreg
cud6oq ll rotdtronoto0v prto povodtr<r1 npoltetvq oe -Lo neptoltl tou twpov(ou nSpo on6 tov t<otol'uttx6 .upqvo -or. U tuntrrl npcotetVq nept5XeL prto evepy6- :'tto N-teLtxrlq ow(orpogrlq UeroYpoq6onq, pto Kev- .crxr1 neptolq nou oXeti(etot Ue Uto entxouptt<t1 :,,,eoy6tqto q ono(o unoBorl0d to 6(nLotrro rou tvrpo- , rou orrlv evepy6 6opr1 tou (xoeitot protoupdorl' -aturase - BLSne Ev6tqra 23'7), ptoneptoXll np6ode- rtg oro DNA r<ot prto C-retxr1 neptoX( p' evepy6tqto : ,,5ovouxtredorlq. H evdovouxl'edor1 (erwd qv ow(6poor1 qg pret60e- :-"q r<L 61et tov [6to p6tro mov enotrtop6 6no:q q ow(- -cLp1 evdovouxledoq tou twpoviou qq oprd6oq l' H :.rio-rpo<Pn pieroypotpoorl nopdyet 6vo DNA owtypotpo :; rwpov[ou nou evooUoro0qr<e orrl 06o11 enotrtopo0' - evdovout<tredor1 en(oqq droond oe notr'0 loprql'6repq ;-;ydqto 05oetg-or61ouq nou protd(ouv, 1op[q vo ei .:.'novoprot6tuneq, Ue q 05on enotrLopori, oSqyowog ; ivor-opdrool rou twpov(ou oe v6eg 06oetq' H ,,.,, '. :i. nopoUotd(et rqv ow(6poor1 petdoe- :-c ev6q tunrrorl twpov(ou rqq oprd6oq ll' flptrrtov, 11 ., lovout<tredon npoYUoronote( prto 6(xl'ovr1 ewoprrl r-3'"' owtouv0ettr<6 (owcilvul-to, antisense) xtr<rlvo' H - :rroo1 tou ouvOetrrori x.I.o-rvou enttuyXdvetot Ue Uto :...5poor1 owiorpoqrlq ouppo<pqq, rord qv ono(o ro ="A Lwp6vto evotrll.tottlverot ovdpeoo oro efrovto tou l'.A Aur6 to 6v0eto RNA twp6vto evepye( nL5ov tr:g .-*cye(o Yto rrlv ow(mpo<pq preroypoqdoq lXe66v - r rc twp6vto rrlg opd6oq ll dXouv evepy6qto ow(- - :cof g pretoyporpdoqq nou e(vot et6txrl yto to twp6- : tt ow(orpoQrl UeroYpoqdoq drlprtoupye( 6vo DNA -. .!'poqDo tou twpov(ou pe tefur6 onot5Leopto rqv ev- - --oroon tou twpov(ou orq 06o11-o161o un6 Uopqtl - 't.-rvou DNA. 23.7 Optap6vo AurooupponT6pEvo lvTpovlo Anotro0v MorouPdoeq * T' ouTooupponrogevro tvtpdvto pnopei vo Xpeto(ovrot evep- yoqreq poroupoorlq nou rodtronotouvrot on6 ro tvrpovto rot porl0otiv oro Dintrorpo rou o' evepyo xorotrurtrtl 6opfl' l-1op6tt ro twp6vto ttov opddtov I xot ll 6Xouv rrlv tKo- v6rqro ouroouppogtlq in vitro, un6 rovo.vtt<5g ouvOr1- xeg ouvr'10toq Xpetd(owor BorlOeto on6 npo:tefveq' Koi ot 60o rUnot twpov[ov (ocog t<trl6tronoto[v evtp- y6trlteq poroup6oqg, nou unoBoqOo0v rqv ow(6po- on rnq ouppoqrlq. ' H'evepy6tlto protoupdoqg e[vot rgnpo tou 1'tovo6t- xori ovotXrof n].oto(ou ovdyvtootlg nou t<<o6tronote(tot on6 to Lwp6vto. Ito nopd6erYpo rov twpov(u:v nou ror6tronotoriv evdovoux}'edoeg enotrtoptoti, to ;-tovo6t- 16 npo.lteivtt<6 npoi6v 51et evepy6trlto t6oo ev6ovou- rledoqg doo xot potoupdorlg. H ovdtruoq getotr'- Ld[eotv 6ei;yet 6tt ot 60o evepy6tqteg e(vor ove[dprq- ."g. U 6oprtr<r1 ovdtruoq entBeBorc6vet to 5e6opt6vo tcov pretoMd(eolv t<ot 6e(Xvet 6rt ot evepy6tqteq qg ev6o- vouxtredoqg Kot rnq protoupdorlg nop6Xowot on6 6to- qopettrd evepyo rdwpo tqg npo:relvr1g,.r<o06vo on6 to'ono(o xtodtxonotekot ond 6tocpopettxq neptoXrl H ouvOnop(r; tov 6[o evepyorlto-lv oqv (6to nptoteivrl unodqtrutvet qv e(etrtrcrtrrl nope[o tou twpov(ou' H ": , :' ', uno6ntrovet 6tt to twp6vto e[e]'(10r1re on6 6vo ove[dptrlro ouroouppon-r6prevo orotXe(o' Etr<d(e- ,'n?Vr0don-oroxoq ,72rru, I IMeroypoQn Evdovourltedon / Awi'rporpn UetoYPotP0on ev6ovoure6ori 5'& 3', EvorLlgortrloq "rou RNA tvtpoviou nou orotrou0elrot ond ovr(orPoql UeroYpoQn Yto ro oXrlPonogo cDNA . H .vTioTp.qrl ueroyporpoon/evbovourtreoorl nou rootronote[rot ono 6vo tvrpovLo enrrpdnet o'6vo ovr[ypotpo rou RNA vo , , *:oro.l0e[ oe Pto 050r1-otoXo. ?t.: 0proU6vo Aurooupponropevo lvrp6vto Anotro0v Moroupdoeg 661
*6.&{4 *$ffiW,t 6{, I ,-ll.q*L;* I - rwoo*o*#%,-l ITo yovidto rnq w ; EvDovouxtreoorl evdovourtredorlq I evotouur6veror I oro tvrprivto * I, Torwpdvrorrrr, JLill-Aqvevoovourieoqff-l To yov[6to rrlg goroupdo4g evour gordvetor oro rvrpdvto W I I * To rvrpovro rpeper ev6ovourtreoorl K0r uoToup0orl '' , . ) To tvrpovlo lerlv1oe otg ove[opqu1 otrtr1trouXio, nou xodrronorouoe ovo ourooupponrogevo RNA. H evotoporool qq otrtr1trouXLoq qg evdovourtreoolg drlproupyloe 6vo rvrp6vro enorxropou nou 1rov rrvqro. Iq ouv6Xelo, 1 evourgdrrrlorl qg otr- trltroux(oq rrlg poroupoorlg evloXuoe qv rxovoqro rov o],trqtrou- Xr6v rov rvrpovlov vo 6rnl6vovror orrlv evepyo doUrl yLo rn ouppoqrl roug. tor 6l nop6goro e(vor r; 6tepyoo(o Kor yro to rwp6vro qg oprd6og ll, H evoopdtoorl oe our6 ro ororXe(o puog oL).r1LouX(og nou rol6rr<onore[ ev6ovour].edoq rou npoo6(6er xrvrlrtx6t4ro. Oo16oo, 4 evotrrpdro.ron Uno- pe( vo droropd[er qv lrov6rqro qg RNA oMq],ou1(og vo 6tnLrrtveror mrlv evepyq 6opr1. Aur6 0' ooro0oe n(e- orl yro Boq0eLo on6 nporetveq nou 0o pnopoUoe v' onororoorrloouv rnv rrovdtrlro ovod(n).oor1g. H evoot- p6roor1 truog tS,rotog oMrllouX(og oro rwp6vro 0o dro- rqpotoe rqv ove(oprqo(o rou. Opro;r6vo twp6vto qq ogddoq ll, oro16oo, dev rco- dLronororjv potoupdoeg, To ouyrerprprSvo rwp6vLo [oo:q Xpqorpronotoriv nporeiveq (roo6rivopreg Ue rrq Uo- roupdoeq nou xr.odrronororjwor on6 to rwp6vto) nou r<ro6 tronoro tjwor on6 o).).qlo u1(eg ro u yovLdtco proto g rou [evtorr1. To yeyov6g ow6 uno6r;].rrtver pro nrOovrq e[e].trrrxr1 nope(o yro rouq yevrr<oig nopdyoweg oup- poqnS. O nopdyowog (or,-rg [er<[vr1oe otg proroupdorl nou Boq0oUoe eLdLxd orn ouppoerl ev6g ouyxexprpd- vou rwpov(ou. H xro6rr<onorqtrrq o).trr1lou1[o onoUo- voOrp<e on6 ro rwp6vro oro yovLd[ropro rou (evtorq r<or 662 KE0MA|0 23 Korotrunro RNA orrl ouvdlero e[e].iX0qr<e dlore vo ].enoupye[ o' eupG repq xl,(prorq unoorpoUdrtov oe oX6on Ue rlv ouOev- rrr4 oLl,qloul(o rou rvrpovfou. O roro].urrr6g nuprlvog rou rwpov(ou 0o prnoporioe vo 6Xer e[eh1Oe( o'6vo snRNA. 23.8 H KoroAurrrfi Evepy6rnro Tnq RNAdonq P OgeiAeror oTo RNA * H prpovoux),eoo1 P (RNAooq P) eivor pto ptpovour),eonpot- refuq orrlv ono(o ro RNA 6Xer rorotrurtxl evepydrrlro. e H RNAoon P e[vor onopo(rrlTr] 0To porrrlpto, r' opyolo xo. roug europudreg. a H RNAoon MRP rtlrv europuor6v oXer((eror pe qv RNAoor P ror epntr6reror orqv ene[epyoolo rou rRNA ror qv ono6o- Urlorl Tou mRNA qg rurtrrvqq B. Mto on6 tg np6reg ono6e[[erg trrtv roro].unr<lrv 6u- vororqtorv rou RNA 6d04re Ue rrlv ovdl,uorl rnq pr- Bovour<i.edoqg P (RNAdor; P) rou E. coli. Evc6 opXrxd ovoyvrop[orrlKr oro Bor<rrlpro, q RNAdoq P rouronoL- qOrlxe <og Boorr<4 evdovour<Ledol nou epnLdreror orqv ene[epyoo[o tou IRNA oroug neptoo6repoug, ov 6Xr 6Loug, rouq opyovro;rotig, Borcrqpro, oplo(o xt euKopuoreg. Iqv oni.oUorepn UopAn qg, 11 Borcrrlproxrl RNAdoq P 6r[orotor oe 66o ouorortrd: dvo RNA 350-400 voux.tre- ottS[ov Kor l-uo npcoreivrxrl unopovd6o. H RNA unopovd- 6o trov Borcrqpfrr;v, 6rov onopov6verot in vitro, etrrgovl(er r<oro).urtrr1 evepy6qto. H RNAdon P ttrtv opXo(ov Kor rov euKopuorotv onoreekot on6 6,vo provodrr<d RNA 6opur<d ouyyevSg pre ro RNA rtov Borcr4p(rov, oMd 61er geyoL0repq neprernr6r4ro oe nporelvr1. To RNA 61er e).d1rorr1, ov 61r roOd).ou, r<oro).ulxr1 evepy6qto, 6rov pe).erdror in vitro. Tunrxd, n RNAdon P tcov oplo(ov o.tr- LrlLenrdpd pre rSooeprg nptorelveg evo tr;g (6p49 pre ev- v6o ror rou ovOpolnou trle 66ro. Ie 6Leg rrq nepr- m6oerg, to npcoreivu<6 ouorolx6 e(vor ovoyro(o yro r1v evepy6qro qq RNAdorlq P in vivo. MeroMd[erg e(te rou yovr6fou rou RNA eke rou yovrdfou rqg npcorefurlg odpovonororjv qv RNAdoq P in vivo, enoprSvtoq yv<r-lpi (oupe 6tr rot ro 66o ouororu<d e(vor ovoyr<o(o yto rn,i evepy6rqto rou ev(6prou. ApXu<d e[Xe unora3:. :; - npo.lrelvq nope(1e rqv xoro].urrrr1 evepyorn:c . , - -: RNA eiXe rdnoto entxouprr6 pdlo - n,X, ponOc-:: --np6o6eoq rou unoorpc,-rUoroq, enetdq neotoit. -a:..:: BpoXe(eq oMrlLouX(eg oupnr1po-lpolKrq ur :.1 i.-.i. p6veg nepLoXSq rou DNA. Auro( or p6lor, rr.rr::: :,-. orpdgr;xov r<oorrtg orn npoyUortxdqro ro B,1 I ::-: rlv xoro.trulr4 evepy6qro, evol or nporervoc rtJt, t - 6optxrlornpr(n. Avo),riowog r' onoteL5oporo [-le qv nop:-]:- t- ro RNA e(vor 5v(upro, Kdee "6v(upo" roro.trua. :- : : Boorxeg ewong
onoon notr 6v unoorpoudrov. nop6rr n KorolurtKn evopy6rrlro Bp[oretor oro RNA, ro nproteivu<5 ouorort- r<6 ou(dver nol0 r4v roXfqro qg ow[6poor1q, 6nolg go(vetot on6 tqv ori[4oq tou optO;toti ovorrix].rooqg (BL6ne Ex6vo 23.9). Err6g on6 rqv RNAdoq P, or europurirreg nep6- Xouv Uro dltrq onopokrlnl ev6ovouxLedorl Booropr6vrl mo RNA, qv RNAdoq MRP (mitochondrial HNA pro- cessing, ene[epyoo(o puroXov6proro0 RNA). Auti q ev- douvouxl.edoq onoteLefuor on6 6vo 6opmd ouyyev6q rqrolulr6 RNA ror ;rorpd(eror nol.tr5q npo:relvrr6g unoprovddeg Ue rrlv RNAdorl P. Evd: opXrxd ovoyvop(- orrp<e on6 ro p5l,o rnq ornv ene(epyoo(o rou prroxov- Eptororj RNA, 4 RNAdorl MRP rup(rog tretroupye[ orov nuprlvo, 6nou ene[epyd(eror ro np66po;ro prBooropur6 RNA. H RNAdoq MRP (orog en(or1g no((er o4;rowu<6 p6Lo orq pU0prorl rou KurrqprKo0 xUr.trou, ro069 ou1.r- ger6Xa orrlv onoS5prnon rou mRNA tr'19 r<urd.fuqg B. H ovoyvriprorl r4g RNAdoqg MRP e(vor npox.trr1nrf, ro- Oci:g go(verot 6tr ro npoteivrr6 oumonr6 ourrqpe(tor oe preydlo Bo0p6 pero[0 RNAdortg P r<ot r1v RNAdorl MRP, evri q peroBoLrl r4g er6x5qroq oq npoq ro un6- fipogo ogeftreror mqv owotr].oyr1 rou roroLurtroU RNA. 2 3,9 To loerOfi 'Exouv KoroAurrrfi Evepy6rnro * To LoeLdrl ror ro roooetd4 oXrlpot(ouv 1uo orpupor5qoin 6oUn p' evepyoqro ourodroonoorlg. o l-lop6uoLeq dou6q unopei vo 6q;uoupyq0o0v (euyoprirvovrog 6vov rLdrvo-undorptrllro nou 6toonoror qnd 6vo ev(ugrrd rtr6vo. a '0r0v o ev(uprr6g xkovog erooyeror oro rUrropo, (euyopdrver p' 6vo rtr6vo-ordXo, o ono(og oul ouveXero droondtor. 'Evo or6prq nopddery;ro qg u<ov6rqroq rou RNA vo Lemoupye( og evdovouxledo4 nopSXeror on6 trreprxd grrpd (-350 nt) <pulrd RNAs nou drexneporcbvouv pto ow(6poor1 ouro6r6onqo4g. 'Onog Kor orrlv nep(mat- orl rou rwpov(ou opddog I rou Tetrahymena, e(vor nt- Oovrl q rotooreu( Soprc0v nou prnope[ vo ],erroup- yrloouv o' e[coreprrd unoorpd:poro. Aurd ro prxpd gutrxd RNAs rorordooowor oe 6Uo yevu<69 oprddeg: ro roer6f Kor ro toooer6{. To roer6ri (viroids) e[vor ].ot1.roy6vo pr6pro RNA nou Lenoupyo0v ove(dptrlto Xcopfg vo neprBdMowot on6 rdnoro npo:- reivx6 5).urpo. To roooer6{ (virusoids - ;repu<69 rpo- p6g roloriwor dopuEripo RNA, satellite RNA) rilouv nop6poto opydvrrloq, oMd neprBdtrI.owqr on6 nprore(- veg <purrrdrv urlv r<o0clq ouoreud(owor pro(( pr' 5vo rx6 yovr6(orpro. To roooer64 6e pnoporiv v' owLyporporjv ove[dpqro, xo06g Xperd(owot t4 Bori0ero trr:v rcilv. To roerdq Kor ro toooerdq owrypdgowot 1.uloo ru- l,r6pev<ov r0xLrov (BI6ne Err6vo 12.6). O xAtiryoq rul RNA nou ouoxeud(eror orov 16 roleko xkitwg qr (plus strand). O ouprntrqpcol.torr6q xLtivog nou nop& yeror xord rlv owrypogrl tou RNA rol,ekot xArbvog ilqv (minus strand). Yndplouv nouprepri rot ttov 60o r<trc[vrov, ouv Kor n].4v. Kot or 5Uo t0not provol.teporiq 6qpuoupyoriwor pe drdonoon rnq oupdg ev6q xuht6pre- vou r0x,trou Q. To rurlrrd povopepf rou xl.<itvou ouv nopdyowor ouvddowog t' dr<po ev6g eu0UypopUou provopreporig, T6oo ot xLrivor ouv 6oo rot ot xtrrivot ntrr1v rov toet- dc6v xor rtov toooer66v ug(orowot outo6tdonoorl rn vlf- ro. Opto;r6vo RNA droonoUv in vitro un6 rovovtx6g ouvOrjr<eg. l.trLo 6roonoriv pr6vo prerd 6vo r0rl,o 06p- Uovonq xor rfri(r'1q: our6 unodrlAdlver 6rr ro onopovo- ;r6vo RNA 6Xer oxordtrtrr1lr1 6ropr6pqrrloq, oMd ;rnope( v' onorcrloer evepy6 6ropr6pqtooq ov ono6rqroX0e( xt enovo6rorolOe(. To roer6ri xor roooerdti nou ug(orowor outo6rdono- or1 qlqpot(ouv Uro "oqupoKdeclln" 6ewepotoyn 6o- Uri mn 06o11 drdonooqg, 6nog qo(vetor mo ndvo U6poq rnq ElKOllAI23.14. To o<pupor<5goLo prBodv(ugo ouirouv oe Uro ou<oy6vero prBoev(Uptov nou nepilo1.r- Bdvouv tov 16 qg qnor(n5og 5 (HDV), ro prBo6v(u;ro goupr<drog Kor ro 6opurpoprr<6 ptBo5v(upro Varkud (VS). Aenoupyu<d, o HDV onone( 6ro0evr1 geroMu<d ron6wo yro vo npooOfoer q drdonoorl, evcb to ogu- pox6rpo).o prBodv(upro Kor ro ogupordgol,o gouprdrog 6e Xperd(owor p6rotr.o. H orlproo(o rov perdMorv orrl 0r ouv0werrr6g xegol6q dxouv tperq p(oXoug-filtrdg xor ouwrlprlg6veq pdoerg M(oXoS 3 et E, Arfmoon cG / ^f u^,y' GNCGA IINNNNN cNGA ..cNNNNN Mtoxoq 2 u*ou' vrioxos r Korotrurtrd x6vrpo 0r rerpotr6g drlptoupyorjvror $ o}.$trenibposrl ovdpeoo oe drjo oupnhrlpol-ronrd p6pro RNA Ktrovoq-unoorpouo 8tg ,auonooQ I C6 / aal Y"( PPP G,..G"C"CGA "U.E-GAGCOH H0 CG"6Ga ^AGCUCGGppp^G.- ^utI -u"G Ev(ugutiq xtrrrlvog n fl(0f{A 2*.14 0r neproX5g ourodtdonoolg rorv roerddrv Kor Tov toooetddrv dXouv 1uo ouvorveruq otrtr4trouX(o ror oXrlpor[(ouv gro oQupordQotrn beureporoyrl doUrl U6oro evdopoproro0 (euyoprir- poroq. 0r reqotr6g enioqq drlptoupyouvror Ueoo (euyopdrgoroq ovdgeoo orov rtrdlvo-unoorptrllto Kor Tov (ov(uprro, rtrdrvo. H t*.S T0 loerdq'EXouv Korotrulrq Evepvo:-.: 663 Boorxeg evvorcq
6r6onoor1 rou prBoev([prou VS e(vot or<dprq ooorp4g. 'Oi.' ourd ro prBo6v(upo, oo16oo, npoyUororroto0v gro 6rdonooq nou rorol,o(ner 6vo 5'-OH dxpo rc 6,vo 2'-O'- r<uxAr6 goorpodteorepr6 dr<po. Ze ow(Oeoq pe 6l.o ro un6lorno prBoriv(upro nou riXouv ovoyvrrlprme( pr61pr o4prepo, ro oeupor<dgo).o prBodv(upro rr dLl.o pr6l4 qg ou<oySvetdg roug 6e Xper- d(owor nporeivx6 ouqrour6 yro vo ).eroupyrloouv rn rzrVo, xoOrilg q oMqLouX(o oufig rqg doUnq enopre( yro rq 6rdonoo4. l-ro ro ocpupor6cpoLo prBo6v(upro ro evepy5 r<6wpo e[vor 1.uo oMrl].ouX(o p6].rq 5g vour<].eo- n6(ov. H rerpo).r1 neprSler rpetg neproldg p(oXou-O4- Ltdg, rcov ono(olv n eSon r<or to prdye0og e(vor oroOepd, roOrilg xor 13 ouwqp4pr5vo vouxleor(dro, r<up(og ong neproXSg nou ouv66ouv ro rulwpo r4g 6opqg. Or ouv- rqpqg6veg Bdoerg ror or 6rnlo( g(oXor ouv0drouv 6,vo RNA p' ev6oyevrl n<ov6r1ro 6rdonoong. Mro evepy6g regol4 en(onq dqptoupye(ror on6 ro (euydpopro ev6g RNA nou owrilpooone0er puo nleupd qg doprig trr' 6vo RNA nou owrnpooonerier qv dl,Lq nLeupd qg. To rdrrrr pr6pog t4g Err6vog 20.14 nopou- otd(et rivo nopddery;ro r;g r<ego).rjg nou drlpuoupyefuor on6 rov uBpt6ropr6 ev6g propfou 19 nt g'5vo pr6pro 24 nt. To uBp(6ro trlpe(ror q ogupor<6goLq 6oprr1 pre rrl Srogopd 6n Ae(nouv or oyxriLeg I r<or lll. 'Orov ro RNA 19 nt npom(0eror oro RNA 24 nt, 6rdonoorl ouprBoiver orqv rordMrl).4 05or1qg re<pol,r]g. Mnoporipre vo Oeco- pr;oouge 5rt o ndvco (24 nl) xLcbvog ourotj rou uBpr6(- ou e(vor ro 'un6orpopro' evci o rdro (19 nt) xlrivog ro '5v(upro'. 'Orqv ro RNA 19 nt ovoperyvrjeror Ue nep(o- oero RNA 24 nt, 6roonc6wor noMon).d ow(ypoeo rou 6eUrepou. Aur6 uno6ql,c6ver 6rr undpXet dvoq xrlxl,oq (euyopriprorog 19 nt-24 nt, 6rdonoorlg, 6rdorooqg rov 6roonoopuivrov xLoo;rorov on6 ro RNA 19 nt ror (euyo- p6poroq tou RNA 19 nt p'5vo v6o un6orpopro 24 nt. Iuvencbg, ro RNA 19 nt e(vor vo prBoSv(upo pr' evepy6- t4ro evdovouxLedoqg. Or nopdprerpor ulg ow(6poor1q efuor nop6proreq Ue lq ow(ororleg dtrtrcov owrdpdoeov nou KotqAUowor on6 ro RNA (Fl,5ne Erx6vo 23.9). 'EXer 11611 ovo).u0e( 11 r<puoroMu<ri dopq ev6g eLdXr- orou oeupor<5<po),ou pgoev(riprou. Irrlv eLdlrmq 6o- p4, coor6oo, rl opxffeKrovxrl rou evepyorj r5wpou 6ev ro0toroUoe oocp5g ncirg 0o pnoporioe vo npolrop{oer 11 rcordLuoq. l-l p6ogoro, ovolri04 r<e 4 rpuoro..f.rxr1 6oprl rou nLrlpoug o<puporSgol,ou prBo6v(uprou on6 ro Schlsfosorna mansoni (6vo nopdotro), yeyov5g nou 6rotpdrnoe rlv rordl,uoq. Aur4 q 6oprr1, nou nopouord- (etor opprorrxd mqv EIK0NA 23.15, onoro).Untet pro r<p(otprq rptroroyq oAlrl).enidpoorl ovdgeoo oe pro npoe[oXrl rou pr(oXou I ror tr1g 0r1]ldg rou ;r(o1ou ll. H oMq).en[6pooq oroOeponoe( ro evepy6 xdwpo oe rd- roro dtopr6ptpooq d:ore q G12 v' orporpe( npor5vro on6 ro 2'-OH qg C17, 6n^.o6n qn6 ro 6eopr6 nou 6roond- ror eUKoLq, rot vo 6qproupye(ro enr0eur6 2'-o[uy6vo. Me q oepd rnq, n G8 nop5Xer u6poy6vo yro tr1 oro0e- pono[4oq rou veooxnponopuivou 5'-OH drpou mo 3' npoi6v drdonooqg. Elvor 6uvor6g o oXedroqr6g ouvduool.uirv ev(riprou- unoorpdlporog xovciv vo oXrlpor(oouv eLdXroreq oeu- 664 KE0AAAI0 23 Korohulxri RNA IVlioXoq I [4(oxog Ill Ell(0NA 23.15 H dop4 rou orpuporeqotrou prpoev(rjgou ouyKpo- re[ror o' evepy6 rpuoroy1 dLop6prpoo4 pSoto otrtrqtrentdpooeov pero[u trtv luoXrltirv oyrutrtlv, nou enrorlgo(vovror ond ro petr1. H 06orl qg dLoonoo4g enroqpo[veror pe ro r6xxrvo petroq. l-lpo- ooppoog6vo on6 M. Maftick and W. G. Scott, Cel/ (126): 309-320. pordqotreq 6op6g. Aur6g ot 6opr6g rilouv Xpnoqonor- r10e[ yro vo 6e([ouv 6n r; etooytoyrl rov rordl.q],cov ;rop[cov RNA o' 6vo r<rirropo prnope( vo ennp6rper tr1v ilpoyporonofio4 ev(uptxc0v owr6pdoecov rn vivo. Ztrlv npoyponr6trpo, dvo prBodv(upo oXedtooprdvo Kor' ou- r6v rov tp6no e(vor 1.uo e[orperxd er6u<q ev6ovoux].ed- on ileproprollof nou roreu0[veror evovr(ov ev6g RNA-m6Xou. Av ro pgoSv(upro reOe( un6 rov 6).ey1o ev6g pu0pr(6Uevou unoKrvrlrrl, gnope[ vo Xp4orpronor- q0e( 5ncoq n.X. ot antisense rorooxeu6g yro v' o6povo- nouloet erdxd tqv 6r<ppoorl evdg yovtd(ou-or61ou un6 ro0opro;.r6veg ouvOrlxeg. 2 3.'l O Ene{epyoofo rou RNA lupBoiver oe Mepovtop6veq Bdo€tq * 0r uno0oxelq qg onotrtnonporelv1g-B xor rou ytrouroprrou utp(orovror erdrreg onogrvooerq nou roro),uovrol ono 0eopr- vooeg qg ruld[vr1g ror qg odevoolv1g xor otrtro(ouv rlv Ko- Drronoullq otrtr1trouXio. 'Evo Boou<6 o[(iilpro qg propror<qg BLo].oy(og e(vot 6l 11 oLAq).ou1(o ev6g mRNA owrnpooonerier p6vo 6,rr e(vor rro6rr<onorqprSvo oro DNA. To r<ewprr6 66ypo npodBLene puo ypopprrxrl oX5on, onlv ono(o pro ouve- plg oMrlouX(o DNA preroypdgeror oe Uro oMrlLou- X(o mRNA nop Ue rq oerpd rnq Llerogpd(eror oe no).unem[61o. H Unop(r1 Sroxexoppr6vov yovrdicov r<oL 11 oqo(peo4 rcov rwpov(tov pr6oir.t rrlq ouppoerlg rou M(oXoq ll < )'--+ c'.G 1 n- G,"A t ^"n:]*C.;--"Ai/s A-J G.---"U G A JC FI QI Boolxeq eworeg
RNA eroqyoyov 6vo entnL6ov p(Uo orn drepyoo(o rqg Srgpoo4g tcov yovtd(tov (Bh6ne Ke<pd).oto 21). Iuvo- mr6, 11 ouppoqq npoYporonote(tot oto en(ne6o tou RNA r<r 6Xer og onor5leopo trlv o<po(peoq pr1 rodtro- norqlrrilv oLlqAouXrdrv (twpov(<ov) nou 6tox6nrouv -ng ro6rronorqtrr6g oMql.ouX(eg (e[c0vto) ndvo orqv oMrlLoufo tou DNA. H 6tepyoo(o efvot 1.tto UeroQo- pd nlr;po<poptcilv, oqv ono(o q npoyprottrrl r<r:6tro- notrlrrrrl ollrlloulio tou DNA nopogdvet opetd- 9rnq. AL).oy6q ttr;v ntr4 porpo p tcirv no u rcodtrono toUwot oro DNA npoyporonotofwot oe optoprSveg e(otpenr<5q neprmdoerg, xup(tog t<otd t4v nopoycoyrl v5rov oMq- l.ouXuilv nou rcodtronotoUv ovoooo<porp[veg oro 0qtro- onrd ror ro m4vd. Tdtoreg oli.oy6q ouprBo(vouv et6xd oro ooponKd rrlrropo (B Leg<por<r3rropo) aro ono(o ouw(0ewot ot ovooooeotp(veq (B)r5ne Kerpdtroto 18). N5eq ntrqpogop(eg nopdyowot oro DNA ev6q ot6ptou rot6 11 dtdpreto rnq ovoKqroor<eurig ev6g yovt6(ou ovooooqorpfvqg rot ot n].r1po<pop(eg nou efuot t<codxo- noqp5veg oro DNA preroBdMowqt on6 oroprotx6g pre- toLl,d[etq. Or nLrlpotpop(eg tou DNA ouveX((ouv vo pretoypdgowot ntord oro RNA. H enefepyoo(o tou RNA (RNA editing) e(vot 1.tto 6tepyoo(o mrlv ono(o ot nArlpoEopbq aMd(ouv oro en[ne6o rcu mRNA. Aw6 onoro],0me-rot on6 xotqord- oerq orrq onoteq 11 rodtronotqrtr<ri oLlrll'ou1[o ev6q RNA 6toq6pet on6 qv otrtrql,ou1(o tou DNA on6 qv ono(o peroypdqrp<e. Ene[epyoo(o tou RNA ouptBo(vet oe 6rio Drorpopetrr6g rotoordoetq, 11 r<oOepr(o pre 6to- gopetrrd ofttq. Iro x0nopo ttov orproortrdrv, undp- Xouv neprmciloetg 6nou owtrordmoorl ouprBo(vet oe ;uo Bdo4 rou mRNA pretoBdl,Aowoq rrlv otr).r'1}'ou1(o rou ro:srronoroUpevou nol,unemt6(ou. Aurrl n owtKo- rdmoorl e(vot ro onotStreopo t4g onop(vcoorlg e(te qg o6evoo(vnq npoq tvoo(vq efue qq r<un6fvqq npoq oupr6(v4. Iro puroX6vdpto tou tpunovootitl-toroq, eu- prirepeg oMoy6g ouprBo(vouv oro pretdypoqo noMdlv yovr6[trlv, 6rov oum4pottrd npoor(0ewor 11 onotro(- qowor B6oerg. H EIK0NA 23.16 ouvor!((er ttg oMr;Lou1(eg rou Yovt- 6(ou r<or rou mRNA u1q onol.tnonpotetvqg-B (apo-B) oro 6wepo xor ro qnop trov Or1troortxdlv. To yovtd(copto nepr6ler 6vo provoSxd dtor<eropptdvo yov(6to, tou ono(- ou 4 ol"qtrouX(o e(vot novopot6runq oe 6Loug toug roroig, Ue pro rodrronorqtrxq neptolq 4563 t<todtro- v(o:v. Aur6 to yov(6to preroypdqetot oe mRNA nou Ue- rorppd(eror oe Uro npcoretvq 512 kDa, q ono(o owrnpooonef er qv nln pn xo.t6txonotqtu<r1 oM4Lou- 1(o mo 4nop. Mto Bpolftepq Uopqn qg npoteivrlg (-250 kDa) ouw(0erot mo 6wepo. Auq q nptoteivrl onoretrefror on6 ro N-te].tr6 plo6 rrlg n]rlpoug npo- re[vr;q. Meroqpd(erot on6 6vo mRNA, tou ono(ou t'1 oM4l,ouX(o e[vot novopor6tunrl Ue rrlv ow(orot1r1 tou qnotog g' e(o(peor1 puo oMoyrl on6 C oe U oro rtrl6t- r6vro 2153. Autrl r1 owxotdoroo4 oMd(et to ro6u<5- vro CAA qg ylouropr[v4g oe ro6tr6vto tepprortoprof UAA. Enerdrl 6ev undpXet evoMorrtr6 yov(6ro { eE6- vro oro yovt6(co;ro yro vo rto6xonotqoet q v5o otrtrq- To yov(6to qg onotrtnonporeivqq B 61et 29 e(drvro .!!.ffi ro xtrldtrovto 2153 rto6tronote(q ytroutopfurl t c AA Ene{ePlooto uAA :: I o&rororyP To ouppopdvo mRNA mo rlnop xtrloxonote( pto nept5XeL ro xtldtxdvto UAA npote'tvn 4563 rotdointov nou repuotl(et rn o0v0eoq qg nporetvnq oto rotdtrotno 21 53 EIK0NA 23.16 H oltrn)ouxio tou yovt0[ou qg apo-B eivot [6ro mo 6vrepo roL to r1nop, oilro n otrtrnhou1fo tou mRNA Tporottot- e[ror pre otrtroyn Urog Foonq, n onoio OlptoupYei 6vo rol0txovto reppolopoti oro 6vrepo. Loul(o ror de Bp60ry<e r6noto oMoyri mo np6tuno rnq ouppoqrlg, ovoyroortrd ouptnepo[voupe 6tt rd- noro oMoyrl o'uv5Bq d;reoo orrlv oMrlLoul(o rou RNA petoypd<pou. 'Evo or6prq nopddetypo e(vot ot uno6ole(g tou ytrouropur<o6 orov eyr<6goAo rov opoupofrov. H ene- (epyoo[o oe pto 0do4 ;retorpdnet ro rro6tr6vto t4g yl.owop(vqg oro DNA oe rodtr6vto rrlq opyrvfuqg oro mRNA. H oMoyrl on6 y)routoprtvrl oe opyw(vq enrlped- (er t4v oycoyrp6rqro tou StoULou n enop6vtog 51et o4- prowu<r; en(nroorl mq pri0;uoq rnq po{q rrov t6wtov dropSoou tou 6rqUlou. Ie dtrq 05oq tou uno6oX5o, 6vo ro6rx6vto opytv[vqg perorp6netot oe rodtr6vto ylur<rvr;9. To ou;rBdwo nou neprypdenKov yto rouq uno6o- 1e(g qg apo-B ru rou yl.ouropKoU e[vqt to onorStre- opo anastwritoe@v onq ono(eg oqotpe[tot q optvoptd6o rou vour<Leorr6rro0 SoxruL(ou. H ene[epyoo(o qg apo-B nporo),e( oMoy4 rrlS Czrse oe U, evci xot ot 6tio olloy6g orov uno6oX6o tou yloutoptro6 e(vot A npoq l(voolvq). Ot onogwcioetg tqg onoltnonporetvqg B ro- tol0owor on6 q deopuvdoq t4g rutt6(vqg APOBEC (apolipoprotein B mRNA editing enzyme complex, otil-t- nlor<o ev(Upro:v ene(epyoo[og tou mRNA t4g ono].t- nportefvrlg B), evdr ot onopwrioetq orov uno6oX5o tou yAouropuxorl on6 ng 5eo;uvdoeg qg odevooivqg nou 6pouv oro RNA (roLotiwot ADAR, adenosine deami- nases acting on RNA). Aut6q o tUnog ene(epyoo(og go(vetor vo ouprBo(ver r<up(tog oro veuptr6 o0mqpo. Yndplouv '16 (duvqrtr<o() or6Xot rcov ADAR otq Drosophila melanogaster ror 6lor e(vot yov(6to nou eU- nL6Kowqr mr1 veupo6roB(Boo4. Ie noltr6g neptmdl- 23.10 Ene[epyoo[o rou RNA Iuppo(vet oeMegovtop6veq Bdoetq 665 I wTo mRNA tou evr6pou
EIK0NA 23.'17 H ene[spyoo(o Tou mRNA npoypoTorore[Tur 6T0v pro deopLvdorl snrdpd oe pro odeviv4 grog drilootoop6vlg nepLo- Xrlg ev6g ore)drg (euyoprop6vou RNA. oerq, n ene[epyoo(o o].].d(er 5vo optvo[ri oe pro ].er- roupyu<d oqprowrxrl 06oq qg np<otetvrlg. To 6v(upro nou TrpoyUoronoroUv rrlv onopr(vooq ou- yyd 61ouv eupe(o er6rr5rpo - n.X. n r<oLwepo preLeq- ;r6vr1 6eo;rrv6o4 rqg o6evoo(vqg enrdpd oe rdOe rordLorno A ;uog 6rnLooroql5vqg neproXqg rou DNA. Om6oo, 11 onopr(vooq qg o6evoo(vnq Kor rqg ruu6(- vnq oro RNA ep<pov(er er6r6r1ro. To dv(u;ro ene[ep- yoo(og oXer[(owor Ue rrq yevrKSg 6eopuvdoeg, o].].d nepdXouv dLl,eg neproX6g q enrnp6oOereg unopovd- 6eg nou puOpr(ouv t4v et6u<6qrd roug. Irrlv nep[mro- on rnq ene(spyoo(oq rnq apo-8, 11 rorolurrrrl unoprovdSo ev6q ouprnL6r<ou ene[epyoo[og qXeti(eror pe rr1 Bomqpror<4 6eopuvdoq qg xurtd(vqg otrld 6Xet pro enrnp6o0eq neproXrl np6o6eoqg rou RNA nou Bo40d orqv ovoyvrilpron rnq erdu<rjg neptoXqg-or61ou npog ene(epyoo(o. Mro er6rrci 6eo;rwdoq qg odevoo(- vrlq ovqyvop((er rrq neproldg-or6Xoug mo RNA rou uno6o16o rou ytrouropu<oti, evc0 nop6proro ouprBdwo Trpoyporonoto0wor oro RNA rou uno5o16o rnq oepo- rovfuqg. To orjpnl,oxo (orrlg ovoyvrop(er puo et6xr1 ne- proXq qg 6ewepotoyorig 6oprig xord rp6no ovdl,oyo ge ro 6v(upro rponono(r1oqg rou IRNA 11 0o prnopo0oe v' ovoyvrop((er oneu0e(og pro vourl,eorr6rxrj oM4l,ou- X(o. H ovdmu[r1 ev6g in vitro ouorriprorog yto rnv ene- [epyoo[o rqg apo-B uno6r1].6ver 6rr rord].].4].og or6Xog e(vor ;uo qXeru<d purp4 oMqLou{o (-26 vou- rtreotbto) nou neprBdLl,er rnv neproXrl ene[epyoo(og, H EIK0NA 23.17 6efiyer 6rr orqv nepfmooq tou RNA yro tov uno6o15o rou ylouroprro0, ovdpeoo ornv e[epyo- op6vr1 neproXq rou e(ov(ou Kot Uro ou;rn).4pcogotrrrl oM4LouX(o rou xo0odrro0 rwpov(ou q4rlgod(eror pro 6(xl,ovq neproXq, yvomrl og GluR-B, nou e(vor ovq- yxo[o yro qv ovoyvr6prorl rou or6Xou. 'Evo ].dOog (eu- ydprol"ro pr6oo orq 6(xlrovrl neproXri e(vor ovoyro(o yro er6u<ri ovoyvcbpron. Iuven6g, drorpopeurd ouonjproro ene[epyoo(og prnope[ vo 6Xouv Srorpoperu<69 onorq- oerq ovqgoprrd pe qv erdx6rrlro yro qv oMrll,ouXio rov ufloorpoprdtcov roug. 23.1 I H Enefepyooio Tou RNA Mnopei vo Ko0o6nyeiror on6 O6nyd RNA * Iro Utrox6vdpr0 Tou Tpunovoorogorog ouppolver eKTeTopsvrl ene[epyoo(o rou RNA p'ev0eoeLg r1 onotrorqeg oupLdivrlg. e To RNA unoorporpo (euyoprirvet U' 6vo odnyo RNA xot ono ttg 6uo ntreupeg qq nepioXqg 1 ono(o nporeuor vo unoorel eneIepyooro. * To od1y6 RNA nopdXerro er;loyefo yro Tlv npoo0rtrn (n onovrorepo qv onotrouprl) ouprdrvrirv, * H ene[epyoolo rorotruerot ono ro otopor[dro ene[epyoolog, evo oupntroro p' evepyorrlro evdouvourtreoolg, e[ovouxtre- ooqq, retrurlg ouptdu),orpovotpepoorlq xot RNA trtyrooqg. 'Evog 6togopelr<69 rlinog ene(epyoo(og onoro).Ume- tor on6 lq ewunoorox6g o).).oydq qq oMrlouX(og oro npoi6wo noMdrv yovr6f<ov rrrlv Urroxov6p(olv rou tpunovoocitllorog. Irqv npotq nep(moloq nou ovoKo- ).Urp0r1re, q oMq).ouX(o rnq unopovd6og ll rrlg o[er6o- orlq rou r<uroXpcbporoq 5Xer Uro eooreprK4 prerot6nrorl noro(ou nou 6ev npoBAdneror pe Bdorl q voux.treort- drr<r1 oMr1l.ou1(o rou yovr6(ou coxll. Ot oMrl).ou1(eq rou yovL6(ou Kor rnq npolre'ivr1g nou 6(vowor orqv ::....i, ..:.: , " ouwr]poUwor oe noMd e(6q rpunovooropro_ rog, ouvenolg 11 p60o6og ene[epyoo(og tou RNA 6ev e(vor onor.treronxrl yLo 6vo ;r6vo opyovropr6. H ooullgrov(o ovdpeoo orqv oMrlLoul(o rou yovt6(- ou coxll Kor ro nporeivrx6 npoi6v ogeil.etot orrlv ene- (epyoo(o rou RNA. To mRNA tou coxll neptSler ;uo Adevoolvr1 lvoo(vr1 NHI C I-rl * -c -6 -N, HN -C .c -N.. I tr cH-> I lt cH HC --n, -C -g/ HC --n, -C -g/ 'r | ,, L Ktodrronotl96vo oqv otrLr1trouX(o rou yevtogworl DNA Metor6nrorl tou ntroto[ou ovdyvoorlg Atr)trftouXio RNA /UtrqlouXio nporeivrl g ouloro ntro[oto ovoyvtoolg nou noporr;pe[Tot oTrlv npolreiVl drlptoupyeuor ono rqv evoorporooq reoooptov vourtreoldlrrrv ouprdrvrlq. 666 KE0A Al0 23 Korotrulxo RNA I S S L G I K V E N i:L€ V C AUA UCA AGU UUA GGU AUA AnR GUA GAG MC eUGJGUA GGU AUA UCA AGU UUA GGU AUA AAA GUA GAU UGiJ AUA CCU GGU AGG UGU MU ISSLGIKVDCIPGRCN Boorxeg eworcg
U UAUAUGUUUUGUUGUUUAUUAUGUGAUUAUGGUUUUGUUUUUUAUUGGUAUUUUUUAGAUUUAUUUAAUUUGUUGAU ru @9AAUACAUUUUAUUUG UUU-GUUAAUUUUUUUGUUUUGUGUUUUGGUUUAGGUI @ UUUUUUGUUGUUGUUGUUUUGUAUUA TUnUo qg otrtrr1trouXrog mRNA coxlll rou T. brucei.l-lotrtr6q oupL6rveg Dev rodtronorouvrot oro DNA (rorrtvo) n oQotpouv- ror ono ro RNA (olUo(vovrot pe T). 6v0eor1 rcoodpov vour.l.eott6(orv (ouptd(veg) y0po on6 qv neproXri rqg petot6ntorlg nLoto(ou. H 5v0eor1 ro- Oop((er ro rotdMqlo ntro(oto ovdyvtloqg Yto rnv npo- retv4. Aev 61er ovorolug0e( dtrLo yov(6to coxll nou vo q6per q petot6ntoq nl.oto(ou: 6rot ouvdyetqt 6tt ot enrnp6oOereg Bdoerg etq4opo0v rotd rr1 6tdpreto rqg preroypogr1q rj petd on5 owrl. Mto nop6ptoto ooupqo- v(o ovdgeoo oro mRNA Kot rrq yevulptr6g oMrltrouX(eq Bp(oreror oe yov[6to to6 nopo;ru[oioU SVS xqt tou toti t4g rLopdg rot o<peil,erot oe npooOrlrrl xorolofutov G oro mRNA. l-'lop6proto ene[epyoo(o oM4].ouXtcilv tou RNA oupt- Bo(ver ror oe dMo yov(dto xot nepil,oprBdvet t6oo ono- ).o1969 6oo xot npooOrireg oupr6(vqg. H oouvti0tmq nep(nrrooq rou yovr6(ou qq o[etddoqg lll tou xuto- Xpc0prorog c (coxlll) :rou Trypanosoma bruceiouvotf ((e- ror crrnv EIK0NA 23.19. fdvu an6 ra ptod xatd^olfta rou nRNA e[vu oupt[veq nou 6ev xadxonoolwat oro Yo- Iovt6[togo fl poeneqepyoou6vo AAAGoGGAGAGAAMGAAA RNA mRNA Guide RNA v[61o. H o(yr<pro'q ovdpeoo oro yevoptK6 DNA rot ro mRNA deftver 611 6ev undpXet rpnpo rou mRNA ndvo on6 7 vouxl,eodSto nou vo prqv 6Xet unosre( oLLoyti ror 6u erodyowot ouorotl(eg oupr6lvqg pr{rouq 5og 7 roroLo(nov. Or ntrlpogop(eg yto rqv et5xrl 6v0eoq tov oupr6tvri:v nopSXowot qn6 6vo o6qy6 RNA (guide R- NA). To odr1y6 RNA nepdXer gto oLqLoufo ouprntrrlpo- prorrr{ pe ro otrlord ene[epyool.tdvo mRNA. H EIK0NA 23.20 6e(tyet 6vo prow6lo yto t1 6pdoq rou oro yov[6to rou r<uroXpcilgotog b tqg Leishmania (6vo dMo nopdor- to). H oltrqLoufo oro ndvo prdpog qg ex6vog defiyet to qu0ewu(6 prerdypogo 11 ro getdypo<po 6ncog e(vot npw on6 qv ene[epyoo(o rou RNA. Tq revd defiyouv nori ero6pXowor Bdoerg rord t1v ene[epyoo(o, Orcrc0 ouprd(veg np6ner vo etq(oprioouv oe ouri qv neptoXti 6ore vo 6rlpuoupyq0s( puo dyrup4 oL}'4l.ou{o mRNA. To odqy5 RNA e(vor ouprntrqpoporxS pe to mRNA yto AAAGCGGAGAGAAAAGAAA A G G CTTTAACTTCAGGTTGTfiATTACGAGTATATGG !t,,;t1,';g'pr * A G G C UUUMCUUCAGGUUGUUUAUUACGAGUAUAUGG I Zeuvupr,lucvo pe ro o0qy6 RNA t llpoene[epYoo[6vo Rpvq66664qAGAMAGAM AG GcUUUM6UU6AGGUUGUUUAUUA6GAGUAUAUGG RNAmm*83ilXmm$$m$XXXX OOny6 RNA AUAUUCAAUAAUAAAU UUUAAAUAUAAUAGAAAAUUGMGU UCAGUAUACACUAUMUMUMU I rvor,rpotrooq ouptdwrlv v AAAGCGGAGAGAAAAGPAA UUUAUGUUGUCUUUUAACUUCAGGUUGUUUAUUACGAGUAUAUGG il illllil lHII}I ltil Ilil lIAUAU UCAAUAAUAAAU UUUAAAI.'AUAAUAGAAAAUUGAAGU U CAG UAUACACUAUMUMUMU I t--', ;-.1;; :' .. .:j ; - I mRNA AAAGCGGAGAGAAAAGAAA UUUAUGUUGUCUUUUAACUUCAGGUUGUUUAUUACGAGUAUAUGG ,;:rr. .' ,r:', lr ,r.r To npoene[epyoop5vo RNA (euyoptirver p' rlvo o6qyo RNA ror on6 rtg 6uo ntreup6g qg neptoXqq nou nporettot v0 uro- orei ene[epyooio. To odnyo RNA nopdXer dvu erpoye[o yro qv evourlroroorl Tov ouptdtvtirv. To mRNA nou nporuntet ono qv evotlporol- o1 e[vot oupn],r1pogolr6 tou o61you RNA. 3;.'!i H Ene[e pyoo(o rou RNA Mnope(vo Ko0o6lyeuot ono 06qyo Rri,+ 667
MURF3(5') CyB-2 O6ry6 RNA MURF2-1 Coll-FS MURFII-2 CyB-1 l:.:.;i7:,r::125 95 MURF3 CoillCytsMURF4 tMURFl NDlCoilMURF2Cot ND4 ND5 :r,..:i,::: 'l i .': To yovLd[oU0 Tnq lelshrnarla nepr€Xri y0v[6r0 nou KodrKonorouv np0ensqspy0og6v0 RNA ovoperKTo Ue UOV06eq, n6u K0]- dtronotouv t' onopo[qro odrlyo RNA yto q onuroupy[o rov ooortirv mRNA otrtrr1trouxrdv. 0prourvo yovroro eXouv notrtrontrd o61yo RNA. CyB e[vot T0 nporeEepY0opevo yovisLo tou ruroXptirlrorog b, evdr CyB-1 roL CyB-2 ro yov[6r0 ro]v 06ly6v RNA nou epn],6rovror orrlv sneqepyoo[o Tou. o41-rowm{ 5rcrooq, nou nepil.o1lBdver xor nepgdtrtrer rqv eile(epyooUdv4 neproXrl. Tunu<d, q ou prnLq po proru6r4- rq e(vor nro ercrevrlg ornv 3'n).eupd tqg ene[epyooprd- vnq neproxnq, Kor. ilro BpoXe[o mqv 5'n].eupd. To (euydpo;ro ovdpeoo oro o64yd RNA ror ro npoene[ep- yoopr6vo RNA og4ver revd 5nou o(euydpcoreg A rou o6rlyoti RNA 6e Bp(orouv oupn).qproporu<6g Bdoerg oro RNA nprv on6 qv ene[epyoo(o. To o6r1y6 RNA nopdXer 5vo erpoye(o nou enrp6ner r4v evocoprdroon rov Koro- Aohcov U oe our5g ng O5oerg pe puo 6epyoo(o nou 0o neptypoqe( orq ouv61ero. Otov oloxL4pto0e( r; owi dpoon, ro o64y6 RNA onoondrot on6 ro mRNA nou e(- vqr nL5ov 6ro06orpo yro ;retdrppoorl. H e[eL6[r<euo4 qq re].rxr1g ene[epyoop6vqg oMq- ).ouX(og e(vor opxerd nol,rin),oxr1. Iro nopddetypo rou rwolp6prorog b rqg Leishmania, pto prorprd neproXrl tou perdyporpou urp(ororor ene[epyoo(o pr' 6v0eoq ou- voLrrd 39 roroi.o(ntov U, 4 ono(o go(veror 6u onone( 6Uo o6qyd RNA nou 6pouv oe yenovtr5g 06oerg. To np6ro odr1y6 RNA (euyop6ver pe rrlv mo 3' 06o4 ru 6nemo 11 ene[epyoop6vq o].].q].ouX(o yfverot ro un6- orpopo yro nepon6po ene[epyoo(o on5 ro end;levo o6qy6 RNA. To o64yd RNA xrodrxonoroUwot oq ove- [dprrlreg preroyporprxdg povd6eg. H E!K0NA 20.21 no- pouord(er 5vo 1dprr1 rrlg oXerrxrlg neprol4g rou 1-utolov6proxod DNA qg Lershmanra. O Xdprrlq ilepr- troprBdvet ro yov(6ro rou rurolpriprorog b, to onoio r<co- drronore( qv o).).4Aou1(o nprv on6 rrlv ene[epyoo(o, ror 6Uo neprolSg nou r<o0op((ouv ro o64yd RNA. To yov(6ro trrlv r<tiprrov r<todrronotrlnr6v neprolcbv e(vor ovdperrcro pre ro yov(6to trov ow(ororXcov o6qyc0v RNA. Oe<op4nrd, pro ;rerdMo[r1 e(te rou yovt6(ou e(te ev6g on6 to 6rio ow(ororlo o6r1yd RNA 0o ;rnopoUoe v' oLL6[er rrlv npororoy4 oM4IouX(o rou mRNA dpo Kqr rnq npolrefv4g. Me yeverrrd r<pn4pro, xoOep(o on6 ourrig ng provd6eg 0o prnoporioe vo 0ecop40e( 6tr ono- reLe( rpqpo rou yovr6(ou. Or provddeg ergpd(owor ove[6prr1ro, on6re oogoA6g 0o dnpene vo vo e(vor ougnLrlpcolrorx5g in frans. Iuvenc0g, ov unqplov 6ro- 06orpreg preroMd(erg, 0o dronrm6vopre 6rr yro vq Ko- 6rrono140e( 4 npororoyrlg o).l.r1LouXio tlroq 668 KEOAAAI0 23 Korotrurud RNA provodu<4g nporeiv4g Xperd(owor tperg ou;rnLqpoJUo- ttr6g oprddeg. O loporrqpro;r6g tolv ev6ropuiocov nou u<p(orowor pepr4 ene(epyoo(o uno6r1).cirver 6l q ow(6poon ilpo- lrope( rotd ;rrlrog rou npoene[epyoopr6vou RNA oe r<o- re[Ouvor1 3'-5'. To o64y6 RNA r<o0op((er rrlv er6x6rrlto torv ev06oeov oupt6ivrlg prSoot tou (euyopdrprot6g rou Ue ro npoefle(epyoopSvo RNA. H ene[epyoo(o rrov ouprdwcbv xoro].Ueror on6 6vo 20S ev(upur<6 ofpn).oro nou xol.efuor oapailo eneep- yoolaq (editosome), onore].efuor on6 nep(nou 20 nprrl- -retveg rcor neptSler pro ev6ovouxl.edoq, pro re),rr4 o u p 16 uLotpovogepdorl [f UTdorl), puo 3'-5'U-er6u<r1 e[o- vour<Ledorl (eEcoUdofl Kor Uro RNA Lryr<do4. 'Oncog <po(- veror. orrlv EIK0NA 23.22, to oropor(6to ene(epyoo(og npoo66ver ro o6qy6 RNA r<or ro Xprlorpoilore( yro vo (euyopc0oet Ue ro npoene(epyoo;r6vo mRNA. To RNA un6orpopro 6roondrot oe Uto nepropl nou (nr0ov6v) ovoyvtop[(eror on6 rlv onouo(o (euyopuiproroq Ue ro EI 3', *"q&ff I rvoovourreoon ry&ff . TetrLrrl I f oupr0utrorpovoqepoorl '(TUTdon) :ry J **o ^,y*ooqY .. .,, , l1poo01r1 :l onolouprl roro),0[nov U npoyporo- nole[ror pe Droonool rou RNA, otporpeoq 11 npooOr1u1 U ror ofv- deol riov orptov. 0t ovlDpooerg roro),0ovror ono evo oupntroro ev(uprrrv uno qv ro0odrlyloq rou odrivou RNA. 5',
o64y6 RNA: truo oupr6ivrl evool.rotriveror 11 onolo(rperot yro vo (euyopcioer pre ro o6qy6 RNA r<t 6nerq ouv6Serqr ro RNA un6orpo;ro. H tpupargopuq oupr6fur1 (UTP) ei- vor 11 n4yrl rorv r<orol.o(nov oupr6fur1g ror npoor(Oeror pe q 5p6o4 rrlg TUTdorlg. H onoi,orqri rov rorolo(nrov U 6ropreooLoBefuor on6 pro e[oUdoq, q ono(o ouvepyd- (eror pre pto 3'gtoogor6oq enmp5nowoq oro RNA nou unoBtrrlOqr<e o' ene[epyoo(o vo enovoouv6e0e(. Or 6opdg ;rop(cov nou 6Xouv unoBLq0e( oe prepu<r1 ene[epyoo(o unodrltrdrvouv 6l ro r<otd].orno U npoor(- Oewqr 6vq q rpopd nopd r<ord o;rddeg. l-10ov6v 11 ow(- 6poon npoXrrrpe( 6roprSoou 6ro6o1rrd:v rfrtrcov oroug ono(oug npoor(Oewor r<otdtrorno U, 6ort;rd(owot trlg npoq rn ouprntrrlpoprorx6rrltd rouq pe to odqy6 RNA ror 6ror4po0wor ov e(vor ono6errd evdl orpotpotiwot or4v owi0erq nep(mcooq: ouven6g, 4 rotooreur] rqg ooord ene(epyoopr6qg oMqtr ou(og ouptBoiva orodto- xd. Ae yvop((oupe ov or (6tor t6nor owt6pdoerrrv eprntr5- Kowor. orr;v ene[epyoo(o nou npoo06rer rord.trorno C. 23,12 H tuppogfi llporeivdlv Elvor AuroxoroAurr16 r Mro rvretvrl 6Xer rr;v rrovdqro vo rorotruer rrlv orporpeon rnq on6 pro npolre[vr1 rord r6roro rpono tiore vo ouv06ovror or e[reiveg rou Trlv neptpotrtrouv. r H ouppoQn rov nptoreivdrv roroLferor ond qv twelVrl. t 0r reproodTepeg rwetVeg 6Xouv 6rio ove[opqreg evepyoq- req: ouppoqn nporciVtiv rt evdovourtredorl enotuqrou. H ouppogri rov nporeiVdrv (protein splicing) SXer to (6ro onordLeopo Ue rrl ouppogrlrou RNA: puo o).- L4l,ouX(o nou owrnpooonerjeror fio eooreptr6 ev6g yovr6(ou 6ev owrnpooorne6eror orrlv nporeivq. To r;rq;roro rqg nprore(vqg ovoprd(owqr Kor' ovoLoy(o pe rn ouppogq tou RNA: elreiVeg (exteins) e(vor or o),).4)rou1(eq Trou clwrnpoo(l)neJowor mnv drptprl nporetvrl, evri rweiVeg (inteins) or oMrll.ouX(eg nou ocporporiwor. O prqXovropr6g yro qv o<po(peon rrlq tv- retvqg e(vor eweLtirg 6torpoperu<69 on6 rov ow(oror- Xo rnq ouppo<pqg rou RNA. H EIKONA 23.23 6e(yyet 6tt ro yov(6to ;rerorppd(eror oe Uro np66poprr; np<oretvrl nou neprdXer trlv rwelvq xr 5nero q rwei4 orporpefuor on6 r4v nproref,rr1. fldvo on6 350 nopodetyporo oup- pogns npoteivdlv efvor yvoord ror 6rodedopt5vo oe 6l,eg ttq r<oqyop(eg rov opyovroprc0v. To tuntx6 yov(- 6ro rou ono[ou ro npoi6v ucp(ororor ouppogti 6Xer 1.uo l.rovodrq rweiwl. H npcirq rwetvq ovor<olriq0qKe oro yov(6to rnq D- NA noLuprepdoqg ev6q opXo(ou un6 poprp{ Uroq no- pepBoM6pevrlg oMrll,ouX(og oro yovbro q ono(o 6ev or<oLouOe[ roug r<ovdveq yro ro rwp6vro. Irrl ouvdlero onodeffiqre 6n 11 roOop4 nporeffi prnopef v' onoMo- nporeivnefu ; Exioprl n1g . * rwervnq ono ' Trlv fipolervn w4EIK0NA 23.23 Koro tr1 ouppoqn nporeiVdrv, ot e[retveg ouvD6- ovrot orporptirwug qv tvre[vr1 ond r4v npotelVq, ye( on6 ourrl qv oMql.ouX(o oe Uro outor<oro).uttr4 ow(6poor1. H ow(dpoo4 6ev onone( npoo0rp<r1 ev6p- yeroq Kor npoyUoronoefuor drogSoou rnq oepdg ovo- drord(etov beoprcbv nou nopouotd(etot mrlv EIKONA 2X.24. H ow(6pooq drernepouiveror on6 qv twetvq, prolov6n q onoreleopotrr6ulrd rrlg enqped(eror on6 rrg e(teiveg. H np6tr1 ow(6pooq e(vor 1.uo en[Oeoq on6 pto n].eu- pu<r1 oLuo(6o -OH n-SH tou npritou o;tvo[5og rrlq rv- reivrlg orov nentr6rx6 6eo;r6 nou ro ouvdSer Ue rnv npc0rr1 e(retvq. Aurn q ow(6poor; preroqdper qv e{tei'- vr1 on6 r1v o1.uvotetrrrr1 oprddo qq rweivqq o' 6vov qru- Lrr6 deopr6 N-O { N-S. 'Enerro, our6g o 6eopr6g dSXeror en[Oeor1 on6 tr1v -OH n -SH n].euprr4 qluo(6o rou npdlrou oprwo[6oq rqg 6e[repqg twetv4g. To ono- rSl,eopro efvor 11 prerogopd qg e[teivrlg 1 or4v n].eupt- 14 oluo(6o rou ogrvoreLu<o0 o(6o9 rrlq e[teivrlg 2. T6Log, q C-re}'l<r1 oonopoy[vq qg rwetvrlg xur.l.onore(- ror, evoi 11 rel,u<r1 NH qq e[retvqg 2 enn(Oeror mov or<uLrr6 6eopr6 c6ore vo rov qwrKqroorrloer pr' 5vo ou1.t- Borrr6 nemrdx6 6eopr6. KoOepr(o on6 outSg rrq owr- 6pdoetg prnope( vo ouprBe( ou06p;rqro pre noLri XoUnIn roXrir4ro. l-ro vo npoyproronor4Ooriv ouwovropSvo ror oprerd ypnyopo rirore vo entteulOe( 11 ouppoqrl qg nptotetvqg Xpetd(eror rordluorl on6 rqv rwefvq. Ot tweiVeg 51ouv Xoponrqprcrxd yvop(oproro. Bp(- oKowor cog ev06oerg ew6g nl,oro[ou oro eooreptx6 ro- 6rronor4rrrrilv oMrllou1rc6v. Avoyvrrlpf(owor on6 t4v 0nop[r1 o;r6],oycrlv yovr6(rov nou 6e 6to05touv rn ouy- rer<pr;.r6vr1 6v0eoq.'E1ouv puo N-refuxq oep(vn r1 ruorei- vr1 (nou nop6let Uto -XH nl,eupu<ri oluo(6o) Kor Uro C-relrrq oonopoyfvrl. H tunrxq twetwl 51er pto otrl.q- l,ouX[o -150 oprvo(5ov oro N-re].u<6 dr<po r<or -50 o;u- vo[6cov mo C-retrx6 drpo, or ono(eg epntr6rowor orrlv xordl.uorl rnq ouppoqrlq rov nproreiv6v. H oMqlou- {o oT o r6wpo qg twetvrlg 6Xer dMeg }.emoupy(eg. 'Evo oouvq0roro XopoKrqptoru<6 notrtrriv rweiv6v eL ?* ii H lupporpf l-lporeivorv Elvor Auroxorotrulrl 669 Boorxfg fvuoreg E[retv1 lvTsivn Elrerwl reDNA IBNA
X=Sor0 vor 611 6Xouv evepy6rqro ev6ovouKledonq enoxrol.rori. Mrq evdovouxledoq enoxrol.rori 6roond ro DNA-or51o 6q;uoupyriwog eror puo Odo'q 6nou prnope( vo eroX<r:pr1- oer q oMql,ouX[o rou DNA nou ro6xonore[ t4v rwefvr1 (BI5ne Eu<5vo 23.11 orqv Ev6trya 23.5). H ouppog4 rurv npcoreiv6v r<r 11 evepy6rqro rqg evdovouxtredo4g enou<to1-rori e(vor ove(dprqreg treroupy(eqrnq rwetvrlg. Aev dXoupe xorovoqoer nLrjpcog ndrg ouv6Sowor or 6rjo our5g evepy6rrlreq p6oo oe pro rweiv4, oor6oo 61ouv nporo0e( 6Uo rUnor prow6lcov. liprgovo Ue ro nprilro, opXu<d unrlple rdnoro e(6og oriv6eo4g ovdpre- oo orrq evepy5rqreq, oMd opy6repo Sytvov ove[dpr4- reg xr 6tor x6noeg rweiVeg 61ooov qv evepy6rrlro rnq ev6ovouxLedonq efior.KtoUo0. IUprgovo pe ro 6e0repo prow6lo, or rwe'fveg (orog [er<(vqoov rog pov66eq oup- pqeiq nporeivriv Kr 6nerq (or nro noMdg) 66X0r1r<ov etoBo).r1 on5 ng ev6ovoux.l,e6oeq enorKroUori. H un60e- or1 ourri e(vor ouprBor4 pre ro yeyov6g 6n or ev6ovou- rledoeg enorrroprotj go(veror 611 6Xouv en(oqg eroBdMer Kor oe dMoug r0noug provd6cov, xup(rog oe rwp6vro qq o;rd6og l. 23.13 llepiAn$n H ouroouppocp{ e(vot 1616111ro 60o oprddov rwpo- 670 KEOA^AI0 23 Korotrurx6 RNA EIK0NA 23.24 0r 6eopo[ ovodrqroooovror p6oo ond pro oxotrou0fo frans-soTeponoqoetov olg ono(eg ougper6- Xouv or -0H ogddeg qq oepiv4g 4 qg Opeov(vr1q Kor rl- SH opo6o rrlg ruoretvrlg 6tog 6rou ouvde0o[v or e[reiVeq pr' 6vo nemrdxd Deogd xr one],eu0epo0e(11 rv- Te(vn U' 6vo rurlonotrlpdvo C-relud 6xpo. v(tov, ro ono(q e(vor 6ro6e6oprdvo oe povor<rit- topoug/o)lyor0rropou g euxopudlreg, npoKo- pucorrrd ouorrlproro xor 1.rrroX6v6pro. Or ovoyr<o(eg nLrlpo<pop(eq yro rnv ow(6poor1 Bp(oxowor orqv oMr1l,ou1[o rou rwpov(ou, 1-ro- l,ov6rr, in vivo, srnv npoypotrx6r4ro q ow(- 6pooq unoBoq0ekor on6 npotrefveg, l-ro ro rwp6vro 16oo qg otrrddog I 6oo rqr qg otrrd- 6o9 ll, 11 ow(6poo4 onorre( ro oXrlprorropr6 grog er6trqg deweporoyorjg/tpnoroyorig do- Unq Ue BpoXeieg ouvorverrr6g oMqLouX(eg. To RNA rov rwpov(cov r4g opd6og I 64trloupye( ;uo 6opul oqv ono[o 4 oM4].ou1(o-un6mpr,r;- Uo ouyKporefuor pr5orr: r4g neptoX(g IGS rou twpov(ou, evol dMeq ouwrlpqprdveg oM4).ou- X(eg 64;rroupyoriv prro neproX4 np6o6eorlg vouxLeorr6(ov youov(vr1g. Aur6 ouprBo(ver pre trans-eorepono(r1oq orqv ono(o ou pprer6Xer co g ouprnopdyowog 6vo xord).orno youvoo(v4g. Aev qnotre(ror nopop; ev6pyerog. H youovoo(- v4 6roond ro 6eo;r6 ornv 5' ouprBo).r1 ;rero[ri e[cov(ou-rwpov[ou. 'Eneno, ro udpo[[].ro mo eLef0epo dr<po tou e(cov(ou entr(Oeror orrlv 3'ouprBo- L4 e[rov(ou-rwpov(ou. To rwp6vro rur].onorefuor ror ldver 14 youovoo(vr1 Kor rrq retrewo(eg 15 Bdoerg. Mro oerpd ouyyevcilv owr6pdoeov xoro,l.riowor ;16oro ent0Soeov qro uq eocoreprro ri g qrrlogo6remepu<oUg deoprorig on6 ro rsfur6 G-OH rordlorTro rou rwpov(- ou. Me qv nopoXrl tcov xordMql,rov unoorpco;rdrr,lv efuor duvorri 4 xorooreu{ prBoev(riprov nou npoytlo- ronoroUv norrfi.eg roroLurrr6q owr6pdoerq, mtg ono(eg neprLoprBdvsror Kor 4 evepy6rqro rnq rpov- o<pepdo4g vou xi.eor6(ov. Opro;rdvo ;utolovdpror<d rwp6vro qq oprddog I xor ll 51ouv ovorXrd ntrq(oro ovdyvo:or1g. Or nporer*veg nou t<to6txonoro0wor qn6 ro rwp6vto rrlg oprddog I e(vor ev- dovouxledoeq nou npqypqronororiv 6(xtrcoveg ewopr6g oe 06oerg-or61oug rou DNA. H ev6ovour<.l,eoLunrri 6rd- onoon nupodore( pto 6repyoo(o yovr6rorqq Uerorpo- m'19, or4v ono(o 11 oMrl),ouX(o rou (6rou rou rwpov(ou owrypdgeror orq 05or1-m61o. Irrg nporetveg nou Ko- 6xonotoriwor on6 ro rwp6vto qq oprd6og ll oupnept- l,oprBdveror pro ev6ovourledorl, 11 ono(o (ar<rvd 14 perdOeoq, Kor Uro ow(orpogq preroyporpdoq, nou eTrr- rp6ner rnv owrypoe{ ev6g RNA owrypdqou rou rwpo- v(ou m4 06oq-or61o. Auto( ot rUnor rwpov(cov (ocoq nepd.oprBdvouv evepy5rrpeg ;roroupdorlg nou ouv6p6- Uouv ornv ogo[peor1 rov twpov(rov oro0eponorcirwog il a _tu I .HX QHz l I H Yn2-u:',; -N -i { rr.rivnz l________J (i,, -In-iY ilHn XH CH, -J;g zHN il L ir;),, ; *, ---=
ro oxn Uortop6 rnq dewepotoyoti g/tproroyo 0S 6o U( S tou evepyori rSwpou. Kotoluttrdg owrdp6oetg 6texnepotrirvowot on6'ro RNA oumottx6 qq RNAdorlg P. Tq RNAs rtov toooet- 6cirv npoyprotonoto(v ouro6tdonooq Utqq oocpuporS- qol.r1g" 6opriq. Igu por6rpoLeq 6opr6g oxq got(owot ovdpeoo oe 6vo RNA un6mpopo Kot to RNA ev6g pt- Boev(Upou, yeyov6q nou xo0odqye( et6tr<6 q 6tdono- orl oe ouyrerptprdveq oLl,rlLoul(eg. Aut6q ot ovrt- 6pdoetg mrlp((ouv tqv dnorlrq 6tt ro RNA oXrlpot((et erdrrd evepyd r<6wpo nou 51ouv roroLwtxrl evepy6- rnro. H ene[epyoo(q rou RNA oMd(et qv oM4].ouX(o ev6g RNA rotd q pretoypotpri 11 rot pretd on6 ouq. Ot olloydg e(vot onopo(tqteg d:ore vo 64prtoupy40e( pto rro6xonotlnrci oMrll,ou{o ;re v6r1 go. Awtt<otoor6oetg gepovcopdvtov Bdoerov ouprBo(vouv oro Oqtroottrd rot no(pvouv rn Uopqf onogtv6oetov, ortq ono(eg eke 11 C perotp6netot oe U e(te 11 A petorp5nerot oe l. Mto t<o- rotrwrrtj unopovddo nou oXet((erot Ue rn deogrvdoq r1g r<wr6(v1q n rnq odevoo(vqg Letroupye( og tprlpo ev6q preyol.0tepou oupnl,6rou to ono(o eggov((et et6t- x6t1ro yto Uto ouyrer<ptpr6vq dLqlouX(o-or6Xo' I-1 poo0rjr<eg rot onotrotg5g (ouly6repo ouptdiv4g) oupBo(vouv oro puol6v6ptq rou tpunovooritrloroq Kot orouq nopq;ru[oioti g. Ercretop6veg owtdpdoetg ene- [epyoo(og npoyporoTloto tiwot qrq rpunovootill.toto, sro ono(o oXe66v ot prtodg Bdoetg ev6g mRNA npo5p- Xovror on6 ene[epyoo(q. H ow6poorl qg ene[epyo- o(oq Xpqorpronote( dvo erproye(o nou onotetre(tot on6 5vo o6qy6 RNA ouprntrqpcoporrr6 qg oLr1l'ouX(og tou mRNA. H ow(dpoorl rfiol0etot on6 ro oopotbto ene- (epyoo(og, 6vo otipnLot<o ev(riprov nou nepfioprBdvet gto ev6ovouxledoq, pto e[ovoux].edoq-te]'u<r1 oupt6u- l,orpovorpepdoq xot qv RNA ).tyr<doq rot lp4otpronoe( el,e[0epo voutd.eot(6to oq flnYn Yto rrlv npooOrlrq ti oneLeu0epdlvet 6toonoop6vo voutd,eot(6to pretd tqv onoLotgrj. H ouppogri rcov npoteivcov e(vot ouroroto)r,wtxr1 ow(6poo4 nou npoyporonote(tot ;16oo owr6pdoeov prero<popdq 6eopd:v, evcil 6ev onome( nopoXri ev6pyet- og. H rwetvrl r<ffiolUet qv onoxonf rnq on5 ng e[tei- veq nou rrlv neptBdll,ouv. IloLL5g tvtetveg 6Xouv evepy6qto ev6ovourtredonq enoKroUo0, 11 ono(o e(vot ove[d6r1q on6 qv evepy6r4to rnq ouppoqnq npco- reivcbv. BrBAroypogio 23.2 To lvrpdvto rnq 0Ud6oq I AutoouppdnrovTot Ue trans-eorepono[qor1 ii,ti.t,iL{:1. +l Cech, T. R. (1985). Self-splicing RNA: implications for evolution. lnt. Rev. Cytol. 93, 3-22. Cech, T. R. (1987). The chemistry of self-splicing RNA and RNA enzymes. Science 236 1532-1539. Vicens,0. and Cech, T. T. (2006). Atomic level architecture:' group I introns revealed, Trends Biochem. Scr. 31 , 41 -51 . 1.r:*::'.,tt Been, M. D. and Cech, T. R. (1986).One binding site determines sequence specif icity of Tetrahymena pre-rRNA self-splicing. transsplicing, and RNA enzyme activity. Cel/ 47 ,207-216. 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Tase): the key enzyme in U-insertion/ deletion RNA editing. Cel/ 1 08, 637-648. 3enne, R., Van den Burg J., Brakenhoff, J. P., Sloof, P., Van Boom, J H., and Tromp, M. C. (1986). Major transcript of the frameshift- ed coxll gene from trypanosome mitochondria contains four nu- cleotides that are not encoded in the DNA Cell 46,819-826. 3 um, 8., Bakalara, N,, and Simpson, L (1990). A modelfor RNA editing in kinetoplastid mitochondria: "guide" RNA molecules transcribed from maxicircle DNA provide the edited information. Cel/ 60 1 B9-1 98. :eagin,J, E,, Abraham, J IV and Stuafi, K. (1988). Extensive edit- ing of the cytochrome c oxidase lll transcript in Trypanosoma brucei. Cel/ 53, 413-422. ',lemann, M, Kaibel, H., Schluter, E. Weitzel, K., Brecht, M., and Goringer, H. U. (2009). Kinetoplastid RNA editing involves a 3 nucleotidyl phosphatase acliuily. Nucleic Acids Bes. advanced access, published on February 3, 2009. Seiwert, S. D., Heidmann, S., and Stuarl, K. (1996). Direct visuali- zation of uridylate deletion rn rllro suggests a mechanism for kinetoplastid editing. Cel/ 84, 831-841 . 23J2 H lupporptl llporeivt0v Efvor Aurororo]runnl ivi:trr:;I'ltil; Paulus, H. (2000). Protein splicing and related forms 0f proteri a-- toprocessing. Annu. Rev. Biochem. 69, 447-496. Saleh, L. and Perler, F. B. (2006). Protein splicing in cis and in trans. Chem. Rec. 6:183-193. llit,:tti:l Derbyshire, V. Wood, D. W., Wu, W., Dansereau, J. T. Dalgaard l.2., and Belfort, M (1997). Genetic definition of a protein-s- plicing domain. functional mini-inteins support structure predic- tions and a model for intein evolution Proc. Natl. Acad. Sci. USA 94,11466-11471 Perler, F. B. etal. (1992). lntervening sequences in an Archaea D- NA polymerase gene. Proc. Natl. Acad. Scl. USI 89, 5577- 5581. Xu, M.0., Southwolrh, M, W,, Mersha, F. B., Hornstra, L. i., and Perler, F. B. (1993). ln vitro prolein splicing of purified precursor and the identification of a branched intermediate, Cell 75, 1371- 1377. Brptrroyporpro 673
MET6q>paon Avonopliyno1 on6: I. L. Mainprize, et al. , Molecular Biolo- gy of the Cell 17 (2006):5063-5074. Copyright 2006, Ame- rican Society of Cell Biology. H (j)W10ypo(j)[o nopoxwp~8T]Ke onci mu~ David W. Andrews , McMaster University KOL lain Mainprize, University of Guelph. E:rttµileta Ke<palaiov: Cheryl Keller Capone 61) Etoaywy!'] flB H Mnaq>paon t.LaKp[v£TOL 0£ Tp[a I1Mta: 'Evap~Tl, Em- µ!']KUVOll KOL Tepµanoµ6c; • To pt~oowµano nepLEXEL 1p£L<; neptoxec; np6o5wric; TOU tRNA. • 'Eva aµtvoaKuAo- tRNA £LOEPX£TOL OTll 8!:'.ori A. • To nrnn5uAo- tRNA npooMv£TOL 011181::011 P. • 'Eva aµtvo~u npoo1f8£TOL mriv no>.unrnn5LK!'] a>.uo[5a µ!:'.ow µnaq>opac; 1ou no>.unemt5[ou an6 TO nrnn5uAo- tRNA 111<; Stone; POTO aµLVOaKUAO-tRNA Tll<; Stone; A. OI) EL5LKO[ Mnxavtoµo[ EAEVXOUV TllV AKp[~£La Tll<; Mnaq>pa- 01']<; • H aKp[~£La TT']<; µ£TOq>paoric; £AEVX£TQL an6 £L5LKOU<; µ11xavtoµou<; 0£ K08£ OTMLO. 8 H 'Evap~T'] om BaKT!']pLa Xp£LO~£TOL 30S YnoµovME<; Kl EmKOUPLKOU<; napayovT£<; • H!:'.vap~n Tll<; µnaq>paonc; anOLT£[ ~£XWPLOT£<; Pl~OOW- µLKE<; unoµovM£<; 30S KOL 50S. • Enforic; anm1ouvrnt ot napayovrnc; !:'.vap~ric; (IF-1, -2 KOL -3), OL ono[OL npo0Mvov10L one; 30S unoµovMec;. • Mta 30S unoµovMa nou µnaq>!:'.p£L napayov1ec; !:'.vap- ~11<; npooMv£TOL 0£ µLa 8£01'] !:'.vap~T']<; OTO mRNA OXll- µm[~ovrnc; tva ouµnAoKo !:'.vap~nc;. • 0 IF-3 np!:'.n£L v' an£A£U8£pw8e[ WOT£ OL 50S unoµo- va5ec; va npoo5e8ouv 010 ouµnAoKo 30S-mRNA. 674 KECl>AAAIO 24 Mnaq>paon - H'Evap~T'] nepLAaµ~OV£l Zrnyapwµa B00£WV Mna~u TOU mRNA KOL TOU rRNA • LTO ~aKTT']pLaK6 mRNA Tl 81::011 !:'.vap~nc; anoT£A£lTOL an6 TO KW5LK6vto !:'.vap~nc; AUG, 10 ono[o aKoAou8£f -10 ~(Joete; µe10 10 e~aµep!:'.c; nouptvcilv Shine-Dalgar- no. • To rRNA 30S Pl~OOWµLK!']<; unoµovMac; TWV ~aKTT1Pl4JV 5ta8!:'.m µLO ouµn>.ripwµanK!'] ann>.ouxra nou ~rnyapw- V£L µ£ TllV aAAT']Aouxra Shine-Dalgarno KQTO TllV !:'.vap~T']. 8 'Eva EL5LK6 EvapKT!']pLO tRNA ::£KLVO TT']V no>.unrnTL5LK!'j AAuo[5a • Hµnaq>paori ~£Ktva µe tva aµtvo~u µ£8£tovrvric;, nou ouv!']Swc; KW5LKonOL£tTOL an6 10 AUG. • t.Laq>oprnKd tRNA µ£8£tov[vric; eµnAEKOVTGL 01riv !:'.vap~n KOL TllV emµ!']Kuvon. • To evapKT!']pLO tRNA 5La8!:'.m µova5LKO 5oµLKO xapa- KTT']PLOTLKO, rn ono[a 10 5taq>oponmouv an6 6Aa rn un6Aoma tRNA. • H oµMa NH2 TT']<; µ£8£tovrvric;, nou npooMvnOL 010 ~aKTllPLaK6 £VapKT!']pLO tRNA, £[VOL q>opµUALWµEVT']. 8 HXp!']on TOU fMet-tRNAf EAEVX£TOL an6 TOV IF-2 KOL TO PL- ~oowµOTLO • 0 IF-2 npooMm TO evapKT!']pLO fMet-tRNAf KOL TOU rnt1p!:'.n£L va £LO£A8£L OT1l µepLK!'] 81::011 P111c; 30S uno- µovMac;.
&!) Οι Μικρές Υπομονάδες Αναζητούν Θέσεις Έναρξης στα Ευκαρυωτικά mRNA • Οι ευκαρυωτικές 40S ριβοσωμικές υπομονάδες προσ­ δένονται στο 5' άκρο του mRNA και το «σαρώνουν» έως ότου εντοπίσουν μια θέση έναρξης. • Μια ευκαρυωτική θέση έναρξης αποτελείται από μια αλληλουχία δέκα νουκλεοτιδίων η οποία περιλαμβάνει ένα κωδικόνιο AUG. • Οι 60S ριβοσωμικές υπομονάδες συνδέονται με το σύμπλοκο στη θέση έναρξης . &11) Οι Ευκαρυώτες Χρησιμοποιούν Ένα Σύμπλοκο Πολλών Παραγόντων έναρξης • Οι παράγοντες έναρξης απαιτούνται σε όλα τα στάδια της έναρξης , μεταξύ άλλων η πρόσδεση του εναρκτή­ ριου tRNA, η πρόσδεση της 40S υπομονάδας στο mRNA, η μετακίνηση κατά μήκος του mRNA και η πρόσδεση της 60S υπομονάδας. • Το ευκαρυωτικό εναρκτήριο tRNA είναι ένα Met-tRNA το οποίο διαφέρει από το Met-tRNA που χρησιμοποιεί­ ται στην επιμήκυνση, ωστόσο η μεθειονίνη του δεν εί­ ναι φορμυλιωμένη . • Ο elF2 προσδένει το εναρκτήριο Met-tRNAi και GTP σχηματίζοντας ένα τριτοταγές σύμπλοκο , το οποίο προσδένεται στην 40S υπομονάδα προτού αυτή αλλη­ λεπιδράσει με το mRNA. • Ένα σύμπλοκο πρόσδεσης της καλύπτρας προσδένε­ ται στο 5' άκρο του mRNA προτού αυτό αλληλεπιδρά­ σει με την 40S υπομονάδα. m Ο Παράγοντας Επιμήκυνσης Tu Φορτώνει Αμινοακυλο­ tRΝΑ στη Θέση Α • Η EF-Tu είναι μια μονομερής πρωτεΊνη G, της οποίας η ενεργός μορφή (προσδεδεμένη σε GTP) προσδένεται στο αμινοακυλο-tRΝΑ. • Το σύμπλοκο EF-Tu-GTP-aμιvoaκuλo-tRNA προσδένε­ ται στη θέση Α του ριβοσωματίου. fIID Η Πολυπεπτιδική Αλυσίδα Μεταφέρεται στο Αμινοακυλο­ tRΝΑ • Η 50S υπομονάδα έχει ενεργότητα πεπτιδυλοτρανσφε­ ράσης που παρέχεται από ένα rRNA ριβοένζυμο . • Η νεοσυντιθέμενη πολυπεπτιδική αλυσίδα μεταφέρεται από το πεπτιδυλο-tRΝΑ της θέσης Ρ στο αμινοακυλο­ tRΝΑ της θέσης Α. • Η σύνθεση των πεπτιδικών δεσμών παράγει αποακυλιω­ μένο tRNA στη θέση Ρ και πεπτιδυλο-tRΝΑ στη θέση Α. &! Η Μετατόπιση Μετακινεί το Ριβοσωμάτιο • Η μετατόπιση του ριβοσωματίου μετακινεί το mRNA διαμέσου του ριβοσωματίου κατά τρεις βάσεις. • Η μετατόπιση μετακινεί το αποακυλιωμένο tRNA στη θέση Ε και το πεπτιδυλο-tRΝΑ στη θέση Ρ, ενώ αδει­ άζει τη θέση Α. • Σύμφωνα με το μοντέλο υβριδικής κατάστασης, η μετα­ τόπιση συμβαίνει σε δύο στάδια, στα οποία η 50S με­ τακινείται σε σχέση με την 30S και στη συνέχεια η 30S μετακινείται κατά μήκος του mRNA ώστε ν' αποκατα­ σταθεί η αρχική διαμόρφωση . fm} Οι Παράγοντες Επιμήκυνσης Προσδένονται Εναλλακτικά στο Ριβοσωμάτιο • Η επιμήκυνση απαιτεί EF-G, που έχει δομή παρόμοια με τη δομή του συμπλόκου αμινοακυλο- tRNA-EF-Tu-GTP. • Η πρόσδεση του EF-Tu στο ριβοσωμάτιο O!eJ·.lΞ~ -: , EF-G και το αντίστροφο. • Η μετατόπιση προϋποθέτει υδρόλυση του GTP, πυροδοτεί μια αλλαγή στον EF-G, που με τη σειρά ~~ πυροδοτεί μια αλλαγή στη δομή του ριβοσωματίου . m Τρία Κωδικόνια Τερματίζουν τη Μετάφραση • Τα κωδικόνια UAA, UAG και UGA τερματίζουν τη μετά­ φραση . • Στα βακτήρια, συνήθως χρησιμοποιούνται με σχετική συχνότητα UAA>UGA>UAG. S Τα Κωδικόνια Τερματισμού Αναγνωρίζονται από Πρωτεϊνι­ κούς Παράγοντες • Τα κωδικόνια τερματισμού αναγνωρίζονται από πρωτεϊ­ νικούς παράγοντες απελευθέρωσης, όχι από αμινοα­ κυλο-tRΝΑ. • Οι δομές των παραγόντων απελευθέρωσης τάξης 1 μοιάζουν με τις αντίστοιχες δομές του συμπλόκου αμι­ νοακυλο-tRΝΑ-ΕF-Τu και του EF-G. • Οι παράγοντες απελευθέρωσης τάξης 1 απαντούν σε ειδικά κωδικόνια τερματισμού και υδρολύουν το δεσμό πολυπεπτιδίου- tRNA. • Οι παράγοντες απελευθέρωσης τάξης 1 υποβοηθούν­ ται από τους παράγοντες απελευθέρωσης τάξης 2, οι οποίοι εξαρτώνται από GTP. • Ο μηχανισμός είναι παρόμοιος στα βακτήρια (τα οποία διαθέτουν δύο τύπους παραγόντων απελευθέρωσης τάξης 1) και τους ευκαρυώτες (οι οποίοι διαθέτουν μόνο ένα παράγοντα απελευθέρωσης τάξης 1). 8 Το Ριβοσωμικό RNA Διαπερνά και τις Δύο Ριβοσωμικές Υπομονάδες - - - - - • Κάθε rRNA φέρει πολλές διακριτές περιοχές που δι­ πλώνονται ανεξάρτητα. • Σχεδόν όλες οι ριβοσωμικές πρωτεΊνες έρχονται σ' επαφή με το rRNA. • Οι περισσότερες επαφές μεταξύ των ριβοσωμικών υπομονάδων πραγματοποιούνται μεταξύ 16S και 23S rRNA. Τα Ριβοσωμάτια Περιέχουν Πολλά Ενεργά Κέντρα • Οι αλληλεπιδράσεις στις οποίες εμπλέκεται το rRNA είναι κρίσιμο μέρος της λειτουργίας του ριβοσωματίου. • Το περιβάλλον των θέσεων πρόσδεσης του tRNA κα­ θορίζεται κυρίως από το rRNA. Το 16S rRNA Διαδραματίζει Ενεργό Ρόλο στη Μετάφραση • Το 16S rRNA διαδραματίζει ενεργό ρόλο στις λειτουρ­ γίες της 30S υπομονάδας. Αλληλεπιδρά άμεσα με το mRNA, την 50S υπομονάδα και τ' αντικωδικόνια των tRNA στις θέσεις Ρ και Α. Το 23S rRNA Έχει Ενεργότητα Πεπτιδυλοτρανσφεράσης • Η ενεργότητα της πεπτιδυλοτρανσφεράσης εντοπίζε­ ται αποκλειστικά στο 23S rRNA. Οι Ριβοσωμικές Δομές Αλλάζουν Όταν Συνδυαστούν οι Υπομονάδες • Όταν σχηματίζονται πλήρη ριβοσωμάτια, η κεφαλή των 30S υπομονάδων περιστρέφεται γύρω από τον αυχένα. • Το ενεργό κέντρο της πεπτιδυλοτρανσφεράσης στην 50S υπομονάδα είναι πιο δραστικό στα πλήρη ριβοσω­ μάτια συγκριτικά με τις μεμονωμένες 50S υπομονάδες . Περίληψη 675
24. l E1oaywyn 'Eva mRNA m:pt8X8L µta 08tp6 KWOLKov(wv nou aMri- A8mopouv µ8 TO OVTLKWOLKOVLQ TWV aµLVOQKU/1.0-tRNA 8TOL WOT8 µLa OVTLOTOLXJ'"] 08Lp6 aµtvosewv va 8Vowµa- TW88l 08 µta no11.un8ITTLOLKfl OA.uo(oa. To pt~oowµano nap8X8L TO n8pl~CI/I./I.OV YLO Tr] pu8µLOJ'"] Tr]c; aMr]A.8n(- opaOr]c; µ8TOSU TOU mRNA KOL TOU aµLVOQKU/1.0-tRNA. To pl~OOWµCITLO ouµn8plcp8p8TOL OQV 8VQ µLKpO µ8Ta- VQOT8UTLKO 8pyoOT60Lo, TO ono(o Tasti58U8L KOTa µfi- Koc; Tou 8Kµaydou ot8s6yoVTac; Taxdc; KUK11.ouc; OUV880r]c; n8ITTLOLKWV 08oµwv. Ta aµLVOQKU/1.0-tRNA 8KTOS8UOVTOL µfoa Kl esw an6 TO µ6pto µ8 an(OT8UTO µ8yC1A.rJ TOXUTrJTO 8VW nap6Mri11.a 8vano8eTOuv aµtvo- sfo. En(Or]c;, napayOVT8c; 8nLµr]KUVOJ'"]c; KUKA.LKQ aMr]- /l.8nLOpOUV µ8 TO pt~oowµano KOL ot'(OTOVTOL an6 auT6. Mai'.;( µ8 TOuc; 8mKouptKouc; nap6yovr8c;, TO pt~oowµ6- TLO nap8X8L TO n11.fip8c; 8Upoc; TWV opOOTr]pLOTr]TWV nou anOLTOUVTOL yta 611.a Ta ~fiµma TJ'"]c; µ8Tacppaoric;. H EIKONA 24.1 napoumai'.;8L nc; oX8TLK8c; OLQOTCI08Lc; TWV OUOTQTLKWV Tr]c; npwTe'Lvoouv88TLKr]c; OUOKWr]c;. To PL~oowµano anOT8/l.8LTQL an6 Mo unoµov6o8c;, Ol ono(8c; otaopaµm(i'.;ouv 80LKouc; p611.ouc; OTrJ µ8Tacppa- OrJ. To ayy8A.LOcp6po RNA OUV088TQL µ8 Tr] µLKpr] uno- µovaoa: 08 Ka88 xpovLKfl OTtyµfi dvm npooo808µev8c; -35 ~CI08Lc; mRNA. To mRNA OLOTP8X8L Tr]V nopda TOU KQTCI µfiKoc; Tr]c; 8mcp6v8LOc; KOVTCI OT!'"] ouµ~0/1.r] TWV unoµovaowv. .6uo µ6pta tRNA dvm n6vroT8 8V8py6 OT!'"] µ8Tacppa0r]: OUVWWc;, r] 8nLµr]KUVOJ'"] TOU nOA.Un8- nTLOLOU n8pL11.aµ~aV8l OVTLopao8Lc; Ol ono(8c; ouµ~a(- vouv 08 µ611.tc; Mo an6 Ta (aop6) i58Ka KWOLK6vta nou KaMmovrm an6 TO pt~oowµano. Ta Mo tRNA 8Loep- xovrm a' WWT8plK8c; 8808Lc;, nou 8KT8LVOVTQL KQTCI µfi- Koc; TWV unoµovaowv. 'Eva TplTO tRNA (owe; napaµev8L navw OTO pt~oowµ6Tto acp6TOu xprimµonotri8d mri µ8- Tacppaori, npOTOU QVQKUKA.W88l. H ~QOLKr] µopcpfi TOU pl~oowµm(ou 8X8L OUVTr]pr]88l KOTa TrJV 8se11.LsrJ, wOT6oo unapxouv oacpdc; napaMa- yec; we; npoc; TO OUV0/1.LKO µey88oc; KOL TLc; QVQAOYL8c; RNA KOL npwTe'Lvwv OTO pl~oowµana TWV ~OKTr]p(wv, TOU 8UKOpUWTLKOU KUTiapon11.6oµmoc; KOL TWV opyavL- o(wv. H EIKONA24.2 OUYKPLV8l TO OUOTQTLKCI TWV pl~OOW- 220 A /VVVVV. mRNA 35 ~GoEwv o<( 0 0 N EIKONA 24.1 IuyKp(ow; µf:y£8wv 0£1XVOUV 6n 10 PL~oowµcmo £(VOL apK£16. µ£yo.Ao wo1£ va npooi'iev£L tRNA KOL mRNA. 676 KE4>AAAIO 24 M£To.(j)paol'j Pljloowµdna rRNAs npw,EivEc; r Bmrn')pta (70S) 50S 23S = 2904 ~GOEi<; 31 MG½a: 4,2 MDa 5S = 120 ~Goe1c; 60% RNA .._30S 16S = 1542 ~GOEi<; 21 28S = 4718 ~ci.OEI<; 49 Bn>..aOTLKG (BOS) '9 60S 5,8S = 160 ~ci.OEI<; MG½a: 4,2 MDa 5S = 120 ~ci.OEI<; 60%RNA .._40S 18S = 1874 ~ci.OEI<; 33 EIKONA 24.2 Ta Pl~oowµo.na £!VOL µ£yo.Ao pl~OVOUKA£0TIPWT£l- VlKO. owµOTlOLa, nou n£pLexouv n£pL0061£po RNA napo. npw1tlvrJ KOL OL0Kpfvov10L 0£ µ£yo.A£~ KOL µLKP£~ unoµovdo£~. µm(wv OTa ~aKTfipta KOL TO 8ri11.aOTLK6. Km TO Mo d- vm pt~ovouKA.eonpwTe'LVLKa owµm(ota, TO ono(a n8pte- xouv n8ptoo6T8po RNA an6 npwTdv8c;. Ot pt~oow- µtKec; npwrnTv8c; dvm yvwOTec; we; npwTdv8c; r (r-pro- teins). K688 pL~oowµLKr] unoµovaoa n8pl8X8L eva KUPLO rRNA KL optoµev8c; µLKpec; npwT8'(v8c;. H µ8ya11.ri unoµo- vaoa µnope( en(oric; va n8pl8X8L µLKPOT8pa RNA. LTO E. coli, ri µtKprJ unoµovaoa (30S) anoT8A.8LTOL an6 TO 16S rRNA Km 21 npwrnfv8c; r. H µ8y611.ri unoµovaoa (50S) n8pl8X8L 23S rRNA, TO µLKp6 5S RNA KOL 31 npwTdv8c;. M' 8S0lp80r] µLa npWTe'fVr], nou ~p(OK8TQL 08 T8008p0 avr(ypacpa ava pt~oowµaTLo, un6pX8L µ6vo eva avr(- ypacpo K688 npwTdvric;. Ta µdi'.;ova RNA ouvtOTouv TO KUplO µepoc; TJ'"]c; µ61'.;ac; TOU ~QKTJ'"]PLOKOU PL~oowµm(- ou. Bp(oKovrm navrou KOL, µ6Mov, OL n8pt006T8p8c; PL~oowµLK8c; npwTdv8c; OTJ'"]V npayµaTLKOTJ'"]TO 8PXOVTQL o' enacpfi µ8 TO rRNA. Iuv8nwc;, TO µ8(1'.;ova rRNA crxri- µm(i'.;ouv au16 nou µ8pLKec; cpopec; 88Wp8LTOL we; OK8A.8- T6c; KCl88 unoµovaoac; - 8VQ OUV8X8c; vfiµa nou KUpl0p- X8l OT!'"] ooµfi KOL Ka8op(i'.;8L nc; 8808Lc; TWV PL~OOWµLKWV npwrn'(vwv. Ta pt~oowµana OTO Kunap6n11.aoµa TWv euKapuw- Twv dvm µ8yaMT8pa an6 T' avr(OTotxa Twv ~aKTrJ- p(wv. H ouvoA.LKfl n8pL8KTLKOTJ'"]T6 TOuc; T6oo 08 RNA 600 KOL 08 npwTdvri dvm µ8yaMT8PrJ: TO µdi'.;ova µ6- pta RNA dvat µaKpuT8pa (Ka11.ouvTOL 18S KOL 28S rRNA) Km unapxouv n8ptoo6T8p8c; npwTdv8c;. To RNA 8SOKOA.ou8d v' anoT8A.8l TO Kup(apxo oumanK6 an6 TJ'"]V anOljJr] TJ'"]c; µ6/'.;ac;. Ta PL~oowµana TWV µLTOXOVop(wv KOL TWV x11.wpo- n11.00TWV OLOKPLVOVTQL an6 TO PL~oowµaTLQ TOU KUTIQ- pon11.6oµmoc; KOL na(pvouv nOLKLA.8c; µopcpec;. Optoµe- V8c; cpopec; 8XOUV TO µey88oc; oX800V TWV ~QKTr]plOKWV pt~oowµm(wv Km n8ptexouv 70% RNA. 'NJ...orn, dvm µ6vo 60S KOL n8pl8XOUV <30% RNA. To pt~oowµano n8pL8X8t noM6 8V8py6 Kevrpa, Ka- 8eva an6 TO ono(a KQTQOK8UCl/'.;8TOL an6 µLO oµaoa npwTe°LVWV nou OA.A.r]/.8nLOpOUV µ8 µta n8pLOXrJ TOU pl- ~OOWµLKOU RNA. Ta 8V8py6 K8VTpa anOLTOUV ciµwri ouµµ8TOXrJ TOU rRNA o' eva ooµLKO Tl QKOµr] KOL KQT0- /1.UTtKO p611.o. Optoµev8c; KOTa11.uTtKec; 11.8LToupy(8c; anm- TOuv s8XWPLOTec; npwTdv8c;, wOT6oo Kaµ(a an6 nc;
opaow:; 5£ µnop£i v' avanapax8£i an6 µ£µovwµev£c; npWT£°fV£c; rJ oµ6i5£c; npwTE°LVWV: A£lTOupyouv µ6vo OTQ n>-.aima Tou p1l3oowµmiou. H av&uari n1c; A£tToupyiac; Twv 00µ1Kwv oumm1- Kwv Tou p1l3oowµmiou µnop£i va y(v£1 µ£ Mo n£1paµa- T1Kec; µ£86oouc;. H av6>-.uari µ£TaM6~£wv avayvwp(~£1 £ti5tKec; p1l3oowµ1Kec; npwT£°fv£c; 11 l360£1c; TOU rRNA nou ouµµ£T8XOUV 0£ ouyK£Kptµev£c; avnop60£1c;. H ooµtKl1 av6>-.uori, nou ouµn£p1>-.aµl36v£1 6µwri Tponono(riori TWV OUOTQTlKWV TWV p1l3oowµm(wv KQl ouyKpl0£tc; npOK£lµ8VOU Va TQUTOnotr]80UV OUVTf]pr]µeva xapaKTr]- plOTlKO Tou rRNA, rnuTonm£i TrJV £VT6mari Twv ouma- TlKWVTa ono(a £µnMKOVTQl 0£ OUYK£Kptµev£c; A£lTOUp- y(£c;. 24.2 H METOcppaon ~IOKpiVETOI OE Tpia ITOOIO: 'Evap~n. EmµnKuvon KOi TEpµaT10µ6c; BomK&~ &VVOlt:~ • To p1~oawµdno n£p1tx£11p£1c; n£p1oxtc; npoaowric; 1ou tRNA. • 'Eva aµ1voaKuAo-tRNA £1atpx£10t 0111 Stari A. • To nrnTlOUAO-tRNA npoaOEV£TOl OTll 8tari P. • 'Eva aµ1vo~u npOOTf8£TOl OTllV no;l.unrn1101Kn a;l.ua(oa µtow µnaQJopdc; TOU no;l.unrn11o(ou ano TO n£m1ou;l.o-tRNA 111c; Storie; p OTO aµ1voaKUAO-tRNA TllC: Storie; A. 'Eva aµtVO~U µ£Tacpep£TQl OTO ptl300WµOTlO µeow £v6c; aµ1voaKu>-.o-tRNA. H npoo811KrJ mu mriv au~a- v6µ£Vf1 npwT£°ivtKrJ a>-.uo(oa npayµmono1£iTm µfow µlac; aMri>--w(opaaric; µ£ TO tRNA TO ono(o £LX£ µ£Ta- cpep£l TO nporiyouµ£VO aµ1vo~u. Ka8eva an6 aUTO Ta tRNA l3piOK£TQl 0£ µta OlaKplTl1 8eari n6vw OTO p1l30- owµ6TlO. H EIKONA 24.3 0£l)(V£l OTl Ol Mo 8fo£tc; exouv 01acpop£TlKO xapaKTrJplOTlKO: • 'Eva £to£px6µ£vo aµ1voaKu>-.o-tRNA npooi5ev£TQl OTrJ 8£01') A (A site). npiv an6 Tr]V £LOOOO TOU aµt- voaKUAO-tRNA, rJ 8eori £K88T£l TO KWOlKOVlO TO onoio aVTmpoown£U£t TO w6µ£vo aµ1vo~u nou np6K£lTat va npom£8£i OTrJV a>-.uo(oa. • To Kwi51K6v10 nou aVTtnpoown£u£1 TO mo np6ocpma £VOWµmwµevo aµtVO~U OTrJ V£00UVT£8£tµ8Vf1 nOAU- n£ITTlOlKl1 a>-.uo(oa l3p(OK£TQl OTrJ 8£01') P (P site). H 8fori QUTl1 KQTQAaµl36V£TQl an6 TO ntTITl~u>.o- tRNA, eva tRNA nou µ£Tacpep£l Tr] V£00UVT£8£tµevri no>-.un£mti5LKl1 a>-.uo(oa. H EIKONA 24.4 unoi5£LKVU£l 6n TO aµ1voaKUALK6 6Kpo ou tRNA £VTon(~£Tat OTrJ µ£y6>--ri unoµov6oa, £vw OTO 6Mo 6Kpo To aVT1Kwo1K6v10 aMri>--w1op6 µ£ To mRNA OU £!Vat npooi5£i5£µevo OTrJ µtKPl1 unoµov6oa. LUV£- wc;, Ka8£µ(a an6 nc; 8foe1c; p KQl A £KT£LV£TQl KQTO µ11Koc; Kat Twv Mo p1l3oowµ1Kwv unoµov6owv. npoK£Lµ8VOU TO ptl300WµOTLO VQ OUV880£l 8VQV n£- ITTIOLKO owµ6, npen£L va l3piOK£TQl OTrJV KQTOOTQOr] Kw1itK6vm "n' Kw1itK6vm •n+1• Hee011 PouyKpan:( To aµtVOOKUAO-tRNA TO m:nn1iu>.o-tRNA ElOEPXETOl OTTJ 81:'.0T] A _. M&TOKlVTJOT] p1poowµ01(ou 3' 1 nptv an6 10 oXT]µanoµ6 ne:mt1ilKOU 1ie:oµoti TO nEnTLOUAO-tRNA KOTOAOµ~dve:1 TTl 81:'.0T] P, EVW TO aµtVOOKUAO-tRNA TTl 81:'.0T] A NEOOUVTL81:'.µEVT] a>.uo(1ia - ;~::-,1,... ......... 2rxnµanoµoc; nem11i1Koti 1ie:oµou: 10 no>.une:m(o10 µ£Tmpepe:Tm an6 TO nEITTL1iUAO-tRNA TTlC: 81:'.0T]c; POTO aµLVOOKUAO- tRNA TTlC: 81:'.oT]c; A 3HµETOTOnlOT] µe:TOKLVE( TO pt~oowµdno KOTd eva KW1ilKOVLO/ TOn08ETE( TO ne:mtOUAO-tRNA OTTJ ee011 P/ TO anOOKUALWµl:'.vo tRNA EYKOTOAE(ne:1 µ!:'.ow TTlC: Seonc; E/ T] 81:'.0T] AElVOl di5ELO y10 TO w6µe:vo aµLVOOKUAO-tRNA ........ Kwi5tKOVlO "n+1" / Kwi5LK6vto "n+ 2" EIKONA 24.3 To p1~oawµd.no n£p1tx£1ouo 8ta£1c; y1a npoaowri Q)OpT1aµtvou tRNA. nou (j)QLV£TQL OTO 1311µ0 1 Tr]c; EtK6vac; 24.3, OTQV TO n£- ITTl0UAO-tRNA l3ptOK£TQl OTrJ 880T] P Kat TO aµLVOQKUAO- tRNA OTrJ 8fori A. Ixriµanoµ6c; n£nTlOLKOU owµou ouµl3atV£l OTQV TO nOAUn£ITTtOlO, nou cpep£TQl an6 TO n£ITTlOUAO-tRNA, µ£rn<pep£TQl OTO aµtvo~u, TO ono(o cpepnm an6 TO aµtVOQKUAO-tRNA. AUTO TO 1311µ0 npo- Ta aµLVOOKUALKd dKpa TOU tRNA a>.>-n>-emopouv µeoa OTTJ µe:yd>.T] p1~oowµ1Kl'j unoµovdoa Ta OVTlKW1iLK6VLO npo0Mvov101 OE1iml,.avec; TptnAE:TEC: TOU mRNA OTTJ µ1Kpl'j p1~oowµ1Kl'j unoµovdoa EIKONA 24.4 Ot 8ta£1c; PKOi ATOn08£TOUV TO QUO tRNA nou aA- AllA£mopouv KOTO µnKo<:; KOi TWV QUO pl~OOWµLKWV unoµovdowv. 24.2 HM£TdQJpaari t.1aKp(v£10t a£ Tp[a I1do1a: 'Evap~ri, EmµnKuvari KOi T£pµanaµoc; 677
To aµ1voaKUAO-tRNA El0£PXETOI 011 BEOrJ A To no>.unemio10 µernq,tpnm OTO aµlVOOKUAO-tRNA Hµn01dmo11 µETOKIVE[ TO nEmlOUAO- tRNA OTrJ BtOrJ P EIKONA 24.5 To aµtVOQKUAO-tRNA £l0£PXETQl CJ1TJ 8t011 A, napa>..aµ~dvet TTW no>..unrnnOtKf] a>..uoioa an6 TO nrnnou>..o-tRNA KQl µ£TQ(j)£- P£TQl OTTl Stan Py1a 10v rn6µevo KUKAO emµf]Kuvoric; Ono8eu:1 awm~ rnno8en1ari TWV aµLVOOKUALKWV 6.- Kpwv Twv Mo tRNA µfoa mri µ£y6.t..ri unoµov6.<3a. H auyK£Kptµevri OVT(<3poori KOTOAU£TOL ano Tl'l µ£y6.t..ri unoµov6.<3a TOU pt~oowµm(ou. H µ£TOcpop6. TOU nOAUn£mt<3(ou <3riµ1oupy£( TO pl~O- awµ6.no nou napouo16.i'.;£TOL mo ~~µa 2, onou TO anoa- KUA1w1,,1tvo tRNA, xwpic; KOV£VO 0µ1vo~u. ~piOK£TOl OTTl etori P, £vw eva vfo n£mt<3ut..o-tRNA EXEL <3riµ1oupyri8d mri etori A. auTo TO m:m1<3ut..o-tRNA dvm µaKpuT£po KOT6. eva KOT6Aomo 0µ1vo~foc; ano TO m:m1<3ut..o-tRNA nou ~plOKOTOV OTTl etori P KOTO TO ~~µa 1. To p1~oowµ6.T10 µ£TOKLV£LTOL Twpa KOT6. µ10 TpmM- TO KOTO µ~KOC: TOU ayy£ALOcpopou RNA. AUTO TO m6.<3LO KOA£LTOL 1,J&TOTOTIIOI') (translocation). H µ£TOKivriori £~6.y£L TO anOOKUALWµevo tRNA ano Tl'l efori P, onou £to6.y£L TO nem1<3ut..o-tRNA (~Mn£ ~~µa 3 mriv EtK6va 24.3). To £n6µ£VO KW<3LKOVLO y10 µ£T6.cppoori ~pLOK£TOl Twpa mri Sfori A, eTmµo va uno<3£X8£L TTlV doo<3o £voe; vfou 0µ1voaKut..o-tRNA, oriµeio mo onoio ea ena- vat..ricp8d o KUKAoc;. H EIKONA 24.5 ouvoljJii'.;£L TTlV oMri- t..£ni<3poori µ£TO~U TWV tRNA KOL TOU pl~oowµmiou. To anOOKUALWµevo tRNA £YKOTOA£(m:1 TO pl~OOW- µ6.no µfow Tl'lC: Sforic; E, µ1ac; 6.Mric; Sforic; npoo<3£- oric; tRNA. H etori OUT~ KOTOA0µ~6.V£TOL napo<3LK6. an6 TO tRNA, nou ~pioK£TOL mri <3L0<3tKaoia va £YKOTOA£i- ljJ£L Tl'l Sfori P KOL v' an£A£U8£pw8d ano TO p1~oowµ6.- no mo Kunapont..aaµa. 'EToL, ri po~ Tau tRNA £LVOL µfoa OTTl etori A, <31aµfoou TTlC: etoric; P KL e~w ano Tl'l Sfori E (~Mn£ rn(oric; EtKova 24.28 oTriv fv6r17ra 24.12). HEIKONA 24.6 OUYKPLV£l Tl'l µ£TOKLVl'l0Tl TWV tRNA ...• 0EOrJ E .·...... ..... ....0tm, P •• ••••••••• ~ =~ ·- • a-•.-a---.aaaa a a a aaa a aaaaa I EIKONA 24.6 Ta tRNA KQl mRNA µ£TGKLVOUVTQl OLaµtoou TOU PL- ~oowµm[ou npoc; TTlV 101a KGT£tl8uvori. 678 KECl>A/AIO 24 M£Td(j)paori KOL mRNA, nou £LVOL <3uvmo VO 8£wpri8d we; £VO d<3oc; 0<30VTWTOU TPOXOU, ITTOV ono(o Tl OVT(<3p00Tl K080<3Tl- y£(TOL ano TTlV 0AATlA£n(<3poori µ£TO~U KW<3LKOVLOU KOL OVTLKW<3LKOVLOU. H µ£T6.cppoori <310Kp(v£TOL a£ Tp(a m6.<310, nou na- poum6.i'.;oVTm mriv EIKONA 24.7: • H tvap~I') (initiation) n£pLA0µ~6.v£1 nc; OVTt<3p6.0£1c; nou auµ~a(vouv npornu axriµm1m£( o n£m1<31Koc; <3wµoc; µ£Ta~u Twv Mo npwTwv 0µ1vo~ewv Tl'lC: npwTE'fvric;. npouno8eT£L npoo<3£0Tl Tou p1~oowµa- T(ou mo mRNA, WITT£ VO axriµOTLITTEL £VO ouµnt..o- KO £VOP~TlC: nou n£pl£X£L TO npwTO aµLVOOKUAO- tRNA. AuTo £(vm eva oX£TLK6. ~paM ~~µa mri µ£- 'Evap~11: 11 µ1Kpn unoµovaoa OTrJ 8£011 npdoownc; mRNA ouvoua~e1m µe 111 µeya>.11 unoµovaoa Km npoootvnm 10 aµ1voaKuAo-tRNA EmµnKUVOrJ: 10 pl~OOWµ0110 µ£1aKIV£l1Ql K011l µnKo<:; 1ou mRNA, emµ11Kllvovrnc:; 111vnpwn:fvn µe µernqiopa and 10 nrnnouAo-tRNA 010 aµ1VOQKUAO-tRNA Tepµanoµ6c:;: 11 n0Aunrnni'i1Kn a>.uoioa ane>.eu8eptiJV£10l and 10 tRNAKm10 p1~oowµa110 olima1m and 10 mRNA EIKONA 24.7 Hµ£Tdq>pa011unoo1ap£[TQ10£ Tp[a OTdOlQ.
TC!cppaori KOL ouvfJewc; Kaeopil;el TT']V TOXUTT']TO µe TT'JVonoia ea µeTOcppaOTei TO mRNA. • H &mµ~Kuvo11 (elongation) nept11.aµp6veL 611.ec; nc; OVTtopaoetc; an6 TT'] ouv0eOT'] ,OU npJnou nemtOl- KOU owµou ewe; TT']V npoo0fJKT'J TOU TE:11.WTOLOU aµL- vo~foc;. T' aµtvo~fo npoOTieeVTOL OTT'JV 011.uoioa eva TT'] cpop6: T1 npoo0fJKT'J ev6c; aµtvo~foc; eivOL TO Ta- xurnpo pfJµa TT']c; µeTacppaOT']c;. • 0 T&pµaT10µ6c:; (termination) neplfl.aµpaveL Ta Pri- µma nou anOLTOUVTOL yta TT']V ane11.eueepwof1 TT'Jc; 011.oK11.ripwµevT']c; no11.unemtotKfJc; 011.uoioac;: TOUT6- xpova, TO ptpoowµ6Tto otiOTOTOL an6 To mRNA. ~LacpopenKec; oµaoec; E:nlKOUplKWV napay6VTWV unoPorieouv TO ptpoowµano oe K60e OT6oto. Evepyeta napexeTOL oe otacpopa OT6ota an6 TT'JV uop611.uOT'] TT']c; TptcpwocpoptKrJc; youavivric; (GTP). Km6 TT'JV evap~T'J, ri µLKPrJ ptpoowµLKrJ unoµovaoa pooMveTOL OTO mRNA KOL OTT'] ouvexeta evwveTOL µe µey611.ri unoµovaoa. Km6 TT'JV emµfJKUVOT'], TO mRNA uaKLVeiTOL otaµeoou Tou ptPoowµmiou KOL µeTOcpp6- £TOL oe TpmMTec;. (nap6n ou~ewc; µtMµe yta µeTO- ivriori TOU ptpoowµm(ou KOTC! µfJKOc; TOU mRNA, 8LVOL epLOOOTepo pea11.LOTlKrJ Tl tMa TT']c; e11.~ric; TOU mRNA taµfoou Tou ptpoowµmiou.) Km6 Tov rnpµanoµ6, ri pwTefVT'] KOL TO mRNA ane11.eueepwVOVTOL, evw Ol ~e- wptOTec; ptPoowµtKec; unoµovaoec; OllOTOVTOL npoKet- evou VO ~OVOXPT']otµOnOtT']eouv. 24.3 E101Koi Mnxav1oµoi E.Myxouv mv AKpiBEIO me; METCJcppaonc; 1 BOOIK&I; &VVOl&I; • H aKpf~£ta Tll<; µndcppaoT]<; £A£YX£TQl and £lOlKOU<; µT]xavt- oµouc; 0£ Kd8£ 010010. rvwpil;ouµe 6Tt ri µeT6cppaori eivOL yevtK6 aKptPfJc;, C!pf'] OTT'] ouveneLO TIOU OlOnLOTWVOUµe OTOV K00op(- ouµe TT'JV 0Mri11.ouxia ev6c; no11.unemtoiou. 'Exouv npayµmonotrieei 11.iyec; 11.emoµepdc; µeTpfJoetc; TT']c; ouxv6TrJTOc; M0ouc; in vivo , wOT6oo yevtK6 mOTeue- TOt 6Tt KUµaiveTm oe KMµaKa mu ev6c; 11.68ouc; ova 104 ewe; 105 aµtvo~fo TIOU 8VOWµOTWVOVTOl. /aµpa- VOVTOc; un6ljJT'] 6n ot neptoo6Tepec; npwTdvec; nap6- 0VTOL oe µey611.ec; noo6TT']Tec;, auT6 oriµa(veL 6n ri ouxv6TrJTO M0ouc; eivm noM xoµri11.fJ yta va exet K6- no10 eniopaOT'] OTO cpmv6Tuno TOU Kunapou. ~ev eivot 6µwa npocpavec; nwc; emTuyxaveTOl TOOO xaµri11.11 OUXVOTT']TO M0ouc;. npayµmL, Tl cpuori TWV eµ- n11.eK6µevwv µT']XOVLOµwv OTIOT£11.et Y£VlKO eeµa yt' Op- 8TC! pfJµma OTT']V eKcppaOT'] TWV yovtoiwv: • nwc; TO evc;uµa nou ouveeTOUV RNA avayvwpil;ouv µ6vo TT'] paori nou eivot ouµn11.ripwµmtKrJ µe TO eK- µayeio; • nwc; 01 ouveeTaoec; avayvwpil;ouv µ6vo xa tRNA Kot aµtvo~fo; • nwc; eva ptpoowµano avayvwpil;et µ6vo TO R nou OVTtOTOlX£i OTO KWOLK6vto OTT'] 0eOT'] A; Kaee nepimwori eernt eva nap6µoto np6P11.riµa: nwc; µnope( va otaKpt0ei eva ouyKeKptµevo µe11.oc; an6 oMMrJPTl TT'JV 0µ600, TT'Jc; onoiac; 611.a Ta µe11.ri µrnp6- l;oVTot TO LOLO yevtKC! xopOKTT']plOTlKO. nteav6v OTIOlOOrJTIOTe un60Tpwµa µnope( OPXLKQ VO e11.8et o' enacpfJ µe TO evepy6 KeVTpO KOTC! TUXT'J, aM6 enetTO TO 11.680c; unoOTpwµma anoppimoVTot Kot ano- oeKT6 y(veTQl µ6vo TO KOT0/.1.T']II.O. To KOT0/.1.T']fl.0 un6- 0Tpwµa ppioKE:Tot n6VTa oe µetoljJT']cpia (µta an6 Ttc; Tfooeptc; paoetc;, tva an6 Ta eiKooL aµtvo~fo, tva an6 TO - 30-50 tRNA) , ouvenwc; TO KplTrJpta OlC!KptOT']c; npe- neL VO eivot OUOTT']p6. H EIKONA 24.8 ouvoljJ(l;eL TT'] OUXVOTT']TO M8ouc; OTO pfJµma nou µnope( va enripeaoouv TT'JV aKpiPeta TT']c; µeT6cppaoric;. Ta Meri OTT'J µeTOypacpfJ Tou mRNA eivot on6vta, < 1o·6 . np6KetTm yta oriµaVTtK6 OT6oto yta TrJv aKpi- Peta, Ka8wc; K6ee µ6pto mRNA µeTOcpp6l;eTot oe noM6 npwTe"tvtK6 aVTiypacpa. Ot µrixovtaµoi nou e~aocpaM- c;ouv TT'JV aKpiPeta TT']c; µeTOypacpfJc; e~eT6l;oVTot OTO Ke- cp611.oto 19. To ptPoowµ6no eivOL uneu8uvo yta Mo Tunouc; 11.aewv OTT'] µeT6cppaori. Mnopei va np0Ka11.foet µua- T6mori Tou n11.otoiou avayvwaric; (frameshift) napa11.ei- LUXVO-rT)rn MSou~ I ~ /dSo~ ~dITTl I M£TOTOTILOT) I nAma[ouovdyvwari~ /d9o~ I oµtVOOKUAO-tRNA t /dSo~ 0µ1vo~u -/d9o~tRNA I 10-6 10-s 10-4 EIKONA 24.8 LTO 01acpop£TLKd OTOOLa TT]<; µ£Td(j)paOT]<;, Ad8T] ouµ~aivouv µ£ ouxvdTTJTO and 10·6 twc; 5x 10-4. 24.3 ELOlKO[ MT]XOVtoµo( EMyxouv TT]V AKpf~£la TT]<; M£Td(j)pOOT]<; 679
nowoq Uro Bdon 6rov 6roBd(er ro mRNA (rl 6roBd- (owog pro Bdoq 6Uo gopdq -nptirro og teleuro(o Bdoq ev6q ro6rrov(ou rr 6nero o:g npdrrl Bdor'1 rou en5trlevou xro6rrov(ou). Aurd ro l,d0r1 e(vor ondvrq (oup6rqro -10-5). En(or1g trrnope( vo enrrpdqUer (ro- ro)ro(ptoogo ev6g LdOog opuvoorulo-tRNA pr' 6vo rco6rx6vro, pe ouv6nero vo evoroporc0veror ).60o9 o;u- vo[ti. Aw6 prd].].ov e(vor ro oulv6repo Ad0og orn Ue- r6qpoo4 (ouXv6r4to -5 x 10 4). ELdyXeror on6 rrl 6o;111 tou ptBooroprot(ou Kor rnv r<rvqnrf qg 6rdoro- oqg (BL6ne Ev6trpa 25.1 5). Mro ouvOerdorl rou IRNA prnope( vo nporol*ioet 6uo rrlnoug LoO<irv: vo ronoOerloer Ld0oq oprrvo[0 oro IRNA r4g 11 vo goprcioet ro opuvo[ri qg oe ].d- Ooq IRNA (BI6ne Evotrpa 25.9). H evocopdroo4 Ld- 0og opuvo[dog e[vor nro rorvrl, (oolg enerdq ro IRNA npoorpdper peyolUreprl enrrpdvero [e rnv ono(q to 5v- (upro pnope( vo 6rlproupy{oer neptoo6repeq enogdq yro vo SroorpoL(oer er6rr6rqto (specificity). Or ouv0e- rdoeg rov optvooKuLo-tRNA 6ro0rirouv er6rxorig ;rr1- Xovroporjg yrq rnv enrd16p0ooq Lo0cirv npo'roU oneleu0epto0e( 6vo IRNA pre ).dOog <popr(o (BL6ne E- v6tqra 25.1 1). 24.4 H 'Evop[n oro BoKTtipto Ynopovd6eq Kt EntKouptKo0q Xperd(eror 3OS Ilopdyovreq e H evop[I qq peroqpoo1g onoue[ Eextoproreq prpooorgrr6g unopovodeg 30S ror 50S. * En[onq onouotivror ot nopoyovreg evop[1q (lF-1, -2 rot -3), ot ono(ot npoodrlvovror otrg 30S unopovodeg. e Mro 30S unopovodo nou gerorpeper nopoyovreg evop[qg npoo66veror oe pLo 0eo1 Svop(1q oro mRNA oXlpor(ovrog evo o0pntroxo evop[r19. * 0 lF-3 npener v' one),eu0epot0e( tlore ot 50S unogovodeg vo npoode0ouv oro o0gnLoro 30S-mRNA. To Borrrlpror<d prBooroprdtro nou lprlottrlonotoriwor ornv entprlKuvoq Uroq nolunemLdLr<r1g oLuo(6og ug(- orowor rr;g 70S ooll.ror(6ro. Kord rov repprotropr6, one).euOepovowqr on6 to mRNA cr.lg eeU0epo pLBo- o<r.lprdrro r; prBooroprxdg uno;rovddeg. tre ou(ov6pevo Bor<rqpto, ro neproo6tepo ptBoorrtprdtro ouvO5rouv nporelVeg : 11 de[opevrl rotv e].eU Oepo-lv p rBooropror[cov nr0ov6v nepr5ler -20o/o r/ov prBooropror(ov, To ptBooroprdtro r4g eLe60eprlq 6e[opev4g ;.rnope( vo Srooro0oriv oe leloprm6g unopovd6eg: ow6 oq- ;ro(ver 6rL to 70S prBooopdtro Bp(or<owot oe 6uvopu<q Looppon[o pre ug 30S xor 50S unopovddeg. H |vaplrl tqq pefiEpoo4g 6ev elaptotaL an6 oxipata pLBoortt- paua oMa avalapBavetat an6 tq eyuporcq unopo- vddeg, or ono(eg otrtrrllenr6porjv r<ord qv ow[6poor1 rrig 6vop[49. H ri{*i!]ti !;i.} ouvoil((et rov r<0r<}.o rov pr- 680 KEOAAAI0 24 Meroqpool 30SunopovdDeg :eXoptotdg Ae(opevrl 'Evup{n EnqrKuvoq Teppottogtig EIK0NA 24.9 H evop[1 onoLre[ etreU0epeg pgooopLr6q urotto- vddeg. 0rov ro pgooropdrto onetreu0epdrvovTor oTov reppouoprl, or 30S unopovoDeq npoo66vouv nopoyovreg 6vop[r1g ror 6ri- orovTor dlgroupytowoq etre0Oepeg unopovd6eg.'0rov or unopo- vodeg enovoouvduo(ovror yro v' onodtiroouv rivo treuoupyrxri prpooogdno oqv 6vop[1, onetreu0ep6vouv Touq nopoyoweg. Boocoprrdrv unopovddrov r<ord tr1 pretdgpoorl oro Bo- rcrripro. 'Evop(q ouprBo(vet or puo erdrxrl oLlqLoul[o rou mRNA yvroorrl cog neprol{ np6o6eo4g rou plpooo- poriou (ribosome-binding site) (ouprnepr).oprBdver n1v oMqLouX(o Shine-Dalgarno - BLdne en5prevq Ev5rq- ro). l-1p6r<error yro Uro BpoXe(o oLlqLouX(o Fdoeov npw on6 ulv x<o6rxonorqtu<f neproXrlg (BL5ne Err6vo 24.12),4 ono(o e(vor oupnlqpro;rorxrl ev6g rp4prorog tou 165 rRNA (B).6ne Ev6rqro 24.18). H prxpri Kor n Ue- ydLrl unoprovd6o oMrll,entdpo0v orrlv neproXrl np6o- 6eonq rou prBootopror(ou olqpot((owog 6vo d0ucto prBoocopdrro. H ow(6pooq ougBo(ver oe 6Uo BnUoro: . Avoyvcbprorl rou mRNA npoyUaronore(ror 6rov npoo66veror puo prxpri unopovddo oXqpror((owog 6vo olpnAoxo 6vopl4q (initiation complex) orrlv Tre- ptopl np6o6eorlq rou prBooroprot[ou. o 'Enetto prro preyd),r1 unopovddo ouv66eror ge ro o[ pn].oxo oXq poil(owog 6vo n],r1 peg prBooro prdrro. l-iop6n q 30S unopovddo eprnLdreror ornv Svop[q, 6ev enopre[ on6 g6vr1 rnq yro ng owr6pdoetg np6ode- orlq rov mRNA ror IRNA. Anore( entnp6oOereq npto- refveg yvom5g og nop6yovreg 6vop[49 (initiation factors - lF). Tdtorot nopdyoweg Bp(or<owor ;r6vo nd- vo orrq 30S unoprovddeg rr oneleu0eprivowor 6rov or 30S unopovd6eg otrL4l.enrdpo0v pe ng 50S unopovd- 6eg nopdyowog 70S prBootoprdrro. Aurrl 4 6pdo4 6ro- rp(ver roug nopdyovreg Svop{r1g on6 rrg 6opur6g nporetveg rou prBoorollffi[ou. Or nopdyoweg Svop[r1g ooXol.oriwor onox,.eronrd Ue ro oxrlpotopr6 rou oup- nl6xou Svop[qg: onouord(ouv on6 ro 70S prBoorr:pr6- lo Kqr 6e 5to6po;ror((ouv rdnoro p6lo oro ord6to rqg enrpuiruvorlg. H Elf0NA 24.'10 ouvorf((er ro md6ro rqg 6vop[qg. Boorxeg evvoreg llopdyorneq
1 H 30S unouovddo npoo66verot oro mRNA tcail-J G...... A AuvogLrl rooppon[o Ae(ogevrl 70S prpoorilgoriolv EIK0NA 24.10 0r ropoyovTsq 6vop{19 oToeeporototiv nq etreU- 0epeq 30S unopovqdes Kor rrp0o66vouv ro evopKTnpto IRNA oTo oriUrIoKo 30S-mRNA. To Borrrjprq XpnorponotoUv rpetg nopdyovteg 6vop(r19, lF-1,1F-2 Kqr IF-3. Or nopdyoweg outo[ onot- roUwot yro rnv e(oo6o oro oUpnloro 5vop[qg t6oo rou mRNA 6oo rot rou IRNA: . O lF-3 61er noMonl,Sg Letroupy[eg: mo0eponote( rrg (eleri0epeq) 30S unogovdSeg rot ovomSlLet qv np5op4 np6o6eoq qq 50S unoprovddog. Ko0t- md rrg 30S unoprovd6eg trov6g vo npoo6e0o0v ong O5oerg dvop[r1q rou mRNA. En(or1g, tog t1.ui1.to tou oupntr6xou 3OS-mRNA, etr6yXer qv or<p(Bero q q ovoyvri p rorl q ro u n pdlro u ol.uvooxul.o-tR NA. . O lF-2 npoo66ver 6vo erdrr6 evopxuipto IRNA r<t el,6yXer r1v efoodd rou oro prBoocoptdtto. . O lF-1 npooS6vetor oe 30S uno;rovd6eg oq rprlpo rou ntrrlpoug ouprnL6r<ou 6vop[qg. [1poo66vetot rowd orq 06oq A Kor nopeUro6(et qv e(oo6o tou o1lwooxul,o-tRNA. H ew6ntoq tou prnope( eniorlg vo epno6(oer qv np6o6eor1 qg 30S unopovddog pe qv 50S unopovddo. I1o),),59 6opur<59 prel6teg unod4trtilvouv 6tt o lF-3 51er drio 6tor<pm6g, odpd ogotpu<5q neptoX6q, Ue rnv C-reLtrrl neproXrj orrl 06o11 enogrlg yto rrlv 50S ndvo orrlv 30S unopovddo Kor rnv N-teLxri neptoX4 xowd orq 05oq E qg 30S. H npdrr4 l.enoupy(o tou lF-3 eLdylet qv tooppon(o prero(ri tov ptBooco;ru<tilv xoroor6oeov, 6nog Eofuerot orqv El(0t'lA 24.11. O lF-3 npoodSvtrot 01 etre[0epeg 30S unoprovd6eg, or ono(eg oneleu0epdrvowot on6 tr1 Etreu0epeS unopovodeg t- I t I H 30S unopovodo pe rov lF-3 Unopet vo npoode0et I oro mRNA. oxr opoc xot oqv 50S unopovo0o i I 0 tF-3 npener v 'nttrsuospooet yt. v0 unopeost " * . . . ' I vo npooDe0el 11 50S unouovci5o EIK0NA 24.11 H 6vop[r1 onoLtei 30S unopovodeg nou getoq6- pouv lF-3. de(qrevrl tov 70S prBoocopror(ov. H nopouo[o tou lF-3 nope;rno6((er rrlv enovoo0v6eorl qg 30S unopovddog pe Uro 50S uno;rovd6o. O lF-3 Unope( v' oMrlLenrdpd- oer dpeoo pre ro 165 rRNA r<or undpXet orlprowu<r1 ent- rdLur!4 pero[ri rov Bdoerov tou 165 rRNA nou npoorore0owor on6 rov lF-3 Kot rov ow[orotXtov Bd- oeoJv nou npooroteUowot on6 rqv np6o6eor1 qg 50S unogovddog, yeyov6q nou uno6qtrcilvet 5fl nopoxtol,rjet 14 quoxri ouprBoLrl tov unopovddtov. Iuvenrig, o lF-3 ouprnepup5perot cog nopdyowog owt-oltr4lenbpoo4g, nou 6Xer org onot6)reopotnv nopogovrl qg 30S uno- provddog orq de(opevrlrov etreri0eporv unopovddtov. H ow(6poor1 peto[ti lF-3 ror 30S unoprovddog e(vot orot- Xetoprerprx4: 5vo pr6pro lF-3 npoo6dverot ovd unopovd- 6o. Enerdrl 11 noo6rlro tou lF-3 e(vot oXettrd ptrprl, ouvdyeror 6u q 6ro0eotpr6rqrd rou ro0opf(et tov opt0- pr6 ro:v ee0Oepov 30S unopovd6tov. H 6efrepq ].emoupy(o tou lF-3 e].6yXet tqv xov6rl- ro rov 30S unoptovd6rov vo npoode0oUv mo mRNA. Mrxp6g unopovd6eg np6na vo 6to05touv lF-3 nporetp6- vou vo g4rlpot(oouv otipmloxo 6vop(qg pe to mRNA. O lF-3 npdner v' onetreu0epoOe( on6 ro o0pnLor<o 30S- mRNA d:sre vq prnopSoet vo npoo6e0e( 11 50S unopovd- 6o. M6tg oneleu0epo0e(, o lF-3 ovorurl.6vetot opr5ocog Bp(orowog puo dMq 30S unopovd6o. T6Log, o lF-3 el,6yXet t4v ot<p[Beto ovoyvdrptorlg rou nprilrou ol.rloorul.o-tRNA r<ot Fon0d orov npooovoro- Lropr6 rou rpoq rrl 05oq P rrlq 30S unopovd6og. H nprirq on6 owSg rg Lemoupy(eg 6Xer ono6o0e( oqv C-relrnj neproXq tou lF-3 (BL5ne Ev6t4ta 24.7). H N- teltxrj neptopl rou lF-3 e(vot tono0etrlpr6vq drme vo Bor10d fiov npooovotoLtop6 rou optvooKu).o{RNA npoq rn 06oq P qg 30S uno;rovddog onoqpdooowog rq 06o11 E ev6oo o lF-1 onogpdooetrq 05or1 A. O lF-2 dXer 6p6or1 GTPdorlq e(oprc(rprevqg on6 to prBootoprdrto: nopouo(o prBooto;ror(rov npodyet tr1v :i 0 lF-2 geper ro IRNA orn 0don P 3 0r lF oneleu0ep6vovror xor npoo66vetot 11 50S 2&,4 H 'Evop(r1 oru Boxqpro XpeLd(eror 30S Ynopovddeg xt Enxouptro6q flopdyovres 681
udp6Luorl rou GTP one),euOeprilvowoq qv ev6pyero nou e(vor ono04r<eup6vr1 oro 6eop6 uUnInS evdpyerog. To GTP u6poLtieror ;16119 npoodeOe( 11 50S unopovddo yro 14 drlpuoupy(o rou ntrrlpoug prBoocogot(ou. H 6Ld- oroon rou GTP 0o pnoporioe vo epnl,6reror fin Uero- BoI4 qq 6ropr5prpcooqg rou prBoocopor(ou, Stot 6qre or ouvevopSveg unopovd6eg vo perorponoUv oro evepy6 70S prBooroprdrLo. 24.5 H 'Evop[n lleprAopBdv€t ZEUydpoUo Bdoeulv Mero[ti mRNA Kot rRNA '. Iro gorrrlproro mRNA 11 0eo1 evop[1q onore],e[rot ono ro rodrrovro 6vop(1q AUG ro onolo oroiouOei -10 pooerg ge- ro ro e[opep6g nouplvrilv Shine-Dalgarno. ,, To rRNA qg 30S pr$oooprrnq unopovodog rotv Borqpuov Dro0erer pro oupntrrlpo;rolr1 otrtrr1trouX[o nou (euyoprlver pe qv otrtr1trouXLo Shine-Dalgarno roro qv evopil. To orlpo yro rnv 6vop[q prog no].unemrdrrrlg otruo[6og e(vor 5vo et6tx6 rto6u<6vto Svop(4q, ro ono(o orlporo- 6ore( qv oppl rou n).oro(ou ovdyvotorlg. Iuvrl0otg, r<o- 6rx6vro Svop[qq e[vor qtptn}.6ro AUG, toor6oo oro Borcrrlpro XprlorUonororiwot en(or;g ro GUG xor UUG. 'Evo mRNA nepr6ler nol),69 rprnAdreg AUG: nog dpoye ovoyvrop((eror ro r<ro6rxdvro dvop[qg oq oeern- p(o qg perdgpoorlg; Or 0ooetg tou mRNA dnou opX((er q perdqpooq rouronotofwor on6 qv np6o6eoq rou pr- Booorpror(ou oro mRNA un6 ouvOrp<eg or ono[eg ovo- m5).Louv rqv entpqxuvorl, r<o0r1).rrtvowoq ro ptBootr-l- pdrto mrlv o<peqp[o. Av oro r<o04].cop-rdvo oripn].or<o Svop[qg npooreOe( prBovouxtredorl, 6].eg oL neproxeq rou mRNA nou Bp(orowor 5lo ond ro prBooroprolo 0' ono6opr40oriv, evc': or neprolSg nou 61ouv npoo6e0e[ oro pLBoorrtl-rorro 0o Trpoororeurorjv, 6nog qo(verot oqv . To npoororeupSvo r<Ldo;roro ;rnope( v' ovocrq0oJv Kor vo loporcrqptoroJv. Ot oMq).ouXieg Svop[r1g nou npoorore0owoL on6 ro BorurlpLor<d pLBoooprdlo dlouv prrp<og -30 Bdoeotv. Or nepLoXdg np6odeorlg rou prBoooprorfou 6rorpoperL- rurv BorcrrlpLoxrr;v mRNA eprgov((ouv 6io xorvo Xopo- KI1proflKo: o To r<odrx6vro dvoplrlg AUG (n Lrydrepo oupo GUG n UUG) ndwo oupnepL).opBdveror ornv npocrroreu- pdvq otrI.q).ou1[o. o Ze ondorooq 5og 66ro Bdoeov ovo6Lr<d rou AUG Bp(or<eroL puo oMqLouX(o nou owrororXe( oro e[r1g elopep6g 11 oe rprl;ro rou: 5,..AGGAGG..3' Autq q nolunouprvrr<r1 ol.).r1).ouX(o e(vot yvcrlorrl orq oMqlouX[o S h i ne-D al garno. E(vor ou prn].r1 potl.rortr<r1 ;re 682 KEOAAAIO 24 N{sToQpoorl r^r{F, /t' 0Dnyoq KoDrronorrlrtrui neptoxrl Shine-Dalgarno < 1 0 pdoerg ovodLxd'rou AUG Al-li '0tre9 ot neproxdg oro rdv.rpo rou tivop(1q 6to0erouv npoorortuUtvou 6tloouvotvertro xtrdoporoq orotxe[o EIK0NA 24.12 l-leproX6g npdodeorlg rou prpooogorfou novto mo mRNA pnope( v' onopovtoOouv on6 oupntroro 6v0p[rlq. 0r nepo- X6q 0ur6q nepilogpdvouv qv ovodrrrl otrtrntrouxio Shine-Dalgarno ror ro xtodurjvro evop[nq" pto eforpetrrd ouw4p4pSvq o).).4).ouX(o rowd oro 3'drpo rou 165 rRNA. (To eripog qg ouprnlqpcoprorrr6- rqrog drorp5per ovd).oyo ge to mRNA ror pnope( vo ercre[veror on6 r4v xewpu<r1 oMrl],ouXio reoodpov B6- oeov GAGG 6o9 puo oMr'1),ou1(o ewSo Bdoecov.) l-potrr- pr6vr1 oe ow(mpog4 rotef0uvor1, q o].].4].ouX(o rou rRNA efuot ro e(oprepdg: 3'...UCCUCC...s', Apoye, rord rqv np6o6eoq rou mRNA oro pgooro- pduo q oMrlLouX(o Shine-Dalgarno (euyoprirver Us ro oupnl4prllporu<6 qg rRNA; Meto).)d[etg efue orn Uto eke mqv dtri,q oMqlouX(o uno6rll.rr:vouv rr1 oqproo[o rouq orrlv Svop[q. Irlprerox6g preroMd(etg orrlv oMrl- ouX(o Shine-Dalgarno prnope( vq nopegno6(oouv 14 prerdrppoo4 tou mRNA. EnrnLriov, 11 erooyoyrl ;rerotri.d- (eov orrl oupnl.qpolprotu<ri oMqLoul(o rou rRNA e(vor enp),oBr1g yro ro r[rropo r<or pretoBdMer ro np6runo qg perdrppoonq. H xoOoproru<r1 engeBo(cooq qg ow[- 6pooqg rou (euyoprilgoroq rov Bdoerov e(vor 6rL puo perdLLo[r1 orqv o].].4].ouX[o Shine-Dalgarno evdg mRNA pnope( vo roroorotre( on6 pro prerdtrtro(4 oro rRNA nou onoroOrord ro (euydpopro rtov Bdoeotv. H oMqLou{o oro 3'6xpo rou rRNA e(vor ouw4pr;- prdvq prero[U npoKopuord:v xr europucorcbv, Ue rrlv e(o(peor1 6n oe 5loug rouq euKopucirreg undpXer ono- Lorqrl qq oL).4l,ouX(og nSwe Bdoetr:v C C U C C, n ono(o e(vor ouprnLrlpoprorLrrl qg oM4louX(og Shine- L---T----------l npdo0eon rou prpootopor(ou oqv neprcXn SvopEnq tou mRNA npoo0rlKrl vourtredolq yto 6trou rou onpomdreurou mRNA Anopdvoorl rtrdoporog rou npomoreug6vou mRNA Boorxeg 6vuoreg AACAGGAGSAU UACCCCAUG UCGAAGCAA... npoototeu gSvou rtrdopotoq
Dalgarno. Iuvendrg, prdMov 6e ouprBo(ver (euydporpro Bdoeov preto[ri rou euKopuotrxoU mRNA rot rou 18S rRNA. l-1p6r<error yro o4prowu<4 dtogopd oro prrllovropr6 rqg 6vopl4g. Iro Bor<rqpro, Uro 30S unoprovd6o npooS6veror d;reoo oe pro Odoq rou prBooco;rodou. 'Eror, o111prot(- (eror dvo o[pn],or<o 6vop[qg oe Uro o].trr1].ou1(o nou neprBdMer ro ral6u<5vro Svop[r1q AUG. 'Otov ro mRNA efvot no].uormpovrx6, rdOe rcodrronot4rrrrl neprol4 opX((er pe pto 05o4 np6o6eorlg prBoocopror[ou. H rptio4 qq Srrppoolg rov Borcr4prordlv yovrd(otv orlpo(ver 6n q prerd<ppooq ev6g BorrrlproroU mRNA npolcope( 6ro6o1u<d 6rop5oou rov ororpov[rr.rv rou. M6- fug to ptBoocoprdtro npoo6e0o0v orrlv nptirq rrrrdrronor- rpu<r; nepro1rl, ot en6peveg rcodrronorqru<5q neproXSg 6ev 6Xouv or<6prq preroypoge(. M6rg y(vet 6to06orgr1 4 deurepq 06oq rou prBooroproilou, 11 prerdgpooq Bpiore- ror xo0o66v 6rop6oou rou nprirrou otorpov(ou. To ou;rBdwo olg rro6rronoulnx6g neprol6g norri Aouv ovdLoyo Ue ro mRNA. Ilg neptoo6tepeq ilepr- nrrioetg, ro ptBoocoprdrro prdAov npoo6Svovror ove[dptrpo orrlv opXrl rdOe ororpov(ou. H ouq06ore- pq oerpd rrov ou;rBdwolv nopouord(erot mqv EIK0NA 24.13. Otov repporrore[ 11 o[vOeor1 qg npc0rqg npotrei- vnq, ro prBoocopdno eyrorole(nouv ro mRNA r<or 6ti- orovror orrq unopovddeg toug. 'Enerro, rivo vrlo prBoocopdtto np6neL vo ouvopl,rooyq0e( orqv en6trrev4 rodxonorqtu<r1 neproXq xor vo (errvrloer q perdqpoorl oro en6gevo otorp6vro. Ie opropdvo Borcr4pLord mRNA, 11 pretd<ppoorl rcov 6mtrovrov ororpov(rov e(vor ou(euygdvrl, enet6ri to prBo- ocopdrto onorcrofv np6oBooq oro xco6tr6vto 5vop[r1g rou 6e[tepou ororpov(ou r<o0dlg o].ox].r1p6vouv rrl Ue- rdgpoorl rou nprinou. l-to vo ougBe[ ow6, ro peoo6rd- 6rnUo peto[ri rtov 6tio ro6rronorqrrxciv neptolrirv npdner vo efuor prxp6. Mnope( vo e[oprdtor on6 rlv uqnln rontrti nurv6rrpo rcrlv prBootol-ror(cov. EvoMo- rrtrd, 4 nopdOeo4 O5oeo:v repprolo;ro[ xr Svop[r1g enrpdnet vo nopqKqpq0oUv opropr6vo on6 to ouvr10r1 oupBdwo prero(0 rov ororpov(cov. 'Evo pLBooroprdtro Evop(11 l Tepgortogoq 'Evopqn t' r<oLUmeL -30 Bdoerg rou mRNA, ouvrniue :: * ::: -, oe vo 6pXetor towdlpovo o'enocprl g evo <*5.,:. : repportopori Kor Fe rnv en6pevq 05o1 dvoplqe, :.r. vo prlv on6louv nopd ).tyeq p6vo Bdoerq. 24.6 'Evo Er6rr6 Evoprrrlpro IRNA lerrvd rnv lloAunenr6rrfi AAuoi6o " H peroqpool opXi(er p' evo rorotrorno pe0erov[v1g rou ou- v10og rt.rdtxonore(ror ono io AUG. ,, Arorpopelro tRNA pe0eLovrvrlg egntr6rovror orrlv evop[r1 ror rqv enrgrlruvol. - To evoprqpro IRNA dLo0erer povoDrxo boprro XoporqpLort- ro, ro onofo ro dlotpoponorouv on' o],0 ro unotrorno tRNA. '. H ogo6o NH2 rrlt Usoerovivrlq nou npoodeveror oro poxrq- proro evoprrflpro tRNA e(vor rpopputrulpevl. H orjvOeoq 6lrov rcov npcoteivdlv [exrvd pe ro [6ro oprr- vo(rj, rr1 peOerov(vq. To IRNA nou ovoyvop((ouv ro rodtr6vLo AUG preropdpouv ;re0erov[v4, evo 6rio ttj- not IRNA prero<pSpouv our6 ro otrrtvolrj. O 6vog rJnog XplotponoreLror orqv Svop[q r<r o d].].og orrlv ovcl- yvroptoq rcov rrr;6u<oviti-rv AUG r<ord trlv entgrlruvoq. Iro Bor<tqpro, ro 1-rLtol6vdpro r<or toug g.ropon).d- oreq, ro evopnrqpLo IRNA preropdper dvo rot6otno trle- Oetov(vr1g, rou ono(ou 11 oprrvoprd6o 51er rpoppruLro0er cn1r1 por((owog 5vo pr6 p ro N -rpop p utro- peO er ovuho-t R NA. To IRNA efuor yvoord org tRNAlMet, evcb ro opuvoor<ulo- IRNA ouvrlOog 6r1).rr-rveroL pe rr1 oUw;rqor1 fMet-tRNA. To evoprcrrlpro IRNA onorcrd ro rpononorqpr6vo opr- vo[ri rou p-re puo ow(6poor1 60o plUdrotv. l-lproro <pop- troveror ;-re ro oprrvo(0, olqpror((owog MettRNAt. Irrl ouvdleto 11 ow(6poor1 <pop f-iuL(cr-loqq, nou nopo uoro(e- ror orrlv , onorLe(et qv eLe[Oepr1 opuvopro- 6o: dror, ro evopr<rqpro IRNA 6e pnope( vo o'uprprerdoler ,C7' C,t,^ --- '9ch 'oc/-,/)n. .bn, ,, 2,.[ - ruH' -%..8-s$' I U_U I 0 l^l I l- t -I1poiq rodrronorrllrrl neptoxil I U -U I QDtr o 1ls 3L_-'-_lJ Aetireprl xol6rxonoulrixrl neprox4 'EvopErt ouppolver ove[opqro oe roOe olorpovro ,. -. notruororpovrrou mRNA. 'Orov I neproXrl pero[u duo or- ,-::'iirov e[vor porpurepq ono ro prlrog rou prBootrrporlou, l Dro- , rrn 0T1 0eo1 repporropou oro),ou0efroL ono ove(oprlrq - -. e. opEq oTo tttopsvo oroTpovto, IMet-tRN& N-rpopputro- Ue0srovutrotRNA ru-q,upFu^u-.. Terpoijdporpulitrrd TsTpoU6ooQuAIrKd elK0filA 24.14 To evoprrrlpro N-rpopputro-pe0erovutrolRNA (fMet-tRNAt) oXlgor[(erot Ue QopUutr(toon rou ge0erovutrotRNA, orrlv ono(o trrg oupnopoyovToq Xplorporore(ror ro qop;rutro-re- rpoUdporputrtrrxrS. 2{..6 'Evo Erdrx6 Evoprrflpro IRNA :errvd rqv l1otrunenrt0Lxr1 A-uoi0o 083 1 0-rpopUulto- Boorxeg ewocg
qnlv enUnKUVon'rnq o).uo(6o9, lcop(g 6prcog ow6 vo no- pepBo(ver or4v xov5r4rd rou vo exruve( puo npotefvr1. Aur6 ro IRNA Xp4orponorefuor ;r6vo or4v Svop[q. Avoyvcop((et ro xro6rr6vro AUG xot GUG (onopodtxd ror ro UUG). Tq KodrK6vro 5ev ovoyvcop((owor e[(oou ocomd: 11 ouXy6r4ro r4g Svop[4g perciveror nep(nou oro pro6 ov ro AUG owrKoroora0e( on6 to GUG 6n<og en(oqg Kor ov Xcrloryonot40e(ro UUG. To IRNA nou e[vor uneU0uvo yro rnv ovoyv6prorl trov xrodrxov[rov AUG o' eooreprK5q O5oerg e(vor ro tRNAmMet. Aur6 ro IRNA owrororXe( trr6vo o' eoorepr- rd rrrl6rx5vro AUG. H peOerovlvrl rou 6e prnope[ vo eop- ;ruLurl0e(. floro Xoporcqprorrr6 drorplvouv ro fMeltRNAf qg Svop[4q on6 ro Met-tRNA, rqg enr;rrlr<uvo4g; I4proo(o 6Xouv oprqr6vo lopor<r4pronrd rrlg oMrllouX[og rou IRNA, 6nroq ouvotp((eror or4v E|(0NA 24.15. Meprxd e(- vor ovoyxo(o yto vo ilopepnod(oouv rn Xpnon rou eKKr- vrn{ m4v enrpr4r<uvor1, ev6 dtrl.o yro rq },eroupy(o rou orqv 6vop[r1: . H gopputr(oor1 5ev e(vot ono).frog ovoyro(o, ener- 6]i to pq qoppru},uoprdvo Met{RNAr prnope( en(or1g vo Lettoupy4oer cog exrrvrlulg. Om6oo, n gopUu- L(oor1 Betrrrciver rrlv onoreAeoportr<6r1ro pe rnv ono(o Xp4orponore('ror ro Met-tRNAr, enerdrl e(vor dvo on6 ro XopoKrrlptorrrd nou ovoyvrrlpf(owor on6 rov nopdyowo lF-2, o ono(og npoo6Sveror oro evoprcr{pro tRNA. o Or pdoerq nou owu<p((ouv n prq rrlv dMq ornv re- l,eurofo 06o4 rou pr(oXou orov ono(o npoo65veror to o1-uvo[rj efvot (euyoproprdveg oe 6l.o rq IRNA ercr6g on6 ro tRNArrret. MeroMd(etg nou 64puoup- yo0v dvo (eUyog Bdoeu:v oe outrj rr1 06oq rou tRNnu"t rori ennp5nouv vo lerroupye( orrlv enrpui- KUVorl. Iuvenc0g, 11 onouo(o ouroU rou (e0youg no- pepnod(et rn Xpnon rou tRNArMet or4v enrprqruvorl. 0opUUtrrorU6vo oUrvoIri Xop(q (euyopotto Pooeurv 3 (eUyq pooeov G-C r:;';1li;&:*.:i To ftVlet-tRNAt 6Xer povodtro XopoKTrlproTrKo nou ro [eXopi(ouv trlg ro evoprrlpro tRNA. 684 KEOA Al0 24 Meroqpoon En(oqg, ro (eUyog e(vor onopolrr1to yro r4v ow[- 6poon rpoppruL(tooqg. o To tRNAtMet, Kor. p6vo our6, 6ro0ritet pro oerp6 3 (euycirv G-C oro pr[oXo nprv q Oq].rd nou nept6la ro qwrKo6rr6vto. Aurd ro (eUy4 Bdoecov ononoriv- ror c0ore ro fMet-tRNAtM"tvo pnope( vo evoopmu> 0e( oneu0e(oq crrn e6on P. Iro Borrqpro Kor ro l.rrroX6vdpro, to xordLorno 1-rupl.rqyrorof (gop1.rutroprd6o) orqv evoprrrlpro pre0er- ov[vq ogotpe(ror on6 truo er6rx4 onogop;ruldon, o]n- ptor((ovrog 6,vo r<ovovrr<6 oprvorel,rr<6 dxpo. Av 11 peOerovfuq npoop((eror oq ro oprvoreLrx6 opuvo[f rqg npondfig, our6 e(vot ro 1lovodrx6 ovoyro(o Bqpo. l-le- p(nou mrg puoSg nprrneiveg, 11 preOerov(vq rou dxpou ogotpe(ror on6 gro ol.rwonemr6doq, q onofo 6qpuoup ye( 5vo v5o dr<po on6 to Rz (opXrrd ro 6eUrepo oprvo- [6 nou e(Xe evooprorro0e( or4v o].uo(do). 'Orov ror ro 6Uo Bqgoro e[vot ovoyro(o, ouprBo(vouv 6ro6oXu<d. Ot owr6pdoerg orpo[peo4g npoypororrotofwor ypnyopo, ;rdMov 6rov 11 veoouwr0Sprev4 nolunemr6u<r1 o).uor6o grdoer oe prrlxog to 15 o;rwo[6o. 24.7 H Xpfion Tou fMet-tRNAr EAdVxeTot on6 Tov lF-2 Kot To PlBooropdflo o 0 lF-2 npoo06ver ro evopxrrlpro fMet-tRN& ror tou ennp6ner vq ero6tr0er or4 gepLr4 0eor1 P r4q 30S unogovoDog. Kotd q ;rerdqpooq mo Boxrrlpro, n ongoo(o rov rr.u6rrov(rov AUG rqt GUG <e[opr6rol on6 ro ouu- gpo(6pevo,> (context-dependent).'Otov ro r<o6rx5- vro AUG Xpnotgonore(ror yro 6vop[4, droBd(eror og <pop1-rulo-pe0erovivq, ev6 6rov Xprlorponoe(ror gSoo orrlv rcodu<onorrlru<r1 neprop1, oro nolunem(6ro npo- m(0eror preOerov(vq. H oqproo(o rou xrodrrov(ou GUG efoprdror or<6prq neproo5repo on6 rlv ew6nto{ rou. 'Orov e[vor ro np6ro rcodrx6vro, npoor[0eror gopUu- l,o-pe0erov(vq, evril 6rov Bp(oreror oro eooreptr6 ev6g yovr6(ou, npoo6Sver Val-tRNA, 5vo on6 ro Kovo- vxd pui).q rou ouv6Lou ro:v IRNA, nop6Xowog BoLi- v4, oripr<pcr:vo pe ro yeverrK6 r<c66xo. l-1uig epprqveUowor ro "ouUqpo(6pevoD rov xto6xo- v(ov AUG ror GUG; H EIK0NA 24.16 qvodeKvtier rov r<p[- or1.ro p6),o rou prBoor.opor[ou, 6rov evepye( oe ouv6uootrl6 U' enxouprKo6g nopdyoweg. I' 5vo orlprnLoxo 5vop[r;q, oro mRNA e(vor npoo6e- 6ep5vr1 pr6vo 11 purprl unoprovd6o. To r<rrr6rx6vro rivop- [49 Bpioxeror p6oo oto rpqpo rqg 05o4g p nou perorp6peror on6 rq ;rxpr1 unopovddo. To pr6vo otruvo- oruLo-tRNA nou trlnope[ vo roroore( p6pog rou oupr- nL6rou 6vop{49 e(vot ro evoprcr4pto, ro ono(o dro06rer q povodu<q rdt6qro vo ero5pXeror oneuOe(og orn Ue- prr4 06o4 P yro v' ovoyvrop(oer ro r<ro6x6vt6 rou. I q*, i &&{3 & & & * Boorx6g eworcq
M6vo ro fMet-tRNAf etodpletoL orn Uepxl 06oq P n1g 30S Mrjvo ro aa{RNA eto6pletot or! e6on A rou nlrlpouq 70S . : ':,: l/16v0 ro fMettRNAf Unope[ v0 Xpnotuonotrloe( Yto , :-i- ono rtg 30S uno;rovo6egl , A),),0 olttvoorutro{RNA (aa- ': , - rpener v0 Xp1orponotl0ouv yto entpfruvo1 ono ro 70S pt- - , --0Ti0. ','loLrg 11 ;reydr1 unoprovd6o npoooptqOe( oro oO;i- -' :(o, or pepLr<dg 06oetg np6o6eorlg tou IRNA pero- --rrcwor ortg or<Spoteg 0doetq P rot A. To evoprcrrlpto '''::-rRNAf rcoro).oprBdveL q 0dor1 P, evot q 06o11 A 6to- - :.::r yro rlv eioodo rou outvooKuLotRNA nou elvot , --:i.qpo;,-rotLx6 pe to 6e[tepo rco6tr6vto tou yovt6[- : - C npci.rrog nemt6u<6g 6eopr6q crlqprot((etor preto[ri - - - ./opKrrlprou Kor rou en6pevou o1-tLvoot<utro-tRNA. - evop(r1 entrpote( dtov 6vo t<to6tr6vto AUG (n , - 3 Spior<eror gSoo oe Uto 0do1 np6o6eoqg ptBooto- -rr r;, enetorl p6vo to evoprrrlpto IRNA pnope( vo et- , .'iaL or1 preprx4 06orl P nou crlqprot((etot 6tov 11 30S - -: -:vo6o npoodeOe( de novo oro mRNA. H eorotept- ' - r. tryvoJon entrpote( opy6tepo, 6tov to xtrt6tr6vto : - : r ,ror o' enogrl p' 6vo ptBoot,.lporto, ro ono(o ouve- -. .o ;.rerocppd(et dvo mRNA, enet6q pdvo to Kovovt- . r l-.',/ooKUIo- tRNA e(vot trovd vo eto5trOouv srrlv -. - ^- Acnn A! vevrl,. 3. onrroupLro( nopdyoweg e(vot xp(ot;.rot Yto rov : : .-,.i rlq Xpnorlq rov optvooKuLo-tRNA. Otro to o;.tL- - :. -r,c-tRNA oMqLent6porjv pe to pLBootoprdtro prSool -:::-ioqg o' 6vov entrouptrd nopoyowo. O nopd- , ,-1. nou Xprlotponote(tot orrlv 5vop(q eivoL o lF-2 : .-t Evorqta 24.4) rot o ow(motlog nopoyowog - - - '.norponote(tot mrlv entpqr<uvoq e(vot o EF-Tu : .- t Evotrlta 24.10). I :cooyowog 6vop[r1g lF-2 tonoOerc[ to evoprcr4pto : '.: llr 0eor1 P. Iplprot((owog 6vo oUtrrnl'oro etdtrd -. -- ",let-tRNA, o lF-z 6too<po).((et 6tr pr6vo to evoprrrl- : : :l"iA ror rov6vo on6 ro rovovtrd oprtvoor<ulo-tRNA .: :--.rerd1et orrlv ow(6poo1 6vop[4q. Awtorp6goq, o EF-Tu, o ono(og ronoefie[ optwooruhetRltlA orn Atorl A 6e ;rnope( vo npoo66oet fMet-tRNAt, to onoio otrveiliq 6e Xp4orgonotefuor xotd r4v enqlrlxuvoq. 'Evog entnp6oOetog SleyXog Yto rrlv oxpiBeto ilpoyUoronoeftot on6 tov lF-3, o ono(oq oroOeponote( qv np6o6eoq tou evopr,criprou IRNA ovoYvop((owoq ro ooor6 (euydpolpro Bdoeov pre rq 6eriteprl r<ot tpfur1 Bdoq rou ro6xovfou Svop(r1q AUG. H EIK0HA 24.17 nopouotd(er tremoprepdlq rn 6to6oXr1 tov yeyov6rtov, ptSoto tcov ono(ov o lF-2 tono0ete( to evopr,crlpro fMet-tRNAr or4 05or1 P. O lF-2, npoo6e6e;r5- voq pe GTP, oMqtrent6pd pre q 06oq P qq 30S unopo- vd6og. Ie ow6 to o4trle(o, q 30S uno;rovddo ;reto<p6pet 6Loug toug nopdYoweq Svop[4q. To fMet-tRNAt npoo- 65vetot orov lF-2 ndvco orrlv 30S unoprovd6o n Snemo o lF-2 prerorpdper to IRNA pr5oo orq pepu<r1 06on P. 24.8 Or Mxpdq @doerq'Evoplnq Ynouovd6eg Avo(nro0v oTo EuKCrpuoTtKd mRNA r 0r europuortr6q 40S ptFoooutr6q unopovodeq npoo66vov- ror oro 5'orpo rou mRNA rot To (oopdlvouv,, 5oq orou evro- nloouv 1uo 0501 5vop[qq. . Mr0 euKopuonul 0601 evopltlg onoretre(rot onrl pto 0trtrr1- trouX[o d6ro vourtreotLd(trlv 1 ono(o nepiloppovet 6vo rotdt- rovto AUG. o 0r 605 prgooourx6q unopovodeg ouvd6ovrot pe ro ofpntroro oq 06o11 evop[r1q. IUpn)toro 30S-mRNA 0 lF-2-GTP ouvduo(eror ge ro ougntroro IuvdSeroL ro evqpKrnpro tRNA Iuvdriovror or 50S uno- govoDeg rr onetreu0ep6vovrot or nqpoyovreq lFl -3 @lJ. IF-] IF-z IF-3 GDP P; l:iliii"lA il.:l 0 lF-2 e(vot onopo[qrog yro qv npoodeol rou fMet-tRNAt mo oupntroro 30S-mRNA. Mero qv npoodeon rou 50S, otror or nopoyovteg lF onetreuOeptitvovrot rot ro GTP dtoonorot. ,*o. tr a *4.* 0rMrrp6g Yno;rovo6eq Avo(nrouv 06oetg'Evop{lq oro EuropuorLrc -l',: 685 Boorreq 6vvorcg rF-3 ffi rF-l v -.{ I tF-2 rS..,
H Svop[q qg prerdgpoonq oro xurrop6n].oopo rov europuordlv prord(er pre 14 Srepyoo(o rolv Borcr4p(ov, oMd q oerpd rov yeyov5rov e(vor dro<poperxrl ror o opr0p6 g rov eTr rKo u pxrilv nopoy6wtrlv preyoL0repoq. Oprog6veg 6rogop5g oXe{(owor pe ro drotpopetrx6 rp6no Ue rov ono(o or BorcrrlproxSg 30S rr or euKo- puortr6g 40S unoprovd6eq Bp(orouv ug 06oerg npSo- 6eoqg rouq npoKerpdvou vo (er<rvqoouv rq perd- qpqon rou mRNA. lroug europuc0reg, ot pur<p6g uno- povd6eg npciro ovoyvtop((ouv ro 5' dxpo rou mRNA rr 6,neno perorrvoriwor Trpoq rr1 06o4 5vop(qg, 6nou ouvoworjv trg preyd),eg unoprovddeg. (Iroug TrpoKo- pucbreg, or ptr<p6g unoprov66eg npoodrivowor orreu- Oe(og or4 05or1 5vop(r19.) I1e66v 6tro to europurolxd mRNA e(vor l.rovoor- orpovxd, oor6oo r<d0e mRNA ouvrlOcog e(vor nofui po- rpUrepo ond ro ovoyro(o yro l1v r<ro6rrono(4or1 pr6vo tqg nporeivnq rou. To prSoo mRNA oro europuronr6 r<urop6nloo;ro 51er grlrog 1000-2000 Bdoerg, rpSper 1-rLo pe0uLr<oprdv4 roLUmpo oro 5'dxpo r<or 100-200 Bd- oerg noLu(A) oro 3'dr<po. H gq perogpo(6pev4 5' ol,LqLoul(o-o64y6g e(vot oXenrd BpoXe(o, ouvrlOcog <100 Bdoerg. To grjr<og r4g r<todrronorrlnr<r1 g nep roXq g ro0o p((eror on6 to g6ye0o g qg nporeffiq. H Un preroqpo(5;revq B'oupd ouxvd e[- vor oprerd porpr6, preprxSg qopdq 5og -1000 Bdoerg. To npcbro XopoKrnpronr6 nou ovoyvropf(eror r<ord q perdrppooq evdg europurotu<orj mRNA e(vor 4 peOu- Luopdvq r<o).Umpo, 11 ono(o oqpro[ver ro 5'dr<po. Ayye- )lorp6po RNA tcov ono(rov ot r<ol6nrpeg SXouv o<por- peOe( 5e prerogpd(owor onoreLeoporrrd in vitro. H np6o6eo4 40S unopovd6ov mo mRNA onome( noL- Lo6g nopdyoweg Svop(qg, ;rero[ri dMrov npu.neiveg or ono(eg ovoyvop((ouv q 6opuj qg xoLrJmpog. Tponono(4o4 rou 5' dr<pou npoygoronote(ror oe 6Lo oXe66v to rurroprrd ror ru<d mRNA r<or efvor ono- pokrlq yto q perdgpoorl roug oro euKopuotr6 xur- top5n).oqro (evcb 6ev efvor ovoyxo(o oro opyov(6ro). H provodxri e[o(peoq ourod rou r<ov6vo o<popd oe ].iyo u- rd mRNA (n.X. rou noLrolori)ro ono(o 6e q6pouv ro).ti- mpo: g5vo owd r' oouvr'1Ororo ru<d mRNA gnope( vo Ueroepqoroiv in vitro Xop(g xolfmpo. Xp4or;ronorotiv pro evo.trl.orcrrrr1 o66, nou nopor<dpmer rrlv ovdyrrl yro r;v r<o).rimpo. Mrilpr rri:po e[erdoope 14 6epyoo(o rqg Svop[r1g oo vo rltov ndwo 6ro05orpq q 05oq 6vop(49. H 6ro0e- ot;r6rrp6 rqg, oo16oo, ev651eror vo neprop((eror un6 deureporoye(g 6op6g. H ovoyvclprorl tou mRNA onor- re( noMo0g enrnp6o0eroug nopdyoweg: 5vo onpowr- x6 puipog r4g Leroupy[og roug e(vor 4 orpofpeor1 rov 6norov deureporoyriv doprcbv oro mRNA (BL5ne Err6vo 24.22). MepLr5g qopdq, ro r<to6x6vro Svop[qg AUG ewoni (eror oe on6oroor; ;rxp6repq on6 40 Bdoerg qn6 ro 5' drpo rou mRNA, on6rc 16oo q r<o).Umpo 6oo rot ro AUG Bp(or<owor prSoo orrlv neproxq np6o6eoqg rou pr- Booropo{ou. Ie noMd mRNA, o016o0, 4 r<oAUmpo xot ro AUG on6Xouv neproo6repo -oe orpo[eg neprntdl- oerg, 5og ror 1000 Bdoerg. H nopouo(o qg ro).0mpog 686 KE0MAI0 24 Merdqpoorl e[oro].ouOe( vo e(vqr ovoyxo(o nporerp5vou vo oxnpo- lore( 6vq oro0ep5 oUpnLoro oro ro6u<6vro Svop[qg. l-1c09 pnopei ro prBoorrlprdlo vo Boo((eror oe drjo 05- oerq nou on6louv r5oo noLri; H EIK0NA 24.18 nopouord(er ro pow6lo "odpoorlq" (scanning model), oriprqovo pe ro ono[o 11 40S unopro- vd6o opXu<d ovoyvcop((er r4v 5' r<olUnrpo xr 6netro "perovooreUer,, rord prlxog rou mRNA. H odpcoorl on5 ro 5'dr<po e(vor ypoprpur<4 6epyoo(o. 'Orqv or 40S uno- povddeg ooprivouv rqv neptoX4-o6qy6, pnopofv vo nporol5oouv rnqn rov qoupKerdrv qg 6eureporoyorig 6oUnq pe oro0ep6rqro <-30 kcal. 0o16oo, rpoupxdreg peyoLUrepq g oro0ep6qrog noporo).ri o uv 11 ovoorflt- Louv rq perovdoreuoq. H perovdmeuoq oro;rord trr6).rg 11 40S uno;rovd6o ouvowqoer ro xo6u<6vro 6vop[qq AUG. Iuvq0rog, 11 npdrq rprnl,5ro AUG mr1 oerpd e(vor to r<ro6rr6vro Svop(qg. Aurq roOouorr1 r'1rprn).6ro AUG, coor6oo, 6ev enopxe( yro vo oroporloer u1 petovdoreuon: ovoyvoF p(eror rog ro6rr6vro 6vop(r1g ;r6vo 6rov Bp(oreror oro oorm6 nl,o(oro. Or orlpowtr5repor xoOoprmdg rou nloro(ou e(vot or Bdoetq orrg 06,oerg -4 Kor + 1. 'Evq ro- 6tx6vro 6vop[49 ;rnope( v' ovoyvrrlp((eror orrlv our1- LouX(o NNNPuNNAUGG. Or tperq Bdoerg noup(vr1g (A r1 G) nprv on6 ro rcodx6vro AUG r<or 11 G opr6oog pre-rd prnope( vo enqpedoouv rrlv onoreLeoponr6rqro rqg perd<ppooqg r<ord 10 eopdq. 'Orov 11 oMrlLouX(o-odq- y5g e(vor ;ror<prd, enrnp6o0ereg 40S unoprovddeg prno- pe( v' ovoyv<op(oouv ro 5' drcpo npotori 4 npc0rrl unoprovddo eyr<oro).e(r,Uer q 05o4 Svop[qg, drlpuoup- ycirwog puo oupd unopovddrov nou npoxope( r<ord pq- Koq rnq oM4LouX(og-odqyoU npoq rn 06o11 Svop(r1q. YcccIccauccMe0utruog6vr1 xotrunrpo 060r1 6vop(r1g : H puprl unopovodo npooddveror oq Ueoutrrcou6vn ro),unrpo p**f d- otr..'-or' i.r-a. ./1- r:, G C C A C CAU G G 2 H puprl unopovobd perovooreuet orn 06on ,.*f...- -^ .^ ^-?-^ ccc$)cnuce 3 Av rl 0trtrn),ouxio-odnyoq efvor poxpro, 0r unoUovoDsq gnope( vo oXqgor[oouv oupd 8*'*@_,-=.=^- ,.a:*:..."r;.1-"i1 eccfi ccc:liljt To europuonro prpootrlporto ;lerovooreuouv o , to 5' orpo rou mRNA oqv neproyrl 6vop[19, q ono[o nepLtrop3':. vet 6vo rodlxovro 6vop[1q AUG.
Iro nto onoteleopottrd petoqpo(6pevo mRNA prdMov toXUet 6rt ru:6tt<6vto Svop[4g e(vot to npdno AUG. Tt oupBo(vet, <11016oo, 6tov pro rptnLdro AUG ev- ron((erot orqv 5'o;rerdqpoo.rn neptoxn; Yndplouv 60o nrOovo( prqXovtqro( 6toguyqq yo 6vo ptBooopdno nou [errv6 q odprrloq oro 5' 6r<po. O nto rol6g e(vot 6rt 11 odptoorl e;rqov((et dtopporl: our6 orlpo(vet 6tt 6vo pt- Booog6tto pnope( vo ouveXfoet ndpo on5 6vo pr1 evop- rcrqpro AUG enetdq 6e Bp(or<etot mo orom5 ntrq[oto. Irq ondvro nep(mcooq nou ovoYvop(oet to AUG, ev65- Xeror vo (erwrloet q petdEpoorl, oLI.d tep;roil(et npw ro ooor6 rto6tr6vto 6vop[r1q rt 5netto ouveX((et q od- poon. Iroug europud:teg, ro neptoo6tepo oupBdwo 6vop[qq nepil.o;rBdvouv odpooq on6 tqv 5'xo].fmpo' Om6oo undpXet 6vog evoLtrolmtr6g tp5noq Svop{r1q, Trou Xprlotponoteftot t6(oq on6 optqt5vo ttxd RNA, oro ono(o puo 40S unoprovd6o ouv66etot oneu0e(og pe Uto eocDreprrrl 06oq yvomri tog eooreplx{ plpootoplx{ 06oq ero66ou (internal ribosome entry site - IRES). Ie out4 rqv nepfmooq, rdOe rcl6x6vto AUG nou tu- 1o(vet vo Bp(or<erot mqv S'opretdqpoorn neptoXrl no- pordpnretot evteL6g. To yvtrlord ototXe[o IRES epr<pov((ouv neptoptop5vrl otrrotroy(o oq flpoq qv oMrl- LouX(o toug. Atorp(vowot rpetq linot ;le Bdorl qv otr- TrqLenidpoo4 rouq pe rq 40S unoprovddo: o 'EvoS rUnog IRES neprdXet to t<to6tr6vro Svop(4q AUG oro ovo6tr6 6pt6 tou. H 40S unopovddo npoo65vetot oneuOe(og oe out6 Xpqorpronord:wog 5vo unoo6vol,o ttov (6tcov nopoy6wov nou onot- to[wot yto rrlv Svop[r1 oro 5' drcPo. o 'EvoS 6Mog tUnog ewon((etot 6o9 100 vour.tr'eorf- 6to ovodtrd tou AUG rot onqtte( petovdoteuoq truog 40S unopovd6oq, nt0ov6v t<ot nd}'t p6oo ev6g prlxovropori odptoo4g. o 'EvqS ondvrog trinog IRES otov t6 qq qno{ttdog C prnope( vo npoodSoet oneuOeiog prto 40S unopovd- 6o Xrop(q vo Xpetd(etot rov6vo nopdyowo 5vop- EnS. H dtodoXrl tolv oupBdwov e(vot 6to<popetrr<t1 on6 r<dOe dLLrl Svop(r1 orouq euKopucireg. Metd t4v np6o6eor1 40S t<ot mRNA, npoo65vetot 5vo oripnLot<o to ono[o nept6Xer nopdyoweg Svop[4q Kor ro evopKrqpto IRNA. H Xpnon trov orotlefo:v IRES 61er t6tofuepr1 or1;roofo orrl l,o(po[q on6 ntropvoi6, 6nou npolroovor<o],U<p0r1- xe, enet6rl o t6g ovom5Met tq pretd<ppoorl tou [evtorrl xorootpS<powog ttg 6opr6q qg xotr0mpog rt ovoor6L- ).owog toug nopdyoweg 6vopfilq nou npoo65vowot oe out5g. 'Evog t6totog m61oq e(vot 11 unopovd6o elF4G (BL5ne Evorqta 24.9), rl ono(o npoo66vetot oro 5' dxpo rou mRNA. 'Etot, q ).oigo[r1 nopepno6[(et q gerdqpooq tcov mRNA tou fevtot4, evrir enttpdnet oto tuxd mRNA vo UeroqpomoUv enetdrl Xp4otpronotoriv IRES. H np6o6eoq oro0eponote(tot m4 05o4 6vop(r1q. M6).rg q 40S unoprovddo evoOe( Ue Uto 605 unopovd- 6o, to nl.rlpeq ptBootoprdrto ewon[(etot orrl 05oq ne- pro1q nou toutonote(tot pSoco npoo6toproprorj npooroo(oq (proiection assay). H 40S unouovoOo noo- ororerier pLo nepLoltl 6otq 60 Booerg: orov c. 6-3 - -- povddeg npoode0oriv oro oJpnAor<o, n npoorcre - -. .-. neproXrl ouoreMetot xot <ptdvet oro prqroq rolv 3l--- Bdoetov nou XopoKrnp((et roug npoxoputrlteg, 24.9 Or Europudreq XpnoluonolotjY 'Evo I0UnAoro lloAAcbv I'lopoY6vrtov'EvopEnq e 0r nopoYovreq Svop[r1g onotrouvtot oo 6tr0 to otoDto t4g Svop[qq, UeroE0 otrtrtov n npdoDeon tou evoprqptou IRNA, 11 npoo6eon Trlq 40S unopovodog mo mRNA, 11 perox(vr1or1 ro- rd prlxoq tou mRNA xor 11 npdodeol rqq 605 unopovoDog' I To tuKopuolrrl evopxttlpro IRNA e(voL 6vo MeitRNA ro ono(o 6Lorp6pet onri ro Met-tRNA nou XpnotUorote(tot otlv enrpqruvol, toordoo 1 peOetovivrl tou Dev e[vot tpopputrttrlg6- vl. * 0 elF2 npoo66vet ro evoprqpto Met-lRNAi xot GTP oxnuort- (ovrog 6vo tptrotoy6q orlpilroxo, ro ono(o npood6verot onlv 40S unopovoDo nporo0 ourl otrtrr1trenr6pooet ge ro mRNA. r'Evo oUuilroro npoodeorlg qg xohtjntpoq npooD6vetot oro 5' orpo tou mRNA nporou outo olJqtrentDpooet pe qv 40S unogovd6o. H 6vop[r1 orouq euKoputi:rcg 61et to (6to yevu<d 1o- porcrrlptortrd Ue rrlv ow(orotlq Borcrqptor<r1 on6 rrlv 6norlq 6tt Xprlorpronote[ 6vo et6tr6 ro6tr6vto Svop- [r1q r<or to evoprcrqpro IRNA. H Svop[r1 oro rurrop6- nLool-ro rov euKclputotcitv lpqotpronote( cog evoprcr(- pro ro6rx6vro to AUG. To evoprcrrlpro IRNA efuot (e- Xoprm6, o).L6 q preOetov(vr1 tou 6e goppru).ttivetot. Ko).e(tot tRNAfl"t. H droqopd preto(ri tou Met-tRNA qg 6vop[r1g Kot rnq entptqruvorlg 6yretrot qnoKlet- orrr6 oro rUnUo tou IRNA: to Met-tRNAi Xpnotuonot- e(ror orqv 6vop[r1 evri to Met-tRNAm orqv entprlruv- on. TouldXtmov 6tio Xoporrqptmtrd e(vot povo6trd yro ro evopKrrlpto tRNAlM"ttqg (U;rqq: 6to05tet Uto oou- vn0rorn tprotoyrl 6oprl ror tpononoekot Ue qooqo- putr(oor1 rnq 2' 05oqq qq ptB6(49 mq Bdo4 64 (ov nopepnodtme( our4 1 rponono[4on, ro evopr<tqpto IRNA pnope( vo lp4ottrronotqOe( mqv entprrir<uvoq). 'Eror, 11 Boox4 opln rnq 6tdrptoqg ;reto[ti Met-tRNA yro rrlv Svop(q Kor rrlv ent;r4ruvot1 6totqpefuot oroug europurilrcg, toor6oo q 6oprr4 gdon qq 6to96pet on6 rqv ow(orotXrlrcov Borr4p(ov (yro oriyr<pror1, BL6ne Et- x6vo 24.15). To eu roputrlttrd rtirropo 6to05touv neptoo6tepoug nopdyoweg 5vop[r1q on6 to Bormripro -o tp51ov rot6- ),oyoq nepDr.oprBdvet 12 nopdyoweq ot ono(ot onot- ro0wqr dpeoo n SpUeoo Yto rrlv Svop[r1. Or nopd- yoweq 6Xouv nop6proeq ovopoo(eg ge toug qw(mot- Xoug Borcrqptot<otig r<ot d4trtilvowot pe ro np60epo "e" ,.,. :: 0t Europutirreq Xprlotponotouv 'Evo l0pntroro I-lotrtlv [looe', :'. * ' Boorxeg evvoteg
.-Met .0, J#p , 4E 4A,. Ir.-nt.'lt JG 3 t^to l .g'kj.-.:u4 E -i. .: I (eukaryotic: euxopuorx6g). Apouv oe 6Ao rq orddro rrlq 6epyoo(og, prero(ti dMrov: . sxrlpqrrop6g ev6g oupnl,6r<ou dvop[r1g pe to 5' drpo rou mRNA . o)C4porroU6q ev6g oupnl6xou pre ro Met-tRNAi o np6o6eor1 rou ouprnl6r<ou mRNA-nopdyowo mo oUpnl,or<o M ettRNAi-nopdyowo o 6reur6luvorl rou prBooro;ror(ou vo ooprioer ro mRNA on6 ro S'dxpo 6rrrg ro npcbro AUG o ovftveuorl qg np6o6esnq rou evoprcr{prou IRNA oro AUG orq 05oq 6vop[qg r<or o dropeooLdpnon yto rq ouv6vooq qg 605 unopovd- 6qq. H Ell(0NA 24.19 ouvor.p((er ro md6ro qg 6vop[qg r<ot defiyer roug nopdyoweg 6vop[4g nou eprn].6Kowor oe xd0e ord6ro. O elF2 pro(( pe ro Met-tRNAi, rov elFO, tov elFl xor rov elFlA npoo66veror mq 40S prBootopu<ri unopovddo oXrlproil(owog ro 43S oriprnl,oxo npo6vop- Enq. Ot elF4A, elF4B, elF4E xor elF4G npooddvowqt oro S'dr<po rou mRNA oXrl;ror((owog to orJpnAoro npSo6eorlg t4g ro).0mpog. Aur6 ro oUpnLoxo oMrlle- nr6pd pre ro 3'drpo tou mRNA pr6ool rou elF4G, o ono[og pre rr1 oepd rou otr].q].enr6pd ge rqv nporefvr1 np6obeoqg rqg nol,u(A) oupdg (poty(A)-binding protein - PABP). To 43S o0prnlor<o npoo6SveL roug nopdyov- teg Svop[qg oro S'dr<po rou mRNA r<ot ovo(qre( ro Ko- 43S odynloxo npoiuup{4q: elF2, elF3, Met-tRNAi, elFl , elFl A 5 I0ptroro np{oieo6 rqq xulrmpqq + mRNA, eIF4A, B,E,G To 43S oulriloKo npooddveror oro 5'dKpo Tou mRNA To 48S o$griloro oXrlgor[(erot oro roDrr6vto 5vop{49: elF2, elF3, elFl,1A, eIF4A, B, F EIK0NA 24.19 Mepxo[ nopdyovreg evopfig npoodrlvovror orrl 40S prpoooprrq unopovodo oXlpori(ovrog ro 43S oUpntroro npo6vop[4q, ev6 otrtrot npood6vovror oro mRNA. Mritrrq ro 43S oupntroxo npoode0e( oro mRNA, ovo(r1re( ro rordtrdvro 5vop[nq xoL pnope[ v' onogovtoOe( og 48S oupntroro. 688 lcaAitl0 24 Merogpoorl I * IVe0erovIvn brf :::r.!l:;rl .,,.,::l Koro qv evop[1 oroug euxopuotreg, o elF2 oX1- porL(er evo tpLroroy6g oupn),oro Ue T0 Met-tRNAi rot GTp. Auro ro rpuoroyeg oupntroro npoodeverot ong etreuOepeg 40S uno- Uovodeq, or ono[eg npoodevovror oro 5'orpo rou mRNA. Apyo- Tspo oTrlv ovr[6poo1, ro GTP udpo],ueroL ror o elF2 one],eu- 0epriveror uno poprpl elF2-GDP 0 elF2B ovoyevvo qv evepyo u0pal 6rr<6vro Svop(qq. Mnope( v' onopovc,.rOei o:q 4gS or-ip- nLor<o 6vop[r19, H unopovddo elF2 e(vot o r<p(orprog nopdyowog yro qv np6o6eoq rou MettRNAi. Awi0eta on6 ro Bor<rr1- pLor<d lF2, provo;.repr1 nporelvq nou npoo66ver GTp, o elF2 e(voL ereporprUepr1g nporeivq nou npoo66,ver GTP, q ono(o onorele(toL on6 o, B r<oL y unoprovddeg, xotrr(o ond rrg ono(eg 6ev e(vot op6).oyr1 pre ro Borcrrl- pLox6 elF2 (B).6ne Exdvo 24.36 o.irlv Evor7ta 24.15). O elF2 e(vor evepy6g 6rov npoo66vet GTp r<or ovevepydg npoodSver duprrrorpoprxq youov(vq (GDp) . H ;,rr';:-, ,r r iri,t nopouotd(er qv npdo6eoq rou elF2-GTp mo Met-tRNA. To npoi6v prepu<5q eopdq xo.eftoL rprroro- y6g orJprn),or<o (ternary complex) (on6 ro rpfo ouorol- r<6: elF2, GTP r<or MettRNA). H ouvoppro).6yr1oq rou rproroyo0g ougn).dr<ou puOpr((eror on6 tov nopdyov- ro owoMoyrlg voux.l.eorr6(ov youov(vr1g (guanine nu- cleotide exchange factor - GEF) , elF2B, o ono(og owoMdooeL GDP pe GTP entrpdnowoq rlv u6pd).uor1 tou GTP ond rov elF2. H : ii',rilrl iri.: I rtopouoro(et rrlv ronoOdrr;or1 rou Met{RNAi ndv<rl orq 40S unopovo6q ond ro rprroro- y6g oUprn).or<o. Mo(( Ue rouq nopdyoweg elF.l, elFlA rot elF3, to rprroroy6g oUprni.or<o oplprot((er to 43S o0pn).oxo npodvopl4q. H ow(6poor1 6ev e[oprdrot ond qv nopouo(o mRNA. O elF3, nou onor-re[ror yro rr] 6toqpqorl ro.lv 40S unopovd6r.ov oe r<ordmooq 6rd- oroorlq, e(vot rivog notr[ peyd].og nopdyowog pe B-10 unotrrovd6eg. O elFl r<or o elF'lA, nou e[vot opr6],oyog pe to Borrrlproxd lF1, go(verot 6tr evrolJouv rnv svep- y6rrlro 6tdoroo4g rou elF3. H r;'::.'-'lil" *irr i''i nqpggstd(et qv Oprd6O torv nopOy6v- rov nou npoo66vowot oro 5'dr<po rou mRNA. O nopd- yowoq elF4F efvoL 6vo npotreivtx6 orjprn].or<o, ro ono(o nepdler rpeLg on6 toug nopdyoweg dvop(r1g. Aev e(vor oo<p6g ov ouvoplloi.oye[ror otg oripn].or<o nptv npoo6e- 0 elF2 onoretre[ror ono o. B xor y unogovo0eq elF-28 ..+ GDP GTP 0 elF2B nopdyer rnv evepyd [opqn rou elF2
elF3 6torrlpe( els00epeg 40S unogovdOeg &*w0 elF2 npoo06vet Met-tRNA mo 40S -rr@-'Ea* 0 elF2 e(vor Pto GTPoorl 0 elF2B eivot o nopoYowoq ovTotrLoYnq ?GDP??GTP i-qr..i :::i,rr,'r ]]:;.J I 0r nopoyovreq 6vop[r1q npoooevouv 10 svopKTrl- rro MettRNA orn 40S unopovodo oXlpoii(owoq evo 43S otlp- roro. Ie preroyeveorepo orodLo 11g ovri6poonq' ro GTP -,0potrueror rt o elF2 onetreu0epdrverot uno Uopqn elF2-GDP 0 =,F28 ovoyevvo Tlv evepYo UOPQ1. Oei' sro mRNA 11 ov ot unoUovddeg npoor[Oewot [eXolpt- cro yto vo olrlpror(oouv to o0pnLoKo ndvco crro mRNA' O nopdyowog elF4F neptdXer rnv unopovddo np6o6e- orlg rnq r<oLJmpog elF4E, r1v e)'trdoq elF4A t<ot trlv rnopovd6o-*trp(tlpto, elF4G. M6).tq npoo6e0e( orrlv xo- i,ln.rpo o elF4A ietuiyeL onorodrlnote 6ewepo-royr1 5o;u1 Bp(or<erot ortq nprrlteg 15 Booetq. rou mRNA' H :vdpyeto yto ro [et[].ry;ro nopdXetot on6tlv udp6luorl .ou ATP. I-1epott6p<o (et[l'Lypo qq 6oUrtq rotd ptrlxoq :cu mRNA enttuyXdvetot on6 tov elF4A po(i Ue rov no- :oyowo elF4B. O r0ptog p6troq tou elF4G e(vot vo ouv- 5eer oLl.o ouorotLrd tou oupnL6rou 6vop[r1q' H unopovddo elF4E e[voL f-ito eor[o yto ptiOptoq H ivepy6rqto trlg ou(dveL pe Qooqopul(trlon, n ono(o nu- :oborettot on6 epeO(oproro nou ou(ovouv rq pretdqpo- lr-l Kot ovoorpeQerot qn6 epeO(optoto nou rnv Koro- :rdMouv. H unol-tovo6o elF4F 6Xet 6pdon xwdorlg nou :tr-locpopulr<ovtt rov elF4E. H 6to0eotp6trlto tou elF4E reyletot en(oqq on6 npolrelVeg, ot ono(eq npoo66vov- 0 elF4F etvot 6vo steporptUepfq nou qnotele(tot qnd: elF4G, pto nPtrlretvn-trp(oUo elF4E, nou npood6vetot ornv 5'Ue0u)tttoU6vrl rotrtintpo elF4A, [to etrtrdorl nou [etulQet qv 5'0ogr1 0 elF4G npoo66vet 6uo efltnpoo0ttouq nopoyovreq 0 elF4B Dreyetpet irlv elF4A etrtrooq H PABP npood6verot orqv 3'notru(A) oupd ',r,l.i ;4 ,1;: To ereporptpepeq elF4F npoo66verot oro 5' orpo :'l mRNA ro06q rot oe otrhouq nopoyovreg nteovdq oMnlsltt6Pdoetg: 0 elF4G nPoo0dverut mov elF3 To mRNA npooDdvet elF4G, elF3 xot q 40S unoPov66o EIK0NA 24.23 Altrntrent6pdoetg olq ono(eg eptilr6rowot nop6- yoweq 6voplng elvot otlgovttr6q 6tov ro mRNA npoo66ver0t oro 43S otipntroxo. tor oe out6v (rotroriwot 4E-8P1, -2 xot -3), nporeqdvou vo epnod(oouvrq treroupy(qtou ornv 6voplt H nopouo(o t4g nolu(A) oupdq oro 3-6t<po tou mRNA 6ieye(pet ro oxnUqrrop6 tou ou;rntr6xou Svop- inq *o 5'dr<po. H PABP npoo66vetot cnqv nproteivrl- ,nptrUo elF4'G rdvowog ro mRNA vo utoOedloet pto r<ur<).rr<r1 6tdto[4, orqv ono(o ouyrpote(tot t6oo to 5' 6oo Kqt to 3'dxpo (Bl,6ne Etx6vo 24'22)' O o141'tott- o'trr6q owrlg qq rLeror4g OrlLtdq drele(pet rn. peroYpo- en Kot n Fnap onotre(tot yt' outrl qv enidpoorl' H irlitlLtt iapoorl PABP/e|F4G n6vco mo mRNA npo6yet rqv entmpdreuon tou 43S oupnh6rou mo mRNA' 6nog en(or1g Kor rnv np6odeoq qq 605 unoprovd6og' H ElK0]'lA 24.23 6eiXvet 6tt ot oMrlentdpdoetq nou epnLSrowot or4v np6o6eon rou mRNA pe-ro 43S oript- nior<o 6ev 6xouv npoobtoptote(nlrlpog, otrLd tpofvetot vo eprn),.5xouv rouq elF4G r<or elF3, to mRNA Kot rrl 40S unopovddq. H unotrlovddo elF4G npoo'66vetot orov etFi ror peooLoBe( orqv np6odeorl trlq 40S ptBo- oto;ux(g unopov66og mov elF4F, 6po t<ot orrlv eflt- *pa-"rotl rnq fio o6pnLor<o' Ouotomtxd, o elF4F ionogt-ti io, "ir+o Stot tirore vo pnope( vo npooeLrU- oet q prtrpr] ptBootogtr<r1 unopov66o' M6irq'n UtKpn unopovd6o npoodeOe( mo mRNA' 1r"-ouo*rltt flpoq to np6to (ouvrl0coq) rtodtr6vto nUO. H odptrloq unoBoq0efuot on6 roug nopdyoweg elFl rot elF1A. Aurrl 11 6epyoo(o onone( 6ondvq ev6p- yetog un6 Uopgd ATP: ouvendlq, nopdyoweg nou oxe- t((owot Ue rnv u6p6truon tou ATP (elF4A' elF4B rot eti+f1, en[or1g dtodpoprot((ouv rdnoto p6tro oe out6 to B,iUo. H Ell(0NA Za.Za Oefpet 6tt rl.prtr<p-q unoprovddo mogord p6Ltg <pt6oet mq 06o11 Svop[r1.q, on6te to evopr,crlpto IRNA (euyoptivet ge.to rro6x6vto 6vop[r1q AUG oXnpmi(owoq 6vo oro0ep6 48S o0pnloro' l-'lp6o6eoq tov 605 unopovddtov mo otipntroxo Svop(r1q 6e gnope( vo ouprBe( npototi onogorpuvOo0v ot elF2 rot elF3' Aw6 dropeootroBe(ror on6 tov elF5 xot ovoyrd(ettov elF2 vo u6poL0oetro GTP tou' H qv- {6poor1 TlpoYgqronoekot ndvto cnrl prtrpq ptBootopur<q unopovddo t<ot npouno06tet (euy6ptrlgo B6oetrlv tou evoprcrlptou IRNA pre to ro6tr6vto 6vop(qq' 'Otr'ot ot u"iio*b, nopdyoweg nt0ov6v onetreuOepdrvowot pt6- trrg oXrlponore( to nLrlpeg 80S ptBoocopdtro' 689 PABP ?4.S 0t Europu6reg Xprlotponotouv'Evo l0pntroro l-lotrtrdrv [10p0'' o';l'J'
0 elFs efldyer rnv udpdluon Tou GTP 0fl6 Tov elF2 0r elF2 ror elF3 onetreu0eptirvowor 0 elF5B 0roueootrope( unopovd6oq EIK0NA 24.24 0t elFl Kor elF3 pol0ouv ro 43S ougnhoKo 6v0p- [19 vo oopriroer To mRNA 6tog rlrou qrdoer o' 6vo rodtxrlvto AUG. 0 elF2 udpotruer to GTP rou, on6re pnope[ v' onetreu0epo- Oer po(r pe rov elF3. 0 elFSB dropeootrope( orqv 6voor1 605-405. T5).o9, o nopdyowog elFSB entrpdnet mqv 605 unopovd6o vo evoOe( pe ro oUprnLor<o oXqprot((owog 6vo nlqpeg p$ooropdtro, ro ono(o e(vot 6roqlo vo (ert- vqoer qv entprlruvorl. H unoprovd6o elF5B 51et nop6- proro otrqlou1[o pre rov npoKopur,nu<6 nopdyowo lF2, nou en(or1g udpoLrSet GTP (nop6Mqtro pe ro p6),o tou orqv np6o6eon rou evopr,mqprou IRNA). M6Ltq onel,euOepo0oriv ot nopdyoweg, ;rnope( v' oMqLenrdpdoouv Ue ro evoprcr4pto IRNA rot ttg ptBo- oogu<5q unoprovd6eg o' 6vov dMo r<6r<i.o 5vop(qg. H unopovd6o elF2 51et udpotr(oer ro GTP rqg, ouvenc0g npdner v' ovoyewrlOe(. Aur6 ennuyXdvetot on6 tov elF2B, Svov GEF o ono(og er<ron((er ro GDP 6rot c0ore vo prnope( v' owrKorooro0e( on6 GTP. H unoprovd6o elF2 onorcl.e( or51o pti0puorlq. l-1oL- L6g puO;tortr5g rrvdoeg enr6poriv orrlv o uno;rovd6o rou elF2. H rpcoorpopuL(rooq nopepno6((er qv ovoySv- vnon rou elF2B o' evepy6 Uopgn. Aut6 neptopi(et tn 6pdon rou elF2B o' 5vo r0xl.o Svop[qg xt 6tot ovo- qr5trl.er q prerdgpoorl. 24.10 O l-lopdyovToq EilrUfiKUVonq Tu Ooprdlver AprvooruAo-tRNA oTn O6on A -' H EF-Tu e[vor 1uo uovoueplq nporelvr; G, rrlg onotog rl evsp- yoq UopAn (npoode5egevl oe GTP) npoo66verot oT0 0ptv00- rutro- tRNA. " To ouUntroxo EF-Tu-GTP-oUrvooruLo- IRNA npoo06veT0t 0Trl 0eo1 A rou prpoorogorlou. M6)lg cnlrlpottore( to nLqpeg prBooc,rptdtto oro ro6t- r6vro 6vop(r19, 6o e(vor 5ror1.to yto 6vov r<Ur<Lo 6nou 6vo ol.rrvooxuAotRNA ero6pXetot or4 06o11 A rou pt- oqv npoodeorl rrlq Boorogor(ou, rou onofou 11 06oq P r<ototroprBdvetot on6 5vo nemrdulo-tRNA. Onoto6rlnore optvoorulo- IRNA errc6g rou evoprcrqptou pnope( vo eto6tr0et orrl 05oq A. H e[oo66g rou 6ropeoo].oBekot on6 Svov no- p6yovro enrp{xuvoqg (elongation factor, oro Bo- rcrrlpro o EF-Tu). H drepyoo(o e[vot nop6proto orouq europud:reg. O EF-Tu e(vot 1-uo e[otpettxd ouwnpn- ;r5vr1 npcoretvrl oro Borrrlptcl Kot to trrmoX5v6pto, o96- Loyrl pe rov ow(morxo euKopuotu<6 nopdyowo. Aw[Oetq on6 tov opr6].oyo nopdyowo orqv Svop[4 (lF-2), o EF-Tu ouv65eror Ue ro prBoorrlprdrro pt6vo rotd 14 drepyoo(o ero56ou rou ol-uvooKul,o{RNA. 'Otov ro ol.uvooxul.o-tRNA Bpe0e( orq 05oq rou, o EF-Tu eyro- tole(ner ro prBooo:prdrro yro vo epyoore( [ovd pre dMo o1-rwoor<ulo-tRNA. 'Eror er<6qlcbvet qv xurilr4 oUvde- orl Ue ro prBooroprdrro ror 6tdorooq on6 out6 nou Xo- porcrqp((er rouq entroupu<oIg nopdyoweg. H EIKONA 24.25 nopouord(er to p6Lo tou EF-Tu srrlv rono0ftr'1oq rou ogrvooKutro-tRNA or4 05oq A. O EF-Tu, provopeprlg npoteiq nou npoo65ver GTP, e(vot evepy6g 5rov e(vor npoo6e6e;r6vr1 oe GTP r<ot ovevepy6g 6rov eL vor npoodedeprdvrl oe dr<pcoogoprrrlyouovfir4 (GDP). To drpep5q o{pntroro EF-Tu-GTP npoodSvet optwoot<uo- tRNA g4rlpror((owog 5vo rproroy6q orl;rntroxo optvoo- ruLotRNA-EF-Tu-GTP nou npoo6Svetot pdvo orn e6oq A oe prBoodrporo, rov ono[cov n eSon P e(vot 11611 rcnet- InUU6vn on6 nemr6ulotRNA. Aurq e(vot q xp(oq-rq ow[- 6poon nou drootpotr((et 6tt to ol.twoorulo-tRNA rot to nemr6utrotRNA e(vor ocoqrd rono0erqpr6vo yto ro 96- prorropr6 nemtdxo[ deoprori. To o;.rwooruLo-tRNA <poptdlvetot orq 05oq A oe 66o srddro. l-lprinov, ro drpo tou owtrro6txov(ou npoo66- vfior orrl 05oq A qq 30S unopovddog. 'Enero, n ovo- yvcirprorl ro6tr<oviou-ovrrro6rr<ov(ou nupo6ote[ gto preroBol,rl orrl 6rog6prpoon rou ptBootogot(ou. Aurq moOeponote( qv np6o6eon rou IRNA xor npoxolei u6p5luoq rou GTP tou EF-Tu. To CCA dt<po tou IRNA getoxtvefuor nL6ov orn 05oq A rrlg 50S uno;rovd6og. To 6rtriep5g oti pnl.oro E F-Tu -G D P onetreu Oeprrivetot. Awrl n Uopqn rou EF-Tu e[vor ovevepy6g t<ot 6ev npoo66vet onoreleoporrd ro opwooruLotRNA. O nopdyowog owoMoyrlg vouxtreott6(trtv youov[- vrlg EF-Ts 6ropreooLoBe( orqv ovoy6wnon rnq Xpqot- ponotrlpdv4g troperlq EF-Tu-GDP orrlv evepy6 popgn EF-Tu-GTP. ['lprirtov, o EF-Ts ereoni(et to GDP on6 rov EF-Tu oXqpror((owog to ouvduool-t6vo nopdyowo EF- Tu-EF-Ts.'Eneno, o EF-Ts erron((etot on6 GTP eTlovo- oXrlpror((owog EF-Tu-GTP. To evepy6 dtgepSg o0;rnLo- ro npoo66ver oprwoorutro-tRNA, evci o onel.eu0epop6- vog EF-Ts prnopef v' qvoruKlo0e(. Yndplouv -70,000 p6pro EF-Tu ovd Borcnipto (-5% rou ouv6lou tcov Borrrlptorcilv npcoteivciv), nep(nou todpt0po trre ro pr6pro opwoor<utro{RNA. Aw6 unodq- Lcbver 6tt to neptoo6repo opr.vooKuLotRNA nt0ov6v Bp(or<owot oe rprroroy4 ori;rnLor<o. YndpXouv pr6vo -10,000 pr6pro EF-Ts ovd r(rropo (nepinou todpt0pto pe ro prBooroprduo). H rtvrlttr4 qg oLlqtren(6poor1g prero[ri EF-Tu r<or EF-Ts uno64].rivet 5rt to otipn].oxo EF-Tu-EF-Ts u<p[ororor pr6vo nopodu<d, Sror c6ore o EF- 690 KEoA Al0 24 MetoQpoon 0r elFl ror elFl A enupdnouv q oopoorl + 48S afgni"oxa '1-.: Boorxeg cworeg
EIK0NA 24.25 To EF-Tu-GTP ronoOere( To oUrvoo- rutrotRNA oro pgoorogdlo xr 6neuo onel,eu0epo- veror og EF-Tu-GDP.0 EF-Ts drogeooiopei or4v ovrxorooroo4 rou GDP ond GTP. H ovr[6poo4 xoro- votrrlver GTP rr onetreu0ep6ver GDP. To provodrrd ogrvooruhotRNA nou de pnope[ v' ovoyvtoprore[ on6 ro EF-Tu-GTP e[vor ro fMet-tRNAf, tou ono[ou I odu- vop(o vo npoode0er ro egnodr(er v' onovr4oet o' eototepud xtrtdtxtivro AUG ror GUG. Tu yprlyopo vo perorpdneror orn Uopqn nou npoo66- ver GTP r<t 6neno o' 5vo tpnoroydg oripnloxo. O p6Log tou GTP oro rprroroy6g o0prn).oro 61er petrerr10e( pe owu<ordoroo( rou on6 5vq ovdl,oyo nou 6e prnope( vo u6poLu0e(. H 14prrq 6voor1 GMP-PCP 61er pto y5<pupo pre0ul.ev(ou mq 05o4 rou o(uy6vou nou ouv66,a rtg B r<or y eoogoprKdg op66eq qro GTP. I'lopouo(o GMP-PCP, prnope( vo ox1lgorrore( 5vo rpno- roy5g oripnLoKo, nou npoodSver o;.uvoorutro-tRNA oro ptBooroprdno. Om6oo, nentrdrr6g 6eo;r6g 6e ;rnope( vo oxrlpor.ore[, ouvencig 11 nopouo[o rou GTP eivor ovoyr<o(o yro rrlv np6odeoq rou oprvooKuLotRNA orr; 06orl A. Y6p6Luoq 6ev onqrefuor npoq ro nop6v. H xrppopuxiv4 (kirromycin) e(vor 6vo owrBrorrx6 nou ovom6l,Ler rq Lenoupy(o rou EF-Tu. M6Lrg o EF- Tu npoodSoer r<ppoprur<fur1, nopop6ver rrov6g vo npoo- 6dver oprrvoor<uLo-tRNA orq 05oq A. C)o16oo, ro o0pntror<o EF-Tu-GDP 6e prnope( v' oneleu0epo0e( on6 ro ptBooropdrro. H ouvelrlg nopouoio rou nopepno6(- (er ro oX41.rotop6 nemrdrrof 6eopori pero[0 rou ne- rrrr6ulo-tRNA xor rou ol.rrvooxutro-tRNA. 'Eror, ro ptBooroprdtro or<rvrlronotefuor ndvrrl oro mRNA npoKo- )rriwog nofoq rrlg prerdgpoorlq. Auq 11 en(6poor1 rrlq Kppopur<fur1q ovo6errvfer 6rt q ovooroLrl ev6g Bqporog tqg petdEpooqg ovoorStrtret xor ro en6pevo Brlpo. O 6yog e[vor 6n 1 ouvepg no- pouo(o rou EF-Tu nopepno6((er ro opr.vooKu).rr6 drpo rou oLuvooKuLo{RNA vo ero6}.0er o-r4 06oq A qg 50S unoprovd6og (BL6ne Eu<6vo 24.30).Iuvenrig, q oneLeu- 05poor1 tou EF-Tu-GDP e(vor ovoyr<oio riore ro prBo- oalpdtro v' ovoldBet ro oxnporropr6 rrov nenrt6rr<rilv deotrrcbv. H 6ro opXrl nopoqpefuor oe dMo orddro r4g perdqpoorlq: pro ow(6pooq np5net vo oLorlqpo0e[ oomd nporoU ougBe( q en6prevrl. H oll.rllen(6poon Ue rov EF-Tu enronq 6ro6pqroil- (er rdnoro p6tro orov 5Ley1o nor6rltog. Apuvoor<ulo- To ogrvooxutrotRNA erodpxeror mn g6on A rrlg 30S unopovddog To CCE oKpo gerorle[ror u6oo orn 06on A nq 50S unoUovo6oq IRNA erodyowor offl 05or1 A lcop(g vo yvrop((ouv ov t' owu<trl6u<6vrd roug roptd(ouv Ue to xrrrdx6vro. H udp6- uoq rou EF-Tu-GTP efuor oXeru<d Bpo6e[o: 6ropre( ne- proo6tepo on' 6, tr onorrefuor yro rn drdorooq ev6g Ld0og otruvoor<utrotRNA on6 q 06oq A: 5ror, ro nepro- o6repo LdOog optvoorutrotRNA orporporiwor oe our6 to ord6ro. H one).eu0ripcDorl rou EF-Tu-GDP pretd qv udp6Luo4 efuor en(or1g Bpo6e(o, ouvsnrig 6noro ldOog ol.rwoor<ulo-tRNA SXer ru16v enrBrciroer pnope( vo 6ro- oro0e( oe ow6 ro orddro. H Boorx4 opXrl e(vot 6tt or ov- n6pdoerg nou 4rnLdrouv rov EF-Tu ouprBo(vouv opr<erd opyd cbore vo enmpdnouv oe Ld0og ollwoorulotRNA vo 6tooro0orjv nporof noyt6ewoUv orq perdgpoorl. Itoug eur<opurirreg, ro oprvooKuLotRNA gSperor mo prBoorol.rdno en(oqq on6 tov nopdyowo eEFlo oe puo ow(6poor1 11 ono(o neprtroprBdver drdonoorl ev6g 6eopo0 uUnInS ev6pyerog oro GTP. 'Onrog ro ilpoKo- purotx6 opr6l,oy6 rou (EF-Tu), efuot 6g0ovq npcoreivrl. Metd qv u6p6Luo4 rou GTP, 4 evepy6g propqq ovo- yewdror on6 rov nopdyowo eEFlBy, nou e(vot ow(- ororxoq rou EF-Ts. 24.11 H floAunenrr6rrfi AAuoi6o Merorp6peror oTo AUrvooKUAo-IRNA -, H 50S unopovodo 6Xet evepy6qro nenldu),orpovoqepoorlg nou nop6Xeror ono 6vo rRNA prpo5v(u;ro" ,, H veoouvrrOdgevrl notrunenrrdLrri otruof60 peroq6peroL ono ro nenldutrotRNA qg Oeoqq P oro oprvoorutro-tRNA qg 0eoqg A. .:. H ouv0eol ro.lv nenlDrrdv Deopdrv nopoyet onoorutrrtrlgdvo IRNA oq 0eor1 P roL nenlDutro- IRNA oq 05o1 A GTP .'d ?r Tu-Ts -.7 O '....- Ts '. a uot or..t iTpuoroyeq f,S..r otigntroro 5 ;::".".', H fiotrunenldLrrl Atruorbo Meroq6peror 0ro AItv0oKLi,,1-:i',- 691 -W...^ -...r"trr,trt' o.tK' u.. Tu-GTP Boorxdg eworeg
To prBoorr:;rdrro nopopr6ver orn 05crq tou, evri q no- Lunemt6tr<f o].uo(6q entprlr0vetor Ue rn prero<popd rou nol,unenrrd(ou nou e(vor npoo6e6ep6vo oro IRNA orq 06oq P npoq ro ol.rloorutro-tRNA orq 05or1 A. H ow(dpoorl nopouord(eror orrlv EIKONA 24.28. H uneriOuvq eevpy5qro yo q o0vOeon rou nemrdu<ori deoproU rqlefuot nenrdutrorpovogepdo4 (peptidyl- transferase). H nemrdulorpovo<pepdoq e[vor ].emoupy(o qq pre- yo).rtq (50S n 605) prBoocoprrqg unoprovddog. H ow(- 6poon nupodotefuor 6rov o EF-Tu oneLeuOepri:oet ro ollwoor<ulu<5 drpo-rou IRNA rou. Irq ouv61ero, ro opr- vooruLx6 dxpo otopefuor oe Uro rono0eo[o r<owd oro drpo rou nemrSuLo-tRNA. Auri n 06on 6Xet evepy6rq- to nemrdulorpovogepdonq, n ono(o onlv ouo(o 6ro- orpoL((er rqv toXe(o pretorpopd rrlg nentr6rr<49 oLuobog oro oUr.vooKULotRNA. T6oo ro rRNA 6oo r<ot ot nporeiveg rqg 50S unopovd6og e(vot ovoyxoleq yr' out4 rqv evepy5r4ro, <oo16oo 11 (6ro 11 6pdon rrlq Ko- rd).uor1g e(vor t6r6r4to rou prBoorrlpxo0 RNA ror rrlg 50S unoprovddoq (BL5ne Ev6rrya 24.19). H gUoq qg ow6poorlg perogopdg onoroLriqOrl<e on6 tqv rxov6rr;ro rou owrBrortroU noupopux[vr; (pu- romycin) v' qvoor5Mer q ;rer6gpooq. H noupoUuKivn protd(a ;r' 6vo qtvo{0 npoo6e6epr6vo or4v re}.xr1 o6e- voofur1 rou IRNA. H EIKONA 24.27 6efyer 6t 4 noupol.ru- r[vr1 61er 6vo N ow[ rou O nou ouv66er dvo opuvo(ti oro IRNA. To owrBrorx6 owrprerton(etor on5 ro prBo- orr:prdrro og eroepX6;revo outvooKuLotRNA: 6netro, ro noLunem(6ro nou e(vot npoo6e6e;r5vo oro nemt6ulo- IRNA petogSperor orrlv oprddo NH2 rrlg noupoprur<(vr1g. H p((o rnq noupopur<(vr;g 6ev oyrupoBole( orq 06- oq A rou prBooropor(ou r<t 6rot ro npot6v nol,unemr6u- Lo-nou po pu r[vq one].eu0epcbveror on6 ro p $ooro pdrro :: ', IxnUortoUoq nenndrrou deogou ou;rporvrr Ur 0v- r(6poo1 pero(u rou notrunenndlou rou nenrr6utro-tRNA orrl 0eor1 P ror rou olrrvo[dog rou ol.uvooru],otRNA orrl Odort A 692 KE0MA|O 24 lvlsToqpoo1 Agwouxulo-tRNA CHI CH. r,r / lloupoUuxrwl ;:i,1,::r.r, li.::': H noupopur[v1 ptpe[rOt rO OprvOOru],0-tRNA, ro- Orirg lroro(er ge opo.rpotro o;uvo[u ouvbedepevo oe pro p[(o oox- Xopou-pooqq. un6 propcprl noLunentrdu).o-noupol.rux(vqg. Aur6g o np6copog repprotro;r6g qg prerdgpoorlg e(vot uneU0u- voq yro rq BorcqpLorcr6vo dpdorl tou owrBroru<oU. 24.12 H Merordnron MeToKtv€i To PrBoourpdrro r H Ueroronrorl rou prpooropor[ou gerouvel ro mRNA dropeoou tou prpootrlpor[ou roro rperg pooerg. r H Ueror6ntorl peronvel ro onoorutrtop6vo IRNA orrl 06o1 E rot ro nentdutrotRNA oq Oeoq P, evo odero(er q 0eorl A. c luUrptrrvo pe ro povr6tro uppr6rrlg rorooroorlg, 11 peror6ntol oupporver oe 6uo orodto, oro ono(o q 50S perorrve[ror oe oX6oq pe rrlv 30S ror oq ouvdXero q 30S perorrve[ror roro prlrog tou mRNA riore v' onororooro0e( | opxurl droprlptprrt- 0rl. O xUr).og rqg npooOqr4g optvo[5trlv ornv ou(ov6pre- vq nolunemtdrx4 o).uo(6o oAox).rlpcbvrror pe rn 1-te- rat6ntoq (translocation), r<o0rog ro pLBooropdrro t- NHz NHz AgtvoIri Boorreg ivuoreg
I npoxope[ rp(o vouKleot(dro xord UnKoq rou mRNA. H Ell(0l,lA 24.28 napouord(er n6g 4 peror6nrorl ono- BdMa ro ogdpuoro IRNA ond t4 06o11P, 6ror dsare vo pnope( vo eroSL0et ro v6o nemrduLo-tRNA. To pr- Booroprdtro 6ro05rer nL6ov pro d6eto 05oq A 6rorprrl ytq rnv e(oo6o rou o1-uvooKuLotRNA nou owtororXe( oro en6pevo xto6u<6vro. 'On<og 6efiyet q etx6vo, oro Borrripro ro IRNA pero<p6petor on6 rr1 05or1 P orq 05oq E (on6 qv ono(o onoBdtrtreror oneu0e(og oro r<urrop6nloo;ro) . Erou g e u rop u cireg ono Bdtrl,eror oneu0e(og oro rurrop6nl.oollo Xrop[g vo preoo].oBe[ ;uo 06oq E. Or 06oerq A ror P e<prnneUouv 16oo q pre- ydl,r; 6oo r<or rr1 trurp{ unopovddo: r1 06o4 E (oro Bo- rrrlpro) ewon[(eror rup[rog orrlv 50S unopovddo, evcil 61er xdnoreg enog6g Kor ornv 30S unoprovd6o. Or neproo6tepeg on6rgerg qXeurd pe rq peror6nroq ot<olou0orjv to 1tow6,lo uBptixqq xatdoraorlq (hybrid state model), oriprgcovo pe to ono(o 4 peror6nrorl npoy- fu'lrr+ rii pri+trinr:ri. To onooxutrtol.rtlvo tRNA geronve[roL orrl 06o11 E, ro nenrrdutrotRNA;leroxrve[ror on1 06o11 P EIK0NA 24.28 'Evo porqpLoro pgoorrrlrolo 6ro06rer rperg 0ri- oerg npoodeorlq yro To IRNA. To opwoorutrotRNA ero6pXeror orrl 06o11 A ev6g prpootrlgor(ou nou SXer nemrdutro-tRNA oq 06o11 P. H ouv0eol nenndrrou deogou onooxutrr6ver ro IRNA qg Oeor1g P xor ono6(6er nenndulotRNA oq 06oq A. H perorontorl UeroKr- ve[ ro onoorutrLrop6vo IRNA or4 0604 E xor ro nenndutrotRNA arl Oeol P. tRNA EIK0NA 24.29 To povr6)ro qg geror6nroqg npore(vouv 6Uo md- Dro. Ilptilrov, roro ro oX4golopo nenudrrou deopou ro oprvooKu- trrxd dxpo rou IRNA oq 06o1 A enqvoronoOerefror orq 06ori P. Aeurepov, ro dxpo rou ovrxoldrrov(ou rou IRNA enovoronoOe- retuor or4 Odon P. trloronoefuor oe 6rjo ord6ro. H EIK0NA 24.29 6e(1ver 6rr npcbrov ouprBo(ver pro geror6nron rnq 50S uno;rovd- 6o9 oe q(6oq Ue qv 30S unopovd6o, nou oroLouOe[- rot on6 ;lo deftepq petot6ntorl 11 ono(o ouprBo(ver 6rov q 30S unoprovd6o geroxtvr;Oe( r<ord pujrog rou mRNA, onoro0rord:woq rnv qp1Kn 6rop6pcproo4. Bd- oq ouroU tou 1.row6l,ou qrov q nopotrlpqoq 6l ro 1.ro- r(Bo rcov enorpd:v nou npqygqronore( ro IRNA pe to ptBooroprdrro (pretpdrwor pe Xqprcl ono{noor1) pero- BdLLeror oe 6Uo ord6ro. 'O'rov npomeOe( noupogur<(vr1 o' 5vo prBootopdrro pe opwoorufutr:pr5vo IRNA orrl 06- o4 P, or eno<pdg tou IRNA Ue rnv 50S unoprovddo geto- BdLl,owor on6 rq 06oq P orrl 05on E, evdr or enorp6g pe rnv 30S unopovddo nopopr6vouv operdBlqreg. Au- 15 uno6q).drver 6n q 50S unoprovdSo 5Xet perorpone( evri 11 30S unoprovddo 6ev 6Xer peroB).q0e(. H epprrlve(o ourc0v rov onoreLeopdrov e(vor 6rt, nprilrov ro opr.vooKultrd drpo rov IRNA (ewon((owor mqv 50S unopov66o) peronvoriwot oe v5eg 06oerg (evdr ro dxpo rov qwKo6rxov(ov nopopr6vouv npoode- 6e;r6vo or' owrrro6u<6vrd roug orrlv 30S unoprovd6o). Ie out6 to ord6ro, to IRNA npoo6Svowor onoretre- ollotxd oe uBprdrxdg nepto16g, nou onorel,oUwor on6 ng 06oerg 50S E/30S P xor 50S P/30S A. Irq ouv6Xero, q petoxfurloq enenre(veror orrg 30S unopovddeg, Sror cirore 4 neproXrl (euyopdr;roroq preto(ti qwKo6rxov(ou- rcodrxov(ou vo Bp(oreror orq oolorrl 05oq. O nr0ov6re- pog tp6nog yro vq 6qpuoupyl0e( 11 uBprdrrcj rordoro- oq e(vor vo petortvrl0e( 11 puo prBooroptrrl unopovdSo oe oX5oq pernv dM4, r<ord rp6no rilore q peror5nrorl H 50S unogovdOo getorwe(tot oq npoq To IRNA nou onoptrrlfire eyrorohelner Drogdoou qg 06oqg E EroepXdgevo oluvoorutro- I To nenldutrotRNA ppioKeror orn 06on P, To oUrvooKutrotRNA sro6pxer0r mn 06on A I 3.1.'i* H Meror6nLoq Merouvefro Prpooirrporto 693 tlpw q perotdnroq:
vo nepilqppdver nporcru<d 6rio ordbro, Ue rnv rovovu<r1 6opr{ rou prBooroprodou v' onoroOrordror oro 6eUrepo orddto. To ptBoorrlprdno owrperconf(er 5vo evdtoEripov 6(- trnUUo r<ord tq petor6nroq. l-'lp6net vo droondoer noL- L6g on6 rg eno<p5g rou Ue ro IRNA cirore vo enmpone( 11 preror<fur1oq. Tour6Xpovo, roo16oo, np6ner vo 6ro14- pqoer ro (euydpropro prero[ri IRNA rot owrro6rrovfou (6toondlwog ro (euydpro;ro rou onooKuhopr6vou IRNA p6vo rq ooorrl Xpovrrri orryUfl. Mro nt0ov6rqro e(vor to ptBoooprdlo vo peron(nrer prero[rJ evoL].orclrdlv, Stoxptrcirv 6ropropg<6oerov. H evoMoyrl 0o prnopoUoe vo ouv(ororor oe oMoyrig oro (euydpcopo rov Bdoeov rou rRNA. H oxp(Beto r4g gerd<ppoorlq eilnped(eror on6 oproprdveg peroMd[erg or ono[eg enrlped(ouv rrg evoMornr6g 6rord[erg (euyopdr;roroq rov Fdoecov. H nt0ov6reprl eppr4ve[o e(vor 6tr q en(6poo4 6ropeoo],o- Befuor on6 q 6rorpoprrrl roX{ np6o6eorlq rov evoMo- rcnrrilv droprop<pciloeov oro IRNA. 24.13 Or llopdyovT€q EnrUfiKUVonq flpoo6dvovror EvoAAoKTtKd oTo PrBooopdrro .. H entprlruvol onotre[ EF-G, nou 6yer doprl nopolroro pe rr1 6op[ rou oupntrorou ol.uvoorulo- tRNA-EF-Tu-GTP. e H npoodeol rou EF-Tu oro pLpoorolrolo onoxtre(er rov EF-G xor ro ovrlorpoqo, = H Uetoronrorl npouno06rer udpotruorl rou GTP, 1 onolo nu- podorel pLo otrtroyr1 orov EF-G, nou Us Trl oerpcr qg nupodo- rei pro otrtroyq oq doprl rou prpooroporlou. H perordnroq onorte( GTP r<r 6vov enrnp6o0ero no- pdyowo enrprqr<uvoqg, rov EF-G. Aw6g o nopdyov- tog e(vor r<tipro ouoronr6 rou rurrdpou: e(vor noptov oe ouyr<6wprooq '1 owrypdqou ovd ptBooroprdrro (20,000 ;r6pto ovd xrirropo). To ptBootripidlo oduvororiv vo npoo65oouv EF-Tu xor EF-G rout6lpovo, ouvenotg 11 prerdrppoorl orol.ou- 0e( rov r<rjr<l.o nou nopouord(eror ornv 6nou or nopdyoweg evoMorrtrd npoo66vowor Kr one- LeuOepo.lvowor on6 ro prBoocoprdrro, 'Erot, o EF-Tu- GDP npdner vo oneLeu0epo0e[ nporoU npoodeOe( o EF-G r<r Snetro o EF-G np6ner v' oneleuOepo0e[, npo- rorj npoo6e0e( ro orjprnLoKo oUrvooKutro{RNA-EF-Tu- GTP. H rrov6rlro r<dOe nopdyowo entprqruvonq v' ono- rLe(er roug dMoug dpoye Booi(eror oe puo oMomepr- rr1 en(6poor1 orrl ouvol.rr<r1 6ro;r6ppr,ton rou prBoorrrl.rotiou r1 oe dpreoo owoyrr.lvrop6 yto enrro).u- m6preveg Odoerg np6odeolg; H nopouoLd- (et pro oouvrl0torq opordrlro prero([ tcov doprltv rou tpLtotoyoUg oupnLdrou rou oprvooKutro{RNA-EF-Tu- GDP roL rou EF-G. H 6opr1 tou EF-G onoprpe(ror 11 ouvoLtr<r1 6opr1 rou EF-Tu nou npoo66veror mo Bpo- 094 KE0A,A|0 24 MeToQpoon r 1," "' ,a; t-Y -- - ^-- Ydootruon GTP rl-lll-trlr + trE I' nnD optvoorutro-tRNA H rppogurivrl qvoordiler Uv onel,eu06proon IUv0eoq nemr0rro0 0eopo0 I r,r ; * Y5ooLuon cTP EF-G/GTP -- EF-c + GDP . . To rpouorbrxo o(u 4rJ ovootetrtret rn5: . onetreu0eoulofl - 7^iq<t, V l;u";-..q,*l +j "s*e. L J'U .F H npdodeol rov nopoyovroiv EF-Tu ror EF-G evotrtroooetor ro069 ro prpootopolo 6dXovror vrlo oprvooru),0- tRNA, oXqpol(ouv nenldrroug deopouq ror peroronl(ovror. 1(ovo uno6oplg rou opuvo(dog ndvo-l oro opuvoor<ulo- IRNA. Iuvenc6g, eUtroyo ouvdyeror ro oupndpoo;-ro 6rr or ouyKeKprg5vor nopdyoweq owoyov((owor yro rnv (6ro 05oq np6o6eorlg (npo<povcig rowd orq 06oq A). To yeyov6g 6rr o Svog nopdyowog np6ner v' onel,eu- Oepor0e( nporori npoo6e0e( o dMog 6roo<potr((er 6tt ro oup.rBdwo rqg pretd<ppoorlq 0o npoyuoronoul0oriv pre rorcrrx6 rpdno. Kor oL 6[o nopdyoweg enrprrlxuvorlg e(vor 1-rovope- pe(g nporefveq nou npoo65vouv GTP, or ono(eg e(vor evepyoi 6rov 5Xouv npoo66oer GTP r<or, ow[Oeto, ove- vepyo( 6rov SXouv npoo65oer GDP. H tpr<proogopu<r1 Uopgn e(vor onopofunrrl yro qv np6o6eoq oro prBo- or,rprdrto, yeyov6q nou 6too<pol.[(eL 6rr r<d0e nopdyov- rog onorrd np6oBoorl oto prBooroprdtro pr6vo nopouo(o GTP, ro ono(o lpetd(rror vo exn).qpd;oer 14 ).enoupy(o rou. O EF-G npooddveror oro prBoocopdtLo dnou npo- dyer q peror6nrorl rr 6neno one).euOepctlveror perd q pretor<(vr;o1 rou ptBoocopor[ou. O EF-G pnope( vo npoo6e0e( mo prBooroprdrro or<6pq rr 6rov ro GTP ov- ttt<otootoOe( on6 GMP-PCP: ouvenotg q nopouofo ev6g vour<i.eort6(ou youov(vr1g e(vor ovoyr<o(o yro qv .j Boorxfg ewong
Agtvooxutro-tR NA- E F-Tu-GTP H Dopl rou TprToToyo0q oupfltroKou Tou 0Utv00KU- , :RNA-EF-Tu-GTP (oprorepo) poro(er pe q doptl iou EF-G (6e- : . 0r Doprro ouvqpqpeveg neptoXdg rou EF-Tu rot rou EF-G .:vi(ovror rorrrveg rot npoolveg. To IRNA Kot I neptoxl flou -- proro(e|. orov EF-G etrovl(ovrot UtrtB. H tporoYpotp[o nopoXtrl- -:-xe ono rov Poul Nissen, University of Aarhus. np6odeorl, rrror5oo 11 u6p6luo4 tou 6ev efuot onolti- to4 onopo(rnrn yro u1 petot6nto4 (nop6Lo nou rl Ue- rm6nrorl e(vor noLU Bpo66reprl Xcop(g u6p6Luoq GTP). H u6pSluoq rou GTP e(vor ovoyro[o yro r4v one],eu06- p<oo4 rou EF-G. H ovdyxq yt' onetreuO6poon rou EF-G ovoroLti<p01- r<e on6 ng enr6p6oerq rou qouotdtroU o[5o9, ev6g ote- poer6orig ovtrBtortroU ro ono[o 'oupnt5(et' to prpooroprdrto orrlv rotdoroorl pretd q peror6ntoq (plene Err5vo 24.30). llopouo(o Eouor6txoti o[6oq, npoyporonote(ror Svog r6rLoq pretot6nto4g: o EF-G npooddverot oro ptBootrrl.t6tto, to GTP udpoLtieror Kot ro ptBoooprdtto petortve(rqt Kotd rp(o vouxtreor(6to. To gouor6tr6 o(0, oo16oo, oroOeponote[ to otipntroro prpooropor(ou-EF-G-GDP Srot drore o EF-G rot to GDP vo nopop6vouv oro ptBoooprdno ow[ v' one].eu0epdl- vowor. 'Etor, ro prBooroprdrro o6uvote[ vo npoodSoet olrwoorulo-tRNA r<or 6e prnope(, nl.5ov vo npoore0oUv 6Mo o;rwo(5o. H perot6nroq e(vot evdoyevrig t6t6r4to tou ptBooto- pcn(ou, q ono[o npouno0Sret Ueyoln petoBotrrl orn 6o- pr1 rou (Bldne Ev6trlra 24.17). Autrl 4 ev6oyev{g pruror6nrorl evepyonore(rot on6 tov EF-G oe ouvduo- oU6 pe qv u6pSLuoq rou GTP, q ono(o ouptBo(vet nptv an6'rn peror6nroq n enmolfvst t4 ;retor(vr1or1 rou pt- fuotopro{ou. I0;rqo:vo pe rov nr0ov6tepo prqXovtopr6, 4 u6p6l,uoq tou GTP npor<ol,e[ pto pretoBotrrl orq 6o;t4 rou EF-G, r'1 ono(o pre q oetpd rqg enttdooet pto oMo- Vn orn 6o;111 rou ptBootoprot[ou. Kotd rr1 petot6ntorl oupBo(ver 6poorx6g ovonpooovoroLto;tdg tou EF-G. l-lprv r4 peror6nroq, o EF-G npoo66vetot r<ord prrlroq rov 6rio prBooopurriv unopovd6tr:v. Ot neptoo6tepeg mog6g rou Ue qv 30S unopovddo 6qpuoupyotiwot trre pro neproXri nou rotrefuor neptoX4 4 (domain 4) Kot et- oXrrlpe( orq 06on A. Aurn 4 nepropl 0o pmopotioe w efvor uneU0uvn yro rnv em6nrorlrou IRNA. Mer614 p- ror6nro4, 4 nepropj 4 npooovorotrf(etor owiOgro, npoq rnv 50S uno;rovd6o. Eur<opurr;nr6 opr6Aoyo tou EF-G e(vot q npo:refv4 eEF2, q ono(o Lemoupye( rotd nop6ptoto tp6no pe Uto rpovol.ordoq e[optdrprevrl on6 qv u6p6].uoq tou GTP. H 6pdoq qg en(o4g ovoorStrtretot on6 to tpouotdtx6 o(ri. 'Evo oro0ep6 otiprn),oxo rou eEF2 ge ro GTP gno- pe( v' onopovoOe( r<or vo npoo6e0ei oe ptBootoprdtto pe enor<6trou0r1 udp6l,uoq rou GTP tou. Mto povodrrrj r6t6qro tou eEF2 e(vot 4 eun60erd rou onl 6up0epmrxrl roq(vn. H to((vq Xpqotponoe( vxo- rwol-rwo-o6evwo-6wourl,eot(6to (NAD) tog oupnopdyov- ro yro vo perorp6per pro p[(o ptBo(u].o-6r<poo<poptr<r1g o6evoo(v4g (ADPR) mov eEF2. H ou(euypr6v4 Svooq ADPR-eEF2 e(vor ovevepy6q orn pret6gpoo4. To un6- orpopo yro rnv np6odeorl e(vot 5vo oouvtl0toro optwo- [U nou nopdyerot trle tponono(r1oq rnq ron6frnlg: out6 ro oprwo[6 e(vot row6 orov eEF2 noMc0v et66v. H ADP-prBo(ul[cooq e(vot unefOuvq yto tq Oovoq- 96po 6pdor1 qg 6r<p0eprrxrlg to((vqg. H ow(6pooq e(- vot eforpenxd onorel,eoprottxrl: 6mo rt 5vo pr6pto ro[(vqg ;rnope( vo rpoTlonotrloer opr<erd p6pto eEF2 nporotrcilwog ro Odvoro tou rurrdpou. 24.14 Tpio Kro6rr6vto Tepuqri(ouv Tn Merdrppoon o To roDrrdvro UAA, UAG xor UGA reppori(ouv 14 perorppoor;. * Iro Borrnpro, ouvnOolq Xprloqonototlvtot pe oXenml ouXvo- qro UAA>UGA>UAG. M6vo 61 rprnl5reg roOop((ouv oprwo(5o. Or un6lot- neq rperq e(vor ro6rr6vto rtpportoprori (termination codons, stop codons) ro ono(o teppot((ouv q pret6- epoon. H g[or1 outtilv trov tptn].et<itv opltrd ono- deffiqre Ue Uro yevetu<r1 6ot<t1loo(o, q ono(o 66r<prve 6rio tUnou g oq ;rerorcj g pretdMo[qg : o Mto orluetorrl pretdMo(q nou pretoBdMet dvo rro6t- r6vro riore v' owrnpooone0et 6vq 6toEopenr6 oprt- vo[ri xo].e[ran Suouvletury (missense mutation). 'Evo opwo[ri owu<oOtord 6vq dI o orrlv nprote(vr1: q en(6poo4 or4 tremoupy[o rrlg npore(vqq e[optd- tor on6 t4v ew6nroq qg pretdMo[nq Kot rn gUon rqg owrrordoroonq rou oprwo[riog. . Mto onUetox4 prerdMo[q nou petoBdMet 5vo ro6t- r<6vro d4puoupydrwog 5vo on6 ro tp(o ro6tr6vto reppronopori rol.e(rot aouv1enx4 petdMaq (non- sense mutation). l-'lpor<oe( np6opo repponop5 rqq prerdgpooqq oro pretoMoypt6vo rto6u<6vto. Iro pe- roMoypr6vo r<Urropo ouw(Oetot p6vo to npri:ro pr5- poq rnq npo:teivqg. H Lemoupy[o qg npotetv4q Boorxdg 6woreq 24"'i4 Tp(o Kodrrovro Tep;roti(ouv rI MeroQpoon 69S
Unope( vo Xoee( (ovd).oyo, rpuorrd, Fe rrlv ew6nrorl rrlg perdMo[qg rord trrr]rog qg nporefvqg). (Iqger6ore 6u o 6pog oouv0eru<6 xro6m6vro -non- sense codon- Xpnorpoflote(ror preprr6q qop6q yro vo neprypdqer rrg rprnL5reg rep;roropro0. Iqv npoyprort- K6Tlro, o ilpoodroprop6q (oouv0elK6,, pdMov nepr- ypd<per qv en(6pooq truog prerdMoEnq o' 6vo yov6ro nopd rq o4proo(o rou rco6u<ov[ou yro 14 pretdgpoorl. O opoq xa6Lx6vLo reppartopol e(vot ro).[rcpog.) Ie xdOe yov(6ro nou 61et o).Lr1).ouXr10e(, 6vo on6 tq KodtK6vro repporrol.rori Bp(oreror oprSoog perd ro ro6u<6vro nou owrnpoorr:nefet ro C-re].rr<6 oprrvo[ri rrlg oLLqLoul(og dyptou r0nou. Or preroMd[erg lrop(g v5qpo unodrll,civouv 6n onoro6rlnore on6 ro rp(o rco- 6tx5vro rcpprotropori enopre( yro vo repgor(oet rn Ue- tdgpoorl o' 5vo yov[6to. Iuven<ilg, or oM4].ouX(eg rov tptntrerc0v UAG, UAA xor UGA e[vor ovoyro(eg xor rro- v6g vo repporioouv q prerdgpoorl e(re Bp(oxowor guorrd oro drpo tou yovrdiou e(re 6qpuoupyor3wot pt6oo prerdLloEnq prioo orr;v rrodrronorqtrrq oLLq- l.ouX[o. Ito Borcrqpror<d yov(6ro, ro UAA e(vor ro nro rorv6 xo6rr5vto reppotroprorl. To UGA Xprlorilonore(ror ne- proo6repo on6 ro UAG, nop6l go(veror vo ouprBo(- vouv neproo6repo ldOq rord r4v ovdyvroorl rou. (Evo )d0og ovdyv<ooqg ev6g rto6u<ov(ou teppotrogori, 5rov oe our6 owrororyrore( 6vo ol.rwooruLotRNA, 61er og onor6Leopo vo ouvelro0e( 4 petdgpoorl 5og 6vo dMo t<ro6u<6vro reppotroproU ri 5rrrq 6rou ro prBooroprdrto qrdoa oro 3'dr<po rou mRNA, yeyov6g nou ev66Xeror vo nporol6oet dtrl,o npoBLrjporo. l-t' outq qv nep(- oroorl, ro Borrqpro 6ro0rftouv 6vo er6rx6 RNA.) 24.15 To Kto6rr6vto T€ppoTtopo0 Avoyvtrlpi(ovTot on6 npulreTvrro0q llopdyovreq e To rto6rrovro repgortopo0 ovoyvrop((ovror ono nporreiroug nopoyovreg onetreu06ptoor1q, 6Xr on6 optvoorutrotRNA. e 0r dop6q rtov nopoyovrov one),euOepotonq Toqnq 1 ;roLd(ouv pe lg ovrlorolxeq doUeq rou oupn],orou oluvoorutrotRNA- EF-Tu xor rou EF-G. * 0r nopoyovreg onetreu0Sproor;q ro[r1g 1 onovrouv oe erDrrd rodrrovro reppolopou ror u6potr0ouv ro Deopri no),unenlD[- OUTRNA. * 0r nopoyovreg oneLeu06prrlolq ro[r1q 1 unopol8o0vror ono roug nopoyovreg onetreuS6poorlq ro[qg 2, or ono(ot e(op- r6vror ono GTP. * 0 Urlxovrogrig ervor nop6porog oro porr1pto (ro onoio dro06- rouv 6uo r0noug nopoyrlvrolv onetreu0epoolg ro[r1g 1) roL roug euxopudlreg (or ono[or 6Lo06rouv govo 6vo nopoyovro onetreuOepooqs roEriq 1 ). Irov repporropr6 rrlg prerdgpoorlg eprnl6xowor drio ord6ro. H (6ro 11 aw[6paor7 rou repparopori ouv(oro- ror oe oneLeuO6poon rnq nporeivrrqg oLuo(6og on6 ro reLeutq(o IRNA. H aw[6paor1 petd rcv rcppauop6 ouv(ororor oe oneLeu0Sptrlo4 rrov IRNA ror mRNA, xoOrig Kor oe drdo"rooq tou prBoorr:gor(ou mrg uno- ;.rovddeg rou, Kovdvo rodtx6vro repUol.ogoU 6ev owrnpooo- neUeror on6 rdnoro IRNA. Aemoupyo6v eweLrig 6ro9o- pe'nrd on5 ro un6Lornq rco6u<6vro Kor ovoyvop((owor oneuOe(og on6 nporeivrrorig nopdyoweg. (H owi6po- or1 6e Boo(eror oe ovoyvrirproq ro6rrovlou-owrxcodtxo- v(ou, ouvenc0g 6ev e(vor oogSg yror[ onorte[ror 4 oL}.qLoufo puog rprnl,Srog. Ilt0ov6v ow6 owovoxM qv e[5]l[4 rou yeverrKo0 rri6tro.) To rcoSrx6vro repUqrropro0 ovoyvrrlp((owor on6 roug nopdyoweg onetreuoSpcoonq (release factors, RF) rd[49 1. Iro E coli,60o nopdyoweg one],eu06pr* oqg rd(r1g 1 eivor erdrrof yro 6ro<poperrx6g oM4trou- 1(eg. O RFI ovoyvcop((er to UAA r<or UAG, evc0 o RF2 ro UGA ror UAA. Or nopdyoweg 6pouv orrl pgooopr- K4 06orl A xot ononofv notrunemrdu],o{RNA orn eSon P. Or RF undplouv oe nolti Xopr4L6reprl ouyr6wpoorl on6 roug nopdyoweg 6vop[r;g 11 enr;uir<uvoqq: undg louv -600 ;r6pro rd0e nopdyowo ovd rfrropo, noo6 too6rivol.ro pr'6vo pr6pro RF ovd 66xo prBooroprdtro. K6- nore nt0ov6v unrlpXe pr6vo 6vog nopdyowog ondeu- 0dpoor1g, o ono(og ovoyvdlpr(e 6Lo tq ro6rx6vro tepporrol.rori, ror opy6tepo e[e],ffiqxe oe 5uo nopd- yoweq ge er6r161410 yro ouyKer<prpr5vo rco6rr<6vro. Iroug europucbreg, undpXer pr6vo Svoq nopdyowoq onel.eu05proons rd(nq 1 nou roAe(ror eRF. H onorele- oponr5qro pe rrlv ono(o ovoyvop((ouv r' ow(mor1o xro6rx6vto-or61oug ot Borrqproro( nopdyoweg eflrlpe- d(eror on6 rtg Bdoerg orqv 3'06o11. Or nopdyoweg oneLeuO6poonq rd(rlg 1 unoBoq- OoUwor on6 roug nopdyoweg one),eu05poonq rdqnq 2, ot ono(or dev e(vor erdro( yro ouyKeKprprdvo xo6rx6- vto. Ot nopdyoweg td[qq 2 e(vor nptorefveq nou npoo- 65vouv GTP. Iro E. coli, o p6trog rou nopdyowo td(r1g 2 RF3 5yr<enor oe oneleu05pcoorl rou nop6yowo rd- [r1g 1 on6 ro prBoorrlpr&ro. O RF3 e[vor nptore'lv4 nou npoo66ver GTP, 11 ono(o oXet((eror. Ue roug nopdyov- teg enqlrlxuvorlg. l-'lop6t o yevu<5q pr4Xovropr6g rou repUorr.opori e[- vot nop6goroq orouq nporopuc0req Kor rouq europucir- req, or ol,trq).ent6pdoerg prero(ri rcov nopoy6w<ov rd[rtg 1 r<or rd(49 2 5Xouv xdnoreg 6ro<popdg. Or nopdyoweq rd(r1g 1 RF1 xor RF2 ovoyvtopf(ouv ro r<odr6vto repprouoprori Kor evepyonororiv ro prBo- oopdrro oe u6p6Luoq rou nemr6ul,o-tRNA. H 6rdono- on rou noLunemt6(ou on6 ro IRNA npoyproronoefuor pe puo ow(6poor1 ovdloyq pre q ouvq0ro;r6v4 prerogo- pd nenrt6uLoprddov, p' e(o(peo4 6rt 65rcrr1q e(vor ro vep6 ow(rou ol.uvoorulo-tRNA (BI5ne Err6vo 24.34). Ie ow6 ro oqpre(o o RFI r1 o RF2 onetreuOepdrveror on6 ro prBooroprdno ;.16oo rou nopdyowo rd(qg 2 RF3, o ono(og oXer((eror Ue rov EF-G. O RF3-GDP npoo66veror oro prBoocol"rdno npw npoyporonot40e( r1 696 KEOAAAIO 24 Merorppoorl Boorxdg evuorcg
ow(6poon rou repportogoU, ev6 ro GDP owKoetord- tqr on6 GTP. Aur6 enrp6net orov RF3 vo 6tr0a o'eno- q{ Ue to ptBooto;rtr6 r6wpo trlq GTPdorlq, 6nou nporotre( rrlv ondreu0Spoorl tolv RF1/2, 6tov teppott- ore( 11 noAunemdxrl aAuo6a. O RF3 pot6(er ile rtq neptoXdq nou npo-o66vouv GTP orouq EF-Tu rot EF-G, evri ot RF1 t<ot RF2 pe qv C-teltr( neptoXd tou EF-G, to ono(o ;rq'refuot to IRNA' Aur6 uno6qtrrilvet 6tt ot nop6yoweg onetreuO5poonq ipno,uo"o,otiv tr1v (6to neproXq 4 ono(o.lpqottrlonoteG i'1,'onO touq nopdyoweg ent;r{xuvorlq' H E|(ONA 24'32 qvoseKv0et in tsooxn opXrl 6tt 6trot quto( ot nopdyov- reg SXouv t4v'(6to yevtrrl popgo}'oy(o xot rpoo66vov- .o,'*l p$ootoprdtto 6to6o1tr<d orlv (5to 06o4 (Bootxd' orrl 06o4 A ri oe ;rto neptoXq nou entrol'0rrerot eKre- vd:g pe q 06on A). d "rnoprr-rr<6q nopdyowog onetreuO6pcr:oqq to- EnS 1 eRFi e(vot 1.tto provodrrl npcotetvq q ono(o ovo- yvcop((et Kot ro tp(o t<to6tt<6vto teptrrottol'toti' H oLLnLouXfo tou 6e olet((etot Ue rouq Borcrrlproxo6q nop&yo*tq. Mnopef vo tepprotioet q letd<ppoorl in vltio Xop(q'rov nopdyowo td[4g 2 eRF2, nop6tt o eRFz'iivor'qnopofurltog aroug (upro;rUxrpeg in vivo' H 6opr( rou eRFl orotrouOe( pro ou<e(o Booxrl opXrl: 11 El- ioiil za.aa 6eftvet 6tt onoteLefrot on6 rpetg neptol6g' nou gtgo6wot rn 6opri tou IRNA' 'Evo Boorr6 lro.iBo tptdrv opwo(6cov,.GGQ, ercr(0e- rqt offlv r<opu<pri rrlg neptoXrlg 2' H ew6ntoti tou orrl 06on A owimotle( orq ouvr10r1 tonoOeo(o rou qptvo- [5og o' 6vo o1-ttvoorutro-tRNA' Iuvencirg, prnope( vo XcTlotUonoe( rq ytr'outopr(vn (a) Yto vo tonoOete[ to ,epO oe gSon xqrdl 'nLq d:me v' owtro0tmd to o1'tt- EIK0NA 24.33 0 euxoputonr6q nopdYovroq teppottoltof eRFl exer 6oUn nou ptpe(tor ro IRNA. To potipo GGO ornv oxpn rnq fle- pioXRS 2'etvot onopo[rr1to yto qv udpo]ruol qg no]runentdxnq olroiOos ono to IRNA' H rpotoYporpio nopoXtrlpri0r6e ond tov David Barford, The lnstitute of Cancer Research' vo(ri tou optwoot<u)to-tRNA orqv ow(6poor1. trlq Tlemt- dutro-rpovorpep6onq' H EIK0NA 24'34 ouyrpfuet rrlv ov- t(6pooq repUortopoti pre tq ouvri0n ovt(6poor1 geto<popdq nenrt6(ou' O tepgotropr6g petoq5pet gto uopoiuiop66q on6 to vep6, u6potrtiowoq 6tot onote- lleptopl 1 (orpo 0doq P nerntdtml oluo(60 I H-N .-, H-C,-R / H-C;R o"''{ g'''b IRNA r IRNA Inennorrn duo(60 H-N H-C.R ,lH o'" ' N' H-Ctn H ,Co 000 i*,ro ,**o I nemrdrtl atruo(60 H.N H-i-n I o'"'oH CH I cll I uuu I I eRFl r ,'a-l , m #ffnnr v :,rri;! ?,'i.::r H poptorl onopiprl0rl entrp6net oro ougn)'oxo :F-Tu{RNA, otov nopoyovro perorontorlg EF-G rot orouq nopo- , ovreq oneheuOepuronq RFl/2-RF3 vo npoobtivovrot oqv [dto pt- :coo.r1-LLrr1 neptoxrl. 0 RRF e(vot o nopoyovtog ovorurtroorlg rou :Lpootogor[ou (ptr6ne Etrovo 24 35). :1{Sl'JA :4,3{ H ;reroqopo rou nenrtdfou xot o repportopdg e[- voL nopopoteg ovrtdpooetg, ortg onoieg Uto Boorl oro r6vrpo qq nenlDu),orpovoqepoonq nupo0oreI pto ovr[Dpootl trans-eoTep0- noirlorlq peoo dtoonoolq evoq 6eopo0 N-H n 0-H onetreu0epot- vovtog N n 0 Yto vo dLoonooet ro 6eopo ge ro IRNA ?4.15 To Ko0Kovro TeppOrtOporj Avoyvtrlpt(ovrot ono [lptlreivtrou-c l-lopoyovreq 697 5ruilnxuvcI K6vtpo nenrLdutro- lp0voq0poqq Teili-rn'iiti;*r, K6wpo nenttDutro- rpqvogepdonq DUUII' or$r P ).'r$ /,,'.o'0'-'q ,
Aeoporrrd ro Seopd nennd(ou-tRNA (Bl,5ne ELx6vo 24.48 yto to ncbg ),enoupye( ro rSwpo rqg nenndulo- rpovogepdorlg). MeroA.d[erg oro yov(6ro RF e].orrdlvouv rrlv ono- rel.eopotLr6rrlro rou repporro;rori, 6notg Srqnror6ve- tor on6 rqv ou(qprdv4 rr<ov6r4ro ouv5Xtor;g rrlg prerdqpoorlg n6po on6 ro ro6u<6vro repprotroprori. H unepSr<ppoorl rou RF1 4 rou RF2 ou[dver qv ono6on- x6qro rou replrouopro6 oro rodrr6vro oro ono(o enr- 6porJv. Aur6 uno6rlLriver 6rt 4 ovoyvriproq rou rodu<ov(ou on6 roug RF1 r<ot RF2 owoyrov((eror pre r' opuvoorulo-t R NA nou ovoyvop ((o uv ).ov0oo;uivo ro rrodu<6vro repporrol-rori. Or nopdyoweg oneleu05pco- onq ovoyvop((ouv opr<etd onoreleoporrrd trg ow(- ororxeq oMql,o u1(eg-or6xo u g. H owbpooq rcpprorrollori neprLotrrBdver oneLeu06- poorl rou oLoxlr;poprdvou noLunemr6(ou, oMd ro'ro- Lo(ner 6vo onoorulrrogrlvo IRNA r<qr to mRNA ouv6edeg6vo oro prBoorrlprdtro. H Ilt(0NA 24.35 6e(X3rer 6rL 4 6rdoroon rov un6i.orncr:v ouororrrcilv (IRNA, mRNA, 30S ror 50S unopovd6eg) onome( tov nopdyov- ta avaxlxiaorlg rou ptBooaparbu (ribosome recycling factor - RRF). O RRF evepye[ po(( pre rov EF-G oe Uro ow(6pooq 11 ono(o Xprlorponore( qv u6p6Luon rou GTP. Avo<popu<d pre roug dMoug nopdyoweg nou eU- nA6rowor mrlv oneLeu0Spcooq, o RRF 51er nop6proro 6oprl pe to IRNA, oMd 6e 6ro0fter too6rivol-ro pe qv 3' neproXrl np5o6eoqg rou oprvo(rlog. En(oqg onome(tot o IF-3. O RRF enrdpd m4v 50S unopovd6o, evcb o lF-S oqorpe( ro onooruLropr6vo IRNA on6 rqv 30S unogo- vd6o. M6).rg 6roXatprorofv or unotrrovd6eg, o lF-B e(o- :a t: i: ;.:. .:l:tifiiti:,:-,-, :lt i)tia? xol,ouOe( vo Xpeto(eror yro vo nopepnod(oer rrlv eno- voorivdeo4 roug. H ouyr<p(ver rrg opro).oy(eq oq npoq rn Letroupy(o Kor rlv oMq).ou1(o rov npoKopurotrrtrtv r<L europurrltrxrJtv nopoydwrov pretdqpoorl g. 24.16 To PrBootrll.lK6 RNA Aronepvd Kot Ttq Ado PrBootllUtKdS Ynopovo6eq e Ko0e rRNA rp6per notrtr6g droxpn6g neproX6g nou dlntrtirvov- ror ove[dpqro, r lxeddv 6treg or prpooroprr6g nporetveg 6pXovroL o' enorprl ge ro rRNA. e 0t neprooorepeg enoq6g pero[f rov prpootoprrdrv unopovo- Dov npoyporonorouvror UeToEu 165 ror 23S rRNA. To drio rp(ro qg pd(og rou BorcrqpLoxori prBoocopor[- ou onoreLoUwor on6 rRNA. H nro nfulpogopror<r1 trr5- 0o6o9 yro rnv ovd).uoq qg deureporoyo0g 6oprnq rcov preydLcov RNA efvor q oriyrproq rov oMq],ouXuilv ttr;v ow(ororlrov rRNA oe ouyyeve(g opyovroprorig. 'Ooeg neprol6g e(vor oqprowrrcig yro rn deweporoyrl 6oprl 6toqpo0v qv u<ov5rrlro v' oM4trenrdporiv 96- oo (euyopc0goroq rcov Bdoe<ov. 'Eror, ov 5vo (eriyog Bdoecrlv e(vor onopofrr1ro, prnope( vo oxni]orrore( or4v EIK0NA 24.35 0 nopdyovrog onetreu06poor;g (RF) reppor[(er q peroqpooq onetreu0ep6vovT0q Trlv flpo- reivxrl otruo[60. 0 nopoyovrog ovorurtroorlg rou prpo- oopor(ou (RRF) onetreu0ep6ver ro retreuro[o IRNA rot o EF-G onetreu0eptiver rov RRF nporotrdrwog dtdmo- or1 rou prpoorlporiou. q1 -#qD@ -P ? F*-sruF*"6-B< 698 KEOAAAI0 24 l/leToqpool Boorreg ewoleg #
) 1::,ii: r,--.:':1.,':: . ,:,:. tF-1 tE o*t tF-3 flporopu6reg EF-TUt, EF-Gt trE T. EF.GS [6to q4enrcj 05or1 oe r<dOe rRNA. Aur{ q pr60o6og 6Xet enrpSQer vq KorooKeuooroUv l,emoprepq prowSLo 16- oorou 165 6oo Korrou 23S rRNA. Ko0dvq on6 ro r<6pto rRNA prnope( v' ono6o0e( pe pro deuteporoyrl 6opn pre noM6g 6tor<prdg neproX6g. T6ooeprg yevu<69 neprol6g oXq;rot(owor on6 to 165 rRNA, 6nou nep(nou or puo6g Bdoerg qg oM4trou{og e(vor (euyopropr6veg (BL5ne Etr6vo 24.46).'E[t yevr6g neproXdg oglprot((owot on6 to 23S rRNA. Ot [eXopt- m6g 6(x).roveg neproX6q ouvrl0oq e(vot BpoXe[eg (<8 bp). :uXyd or drnl.ooroopr6veg neprol5g 6ev e[vot t5treeg xor neptSXouv npoe(o16q pre o(euydptoteq Bdoetq. Iuy- xp[oqro prow6].o 61ouv oXedtoore( yo to l.utolovdptord rRNA (ro ono(o e(vor BpoX0rcpo rot nept5louv Lty6re- peg neproX6g) Kqr ro euKopuorxd rurroponlool.tolrcd rRNA (to ono(o e(vor ;ror<pritepo xot 6to05touv neproo6- repeg neproX6g). To peyo).Utepo ;rqr<og rov euKopuorrt- xriv rRNA rup(cog ocpeiletor otqv on6rrqoq ol),q- )'.ouXuiv nou owrnpoocone6 ouv enrnp6ooeteg nepto16g. H xpuorotrrrn SoUn rov prBoorollodrov 6e(1vet 6r oe xdOe unopovd6o ot neprol6g tou r0ptou rRNA 6tn).d:- vowor ove(dprrlro Kr 51ouv 6toxptr6g tono0eobg. 'EXouv Fpe0e( 6rotpop5g mqv u<ov6tqto tou 165 rRNA v' owrdpd ;re X4pur<o0g nopdyoweg 6rov or 30S unopovddeg ouyrp(vowor pe rq 70S ptBootopdtto: undpXouv en(o4g StoEop5g ;reto(rj trrlv etre00epcov pt- Booropor[ov rr ere(vov ilou oupperSXouv mq prerd- ------l------..* - r ^ ! elFlA ! Anogpdooet q gdol A ! 0 elFl A ^unopoq0ei Tov elF2 vo npotoOtl_oet nlv npdoOeon tou Met- i tRNAiMet oro 40S. Entonq npooyer rn dtdotoorl Tov unopovo0ov elF2, elF3, elFsB* I ffooOoq rou evop*nptou IRNA | 0 elF2 eivot Uto GTP6on . 0 elF3 oreyerpet ro oXlgolopri rou TprloToyouq oupnldxou, qv npdodeon Tou 0To 40S, r<006q enlolq qv npdodeorl xot oopcoorl rou mRNA elFl, elF4 oripntroro, , lpoo6eon rrlq ptxprjg 0 elF5B epntrSrerot oqv e(oo6o Tou evopKnlptou tRNA t<ot elvot elF3 ; unopovobog oro mRNA ltto GTPdorl i , !--,+ i;=* i ; I eEFlo+ i npooDeon GTP i I eEFlp, eEFly I AwoMoyrl GDP ! i eEF2E I Merordnrol rou prpootogoriou i ' t -_-L-*ffi ---4 i flpoxopudrcq i Europudlreg RF1 : eRFl , Avoyvrirprorl UAA/UAG : : Rtz eRFl Avoyvoprorl UAA/UGA RF3- eRF3 Ardyepon otrtrou Ctov) RF(s) *OL lF-2 xot elFSB dXouv ouotroyio otrtrr1trouXiog -01 lF-2, EF-Tu, EF-G rot RF3 dxouv ouotroy(o otrtrr1trou1(oq '01 EF-Tu roL eEFl o dXouv ogotroy[o otrtrntroux(oq r0L EF-G xot eEF2 6Xouv ogotroy[o otrtrntrouxioq Aeuoupyrreg oUotroy(eq Tov np0Kopuolr6v rr europurrrur6v nopoyovTolv Tnq UeroQpoonq. epoon. MeroBoLdg mqv owrdpoortr6rlto tou rRNA ouprBo(vouv xord rlv np6o6eoq tou mRNA, roJV uno- ;rovd6ov r1 rou IRNA. Oprop5veg ;retoBol6g owtKoro- nrp((ouv puo dpreo4 olL4l,en(6poorl rou rRNA pe to mRNA r1 to IRNA, evdr 6LLeg npoxolotiwot Spgeoo on6 dMeg peroBoLdg orrl 6o;tq tou ptBoool-tor(ou. To r<ripro or1;re(o e(vor 6n 1 dropr6prpcoon rou ptBootol.totl- ou e(vor eudtrucrq rord t1 perdgpoorl, t6[oq qq UtKp]iq unopovd6og, enet6rl np6net vo etr5yXet qv orp(Beto tou (euyopcirporog rr.r:6txov(ou-owtrro6u<ov[ou. 'Evo lopom4ptonr6 rnq npororqyoUq 6oprrlg rou rHNA e(vor 11 nopouo[o pe0u].topr6vov roto].o(ncov. Yndplouv -10 pre0ulogddeg qro 165 rRNA (r<up(tog npoq ro 3'drpo rou prop(ou) t<ot -20 oro 23S rRNA. Ito rUrropo rov 04trootrrirv, to 18S rot 28S rRNA peto- <p5pouv 43 t<ot 7 4 pre0uLopd6eg, owtoro(Xog: ouvencbg -2Y" rav vourl.eort6(ov e(vot preoutrropr5vo (nep(nou rprnldoro noooor5 ouyrprnxd pre ro Borcrrlpto). H peydLq prBooroprr4 unopovddo en(oqg nepulXet 6vo pr6pro 5S RNA 120 B6oeov (oe 6Lo to ptBootoprdtro ercr6g tclv ;.utoXovdpror<cbv). H oMqLouX(o tou 55 RNA e(vor Lry5repo rol.d ouwrlprlpr6vrl ouyrprtrd pre rg otr- Lrlloulieg rov r<ripuov rRNA. 'Ol.o to ;t6pto tou 53 RNA ep<pov[(ouv pro 6o;14 pre (euyopropr5veg Bdoetg oe pe- y6l.r16mooq. Iro ptBoocoprdrro tou euKopuortKo0 xurroponl,6- opqroq, 5vo dMo pu<pd RNA Bp(oreror orq preydtrrl .:,,,r To Pipoorr,rptro RNAAronepvo rotrtgA0o PtpootopLxec Ynl-:,::.: 6gg l-evml Aenoupy(o
AUtroro ntrorioro os RNA Ell(0llA 24.37 H 30S unopovddo 6Xer pro Ksqqtrn nou 6roXop(e_ r0r ono ro orirpo ond 6vov ouX6vo rr en(o4g dLo06ret pro npoe[ri_ Xouoo e[6dpo. unopovd6o. flp6reuor yro ro 5.gS RNA, rou ono(ou 4 oMrlLouX(o owrororle( cro S'dr<po rou npoKopucrlrxorj 23S rRNA. O ptopr6veg prBootrrplr<dg npoteiveg npoodrivowor tqXupd oe onopovo;rdvo rRNA. 7Meg 6ev npooddvov_ tor o' eLeU0epo rRNA, oMd onorrorjv ourrl t4v rrov6_ ulro og6rou npoo6e0oUv dtr)eg nporeiveg. Aw6 uno_ bql,d;ver 6rt 4 6ropr6peoon rou rRNA r<o0op((er ov undpXouv neproX6g np6o6eorlg yro opropdveg nprorei_ veg. Ko0c0g npoo66vetor rd0e nporetvq, endyer peto_ BoLSg 6rop6pqoonq oro rRNA, or onofeg enmp6nouv r4v np6o6eor1 dL),rov nporeivdrv. Iro E. coli, oXe66v 6Leg or 30S prBoocrtpu<dg npoteiveg oAL4lenr6poriv (prolov6tr oe noxfi.oug Bo0potlg) pre ro 165 rRNA. Ot neproxeq npdodeoqq ndvtrr oflg nporetveg e;rgov((ouv eu pe(o no rril,(o Soprr<civ XqpqKrn p rortrcilv, yeyov6 g nou uno6rll,cbver 6rt or ptlxqvropo( ovoyvriproqq npo_ reivc0v-RNA prnope( vo 6rog6pouv. - _lo Z0S prBoocoprdno 6Xer oo0pr;rmpq xoroor<eur1. H Ell(0NA 24.37 nopouord(er pto qXrlpronrrl bop4 r4g iOS unotrrovdSog, 11 ono(o unoErotpefuor oe r6ooeprg nepro_ X6g: r<ego),rg, ou15vog, ocilpo ror e[d6po. H ElKOl,lA Z4.gg nopouoto(er pro nop6proro oneu<6woq qg 50S unopo- vddog, 6nou ro 6rio rUpLo XopoKrrlprourd e(vor 11 xev_ tpxrl npoe(oXn (6nou ronoOerefuor ro 55 rRNA) xor o Urq(oq (ouw(Oeror on6 noMon).d owrypogo rnq npo_ retv4g L7). H EIK0NA 24.39 deipet 6n 4 e[d6po (ptat_ form)_qg UrKpnq unoprovd6og egopp6(er prdoo or4v npoe[op1 (protuberance) rnq ;reyd].qg unopovd6og. Yndpler truo r<od,6r4ro ;rero(rj rov unopovd6cov, r1 ono(o neptdXer opr4r6veg on6 ng oqprowrdg 06oerg. H 6oUn qg 30S unopovddog oro).ouOe( t4v opyd_ voorlrou 165 rRNA pre rd0e 6optr<d Xoporrqproru<d v, owrororle( oe Uro neproX4 rou rRNA. To oripro Foor(e_ tor ndvo ornv 5'neproXrj, q e[d6po ndvo orqv Kewpr_ EIK0NA 24.39 H e[e6po rng 30S unopovodog etpopg6(er p6oo oqv evrogq qg 50S unogovodog oX4por(ovrog ro 70S ptpooor_ goTto. r{ nepropl r<ot 4 rego},4 ndvor u1v 3'neproXrl. H EIK0llA 24.40 6e(yyer 6n 11 30S unopovd6o 61er oori[;rerpn Ko_ rovoprq RNA ror npolreiV6v. 'Evo orlpowu<6 XopoKnlpr_ ortK6 e(vqr 6n 4 e[66po rrlg 30S unoprovddog, q onoio nopdXer q 6rdrpoorl Ue rnv 50S unoprovd6q, ouw(Oetor o1e66v onori.ermrrd on6 RNA. Kowd m4 6rdgoo4 Bp(or<owor ro noLrj 6[o nporeiVeg Bp(or<owor (dvo pr_ xp6 rpr;pro tqg 57 xor (ocog U6poq.rnq S12). Aw6 or;_ pofver 6u 11 orjvdeoq xor 6rdmoo4 rtov prBooropu<cilv unogovddcov np5ner vo e[oprdroL on6 ol,Aq],ent6pd_ oerq Ue ro 165 rRNA. H oM4Aen(6poorl rov unopov6_ 6ov en4ped(eror on6 puo prerdMo(4 o, 6vo Bp61o rou 165 rRNA (or4 06o11 791) nou ewon((etor orq dtdgoorl rov unopovddov. Me nerpdporo rponono(4oqq/nopep_ polnS, ono6erX04re 6l eprnLdrowot rqt dllo voux.leo_ r[dto rou 165 rRNA. Auq q noporrlp4oq unooqp((er qv 165o 6r ro prBoorrtprdno nr0ov6v e(ell10qr<e on6 6vo oropror(6ro onorel,oUgevo gdMov on6 nruR nopd npcore(v4. H 50S unoprovdSo 6Xer neproo6repo ro6trprq xcno_ vo1.r4 ouorortxcilv ouyr<pnrxd pe qv 30S unogovd6o, pe porpd poB6(o 6(r<Arovou RNA nou 6ronepvoriv 14 6opr1. To RNA qXqpot((er puo pd(o ogrXrd ouoKeuo_ opr6vov el(rov. H e[orepu<q enrgdvero onote].efror xu_ p(trlg on6 npcotetvrl, trr' e[o(peor1 ro rdvrpo rqg nemrdul.orpovogepdoqg (F).6ne Ev6r4ta 2a.1 9). Z14e_ t;l{*tiA :4.3t H 50S unopovodo eXet gro xevrptrrl npoe[oXr1 onou evron[(erot ro 55 rRNA ror droXopr(eror pdoo prog evro;u1g ono ro ploXo nou ouvrlOeror ono ovriyporpo rrlg nporeivqg 17. :;i:r':: -: .:,,.:: H 30S pLpOOtogrrl UnopOvodO e[vOr rivo prpovOu- rtreonporeivuo ooporOro. 0r nporefveg errovl(ovror urptveg, H qoroypotp[o nopoXoprl0rlre on6 rov V. Ramakrishnan. Medical Research Council (UK). 700 KE0AAAI0 24 Merorppoorl
1 65 rRNA ir!,i11.1 ic.+l ToorlueiueiloQriqUeToEuTovrRNAevroni(ovror oe60oneproX6groul65rRNAror pLoneptoXrl rou23SrRNA,Avonopo- yerorono: M. M. Yusupov, et al., Sclence 292 (2001):883-896 lhttp://www.sciencemag.org]. H rptrrroyporpio nopoXrrlprl0rlre ono rov Harry Noller, University of California, Santa Cruz. 66v 6Lo ro qrrlproro rou 23S rRNA oMqLenrdporiv pe nprrrtetveg, oLi.d noMdg on5 rrg npcoreiVeg efuot q4err- xd ovopydvoreg. H ou;rBoL4 ro:v unopovddrov oro 70S prBooroprdrro nepiloprBdver enorpdg ;reto(ri tou 165 rRNA (noLl,5g orrlv neproXrl qg e[66poq) ror tou 23S rRNA. Yndp- louv en(or1g oproprd,veg oMr;lenrdpdoerg gero[6 rRNA xdOe unotrrov6doq rot npo:reivcirv qq dlJtqq, roOri:q rr eLdXrmeq enoq6q ;rero[rj nporeivtirv. H EIK0NA 24.41 rouronore( ro oqpre(o enogrlg ndvro mrg 6o;169 rou rRNA. H EIK0NA 24.42 ovoryer tr1 6o;rq (qowoorefue rqv 50S unopov66o neptorpoppSvrl optmep6mpogq Kor qv 30S unopov66o neptorpo;r;rdvq 6e[t6orpoeo yti- pro on6 tov 6(ovo nou uno6etxvUetot orrlv etr6vo) no- pouord(owog r1v ew6ntoq tcov orlprefov enocp4g ndvto orrlv enrgdvero rdOe unoprovd6og. 24.17 Io PrBootopdrro ileprdxouv noA d Evepyd Kdvrpo r 0t olLnLenLdpooeLq orrq onoleq epntr6reroL to rRNA e(vor rp(orpo g6pog rrlg tretroupy(og rou ptpootrtgor[ou. * To nep0otrtrov rov 06oeorv npoodeoqg rou IRNA ro0op((erot ruplog on6 ro rRNA. To Boorr<6 XopoKrnprfitr5 tou ptBootol-iot(ou nou np6ner vo Ouprdror rove(g e(vor 6rL np6rettot yto ou- vepyoru<r1 6oUn, n ono(o e(oprdtot on6 pretoBo).69 ElK$ilA 24,42 Enoq6q pero{rj rtov prpooropLr6v unopovodtov ruproq npoypotonoto0vtot ond ro RNA (etrov[(erot UtoB). 0t enoq6g nou nepilogpovouv nptote[veg euov(ovroL r(rprveg. 0r duo unogovo6eq neptmp6qovrot gorpLo 11 pio on6 qv o]rtr4 onoro],Unrowoq Ttq ert- rpdveeg dnou npoyporonorouvror or enorpdg: onrl 6vo en[ne6o enorprlg ro0ero oqv o06vq, t1 50S unopovodo neptorp6qetot 900 optme- pdorpoQo, evtir 1 30S unopov60o nepLmp6qeror 900 6e[r60Tpoqo (r1 exdvo ug nopouotd(et oe owlorporpo npooovorotrtop6 ond to ouvri0rl). H rptrlroypogro nopoXropl0rlre qno rov Harry Noller, University of California, Santa Cruz. f4.1? To Prpoorr:porLo [ept51ouv tlotrtro Evepyo Kevrpo 701 23S rRNA Boorrdg dworcg - i
oilq ox6oerq Ueroqrl rov evepyci)v Kdwpov rnq Kord rn Uerdepoon. To evepyd K6wpo 6ev e(vor ptr<pdg, 6tor<pn5g neproX5g 6nc,:g ro evepyd r6wpo rcov ev(U- 1.uov, oMd peydLeg neproXSg, trrlv ono(ov q 6opui r<or ot dpdoerg e[oprcitwor 16oo qn6 ro rRNA 6oo ror on6 rrg prBooropur6q nporeiveg. Ot xpuoroLlm6g do- ;r5g rolv [eloprmc0v unopovddov Kor rov Borcr4pro- r<dlv prBooropor(otv npoorpri,pouv pto r<ol.r1 ew6notoq yro rrl ouvolrrrl opydvcooq xor tov[(ouv ro p6].o rou rRNA. H nro np5o<porn 6oUn, oe ovdLuoq 3.5 A, rou- ronoe( ooepcbg lg ewon(oerq rov IRNA xot rrr:v Ler- toupyrxd:v neprolcitv. Mnopotipre nl6ov vo e[r1yr1oou- pe noLL6g ).etroupy(eg rov prBoorol-ror(rr:v on6 u1v dno{.rr1 qg 6opqg roug. Or prBooropur<69 Lemoupy(eg neptorpd<powor yUpco on6 rlv otrl,rllenbpoorl Ue ro IRNA. H EIK0NA 24.43 no- pouord(er ro 70S prBooroprdno Ue lq ewon(oerg rcov IRNA ong rperg 06oerg np6o6eo4g. To IRNA oug 05- oetg A ror P e[vor oXe65v nopdtrtrql.o ;rero[ri toug. Kor to rp(o IRNA eu0uypopp((owor, Ue rouq Bp5Xoug rov owxo6u<ov(cov rouq vo npoo66vowor oro mRNA ;r5oo orqv orjloxo ndvot or4v 30S unopovd6o. To un6Lorno rpritro r<dOe IRNA npooddvetor or4v 50S unopovd6o. To neprB6Mov yripo on6 rdOe IRNA nopriXetor rup(rrlg on6 rRNA. Ie r<d0e 06oq, ro rRNA r<or IRNA 5pXowor o' enorprl toq rgqproro rnq 6opnq, nou yevrrd e(vor ouwrl- pnuSvn. 'Htov ndwore ;rey6).o o(vry;ro vo rorovo{oouge n6g prnope[ vo lopSoouv 6rio oyrcl6q IRNA ro rivo 6(- nLq oro dMo r<ord qv ovdyvotoq yemovrr6v rco6txo- vicov. H r<puoroMu<r1 6opr4 eprpov((er puo r<oprn4 45o oro mRNA prero[ti tov O5oeov P ror A, 4 ono(o enmp6ner Ell(0NA 24.43 To 70S prpootoluirto onoretrefror on6 r4v 50S unogovo0o (oonpo) K00r Trlv 30S unogovddo (UoB) Ue rp(o IRNA nou ewon((ovtor entqovetoxo: r[rprvo or4 06011 A, pntre oqv 06- or1 P ror npdotvo orn eeon E. H rporoypotp(o nopoXtoprl0rlre ono rov Harry Noller, University of California, Santa Cruz. 06oq A d Iumpoqr'1 I ,. r-i r t;' .', .i /', ' 3* t;iI ,] .,:, q:i-.r- ' ':j. + EIK0NA 24.44 To tpio IRNA 6Xouv drorpopertrri npooovorotrrolrd novo oro prpootrlgolo. To mRNA nepLmp6EeroL pero[u rov 06- oetov P ror A enupenovrog 6ror oro ogrvooru),0-tRNA vo npoo- 66vovror oe 6rn)ovo rodrrrivro. H rporoyporp(o nopoXtop404xe on6 rov Harry Noller, University of California, Santa Cruz. oro IRNA vo eEdmowor, 6ntog cpo(veror orrlv qg ElK0- NA 24.44. To IRNA ong 06oerg P r<or A Bp(or<owor un6 y<ov[o 26o pero[ri rouq Ue r<6wpo t' owrro6u<6vrd roug. H orev6rep4 npooSyyrorl prero[ti rcov ore].er6v rcov IRNA ougBo(ver oro 3'dr<po, 6nou on6Xouv g5).q 5 A (rdOero oq npoq ro en(ne6o r1g oel(6og). Aur6 enr- rpSner orqv nemr6u<4 ol,uo(6o vo perorpdperor on5 ro nemrdu).o-tRNA qg 05or1g P oro oUrvooKULotRNA rqg 05o49 A. To oprwoorulotRNA evoo;rordlveror orn 05orl A pe- oo rou EF-Tu ror ro (euydpoprd rou pe ro rro6x6vro e(- vor ovoyro(o 6ore o EF-Tu vq u6polrioet to GTP r<or t' oneleu0eprrr0e( on6 ro prBoorrl;rdtro (Bl,5ne Ev6rqta 24.10). O EF-Tu opXu<6 ronoOere( to ol.rwoorul,o-tRNA orrl prrp4 unopov66o, 6nou ro owu<rrl6x6vto (euyoptr> ver Ue ro rc,r6x6vto. H peror<tvr1on tou IRNA efvor ono- pofuqr4 6ore vo ;rero<pepOe( o).6x),qpo ;rdoo orq 05oq A, 6rov to 3'dr<po rou erodpXetor oro r6wpo rrlq nerrn- 6 ul,orpovorpepdoqq or4 peyo),q unopovd6o. YndpXouv 6to<popenr<d prowSLo yro ro n6g npoyporonoefuor qu- q q 6repyoo(o. liprqovo pr' 6vo on6 oLnd, o).6r<L4po ro IRNA neprorp6geror, 6ro1 6ore o oyr<rivog qg 6o- prlg oXql:orog L, r'1 ono[o oX4pror[(erot on6 roug Bpo- X(oveg D ror TIUC, vo UeroKrve(rot prSoo mo prBo- oolpdrro, enmp6nowog oro Bpo{ovo TIUC vo (euyoprrl- oer Ue ro rRNA. li;r<pcovo p' rlvo dtri.o ;row5Lo, n roo- repx4 6oprl rou IRNA peroBdMeror Xorlorponot6wog ro Bp61o rou owu<ro6u<ov(ou oov oppr6, evr.ri ro un6l,or- no IRNA neptorpdqeror on6 pro 0rio4 6nou oropd(etor orrlv 3'nleupd rou Bp6Xou rou owu<ro6u<ov(ou oe Uro dMn 06on, 6nou oropd(eror orrlv 5'ntreupd. Merd rr; peror6ntoq, o EF-Tu u6poL6er ro GTP enrp6nowog vo ouvelrore( 11 o0vOeoq rou nemr6[ou. H perot6nrorl nepil.oprBdver preydl.eq perorrvqoetg ong 06oerq rcov IRNA pr6oo oro prBoocopdrto. To drpo rou owu<co6rxov(ou rou IRNA perorrveftor -28 A on6 rr1 l 06on E 702 KEOAAAI0 24 MeToqpool
06o11 A orrl 06o11 P xor orq ouv6lero perorrvefuor enr- nLriov 20 A on6 rr1 05on P mq 06or; E. E[on(og r4g ytov(og r<dOe IRNA cDq npoq ro owrrcodrx6vro, o 6yrog rou IRNA peroxrvefuot r<otd no),ri preyoArjtepeg onootd- oetg: 40 A on6 q 06oq A orr'1 05o4 P r<or 55 R on6 r1 06orl P orrl 05or'1 E. Aw6 uno64l.c0ver 6l q preror6nrorl npouno06ra 6poonr<q ovo6ropydvoon rnq 6oUriq. l-ro noMd Xp6vro enrrpotodoe 4 peror6nron Llno- po{oe vo npoyl-loronorrl0e[ p6vo nopouo(o rou nopd- yowo EF-Tu. Om6oo, ro owrBronr<6 onopoopur<(v4 (sparsomycin) (to ono(o ovoor6l,).er q 6pdo4 rnq ne- mr6uLorpovogepdoqg) nupodote( rr1 peror6nrorl. Au- 16 uno64),civer 611 4 evSpyeLo nou npootOe( 14 peror6nror; ornv fl poyponr6rlto onoO4reUeror oro ptBoorr:prdtro oe6rou olrlprorrme( o nenrrdrr6g 6e- op6g. O EF-G ouvr'1Ocog enr6pd oro ptBooro;rdno one- )reu0epri:vowog our4 r4v evdpyero ror ro0ror6wog ro rxov6 vo npoco0foer 14 peror6nto4, roorSoo rl oTropoo- guxftn4 6Xet rov [6ro p6tro. H onopoopur<(v4 ovoord].- Aer rrlv nemr6u).orpovogepdoq pr6oo np6odeorlq fio nemtduLotRNA: our6 qvoordl er qv oM4Len(6poo4 rou Ue ro oprvooKuLo- IRNA. l-I0qv6v 64ptoupye( puo 6rop6pgolo4 nou prord(er ;re t4 ouvqOro;rdv4 6rop6p- gtooq prerd 14 peror6nrorl, q ono(o pre q oerpd r4q npodyer tr1 peror[v4o4 rou nemr6utro- IRNA. To oq- gowx6 orlpreio efuor 61 4 peror6ntorl onorele( ev6oye- vr; rdr6qro rou ptBoocol.ror[ou. ljpr<provo ge to prowSLo rov uBptdu<rilv r<otoordoe- rov, 4 perot6nrorl prnope[ vo ouprBofver oe 60o srd6ro, Ue rn Uro prBoooprrrl unopovd6o vo petorrve[tor oe oxSon Ue qv dtr.q c6ore vo 6r1puoupy40e( 6vo evdrdpe- oo mddro mo ono(o undpXouv uBpr6u<rig neptolSg np5o6eoqg rou IRNA (50S E/30S P ror 50S P/30S A) (Blene Erx6vo 24.29).Iuyrp(oerg r4g 6oprlq rou prBo- orol.ror[ou pero[U trov roromdoeov nprv ror perd tr1 petot6nrorl, r<oOcig xor ouyrp(oerg qg Sropr6peoonq tou 165 rRNA gero(rj el,eU0epov 30S unopovddorv rot 70S prBoocol.ror(rov uno6rllrilvouv 6tr 6op4 SXer 5wovr1 xlqrtr6rrlto r6(cog ong neprol6q qg r<eqo).rig Kor rrlq efd6pog rng 30S unoprovd6og. Mto ev6rogdpouoo 6v- 6er(q yo to 1-row6l,o trov uBpr6u<rilv r<oroordoeov npo- 6pleror on6 ro yeyov6g 6rr noM5g Bdoerg rou rRNA nou epnfuirowor orrl o[v6eoq rov unopovddtov, Bpi- oKowor rowd oe Bdoerg nou epnl.6xowor mrlv oMrl- Aeni6poorl pre ro tRNA. Aur6 uno64lcirver 6rr ot neproXSg np6odeoqg rou IRNA Bp[orowor rowd or4 6t6rpooq pero[ti rov unopovd6otv, ev6 en(o4g unovoe( 6n oMoy6g orrlv oM4Len(6poor; rov unopovddcov 0o ynopo0oe vo ouv66owqr Ue rrl preror<(vqor1 rou IRNA. Meyd),o rUngo qg 6op{g rou prBootrlpor(ou roro- LopBdveror on6 to evepyd r6wpo rou. H oXqpolr<4 onem6vro4 r<ov ptBooropur<riv 06oetov orr'1v EIK0NA 24.45 6efiyer 6tr onoreloriv nep(nou ro 6Uo tpfuo qg 6oUnq rou prBoocol.ro{ou. 'Evo IRNA eto6pXeror orn 06oq A, prrog6,peror prSoco perot6nronq orn 06o4 P rr Snero uyxmo.trefuer ro prBooroprdtro on6 q 05or1 E. Or 06oerq A xot P ercre(vowor rord ;r4r<og Kor rov 6tio prBooro;u- xtiv unopovd6ov: ro IRNA (euyopriver ;re to mRNA E|(0NA 24.45 To prpootoporto dLo06rer notrtro evepyri r5vrpo. Mnope( v' otrtrr;trenrdpo Ue Uro Uepppovq. To mRNA npoyUoTonor- e( gro neprotporprl roOdrg DLepleror ond lg 06oerg A ror P, or onofeg pp[oxovrot un6 yov(o 4 Uro oq npoq qv otr1. H 06oq E pp(oreror n6po ond U 05oI P. H 0eo1 qg nenldutrorpovoqepo- onq (dev onerxovf(eroL) exre[vrrot droprioou rtov xopurptirv rotv OSoeorv A xor P. M6pog rr;g 06or;q 6nou npooD6vovror or EF-Tu/G ppforeror oq pdo4 rov 0tioealv A xor P, m4v 30S uno;.rovd6o, otrtrd ;rerogopd rou nenrtd[ou ouprBo(vet ornv 50S unopovddo. Or 06oetg A ror P Bp(- oKowor 6[n].o-6[n],o, 6reurol.rjvowoq rrl ;reror<(v4oq rou IRNA on6 q puo 05or1 orqv dM4. H 05oq E ewonG (eror rowd orrl 05o4 P, evdr ro r6wpo tqg nemrduLo- tpovoqepdorlq ornv 50S uno;rovddo, xovrd oro ol.rwoorulrrd drpo rov IRNA ong O5oetg A ror P (B).6- ne Ev6trya 24.18). 'OLeg or nporeiveg nou npoodrivouv GTP ot ono(eg Xpnorponororiwor orrl pretdEpooq (EF-Tu, EF-G, lF-2, RF1, RF2 ror RF3) npoo66vowor orqv 6ro 06o11 (pept- 169 <popdg rq^,e(ror r<6wpo GTPdo4g), yeyov6g nou prdMov nupodore[ qv u6p6truor1 rou GTP. Aur4 4 05- oq ewon(eror ar4 Bdorlrou prioXou trlg preydLqg uno- povd6og, 4 ono(o onotel,efuor on6 rtg npotefveg L7 xor 112. (H L7 e[vor nopoMoy4 qg L12 r<or nepdXer pro oretulopd6o oro o;rworetrrr6 dxpo). Er<r5g on6 out4 qv neproXri, ro oUpnl.oKo rnq npcoreivrlg 111 p' 5vo rpqpo 58 Bdoeorv rou 23S rRNA nopdXer plo 06oq yro rrlv np6o6eoq opropr5vov owrBrorrr6v nou enrlped- (ouv qv evepy5r4to rqg GTPdoqg. It4v npoypotrr6- tqto, r<oprd on6 our5g rrg prBoocrlpru<69 6opr5g 6e 6ro05ret evepy6r4ro GTPdoqg, coor6oo e(vor xor or 6rio ovoyxo[eg yr' our4 rr1 6pdoq. O p6].og rou prBooroprot! ou efuor vo nupo6oreirrlv udp6Luoq rou GTP on6 no- pdyoweq npoodede;rdvouq orrlv ow(mo rXq 05or1. H oplrr<4 np6o6eo4 rrr:v 30S unogovd6ov oro mRNA qnore( qv nporeffi 51, 11 ono(o 61er roXupq ouyydvero yro prov6r<trtovo vouxleivrrd o[5o. H nporei'- v4 51 6rorqpe( oe prov6r<l,ovq xotdmoorl ro mRNA nou efuor npoo6e6e;rdvq orrlv 30S unogovd6o. Auq 11 6pdoq efuot onopofrrlrr'1 d;ore ro mRNA vo prlv npoo- ).dBer pro 6rop6p<pcoo4 pe (euydptopo Bdoeo:v nou 0o rlrov or<ordl.L4Lrl yro q pretd<ppoor1. H Sl 5Xer puo e[orpetu<d enrprrlrrl 6opr4 r<or ouv66eror pe trg S18 r<or S21. Or rperg nprrrreiVeg ouvroroUv pto neptopl 11 ono[o Odon A -:-_-*- -- | r.,l:,:5J" I ?:+.'; t Ta Prpooolpono llepreXouv il0trtro Eveoyo (a,, :: 703
epni.6reror m4v oplrxri np6o6eor'1rou mRNA Kor ornv np6o6eor1 rou evoprerlptou IRNA. Aut6 rono0ete( tr1 06o11 np6o6eonq rou mRNA xowd orq oxropr| rnq Ur- rpqg unoprovddog (plene Exdvo 24.3). To 3'drpo rou rRNA, to ono(o (euyopcbver pe rq 05or1 6vop[qq rou mRNA, ewon((eror oe ourrlrqv neproXr;. Or nopdyoweg 6vop[4q npoo66vowor orqv (6ro ne- proXri rou prBoorollor[ou. O lF-3 prnope( vo 6roouvde0e( pe to 3'dxpo tou rRNA, 6ntr;g en(o4g ror pe opret6g pt- Bootopur<69 nporeiveg, prero[ti d],],rov r<or npo:reiveg nou gdMov epnL6xowor onlv np6o6eorlrou mRNA. O lF-3 0q pnoporioe vo moOeponore[-r4v np6o6eorl mRNA-30S unoprovddog: crTl ouv5lero erron((etor, ;16- l.rg ogr1Oe( q 50S unopovd6o. H evoopdtroon rou 55 RNA oe 50S unoprovd6eg nou ouvoppol,oyoriwot in vitro e(oprdror on5 rrlv xo- v6rrlro tpuiv np<rl'reiv6v -L5, L8 ror L25- vo oXqpo{- oouv rivo ororxeloperprr<6 oUpn].oxo pro(( tou. To oUgnl,oro npoo66vetor oro 23S rRNA, r<drr nou rov6vo on5 ro [eXopror6 ouoronrd rou 6e pnope( vo ilpoy- poronorrloer. To o0gnl,or<o Bp(oreror rowd ortg O5oerg P ror A. 'Evo veoouwtOriprevo nol,unem(6ro ercreiveror 6ropr5- oou rou prBoorol.ror(ou, prorprd on6 ro evepyd r6wpo, npoq rnv neproXq 6nou ro prBootoprdrro [orog ouvdmov- rot pe preprBpdveg. Mro nolunemrdxri oAuo(6o ovo60e- ror qn6 to prBooroprdrto 6roprSoou ev6g 6roriLou e[56ou nou o6r1ye( on6 q 06o4 ulg nemrdul.orpovogepdorlg orqv enrgdvero rrlg 50S unopovddog. O 6(oul,og onore- Le[ror rupfrog on6 rRNA. E[vor oprerd orev6q -e0pog 1- 2 nm- rr 6Xer UnKoq -10 nm. To veoouvrr06prevo noi.unem(6ro ovqdUetot on6 ro ptBooroprdrro -15 A pro- rprd on6 to xriwpo r4g nemr6uLorpovoEepdoqg. O 6(- ouLog prnope( vo ouyxporrloer -50 qrwo(6o ror ;rdMov neprop[(er tr1v no].unemrdxrl o].uo(6o riore vo Un pno- pe( vo 6mLo0e( nporo0 eyxorotre(rler rrlv neproXq e(6- 6ou, poLov6rr ev66Xeror vo oxnportororiv l.(yeg deweporoye(g 6opdg. 24.18 To l6S rRNA Ato6popoTi(€t Evepy6 P6Ao orn Merdrppoon c To 1 65 rRNA OrodpoporL(er evepyrl potro olg treuoupyieg rrlg 30S unopovodog. Atrtr1trentDpo opeoo pe ro mRNA, qv 50S unopovodo r0r r' ovlruldrrovlo rorv IRNA olg 0eoerg P ror A. To ptBooropduo opXLrd Oe<rlpr10r;xe toq Uro ou).).oy4 nporreivtov pe norr<(l.eg xoroLurrr5g evepy6trlreg TIou ouyKporo6wqr pSoo oLLrlLentdpdoeov 16oo trreto[rj npcoteivov 6oo ror geto(ri RNA xor npctlrei- vrrrv. Qor6oo, 11 ovor<dlurf4 prop(cov RNA pe rorou- lrrl evepy5tqto (BL6ne KegdLoro 21) oo<pc0g uno- 6qLcirver 6rr ro rRNA (oog 6ro6popror(er nro evepy6 p6lo orq Letroupy[o rou ptBooropor(ou. YndpXouv nl.5ov ev6e([erg 6tr ro rRNA o].Lq].enrdpd pre ro mRNA ( to IRNA oe xd0e orddro qg trrerd<ppoonq Kr 6tr or np<o"retveg e[vor ovoyr<o(eg oiore ro rRNA vo 6rorqpq0e( oe Urq 5oprl orrlv ono(o gnope( vq enme- Le( trg r<oroLutrr6g l,enoupy(eg rou. l1otr).59 otri.qLe- ntdpdoerg nepri,oprBdvouv erdrr<rlg neproX6g rou rRNA: o To 3' dr<po rou rRNA otr4lenrdpd oneuOe[og pre ro mRNA rord qv 6vop[r1. o Erdu<6q neprolSg rou 165 rRNA o).l,r1l,enr6po0v oneuOe(og Ue rrq nsproX6g owrro6rrov(ou trov IRNA 16oo or4 06o11 A 6oo ror m4 06o11 P. l-lopo- pto(og, ro 23S rRNA oMqLent6p6 pre ro CCA drpo tou nemrdulotRNA 16oo or4 05oq P 5oo xor orrl 06ortA. o H qI ntren(dpooq rorv unopovddov nepil.opBdver oLtrrll,enrdpdoetg peto(ri 165 r<or 23S rRNA (9).5ne Ev6rrTta 24.1 6). l-loLLdg nA4pogop(eg oXeru<d ;re ro (eloprord Fri- Uoro rnq perdtppoorlg oro Bolcrlpro npo4l0ov on6 tr1 Xpnon owrBrorxc0v ro onofo ovoor6Mouv t4 6epyo- o[o oe ouyrer<prpSvo orddto. O or6Xog ev6g owtBrorr- rof prnope( vo rouronotqOe( on6 ro oumorrr6 mo ono(o ouprBo(vouv peroMd(erg nou npood[5ouv owo- Xrt oro owrBronrd. Meprrd owtBronrd enr6porlv oe (e- Xrrlprordg prBoorotrur6g npo:reiveg, evtil noLl,d 6pouv ndvo oro rRNA, yeyov6g nou uno6qkivet 6l ro rRNA epnLdr<erot oe noM6g 11 or6pr1 r<or oe 6treg ng l,etroup- y(eg tou prBooroproilou. Or Lenoupy(eg tou rRNA rilouv 6tepeuv40e( pe 6tio 6rogoperu<59 npooeyy(oerg. Or 6o;ur<rig preLSreg 6e(;youv 5tr t6ro(repeg neproX6q rou rRNA ewon((ov- ror oe o4;rowu<59 06oerg rou prBooutl.ror[ou ror 6t or Xqgrr<59 rpononorrloetg ourcbv rrov 06oeov nopoKo- Lriouv ouyxerptpr6veg prBooalpur<69 Leno u py(eg. Enr- nL6ov, or prerd.1,d(erq rouronotoriv Bdoerg rou rRNA onopo(rqreq yro ouyKerptpr6veg prBooropl169 Ler- roupy(eg. H EIK0NA 24.46 ouvor|((er ttg O5oerg mo 165 rRNA nou rowonororiwor Ue rn ouyrerpr;.uivn npo- odyyroq. Mro 5v6er[q yro rrl orlproo(o rou 3' dr<pou rou 165 rRNA nop6leror on6 r4v euoroOqo(q rou oro 0ovorn- <p6po nopdyowo r<ollolvq E3 (colicin E3) nou nopdye- ror on6 opro;r6vo Borrrlpro. H ro)lo(vq dtoond -50 vour<,l,eor(6ro on6 ro 3'dxpo rou 165 rRNA rou E. coli. H drdonooq r<oropye( ewelcirg rrlv 5vop(4 rqg prerd- epoonq. l-1oLf.5g o4prowrrdq Lemoupy(eg onoroUv rqv neproXri nou droondror: q np6o6eon rou nop6yowo lF-3, n ovoyvtilpto4 rou mRNA r<or 4 np6odeon rou tRNA. To 3'dr<po rou 165 rRNA eprn).5r<eror dpeoo mqv ow(6poor1 6vop[qg prSoo (euyopd:prorog pre qv o].].r1- LouX(o Shine-Dalgarno on1 06oq rou prBooropror(ou yro np6o6eon oro mRNA (BL6ne Etr6vo 24.12).'Evoq 704 KEOA Al0 24 Merorppoo1 Boorxeg ewong
flposroteuUevo ond To P{RNA I (tl ltnl ti Kovr6 oqv npdodeon -rou EF-G Kewptxfl nepLolrl IIvdeoq unogovddtov I :.:lr1 opeoog p6Log tou 3' dr<pou tou 165 rRNA mq -::cepoorl ovodetKvUerqt on6 ttg t6r6qteg petol'l'oy- -::Lrv ov0erttrorv mqv KooouYKopur<(vq (kasugamy- : ^ ro ono(o 6e 6to06touv optoprdveq tpononotloetq -:- 165 rRNA. H KooouyKoUUr(vr1 ovoot6L).et tqv . .:clr1 rqq pretdgpooqg. Av0errtrd preroMdyUoro rou -- .ou ksgA eprgov((ouv ovendpreto prtoq preOuLdoqq -: r srodyet t5ooeptg ;reOutroprddeg oe 60o yetrovtt<5q :Ii,,'[veq oe Uro neproX4 xowd oro 3' drpo tou 163 'r"A, H preOul.(too1 drlprtoupye[ qv e[otpettr<d ouwq- :-*evl o).Lq).ou1(o 6-602A-m6zA, 11 ono(o Bp(or<etot -:lc oe npoKopuortKd 6oo r<ot o' suKopuolrrr<d pLrpd ':'rA. H peOuLttopr6vrl ol'r;).ou1(o egnl'6xerot mqv ., *cl rov 30S rot 50S unopovd6cov, q ono(o pe r1 :..:o rqg en(or1q oXet((etot pe tr1 6tottlpnon rou . . :crrrlpLou IRNA mo n).r1peg prBootl;rdtto. H xo- 3'gei(ov nepLoXrl oouyr<opux(vq nporoLe( oneleu 05ptoon rou f Met-tRNAr on6 ro euo(o04to (pe0ul.ttrlprSvo) ptBootoprdrto, oMd t' ovOerrtrd ptBootogdtto e(vot tt<ovd vo dtornpoUv ro evoprcr4pro tRNA. 'Orov ro ptBoo<,Dpdrto dpoorrlptonotoUwot orn Ue- rdgpoorl, ougBotvouv oL).oy6q orrl 6opti rou 165 rRNA, 6nog eo(verqt on5 rrlv npocrroo[o ouyrexptpr6- vtov Bdoeov 6vowt XnUKIiq en(Oeor1g. Ot (eloptor6g 06oetg Kqrordooowor oe Liyeg opddeg, ot ono(eg ouy- rewpc0vowot orrlv 3'etrdooovo Kor rnv r<ewpxti nepro- 1q. MoLov6tt ot evron(oetg e(vot dtdonopreg otrl ypoprpu<r1 otrrlLoufotou 165 rRNA, orrlv npoypronr<6- rnro or 06oerg ttov Bdoeov nou epnldxowot orqv (6to tretroupy(o prnope( Bp(orowot t<owd 11 ;rto orrlv dtr'q finv rprroroyn 6oUn. Optop6veq on6 rtg ol,oy6g mo 165 rRNA nupo- ll lll Bprjxoq llllll ll lll I 2. orn 06on A Kotomotrn 0dor1 492 orn e6on A TNV Ue0utrfoon oro ksgl - IuuntrnpouoTtKo Ue To mRNA - AKp(pero 3'eldooov neproxn : r ;i: MepLr6q 06oeLg rou 165 rRNA npoororeuovror ono Xrlgtro0g ovLyveur6g 6rov oL 50S unopovodeg ouvduooro0v pe rtg :,: lnoUovodeg 1 ro optvoorutrotRNA npoodeQouv oU Oeon A. Atrtreg etvot 0eoetg perotrtro(eov nou enqpeo(ouv q peroqpool. 0t 05- ,. , <oroorotrflq TERM pnope( vo enlpeo(ouv rov repporLopo oe pepLro rt notrtro roDtxovto In[lS. To UeVotro SYXptlUo ntro[oto defivouv - : eoosprq neptolSg rou rRNA. J,.: 1= To 165 rRNA Aro6po;rori(er Evepyo Potro oq Merorppool 705 Korqorotrn TERM
llemduho-tRNA Ilemdtr1 oluoi6o I H.N I H.C.R I ,C. 00I r IRNA I V flenldrxfl dtuo[6o I llemdml duo(60 I H.N I H.C.R iH 0" '11 ' H-i-nI H ,C.. 00'0It IRNA IRNA EIK0NA 24.4S 0 oXlgologog nenldxoU OeogoU npoUno0Srer o[eo-Booxrl KoTotruon, K0T0 Trlv ono[o 6v0 dropo H UeTqqdpeTor o' 6vo pooxo KoTotrorno. 24.2O Or PrBoourprKdq AopCq AAAd(ouv 'Orov luv6uooTo0v or Ynopov66€q & '0r0v oXqpor((ovror ntr(pq pLpootol.rotto, 11 regotr1 rtllv 30S unopovoDtr.rv neprorp6qerol yupo ono rov ouXrlvo. * To evepy6 revrpo qg nenrrDutrorpovotpepoolq orrlv 50S unopovodo elvot nro dpoortro oro ntrr1pr1 prpoorolrolo ouy- rpurro ge rtq pepovrr:p6veq 50S unouovooeq. l-1o).).59 Sprpreoeq evdei(etg uno6qtrtrrvouv 6tt q 6opq rtov felcoptmtrlv unopovd6rov oL).d(er notrU 6rov outdg ouv6uomoUv yro vo oXrlpor(oouv 6vo nqpeg prBooroprduo. Or 6rogop5q crlq npoq rqv euoLo0qo[o rrr.lv rRNA o' e[oyeve(g nopdyoweg e(voL 1-rto on6 rrg toXup6repeg ev6eferq (B].6ne Ev6rrlto 24.18). llto dgeoo, ouyr<p(oerg r<pumoLLrr<cbv 6oprorv uQr;- l.r1q ovdl.uonq rov [eXcoprorcbv unogovddtov pe 14 dopq logqL6reprlg ovdLuorlg o6r<).qpou tou ptBo- orrr po-r[o u uno 6r1].rilvo uv rnv Un o p [r1 oq povrtr<rrrv 6rogopd:v. Aur5g or on6Qerg enrBeBorcb0qrov cm6 1.rro xpumoMrxrl 6opu1 rou prBooropot(ou rov E. coli oe en(ne6o ov6Luoqg 3.5 A, q ono(q enrnAsov rqu- 'ronore( 6Uo 6ro<popelr<69 dropropgcioerq rou prBo- o<r: gor(ou nou (o<.r:g owrnpoo0Jne6ouv dto<popefl r<d orddro orq prerdqpoo'r1. O xpUoroMoq nepr6Xet 6rio prBoorr:prdno ovd govd- 6o, ro xo06vo pre 6rorpopenx{ 6ropr6p<ptooq. Or 6ro<po- p6g ogefi.owor oe oMoydg orq 6tdto[r1 rov neproloiv p6oo oe rdOe unogovd6o. Irrlv nro o41.rowu<ri otroyri, q regotrr] rnq UrKpriq unoprovd6og neprorp6geror rotd 6" yfprrr on6 rqv neproXri"rou ou16vo npoq rn 0dor1 E. En[or1g, Uro neprorporp{ 6" r<ord qv ow(Oerr1 xote6- 0uvoq noporrlpefuot orrg 6o;159 6ogdg rrov ptBoorol.ro- ilrov rou Thermus thermophilus nou npoo66vowor oro mRNA Kqt 6ro06rouv IRNA 16oo orrl 06oq P 5oo r<ot orrl 05oq A, Aur6 unodqlc0ver 6n q regoAq pnope( vo neprorpoqe( ouvoAxd r<ord 'l2o ovd),oyo pe ro orddro rqq gerdrppoonq. H neprorpogrlrqq xego).r1q oxotrou- Oei qv nope(o rov IRNA 6rop6oou rou prBoorotrlo{ou, eye(powog rqv nt0ov6r4to vo puOpr((er q perox(v4oq tov mRNA ror IRNA. Or oll.oy6g r4g 6ropr6pqoonq nou ouprBo(vouv 6rov ouvduomotiv ot unoprovd6eq e(vor notr0 nro 5164- treg orqv 30S nopd ornv 50S unoprovddo. Or oMoy6g ;rdMov oXet((owor Ue rov 6leyXo rqg 06oqg Kor rnq perox(vqoqg rou mRNA. H oqpowrx6repq oMoyrl orqv 50S unogovd6o o<popd oro r<6wpo rqg nemrdul,orpov- oEepdoqq. Ot 50S unopovd6eg e[vor -1000 <pop6q ].t- y6repo onoreLeoporu<5q crrlv rordLuoq qq oUv0eoqq nemr6u<ri:v 6eopuiv ouyr<prxd pe to nLtjprl pgooopd- rro: o tr6yog (oog e[vor 1.uo oMoyrlqg 6opr1g nou ro- noOere( to un6orpopro nro onotetreoportrd oro evepy6 r6wpo rou nLrlpouq ptBoorr:1.ror(ou. 24.21 llepiAnQn 'Evq rodx6vro rou mRNA ovoyvrrlp((etor on6 6vo o1-uvooxulo-tRNA, ro ono(o 6rq06tsr 6vo owu<ro6rr6- vro ouprnlqpcoporrx6 rou rol6u<ov(ou r<ot peroq5per ro oprwolrl nou owrororXe( oro xodrx6vro. 'Evo er6r- r<6 evopr,crjpro IRNA (fMet-tRNAt orouq npor<opuri:- req Kor Met-tRNAi orouq europuciteg) ovoyvrop[(er ro rodrK6vro AUG, ro ono(o XprlorUonorefuor orrqv 6vopEn 6trcov rrov r<o6 r rono rqrrxcilv oLLrlLo uXrrriv. Itoug nporopucirteq Xpnorgonore(ror en[or1g ro GUG. M6vo rq Ko6rK6vrq repgoloUo0 (Xrop(q v6r1- ;ro) UAA, UAG ror UGA 6ev ovoyvopi(owor on6 opr- voorui.o-tRNA. To prBooroprdlo oneleuOepcivowor on6 q pretd- gpoon xor ero6plowor oe Uro 6e[opevrl e].eU0epov pr- Booropor(crlv, ro ono(o Bp(or<owor oe rooppon(o ge [epopror6g pur<p5g xor geyd].eg unopovd6eg. Ot trru<p6g unopovd6eg npoo65vowor oro mRNA r<ot mq ouv6- Xeto evc0vowor trre peydLeg unopovd6eq nopdyowog 6vo nl,rlpeq ptBooroprdrto nou ovoLopBdver q prerd- 708 KEOA Al0 24 Merdqpoorl &Bdon Boorxdg euvoreg
qpoon. H ovoyvc0proq Uroq npoKoputonrqg ogeqp(oq nepil.opBdver rrlv np6odeoq puog oLLrlLouX[og mo 3'dxpo rou rRNA oro pror(Bo Shine-Dalagarno nou Bp[- oKeror npw on6 ro ro6u<6vro AUG (n GUG) oro mRNA. H ovoyvtirptoq ev6g europuolxo6 mRNA nepil.qrBdver np6o6eoq mqv 5' rol.Unrpo: mr1 ouv61ero, 11 purprl unopovd6o perovooreOer orq 06o4 6vop[qg ovo(rpdrv- tog xo6u<6vro AUG. M6Lrg ovoyvrrlp(oet 6vo rordMqLo xtodrx6vto AUG (ouv4Ooq, ro npcirro nou ouvowd), ou- 6ud(eror ;re puo preydLrl unoprovddo. 'Evo prBoorrlprdno pnope[ vo pretorpSper rour6lpovo touLdXrorov 6rio o1-rwooxulo-tRNA: n e6on P rotoLopr- Bdveror on6 6vq noLunemr6u).otRNA, ro ono[o pero- rp5per rqv nol,unenrr6rrrl oLuo(6o nou 5Xer 4611 ouweOe(, evcil q 06o11 A 1pr1or;ronoe(ror yro qv e(oodo ev6g o;uvooxutro{RNA, nou preroqdper to en6pevo o1.u- vo[ri ro ono(o 0o npomeOe( orrlv o].uo(6o. To Bor,rrl- ptord prBooopdrro en(oqg 6ro06touv puo 06o4 E, 6ropr6oou qg ono(og 6rdpXeror onoorutruoprSvo IRNA, nporori oneLeuOepco0e( perd rn Xprlorpono(qor1 rou orrl perdgpoorl. H notrunemrdu<{ o},uoi6o orq 05or1 P petog6petot npoq ro ol.rlooxulo-tRNA rrlg 06oqg A, 6rlproupyc0wog 5vo onoorul,uog5vo IRNA orq 06oq P xt dvo nemr6u).otRNA orq 06o4 A. Merd rq oUv0eoq nemr6xoti deopoil, ro ppooroprd- ro peroron((erot rord 6vo rodx6vro n6vo oro mRNA, perorwcilwoq ro onooKulru:;r5vo IRNA pSoo orq 06o11 E r<or to nemr6ulo-tRNA on6 q 05or1 A orrl 06oq P. H petot6ntorl roroL0eror on6 rov nopdyowo enrpn'1xuv- oqg EF-G ror, 6nog noMd dMo orddro rrlg )reroup- y(og tou ptBoorrrprot(ou, onone( u6p6Luoq GTP. Kqrd rr1 peror6nroq, ro prBoorrlprdno 66p1etor on6 puo uBpt- 6x4 xordmoon ornv ono(o q 50S unoprovd6o peroxt- ve(rot tog npog rnv 30S unoprovd6o. H perdqpoorl efuor 6onovqpq 6upyoo[o. ATP pq- orponore(ror yro nopoXrl ev5pyerog oe noMd m66tq, preto[6 dltrrov ro rp6prrr:1.ro rou IRNA pe ro o1.rwo(ri rou r<ot ro l,eilNypo rou mRNA. Ercrq-rdror 6tr 5trrg ror 90% 6l<ov rrov gop(olv ATP nou ouw(0ewot o' 5vo Bomrlpro nou ou(dver ypnyopo r<orovotrc0vowor orq ouvopprol.6- ynon rov o1.rwo[5rr:v oe nporetvegl Ie rd0e srddro rqg ;rer6<ppoorlg onomo0wqt ent- np6o0eror nopdyoweg. Ko0op[(owot on6 rqv xur].trr1 o0v6eoq ror 6rdoroori roug on5 to ptBootrlprdtto. Or nopdyoweg 6vop[r1g epnMrowor orqv dvop[r1 dvop[q orouq npoKclpurireq. O lF-3 onotte(tot yto rrlv np6o6e- orl rov 30S unopovd6<ov mo mRNA rt en(o4g efuot uneriOuvog yro rn 6rofipr1or; qg 30S uno;rovd6og o' eLe60epq Uopgn. O lF-z onone(ror yro rrlv np6o6eoq qq fMet-tRNAt or4v 30S unoprovddo ru eivot une0Ouvog yro rov onoxtreropr6 6trtrrov o1-uvoorul.o-tRNA on6 qv ow[6poor1 6vop[qq. To GTP udpoLtietot, og6rou to evoprcr{pto IRNA npoo6e0e( qro oU;rn}.oro Svop(qg. Or nopdyoweg 5vop[49 np6ner v' one].euOepo0otiv drore q geyor4 unopovddo vo prnope( vo ouvduoote( pre ro o6prnLor<o Svop(r1q. Iroug europutineg 4 Svop[r1 nepil.oprBdver preyo].rj- repo opt0pr6 nopoy6wrov. Oprop6vot epntr6xowor orrlv op1Kn np6o6eor1 rqg aOS unoprovdEoq orqv xofurintpo rou 5'6rpou tou mRNA, oqpe[o oro ono[o ro arupr<rr1- pto IRNA efuor npoo6e6eprSvo pe dM4 o;166o nqpay6rF tov. Merd on6 ourl qv oplrrrl np6o6eorl, n Utxp{ unopovd6o oopc0ver ro mRNA 5og 6rou ovoyvcopfoer to o<oo16 r<ro6rx6vto AUG. 'Enerto, ot nopdyoweg 6vop[qg onetreuOeprilvowor rr 4 605 unogovd6o ouv- 6ud(eror pe ro o0pnl,oxo. Or npoxopurrlnro( nopdyoweg enrtrr{r<uvoqg epn},6- Kowor. orqv enrprqruvoq. O EF-Tu npoodSver opuvooru- ).otRNA oro 70S prBooroprdrro. GTP u6pol0eror 6rov oneLeuOepc0veror o EF-Tu, evrir o EF-Ts ononefuor yto v' ovoyevrrqOe( q evepy6g Uopgri rou EF-Tu. O EF-G onomefuor orq peror6nroq. Iro ppoocoprdrro npoo66- veror efue o EF-Tu e[te o EF-G, yeyov6g nou 6roorpo],G (er 6n rdOe Bqgo np6ner vo o),oxLqpoOe( nporoU (errv{oer ro en6pevo. O repporroll6g ouprBo[ver oe onoro64nore on6 ro tp(o er6rd ro6x6vro UAA, UAG ror UGA. Or nopdyov- reg oneLeuO6poonq rd(rlq 1, nou ovqyvop(ouv er6u<d to rrrl6rr6vro repprortopro0, evepyonoro0v ro prBootogd- no oe u6p6Luon rou nemr6ul,o-tRNA. 'Evog nop6yov- rog RF rd[49 2 qnore(ror yro rnv one],euO5poon rou nopdyowo RF rd[49 1 on6 ro prBootoprduo. Or npoo- 6e6ep5vor pe GTP nopdyoweg lF-2, EF-Tu, EF-G ror RF3 6Xouv nop6poro 6op(: or 6tio rel,ewo(ot ;.u1.roriwor q 6ogrj RNA-np<oreivcbv rurv 6Uo np6rrov 6rov e(vor npoo6e6eg5vor oro IRNA. 'OLor rouq npoo66vowor orrlv (6to ptBooogrr( 06o1, tr1 06oq np6o6eonq rou nopdyowo G. To prBoocoprdrro efvor otr:poil6ro prBovoux],eonpro- reivdrv, oro ono(o ro preyolUrepo prSpog qg pd(og no- pdXetor on6 ro rRNA. To oXqporo 6Lcov tcov prBo- orrlpo{tov yevu<d e(vor nop6trroro, oor6oo to Borcrrlpro- xd p$ooco;rdtro (70S) dXouv neprypoge( Lemopreprirq. H pxpri unopovd6o (30S) 6Xer oupneopSvo ofnUo, U' 6vo "oriuo", nou nepdXa nepkou ro 60o rp(o rqq pd- (oq rot 6roXrr:p((eror on6 rqv "xego[rl" p6otrr ptoq oXt- oUnS. H geydLq unopovddo (50S) e(vor neptoo6tepo orporprrrj pre 6vo npoe[61owo ";r(o1o" mo de[td t<ot Uto (KewptKli npoe[oX(". Ot ewon(oetg 6ov tcov npol- reivrilv orrl ptr<pq provdSo e(vot yvoor6g rotd npoo5yyt- on. Kd0e uno;rovd6o neptdXet 5vo pr6vo x0pto rRNA, 165 r<gr 23S oroug nporopucbteg, 18S xot 28S oroug europurireg. Yndplouv en[or1g etrdooovo rRNA pe ne- proo6repo o[roorlpre(<oto to 55 rRNA orrl peydtrrl uno- provdSo. Kor ro 6rio xrjpro rRNA e;rrpovf(ouv ertero- ;i6vo (euydpcotrro Bdoeov, rup(cog un6 propgt'1 FpoXd- cov, orelr6g (euyoptog6vtov drnl,drv pr[oXutv Ue Uov6- xLrovoug Bp6Xoug. Iuwqpqpr6vo Xopocnptortxd oto rRNA towonoroUwqr ouyxp(vowog ttg ollqtrouX(eg pe rrg deureporoye(q 6ogdg nou pnope( vo oledto- oroUv yro ro rRNA noxfi.tov opyovtoprcbv. To 163 rRNA 5Xer r6ooeprg 6ror<ptr5q neproX6q, evd: ro 23S rRNA 6[r. To europurottrd rRNA 6ro06rouv entnp6o0eteg ne- pro16g. H rpuoroMu<n 6oUri 6e[Xyet 6tt q 30S unopovd6o 24:i l]eprtrnUn 709
61et ooriprperprl r<orovopui RNA r<or nporeivc0v. To RNA ouyrewpriveror orrl 6rdgoo4 Ue rnv 50S unoprovddo. H 50S unogovddo 6ro06,rer gto entEdvero npulteiVrlq, pe gor<pd poBS(o 6k).ovou RNA ro ono(o 6too{(ouv q 6o911. H oriv6eoq 30S pe 50S nepfiotrrBdver oqpre(o enogqg prero(ri rrov 165 rRNA r<or 23S rRNA. Aopru<5q oL).oy6q ouprBo(vouv Kor orrq 6Uo unopovddeg 6rov ouvdud(owor. ylo vo oXrlpor[oouv rivo n].r1peg prBooro- prdrro. Kd0e unoprovddo 6ro0drer nolLd evepyd rdwpo, ro ono(o ouyrewpdrvowor oill UrroepoqrrKq nepro- pi rou prBoocol.ror(ou 5nou ouw(0ewor or nporeiveg. To no).unenr(6ro eyr<oroLe[nouv ro prBoocoprdno 6ro- pr6oou qg neproXqg e[66ou, q ono(o ;rnope( v' ol,l.4- Lenrdpdoer pe Uro pepBpdvq. To rripro evepyd r6wpo e[vor or O5oerg P r<ot A, 11 05o4 E, or neproX6g np5o6e- orlq rov EF-Tu ror EF-G, q nenrr6utrorpovo<pepdorl r<or 4 06oq np6o6eoqg rou mRNA. H 6rog6prpoo4 tou prBootrrl-ror(ou gnope[ v' o],].d(et oe drdrpopo ord- 6ro r4g gerdrppoorlg: 51ouv ewonrore( dro<poprig oq npoq rnv npooneloorp6rqto ouyrerprg5vov nepro- 1d:v rtov r0puov rRNA. To IRNA orrg 06oerg A r<or P efvoL nopdMq],o peto- [0 roug. Or Bp61ot rrrlv owrrco6rxov(rov npoo6Svowor mo mRNA oe Uro qU^,oKo qq 30S unoprovddog. To un5trorno rpnpo rdOe IRNA npoo66verot orqv 50S unopovd6o. Mro evotrtroyr1 6rop6prpoo4g tou IRNA p5- oo orrl 05or1 A efuor onopofuqr4 c0ore ro ol.rwooruAu<6 dr<po rou vo Bpe0e( 6(ntro mo drpo rou nenrrdul,o- IRNA or4 05or1 P. H 05or1 qg nemr6ulorpovorpepd- onq nou ouv66er rrg O5oerg np6o6eor'19 P r<or A onorcLefuor on6 23S rRNA, ro ono(o eprpov((er xorotru- tu<r1 evepy5rqro nenn6ulorpovoqepdonq, pol,ov6rr yto vo 6r1proupyr10e( 4 ooorri 6opri prdMov onotro0v- ror Kor. npa:reiveg. O evepy6g p6log rtr:v rRNA mr1 pretd<ppoorl uno- 6errvUetot on6 peroMd[erg nou enqped(ouv rq Ler- roupy(o rou prBooro;ror(ou, on6 olLql,enr6pdoerq Ue to mRNA r<or ro IRNA, 6noq en(oqg ror on6 qv ovdy- xq drorlprlonq Eexoprfitirv oMq).ent6pdoeo:v (euyo- prilprorog Bdoecov pre ro IRNA xor ro mRNA. H 3'rehr<4 neprolq rou rRNA (euyopdrver pre ro mRNA crrrlv Svop- En. Or eororeprr6g neproX6g npoyporonororiv r6to(re- peg eno<pdg ;re ro IRNA 16oo orq 06oq P 6oo ror rq 06oq A. To prBooo;ur6 RNA e(vor or6Xog opropdvrr:v owtBronrcbv r1 dMrov nopoy6wrov nou ovoor6trl.ouv rrl prerd<ppooq. BrBAroypagio 24.4 H 'Evop[r1 0T0 BoKrrlpro Xpero(etor 30S Ynogovodeg Kr EnrKouprKouq nopoyovTeq .'1:,;j1;r 1,1 1 I 11;: I IVlaitra, U. (1982). Initiation factors in protein biosynthesis. ,4nnu. Rev. Biochem. 51, 869-900. Noller, H, F. (2007). Structure of the bacterial ribosome and some implications for translational regulation. ln "Translational Contr: in Biology and IVledicine." (Mathews, M. 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24,20 0t Prpootopx6g AoUeq AIId(ouv'Orov luvDuoororjv or YnogovdDeg AvL:gcpdq Schuwilrh, B. S., Borovinskaya, M. A., Hau, C. W., Zhang, W., Vila- Sanjurjo, A., Holton, J. M., and Cate, J. H. (2005) Structu':. :' the bacterial ribosome at 3.5 E resolution. Science 310 82;- 834. Btptrroyporpio 713
Avotun60qre 0nd J. Struct. Bioi., vol.157, K. Y. Sanbonmatsu and C.-S, Tung, High performance computing in biology , pp.470-480. Copy- right 2007, rordniv obeIoq ono Elsevier Ihttp://www.sciencedirect.com/scrence/journall1047B477). H rporoypogro nopoXcoprl0lre 0n6 rcv Kevin Y. Sanbonmatsu, Los Alamos fJational Laboratory. Tou ovTrKodrKov(ou Kor TIq rpiTnq Boorlg rou KodrKov(- ou nportinrer 0rt0 U X0tr0p6repq enurlplorl rou (eu- y0p6U0T0q ono T0 vouK)e0T[6r0 rou rRNA oTri prpoooprKn 0eon A. ffi fo IRNA l-lopoyovror pe Ene(epyooio Meyotrurepov llpri- 6poUtov Mop[rov * 'Evo rirprUo IRNA dlproupye[ror pe ercqepyoolo svdq npo6popou lLopiou. a To S'oxpo Dlptoupye(ror peoo droonooqg ono rr;v ev- dovourtredorl RNAdol P. e To 3'oxpo 6lproupyeiror geoo notrtrontrov ev6ovou- rtreotrunxdrv rt e(tovourLeotrurtrdrv droonooeov, ng ono[eg oro],ouOei q npoo0qrrl rou rorvou rep;lorxof rprvourtreond[ou CCA" ffi fo IRNA l-lepr6Xer Tpononoulpeveq Booerg * To IRNA neprdXouv novo on6 90 rpononotqlreveq po- 0etq. s H rpononolr]on ouvn0oq ouvlorotor oe dpeol otrtroyr1 rotv nporoyevtirv pooeov rou IRNA, oorooo undpXouv oprop6veg e[op6oetg rinou pro poorl otpotpe(ror xr ov- rlro0rororor on6 Uro otrtrn. O I-eveTrK6q I(66lroq Iohn Perona *iHg ttooY(')Yn ffiff9 Iuyyevfl Ko0rxovLo Avlnpoooneuouv Xqgrrog l-loptlpoto A;uvo[60 * EEnvro pto on6 rtg e(rlvro rdooeprg rprntr6reg rtodrro- norouv e(xoor ollvo[eo. e Tp[o rtodrrdvro dev ovrrnpootoneuouv oprvol6o ror nporoLouv repportop6. e 0 yevelroq 166rro9 unoyooe, oe nptrtrl.to orodro rqg e(elt[1q ror elvor orroupevxog. a To neprooorepo ollvo(60 ovlnpooone0ovror onri 6uo 11 neprooorepo rtodtr6vto. * To notrtrontro xrrrdtxovro yro evo oluvo[u ouvqOorg eivor ouyyevrl pero(u rouq. * XrluLrtlq nop6poro olitvo[eo ouXvo rlXouv ouyyevrl ro- 6ixrivto, etroXLoronordrvrog eror rtg enrnr6oetg Tov Ue- rotrtroIeov. ffi H Avoyvoprol KroDrrovrou-Avlxtodrrov(ou llepiloggover Aoro0eLo * l-lotrtrontro xrodx6vro rou ovTrrrpooone0ouv ro [6ro o1u- vo[t1 ouXv6repo drorpepouv oq 06or; rqg rprqq pooqq. e Aoro0ero oro (euyopcolro pero{f qg npdrr4g pooqq 714 KEOA Al0 25 0 l-everuoq Ktil0rxog ,@, ,! lr I I t'tt l, i, i-. ?a' .i F- '-- r-- . *'t::>- * tF4 &- t/ - @t,
ffig* e 0r Tponoroulueveq Pooerq npoo6[60uv ouEnudvn oT0- 0eprlqro oro tRNA ror pu0p[(ouv t1v ovoyvtitptorl rouq ono nporelveq rot otrtro RNA rqq peroqpoorurlg ouoreuflg. 0L Tpononoulpeveg Booerg Eqpeo(ouv ro Zeuyoptolto AvlrolD lrov(o u - Krr:d rrov(o u e Tpononorrloetg oro ovtrrtrldrrrivLo enrqpeo(ouv ro npo- TUno Tou ooro0orlg (euyoptirporog rot ouvendtg e[vol olpovnr6g yro rov xo0optogo rrlg etdtrdrrpog tou tR- NA. YnopXouv InopodrxSg Tpononou;oerg rou Otroupevtrou K6drro e Ie oproutlvo erdn 6xouv ouppei otrtroytlg oto yevenrd rdl6rro. * Aur6q ot otrtroydq e[vor ouvl0eoTepeq oT0 UrToXov- dpLoro yovtduirl-roro, rinou pnope( vo rorooreuoote( 6vo rputroyevertro devrpo yro rLg otrtroy6g. * Iro nuplvuo yovrduirporo, ot otrtroy6g ouvrl0tog enq- ped(ouv p6vo to xoDrrovro reppolopori. Neo Aprvo[60 Mnope[ vo Evotrlgoro0ouv oe 0propdvo Ktrl6xrjvro Tepgortopou * H evootuoroorl qg oetrlvoruoreivrlg oe oplop6vo ror- drxdvro UGA onoue( rr; Dpool evoq oouvl0rotou IRNA oe ouvduoogo Ue notrtr6q nptorelveg. e To oouvrl0roro ogrvo[u nupotruolvr1 ;lnope[ vo evoouo- rol0el oe oprop6vo rtrldrrrjvro UAG. c lro 0treQoptdoQtipo nporo(oo Euplotes crassus, T0 roduovro UGA ro0opt(er rrioo rr1 oetrlvoruoreiV4 tloo rot rrlv xuoretVl. ffi fo IRNA Zeuyopdrvouv Er),exlrd pe Aprvo[60 Meoto Aur- voorutrotRNA Iuv0eroodrv I 0r ourvooKutro{RNA ouvOerooeg e[vor 1uo orroy6veto ev(0Ur,rv, ro ono(u npood6vouv oluvo[eo oro tRNA nu- pdyovrog ol.uvoorutrotRNA oe pto ovridpool 611o pq- pdrtov nou Xprlorgonore[ ev6pyeto onrj ro ATP. * Kd0e outvoorutrotRNA ouvOeroorl oluvoorutruiver dtro ro IRNA oe pro roodernr1 opo6o nou ovlnpoooneUer 6vo ouyrerpq6vo oluvoIu. * H ovoyvdrproq rou IRNA ond rtg tRNA ouv0erdoeg po- ol(eror oe pLo ouyrerpqr6vq ogo6o vourtreortdltov, qv (oud60 tourdqrog, rou IRNA, ro ono[o ouXvo ouy- revrptirvovror oro ot6treXog rou onod6rrrl ror ro ppo- Xo rou ovlrtodxov(ou. ffi 0'AgrvooxutrotRNA luv0erooeg Kororoooovror oe Arjo 0rroy6vereg I 0r ourvooKutro- IRNA ouvOetooeg Dtorp(vovrot ortq ot- roy6veteg ro[qq I rot ll pe pdoq rdrofrepo Uor(00 otrtrn- trouXuirv xot rdtolrepeg 6opxeg neproX6g. ffi O, Iuv0erooeg Xprlorgonoro0v q Ar6p0ooq Aortplov yto vo Be),rtiloouv rnv Arp(BeLo * H etdrrouro rou (euyoptirlrorog pero[f oprvo[6ov xot IRNA puOpi(eror ;.r5oor ovrtdpooetov 6rop0oorlg 6orr- p[ov or ono[eg udpotruouv troOoq oxrlU0logeveg opr- voorutro-odevutrx6g evdroerg ror oluvoorutro-tRNA. ffi fo Koroorotrlro IRNA l-iepreXouv Metotrtroy;r6vo Avrtro- drrovLo, ro 0no[o Aropo(ouv N6o Kodtrovro e'Evo xoroorotrlrd IRNA runrro qeper pLo perotrtr0[r1 oto ovrtraldxovto nou otrtro(ei t0 ro6txov,o J:i : ,:,": onovro. * Otov ro vto ovlrtodtrovto ovrtorotXel oe rol6t<o;r, reppottoltou, evotopordtverot 6vo optvo[0 rot q no],tr- nenl6rrrl otruoido enerre[vetot n6po ond ro xtr.tdtxcl- vro TepUoTtopou. Aurd o61yei oe oouvOertr1 K0T0o-I0- trrl oe Uro 06o11 oouv0ertrnq Uerotrtro[lq 1 oe ovoyvtrt- o1 nepo ono 6vo rovovtrd rotdtrovto repportopou. * Auouv0elu'l rotoorotrrl ouppoivet dtov ro IRNA ovo- yvtop(er dvo drorpopenrrj rtodx6vto on6 ro ouvrl0to;t6- vo, onore 5vo olLtvo[tj ovrtro0tororot ono ronoto dtrtr0. ffi*,} Ynopxouv Aouv0erxol Kotomotrelg yto Kd0e Ko:Dtrovto Teppolopou. s Ko0e runog oouv0ertrou xodtrov(ou roroor6trtrerot on6 6vo tRNA pe perotrtroygevo ovrtrotdtrdvto. * Oprourivo onovto roroorotrlrd IRNA nept6Xouv perotr- tro[erg oe dtrtro rpt1poro tou ltoplou. ffi O, Koroorole(g Mnopei v' Avroyovi(ovror qv Kovovtrq Avdyvororl rou Ktildrro * To roroorotrtrro IRNA ovroyovl(ovror ro IRNA oypt- ou runou nou nept6Xouv ro (6to ovrtrtrldtr6vto yto rr;v ovoyvtrlol rou ovr[ororxou rordrrov(ou (tl 1(l]6txo- vitov). e H onoretreoUonrrl roroorotrfl eivor en$tropr1q, enetdrl odlye[ oe ovoyvtrloq n6po ono r0 rovovrro rodtxrivto reppolo;rori. * To rtrldtr6vro UGA epqovr(er droppon xor dtopo(eror eoqotrp6vo ond ro Trp-tRNA pe ouXvoqro 1 -3%. ffi fo Prpooorgorto Ennpeo(eL rrlv Arpipero qg Merdqpooqg a H 6o[rl rou 1 65 rRNA ong 06oerq P ror A rou ptpooo- por(ou eqpeo(er rqv oxp[pero qq petoqpootlg. ffi: Merordmorl Tou ll),oto(ou Avoyvoorlg Mnopeivo luppei oe 0trrofipeg AtrtrntrouXieq * To ntro(oto ovoyvtrlorlg enrlpeo(eroL ono rrlv otrtr1trou- Xio rou mRNA xor ro prpootolrLr6 nepLpotrtrov. a 0t otrtoOnpeq otrtrntrouxieq enlrp6nouv o' 6vo IRNA vo peroroni(erot roro pro Boon og6rou (euyoprioer pe ro ovlrodrrdvrd rou, petopotrtrovrog 6rot ro ntro(oto ovo- YV(00nq. er H UerdQpool optop6vtov yovtdltov eloprorot ono rrlv roxnx1 npoypopUoTropdvrl perordnroq rou ntrotolou ovoyvtoorlg ffi afio luppovro Enovoxodrrono(norlq: Merorppoortrfl l-lo- poropqr; ror o MlXovrolrdq rou tmRNA yto qv Anetreu- 06ptrro4 rtov Ko0qtrtop6vrov PLpooropot[ov e H noporouqlrl ouvfororor oqv xovtirr[o rou ptpooto- ;ioriou vo reppori(er q peroqpootl, v' onetreuOepdtve- ror ono ro mRNA rot vo (ovopX((et rI Ueroqpootl nep(nou 50 vourtreor6ro ro0o6rro. * To pL0ootouorto nou e(vol xo0r1trotg6vo novo oro mR- NA pero rr; peprrrl ouv0eorl prog nporreiVlg pnopei v' onetreu0epo0o0v ono rr1 6poor1 rou tmRNA, evoq po- vodxori RNA pe Xoporqprorxo rooo rou tRNA 6oo rot rou mRNA. ffi nepfi1qrl 715
25.1 ErooyoVri H ol,i.4).ouX(o ev6g ro6rronor4rrroU r<),c0vou DNA nou droBd(eror oe rore60uvoq on6 Shpog 3'qnore- l,e(ror on6 rprni.6reg vou xl.eorr6(rov (rrrl6x6vto) owi- ororxo pe rnv oMrlLoul(o o;uvo{6rov ptog nporetvrlg nou 6toBd(eror on6 ro o1-rlorelrx6 npog ro xopBo(u- tetrrr6 drpo. H oMq),oriX4orl rou DNA Kor rov npo- teiVciv enrpdner qv dpreoq oriyrproq rov vouxl,eo- rt6xc6v Kor rov opuvo[rr6v oM4].ouXrc0v. Yndplouv e[qwo r6ooepo rco6rx6vro (xoOrivo on6 rq rSooepo nr0ov6 vourtreor(dro prnope[ vo r<oro],6Ber pro on6 rg tpetg 06oetg tou rodu<ovfou, 6qpuoupyc0wog 43 = 64 ntOovdg oMrl),ou1(eg rpwoux,treorr6(ov). Ko06vo on6 ourd ro xo6u<6vro 6Xer er6u<5 v6qpro orq perdEpooq: el{wo 5vo r<ro6rxdvro owrnpoooneUouv ol.uvofdo, evci ro un6l,orno rp(o enr<p6pouv rov repporropr6 r4g perdgpoor;9. To v6qgo ev6g xur6xoviou nou owrnpooone0er 5vo o1-uvo(r3 r<o0op((eror on6 ro ow(mot1o IRNA, evdr to v64pro rcov rcodxovfrov repgonoproU r<oOop((eror dpeoo on6 nporeivu<oriq nopdyoweg. H onorpumoypdrprloq rou yeverrKoU xril6u<o opXt- xd 66er[e 6r or yeverrr5g ilrr1po<pop(eg ono0qr<eUowor und poprpfl vouxLeorr6rxrilv rprnlerriv, om6oo 6ev onoxdLurle noro opuvo[ri xo0op((eror on6 rd0e 11116r- r6vro. [lprv on6 r4v 5].euor1 qg oMq],otixnonq, ro v6r1- Uo rov r<rodrrov(ov npoodtopr(6rov pe Bdorl 6Uo r0noug preLercilv in vitro.'Evo o0m4;ro nou Boo((eror on1 perdqpooq ouvOenxriv noluvou xLeortd(cov eroriX- 0qre ro 1961, 6rov o Nirenberg 66er[e 6n to no],uoupt- 6uLu<6 o[6 [no].u(U)l roreuOrlver rrl ouvoppo].6yr1or1 qg <porvuloLov(v4g oe noluEorvul,oLov(v4. Aur6 ro onor6l,eopo oqpo(ver 6rr ro UUU npSner vo e[vor ro rco6rx6vro yro rrl eorvul.ol,ov(vq. I' rivo 6e0repo, Uero- yev6orepo o0mr1po, 6vo rptvouxLeor(6ro Xprlorpoilor- q0qre yro v' onoprprlOe[ 6vq rcodu<6vro, rdvowog ro ow[mor1o ol.uvoor<u],o-tRNA vo npoo6e0e( oro ptBo- tlpc,tq pdorl UCAG oropdrro. Towonorc0wog ro opuvo(ti rou ol.rloorulo. IRNA prnope( vo BpeOe( ro v6q;ro rou r<ro6rrov(ou. Mo(i, ot 6rio re;yrr6q npoo6r6proov ro v6rlgo 6trcrlv rrov xro6r- rov(rov nou owlnpooconeUouv oprwol5o. E[rlwo 6vo qn6 ro e[rlwo r6ooepo rrrlSrr6vro owt- npooconeUouv o1.uvo(5o. To un6Lorno rpfo enrg6pow rov rspporroU6 rnq ;rerdgpooqg. H on66oo4 oprwo[6- ov oro r<rrl6x6vro 6ev efvor rulo(o, oMd unoyopeUerur on6 q46oerg ong ono(eg q rpfuq Bdorl 51er pu<p6tepr1 en[6poor1 mo v64pro rou r<ro6rxov(ou. Enrn].5ov, ]3p1. r<drg nop6poro oprrvo[6o ouXvd owrnpoooneUowor on6 ouyyevrl ro6u<6vro. 2 5.2 Iuyyevfi Kto6rr6vro AvTtnpoooJnEUouv Xnprrtiq [op6Uoro Aprvo[6o * E[nvro p(o on6 rLq e{qvro r6ooeprg rptntr6reg rtoDrronolouv e[rooL o;-uvo(eo. * Tp[o xroDrrovro dev ovrrnpooulneuouv olrtvo[60 Kor ttpoKo- trouv reppoloprl. * 0 yevelrdq 166rro9 ((r0y00eD oe npdrrpo orodro r4g e[etrr- [1q xor ervor orxoupevtrog. * To neprooorepo oprvo[eo ovrtnpooone0ovrot ono 6uo n ne- ptoo6repo rtrldrrovro, * To notr),ontro xrodtrdvro yLo tlvo o;uvo[0 ouvrl0oq e[vor ouy- yevl gero[u roug. $ XnUrKdq nopo;.roro oluvo[60 ouXvo eXouv ouyyevrl rodtxovro, e),oXroronouirvrog 6ror ng entnrtloerg rov perotrtroIeov. O rdl6u<og ouvotf((eror orr;v IIH*tiA 15."i. Yndplouv ne- proo6repo rto6tr6vro on6 opuvo[5o: dpo, oXe56v 6].o t' oprvo[6o owrnpooone0owor on6 drjo q neproo6te- po r<rodtx6vro. Movodrx6g e[orp6oerq e(vor 11 preOerov(vq Kor n rpurrogdul. To rcodrr6vro ge ro (6ro v6qpro r<o- Lorjwor ouvtirvupo (synonymous). O yeverrx6g r<or6L- Koq ornv npoy;,rotrx6tqto 6roBd(eror ndvo oro mRNA r<r StoL ouvq0og nepLypdgeror oe o16o4 pe lg rSooe- prg BdoeLg nou undpXouv oro RNA: U, C, A r<oL G. To r<co6Lr6vro nou owrnpooolneUouv to (6to n XnUr- r<org nop6proro opuvo(6o ouvrlOotq 61ouv nop6proro otr- l.rllouX(o. IuXvd, n pdon orrlv rp(rr1 05or1 evdg r<ro6tr<ov(ou 6ev e(vor orlpowu<r1, ener6q to rriooepo rr,,.t- 6tr6vLo nou drorpdpouv pr6vo orqv rp(tr1 Bdoq owrnpo- ooneUouv to (6ro opuvo[ri. H ;reulp6vq erdrxdqro orqv relewo(o 06oq e[vot yvotmrl otg exgr3trroq rpir4g B6- oqg (third-base degeneracy). Fro vo perorppoore[ 6vo ro6u<6vro rou mRNA, r] npcD- eiK&l{A :5.1 Otreg or rprntr6req-rtodrrovto 6Xouv vdqpo: e[rlvro gio ov- rtnpoooneuouv o;-uvo[eo Kor Tperq nporotrouv i1i1 (ST0P) FO b.6 =a P= UUU l_Phe ucu I UAU -LTyr UGU -LCys UUC -] F UCC I Ser UAC -] Y UGC -] C UUA LLeu UcA f S UAA l_(rnD UGA SToP uuc _r L ucc _.1 UAG I "'"' UGG lll CUU I CCU I CAU _LH|S CGU I 696 | Leu ccc I Pro cAC I H cGC I Rrs CUA f L ccR f ecnnleln CGA [n CUG -] CcG I CAG -] O CGG -I AUU l ,- ACU I AAU I Asn AGU I Ser AUC l": ACC Im,nncf ru AGCl-s tut I g, ffit l-' titl'il' t3t lT GUU I GCU I GAU I ASP GGU I GUc I va Gcc I Aa GAc f D ccc I GIy cuA I v GCA l- A GAA lGtu GGA f G GUG -] GCG -] GAG -] E GGG -] 716 KEOMA|0 25 0 l-everxdg Ktildrrog Boorxeq ewoleg
rn 96on rou npSnet vo (euyopdroet ge rqv npc0rr; Bdorl rou owtrrodrxov(ou tou ow[orotXou ol.twooru].o-tRNA. Aw6 ro (euydpropro ouprBo(ver pr5oo oro p$oocoprdtro, 6nou 11 oM4Len(6pooq ;reto[ri oupdrqpoptottt<tilv tpt- vouxleon6[tov oroOeponotefuot pSoto e[openrd ourrrl- prlpr6vov voux.l.eott6(tov tou 165 rRNA orrl 05or1 A. H ouorqpn enrnpncnl rou ouvolu<o0 o;q4porog tcov (euyo- polpr6vov Bdoeov on6 to rRNA enmp5net vo npqYporo- noq0e( 51t pr6vo ouprBcnrx6 (euydpopro peto[0 A-U t<ot G-C ong 6rio npcirreg 06oetq tou ro6tt<ov[ou qI d Kt en[- np6o0ero (euyoprilproro or4v rptrrl Bdor1, 6nou ot eTIo- <p6g rou rRNA e(vot Lty6tepo ouprntrqpoprorxdq. Iuve- nri.rq, 5vo ol.rwooruLo-tRNA prnope( v' ovoyvrop((et ndvol on6 5vo r<todx6vto pt5oto rov enmp6o0erov, Ul'l Kovovt- r<drv (euydrv nou enrp6nowot on;v rpkq 06o11. Enmtr6ov, or olrltrenr6pdoetg tou (euyopdr;rotog (oroq enrlped(ov- ror on6 rrl pero-Ueroypogrrrl tponono(qoq rou IRNA, rd(oq ;16oo r1 og6otr:g 6inLo oro owxtodu<6vto. H rdoq Xqpxtirg nop6ptouov oprvo[5tov v' owmpooo- ne0owor on6 ouyyevn rto6tr6vto el.oXtoronote[ lq ent- mcboerg rcov gerotrtrd[eov. Au(dvet rrlv nt0ov6rr[o Uto pr6vo ru1o(o oUoyt Bdoqg vo prlv npoxotr6oet owxotd- orooq oprwo[6og r1 vo o6qyrjoet oe qwKordqroorl nou nepil.oprBdver optwo(5o nop6protou loporcqpo. l-to nopd- 6erypo, 4 predMo[r1 CUC npog CUG 6ev 6Xet xdnoro en[- nroorl enet6rl rorro d[o ro6tx5vto qwmpoooneUouv rq leur(vr1. H perdllo[q CUU npog AUU 5Xa og onor6ls- o1.ro owu<otdoroon rnq treur<(vr1q on6 tooLeurtvq: ourd ro 6rio oprvo[5o e[vor u6p5tpoBo rot nt0ov6v 6todpopro{- (ouv nop6proro p6Lo ornv rtodxonototlprevrl nponetvq. H Ell(0NA 25.2 nopouotd(et puo ypqqtxli nopdoroorl rou opr0proU ttov rordrrov(ov nou owtnpoooneUouv r<dOe oprwo[0 oe oX6o1 pre q oulv6tnro pe tqv ono(o ro ouyKeKprp6vo opuvo(ri Xprlogonote(tot ortg nporei- veg (oro E. coli). l-evu<d, ro ouvq0Sorepo ogtvo[6o ov- nnpooconeUowor on6 6Uo rl neptoo6tepo ro6tt<6vtq. Aw6 uno6qtr6vet 6l ouv6B4 rdnoto Betrttorono(qoq Leu . rou yeverrKoU r<ci6u<o ovorpoptr6 Ue rn Xprlotgototqtrl rrov o1.rwof6tov. To tp(o xcr:6tr6vto (UAA, UAG xot UGA) nou 6an or rnpoorone6ouv o1.rwo(5o XpnotUonototiwot et6xd yto rov repuqnopr6 qg pret6gpooqg. 'Evo on6 our6 to xor6r x6vro rep gorro;.rof (stop codons) orl poto6orc[ to r6Log rdOe ovotXro6 ntroto(ou ovdyvooqq. Iuyr<p[oerg otr],r1].ouXtc0v DNA pe trg ow(motleg notrunemrdx6g o).Lql,ouX(eg onorol,0mouv 5r rq Kto- 6tr6vto 6Xouv ro (6to v5rlpro t6oo qro porrtlpto 5oo xot orouq euKqpurilreg. Iuven6g, mRNA ev6g e(6oug ouvrl- 0og prnope( vo Ueroepome[ ocoor6 in vitro rl in vivo on6 ro UsrogpoortK6 pr4Xovtog6 ev6g 6Mou e(6oug. 'Etor, To ro6rr6vro nou Xpnotponototiwot oro mRNA ev6g efdoug 6Xouv to (6ro v6rlpro yto ro prBootopduo xor to IRNA otrrov er66v. H orroupevtr6rrlro (p' el,dlrmeg e(otp6oetg) tou xci6rro e(vor 5vder[r1 6rt r<o0tep604xe notr6 nprilrpro ro- rd rqv eESLtEn. 'lorog o r<cir6xoq (ex[vqoe ge pro npcot6- yovrl prop<prj, orrlv ono(o Lty6repo r<o6x6vto owtnpo- odlneuqv ouyxpttu<d l,iyo o1.two[6o, (oolg or<6ptr1 rot pt' 5vo ro6rx6vro yro onorodqnore pr6trog ;rtog opr66oq opuvo(6cov. ArprB6orepo vorlproto t<to6trov[tov Kt eTIt- np6o0ero oprwo(6o eroq104r<ov opy6tepo. Mto nt0ov6- r4ro e(vor 6r opXu<6 Xpnoqonotoriwov g6vo 6tio on6 nq rperq Bdoetq r60e ro6u<ov(ou: 11 6tdxproq orrlv tp(- q 06o11 (orog e(eLiX04re 6rPrpo. H e[6h[q tou t<til6xo prnopoUoe vo 6Xet (noY(ioet,, o' 5vo oqpre(o rord ro ono(o rc oUorrlpro eile r<oroore( 16- oo notr0nLoro tiore onotodrlnore ;retoBotrrl oro v6qpro rcov r<rodu<ov(rrlv 0o dtordpoooe rtg undpXouoeg npotei'- veq ;r6ou: prq ono6errcilv ogrvo(u<riv unorcnoordoeov. H oxoupevx6rqro rou rtil6u<o uno6rltrdlvet 6n ow6 np6net vo ouv6Brl oe 16oo npr6tpro orddto riore 6trot ot dprBtot opyovrol.ro( npo6pXowor on6 rqv (6to 6e(opevq opX6yo- v<ov rurrdpov 6nou ouvSBq ro .mdyoUo, tou rc06u<o. Or e[orp6oetq rou otKoupevtxoti yevettxoti rril6tro e(vot ondvreg. Ot oMoySg oro v6rlpro tou np<rltopltroU yovr6uill.rorog ev6g e6oug ouvqOcog orpopotiv oro Ko- 6rx6vto repportopoti. l-ro nopdderygo, o' 5vo My- coplasma, to UGA xtrl6tronote( tpunto<pdvn: oe opropr6vo e(6r1 rov BLeqopt6or6v Tetrahymena Kot Paramecium, ro UAA r<or UAG rro6tronotoriv yLoutqt[- vq. Ium4potu<6q oMoydg tou x66tro 6Xouv ouprBe( p6vo oro prolovdptor6 DNA (BL6ne Ev6trya 25.7). 25.3 H Avoyvtirpron Ko6troviou- Avrrrtrr6rroviou ileprAoUB6ver Aord0eto * l-lotrtrontro rordtrovro ttou ovTrnpooone0ouv ro Ldto ol.ttvo[u ouXvorepo 6rorp6pouv oq 05or; rqq tpiqq poo1q. * Aoro0eto oro (euyoprrrlro pero[0 qq npriqg poolg rou ovrt- ro6trov[ou ror qg rp(rr1g Boonq rou rodtrovlou nporunret ono rr1 Xotropdrepq enu1prlorl rou (euYoptitporog ono ro vourtreor(dro rou rRNA oq ptpootogurl 06oq A. 10 F .= P o =b d d .a P o b o E B Glu . ^ Lvs. o ' lle. ^ Met. g;s. a Cvs. Trp . . AIa . Gly Ser . . Val . Thr . Pro Arg .Aso . Gld 'Asn . 4 Phe. Tyr . a '.rr:;, ri, :, n.o*r0,.il'';o,;H.iJ.,, qs ouxvoqros Xprlolq oprvoE6tov ortg nptorelvsq Us Tov optOpo rtov rotdtxovltrlv, nou ro0opf(ouv ro oprvo(u. YnopXet pto e[o(peo1 Yto T0 optvo- [eo, nou roOopf(ovror on6 duo rodu<ovto, ro ono(o urpiorovrol oe eupu tpoouo ouxvorr[ov. 25.S H Avoyvtirprorl Ko:dLxovlou-Avrxtodrrov(ou I-lepilqUBdvetAm60eto 711 Boorreg evvorcg
UUU UUC UUA, UUG CUU cuc CUA CUG AUU AUC AUA AUG GUU GUC GUA GUG UCU ucc UCA UCG ccu ccc ccA CCG ACU ACC ACA ACG GCU GCC GCA GCG ?iirt_( UAU UAC UAA UAG CAU CAC cAA. CAG AAU AAC *AA AAG GAU GAC GAA GAG 1 r:i:;:r:. lii;:;:::- i;,:1.. t i:i UGU UGC UGA UGG CGU CGC CGA CGG AGU AGC AGA. AGG GGU GGC GGA GGG U,C,A,G U,C,A e I 0r nouprveq droqripouv A n G G _l or0 Trq flupruro[veq U n C ffi Movu0rxrl 0L rpireg pooerg eXouv Tnv r),oxroTn rnppo1 0T0 v0np0 rov KtldrK0v(01v. To roulo unodorKvuouv opodeg rodrro- v[ov, evrog Tov 0n0[0]v I sKQutrron Tnq Tpiqq poorlq e[ooQotri(eL r1Tt ro vrlrlpo 0o nopoperver ro Oro. 718 KEOAAAI0 25 0 levertroq K66rK0q H ).eroupyio rou IRNA orrl petdgpoorl ernLrlpcirvetor pr6Lrg ovoyvopfoer ro r<ro6rx6vro orr'1 prBooroplr<r; 05- oq A. H ol).ql,en(6poo4 preto(ri owtKodrKov(ou ror rro6tr<ov(o u npoyporoTro te(ror USoo (euyopril poro g tcov Bdoeov, oor6oo unoxoUer oe rov6veg or ono(ot eneKrefuouv ro (euydpropro n6po qn6 ro ouv4Oro;uivo (efyq G-C ror A-U. O (6ro9 o yeverr6g r<d:bu<og ono6(6et oprogdvo or1- gowu<d ororle(o yro rn 6repyoo[o qg ovoyvripro4g rcu6rrov[ov. To np6runo r4g exrpUtrronq rnq rpftr1g Bd- onq nopouord(erot ornv EIK0NA 25.3, 4 ono(o defiyer 6rr oe 6Leg oXe66v trg neprmciroerg e(re 4 rpfurl Bdorl e(vor odtdqoprl e(re y(veror 6rdr<pro4 p6vo prero[6 nouprvriv ror nuprl.u6rvcbv. YndpXouv orcr6 orxoy6vereg xco6rrov(<ov, ong ono(- eq Kor rq r6ooepo ro6x6vro nou popd(owor rtg 6rJo npcbreg Bdoerg 6Xouv ro (dro v6r1;ro, pe onor6leopro r1 rpfu4 Bdorl vo pr4 6todpoproil(er oXe66v r<ov6vo p6l,o orov ro0opropr6 tou opwo(6o9. Ie emd (etlyrl ro6xo- v(rov to v6r1po e(vor [6ro oveloprrpotg qq nuprprd(vqg nou Bp(or<eror firlv rpfuq 05oq. Ie n5we (e0yq, to opr- vo[U nou xrodrronoe(ror 6e peroBdMeror 6noto rr ov efuor 4 noup(vr1 orqv tpfuq 06o4. Yndplouv ;r6vo rperg neprmcrioerg rord rg ono(eg ono6(6eror 1.rovo6rr6 v6rlpro ond rrlv nopouo(o Uroq ouy- rerpqlSvrlg Fdonq or4v rpfuq 06o'q: AUG (pe0erov(v4), UGG (tpumo<pdvr) r<or UGA (tep;rotrop6g). 'Eror, or C ror U nord, 6ev 51ouv ;rovo6u<6 v6r1po oqv rpfuq 0dor.1, ev6 q A norS 6ev r<oOop((er 6vo provodu<6 optvo(ri. To owxcodrr6vro e[vor oupn],4pco1.rorrr6 rou ro6rro- v(ou: ouvendlg, 11 npcirq gdoq ornv oMrl),ouXfo rou ov- rtxro6rov(ou, nou ypdEeror ouprBoru<d pe r<oref0uvor1 lprInq Apr0prig rto0wovltov on6 5'npog 3', e[vor ereftr1 nou (euyopd:ver pre rrlv rp[rr1 F6on ornv otrq),ouX(o rou r<ro6xov(ou, 4 ono(o ypdee- ror r<ord rqv 6ro oriprBoorl. Enoprdvcog, o ouvduoopr6g Kcodu<6vro 5'ACG3' Awrco6u<5vro 3'UGC5' ouvqOcog ypdgeror og rcodu<6vro ACG/owtro6rr6vro CGU, 6nou q oM4).ouX(o rou owu<co6rKov(ou droBd- (eror owf0eto ci:ore vo e(vor oupn].r1pcoprorx4 pre-ro ro6u<6vro. l-to v' onoEeuX0e( oriyXuo4, 0o 6roqp4ooupre rr1 ouvr104 oriprBoor;, origqrrlvo Ue rnv ono(o 6).eg or otr-4- ).ouX(eg ypdrpowot oe roreU0uvoq 5'-3', evtir 0o uno- 6etxv6oupe rrg olLql,ouX(eg rcov owrro6rr<ov(ov pe ovdmpoqo B5).og 6ore vo unevOup((oupre q oxdcnl rouq Ue to ro6rx6vro. 'Etor, ro nopondvco (eUyog ro6r- xov(ou/owu<to6rrov(ou 0o ypdgeror ACG r<or CGU<-, owtmorxrog. KdOe r<codu<6vro onore( dpoye 5vo r6tofrepo IRNA p'tivo ouprnL4proporrr6 owxto6u<6vro 11 pqncog ptnope[ 5vo pr6vo IRNA v' owrororlef Kor. oro 6rjo pr6],r1 ev6g (etiyoug ro6u<ov(rrlv (ror' en6rroon, Kor mo oUvoAo rcov pel.cilv, n.1. 4, ;uog orroydverog r<trr6xov[rov); Iu1vd, 6vo IRNA prnope[ v' ovoyvrrlp(oer ndvro on6 6vo ro6rr6vro. 'O).o ro rrodx6vto nou ovoyvop((er rivo ouyrerprpuivo IRNA npdner vo e[vqr rour6oqgo ortg 6Uo nptireg 06oerg. Aw(Oero, n Bdon orrlv npc0rq 06o11 rou owtrodrrov[ou rou IRNA prnope[ vo (euyopciver p' evoMorcnr6g Bdoerg or4v ow(morXq rp(rr1 05oq rou r<ro6rxov(ou. Ie outrl rr1 05or1 ro (euydpr.upro Bdoeov 6ev neprop(eror oro ouvr104 (efy4 G-C ror A-U. Or rov6veg nou drSnouv to np6runo ovoyvriprorlg ouvoqu((owor orqv un<5oeon rnq oordoelog (wobble hypothesis), 11 ono(o 64Lrivet 6rr ro (euydpropro perofri ro6rrov(ou ror owrrcodu<ov(ou orrg 6Uo nprireg O5oerg rou xcodu<ov(ou or<oLouOe( ndwo roug ouvqOerg rovo- veg, oLl,d orqv rp[tr1 05oq ouprBofvouv oouvrlOroreg (oordee€q,. Aord0eto nporfmet ener6rl n 6oUn rnq pF Booopx4q 05o49 A, 6nou ouprBo(ver to (euydpoLlo Ko- 6trov(ou-owrral6rrov(ou, enrpdnet ou[r1;r5vr1 eue].r[[o orrlv npdrq Bdorlrou owu<o6xov(ou. To ouXy5repo prl oupBorrxd (e6yog nou Bp(or<eror oe ourrlq 05o4 eivor ro C-U (E|K0NA 25.4). l-ro nopd6er4ro, ro owrxro6u<6vto UUG oro tRNAGrn ovoyvropl(a ror ro 6r]o r<ot6rx6vto rrlg yl,outopr(v4q CAA ror CAG ror ro qwrKodrr6vro GUG oro tRNAHi" ovoyvcop((er ror ro 66o rco6tr6vro rnq ron- 6rraq CAU rqr CAC. iMo pr4 ouprBorrd (eUyr1 nou e(vor ovemd orqv rp[rr1 06o4 trov r<co6rov(rov nepil.qrBdvow rpononoLqpdveg O5oerg (B).5ne Ev6qta 25.6). Autrl q u<ov6rlro rrlg rp(rr1g 06oqg rov xo6u<ov(rov v' ov5Xowot (eriy4 G-U drlproupye( 5vo pror(Bo (euyo- pciprorog Bdoecov mo ono(o 4 A 6e prnope( nl.6ov vo 51er povo6u<6 v6qpro oro r<ro6u<6vro (enerdrl q U nou rrlv ovoyvop(et npSner eniorlg v' ovoyvrrtpioer qv G). llopopo(cog, 11 C 6ev 51er nl,5ov provo6x6 v64pro (ener- 6n n G nou rrlv ovoyvrop((er npdner v' ovoyvrop(oer en[- orlq Kor rnv U). H EIK0NA 25.5 ouvorf(er ro np6runo rqg ovoyvdr prorl g. Iuvenrri g, l.rovod rd rod rx6vro ovoyvop(- t_ I l0lrn ll00n J ob,o,popn' an 3 14 10 2
H a:.1:,.:.::.:: HC ,C .C lll Zuvn9 (e(ty Bdotov oe dleg Trq Odoslq To IRNA ouvn9oq ouw[Oewor alq npd6popeq ofrrf 6eg pre fip6oeero uAu<6 oro dvo 4 xor ro 5fo titlpo -roug. Irqv EIKONA 25.6 nopouord(owor or enmp6o0ereg otr4i.ouXieg nou oqoporiwor on6 ouvduoopotig w- dovo u r<.tr eol.uru<cilv r<or ef ovo u t<.),eoLurtrciv 6 p6oeov. To rp(o vourLeo{6ro oro 3' drcpo, nou oXe66v ndwq e(vor q rptntrSro CCA, prepu<5q qop6q 6ev rto6tronot- ofwot oro yovr6(copro. Ie ow5g ttg neptmcioetq, npo- or(Oewor oro ntro(oro rqq ene!,epyoo(oq tou IRNA. To S'dr<po rou IRNA drlpuoupyefuor p5otrl dtdonooqg 11 ono(o r<orol(erot qn6 ro ptBovourtreonpcoteivtr6 6v- (upro ptBovouxhedo4 P. Aw6 to dv(upro ovoyvop(et r1 ogoprxrl 6oprq q4(trrorog L rou IRNA xot u6poLUer erdt- xd ro groogodreorepu<6 6eqr6 nou onel,euOepcivet to cbprpro S'drpo rou ;.top[ou, xototro(nowoq Uto 5'-Qo- oqopx4 opr66o. Zto E. coli, q RNAdo4 P onotelefuot on6 6vq RNA 377 voux..eott6(cov Kot Uto nptotetvrl 17.5 kD, evri ro evepy6 xSwpo qg ouw(Oetot on6 RNA. /n vitro, ro RNA prnope( p6vo tou vo r<otolfer tr1v ow(- 6pooq ene(epyoo(og tou IRNA. (Aur6 onoreLe( 5vo no- pddery;ro p$oev(l1tou; BLSne Keqdl,oto 23.) H l,enoupy[o qg nporreivrr<r1 g unopovddog ouv(motot mr1 mo0epono(4or1 prog dropr6pqoonq tou evepyori To npddpogo tRNA neprdXet erurpdoeereq 5'roL 3'q),trnloux(eq Ev6ovoudedo1 To 3'oxpo tfrotrrDi(eror -N. H. "0 ll -r tc -6 -N til 'H_N.ffi I H * [ouov[wl :CH ? tdKXopo cH" ouooKrtrn nc{h+20.. IdKXoPo 0 H.N,H ,Mrou"''n"rsft,*:! . IoKXopo t;;,.:li,;: i_t 1,1*:: lr !i:,.J1,,:,.t:t ri.C.Jt: .iit.j :::..,,:i ,1aai, 7.1.:;. tilr::1.r.C':,::ii H un 'rC 'n ..0 {. "., ,'Y Y /t-fr-r-r...0 roKxopo r...n_n,.d.._r n *,[.-r-i -,'u),n t" ).0Kx0p0 - 'r ..,, r. .l r H ooroOeto oro (euyoprrrlro rov pooeolv enttp6ner ro oXlpolopo (euytirv G-U UeroE[ Uq tpiqg pooqq rou xodtro- v(ou rot qg nptlqq poolq rou ovlrtobrrovlou. (owor ;rdvo 6rov orrlv rp(-r1 06oq Bpior<erot G rq U. 0o16oo, p6vo ro UGG rot AUG onoteLoUv nopo6ery- Uoro Uroq rdrorog provodu<r1 g ovoyvrrrprorl g. ffi ITo 5', 3'dKpo DroofldTor 25.4 lo IRNA llop6yowor MeyoA0repov np66poUov U' EneEepyooio Mopitrlv + 'Evo rirprUo IRNA dlproupye[ror p' ene[epyooio evog npddpo- l.rou lrop[ou. * To 5'orpo 0qptoupyeirot ptlotrl dtoonooqg ono rrlv ev0ovou- rtreooq RNAoort P. " To 3'orpo drlgtoupyeuot geoo notrtrontr6v evdovourtreotrul- r6v rt e(orvourtreotrulrtiv Droonooeorv, ttg ono(eg orotrou- Oet 1 npooOlq rou rorvo0 repprolro0 rprvourtreortd(ou CCA. **;1 *tii';':t ?iy;rs *i* E+fl ,-*i_q) rtl *vc'lv,*f i(eiti, c ""i,t,;ri *rix*5iit*,:icx 6{1! t3":l i;C! r0! (iraid;ic! To s'dKpo Droondror Ev6ovourtreoorl (RNdon P) * ffi 5',3', tlpoor(0erot CCA 3''Ev(uuo(o) fl npoo0rlrrlq 5'I rou 3'drpou EIK0NA 25.5 To (euydporlro xtrl6lrov(ou-ovlrodrrovtou eprntr6- ret qv ooro0eto orrlv rptq 06o4. ,. . To 3' orpo rou IRNA dqproupye[ror prioo ovldpo- oetov erro;.ri1g (ev6ovourtreotrurrrdv) ror ilotrrdiouoToq (e(orvou- rtreotrurtr6v), nou orotrouOouvror ono qv npooOflr1 CCA ro 5' orpo dqproupye(ror peotr.r luog orprgoug evdovour<),eotrulrqg Dro- 0n00rlq. U C A G An G G Uovo U Uovo cnu ?i.;i To tRNA I-lopoyovror p' Ene[epyoo(o lVleyotrurepov tlpoDpol.u,.rv ["lo0l,-:', 719 Boorxfg eworeg
Kdwpou rou RNA n ono(o e[vot ouprnLrlpoporrr{ rou np66poprou IRNA. Oo e(eroore( nepor6pto anlv Ev6- uyto 23.8. To 6v(upro nou ene[epyd(owor ro 3'drpo 61ouv ne- prypoge( roLrirepo aro E. coli,5nou puo ev6ovoudr,e6- or1 nupo6ore[ qv owbpooq 6roonrilwog ro np66popro pr6pto r<o0o6u<d r<r 6nero noMSg e(ovouxi.edoeq Uoh- 6[(ouv ro dxpo oe roteU0uvoq 3'-5'. H ene[epyoo(o rou 3'dr<pou rou IRNA orouq euKopurireg en(oqg neprl,op- Bdver noMd 6v(u;ro. H npooOrirrlrou 3'-CCA r<orol0e- tor on5 ro 5v(upro IRNA vouxi.eon6ul,orpovorpepdorl, nou l,emoupye( o4 RNA noLupepdo'q ove[dprqrq on6 erproye(o. Me dMo L6yro, ro 6v(u;ro npoo0Srer er6rd C, C ror A cnnv oMqLouX(o 1op(g vo (euyopriver qv r<utoo[vr1 Kot rrlv o6ev(vr1 pre rrg ouprn].qpoporu<69 Bd- oerg youov(vqq Kqr oupoKiLqg ndvo o' 5vo erpoye(o. Aw( yt' ou"r6, 4 6ro 4 6opri rou ev(6prou enopre( yro vq qXq godoet 6ro6oXx6g oupntr4 prrrporx6q O5oerg np5o- 6eor1q yo rtg C, C ror A. Ko0c0g npoor(0ewor rcl vou- r.treor(6ro, ro oOpntror<o ev(Ugou- IRNA peroBdMet tr1 dtopr6pgroo4 rou rilore vo xoroore( oupnArlpcol.ronr6 pe xdOe 6ro6oXtx6 vour<Leor(6to. To rp(o voux.Leor6tq npomiOewor on6 r4v IRNA vourLeorr6uLorpovogepdoq, 6rov 6ev rodrronororjv- ror orn yovr6torci oM4Loul(o rou IRNA. E(vor evdrotp6- pov 6tr ro 5v(upro 6ro6popor((er en(o4q Boorr6 p6Lo orqv enrdr6pOoorl roreorpoprpr6vcov 3'dxprov rou IRNA oe opyovroporig, 6ntog to E. coli, nou xolduronorcAv to CCA. Ie owo0g rouq opyovropro0g, ovoyvropi(owot rp(o 6roqoperrrd IRNA unoorpc0proro: lrop(g CCA, Ue 31C i ue 3'-CC. Or IRNA vourcl.eonduLorpovogepdoeg 6rorp(vowor oe 50o roqyop[eg nou 61ouv oqprowrrrj opror6qro o1.rwo[6rov pr6vo ortg neproldg rov evepy<ilv rdwprov. To 6v(upro rnq Kornyop(og I Bp(or<owor or' opXo[o, evdr ro Borrqproxd rr europuorrrd 6v(upro ouvonotetroUv q defrepq roqyopfo. Ie opropr5vo Bor,cripro, n npo- o0qrq r4g CCA xorql0eror on6 6Uo orevd oXetr(6pe- vo 5v(upro roqyop(og ll: ro 6vo on6 ourd ro 6v(upro npooO5rer -CC, evdr ro dMo npoo06rer qv 3'-re).rxrj A. 25.5 To IRNA tleprdxer Tpononolnpdveq Bdoerq * To tRNA nept6Xouv ndvo ond 90 rpononoulp6veg pdoerg. r H rpononoulon ouvn0toq ouv(ororor oe ogeorl otrtroyr| rtov nporoyevtirv Bdoeov rou IRNA, oordoo unopXouv oprogeveg e[orpeoerg onou pto pdon oQoLpe(rot u ovnxoOrordror on6 pro 0trtr1. r 0t rpononornp6veq pooerg npood0ouv ou[nUdvn oroOeporrl- T0 oT0 IRNA rot pu0p(ouv qv ovoyvtitpro1 roug on6 npoTsl- veg ror otrtro RNA rqq peroqpoorxrlg ouoreurlg. To perogoprr6 RNA e(vor 1.rovo6u<6 ;rero{ti rov vou- 720 KEOAMI0 25 0 levelrdg Kdrdrrog rLeivrrrilv o(6ov oq npoq rrlv nepreKnr<6qrd rou oe rpononorrlp6veg Bdoerg. Mro tpononor4;r6vr1 Bdo4 e(- vor onoroo6rlnote dorcrritrrog noup(vqg 11 nuprpu6ivqg em6g rov ouvri0cov A, G, C r<or U, on6 rrg ono(eg ouw(0ewor 6l.o rq RNA. 'Oeg or dtrLeg Bdoerg nop6- yowor on6 peropetoypoqu<4 rpononoirloq puog on6 trg rdooeprg Bdoerg, orp5rou our4 evoo:1.rorro0e( oqv notruprBovouxi.eotr6rrrl qluo(6q. To odxlopo rnq pr- B6(119 oprop6vcov vour<.l.eol6(tov rou IRNA en(oqg pre- 0uAr6veror oro 2'-udpo(rjro o6qyri:vrog 6ror oe 2'-O-Ue0u(coon. Evcir 6),eg or xorqyop(eg rou RNA eprgov((ouv rd- noto Bo0pr6 rponono(r1or1g, to eUpog rrov l4prrciv pe- toBoLriv orrg Bdoerg e[vor notrrj preyo].Jrepo oro IRNA Ot tpononorqoetg rupro(vowor on6 on).rl pre0utr(rr:oq 5cog yevtrri ovo66;rr1or1 rqg Bdorlq. Tpononorrloerg oupBo(vouv oe 6Lo ro ttrrrlproro rou tRNA. l-lorril,ouv no).rj rog npoq rrlv 5r<roor1 rrlg ouwqp4oriq roug prero- [ri rcov 6rdgopr,:v IRNA, 6n<og en(oqg Kor oq npoq rrlv ew6nroq roug pr5oo oro ;r6pro. Tpononot4oerg er6txdg yro ouyKeKprpr6vo IRNA q prrpdg unoo;rddeg 1RNA yevr- xd efuor ondveg. Yndplouv en(oqg opro;r6vo np6runo rpononorrloeov erdrrd rou e(6oug. IuvoLtrd, ro IRNA neprdXer ndvco on6 70 6rorpopertroUg rpononorqpSvrov Bdoecrlv-r6noug. Kd0e IRNA rpononoe(ror rotd p6oo 6po oe noooor6 15-20y. rcov Bdoecivrou. To tpononorrlpi6vo vourLeoo(dro ouw(Oewot on6 erdu<d 6v(u;ro rponono(r'1onq rou IRNA. To ou0ewrrd voux.treoo(6ro rdOe Orioqg prnope( vo xoOoprore( eke pre oriyxpro4 r4g oMrll.ouX[og rou IRNA ge r;v oMrli.ou- X(o rou ow(ororXou yovt6(ou r1 ().ry6repo onoreleopo- lxd) pre onop6vcoorl np66popr<ov prop(r.ov Xop(g rdnoro r1 6),eq rrq rpononouioerg. Or oAlqtroul(eg r<ov np6- 6popur:v 1.rop(rov deiXvouv 6tt 6ro<popelxdg rpononoL- 4oerg erodyovror oe 6ro<poperrxd otd6ro xord r1v rr:p(provorl tou IRNA. To no).).d 5v(upro tponono(r1onq rou IRNA norx(- l,ouv noLU oq npoq r1v erdrr6r4to. Meprrdg qop6q 5vo tiv(upo nporoLe( Urq ouyKeKprp6vrl rpononofqor1 oe Uto pt5vo 05oq. 'AMore, 6vo 6v(upro prnope( vo rpo- nonorrloer Bdoerg oe oprerSg 6rogoperrrd,g 06oerg-ord- Xouq. Opro;r6vo 5v(upo ovo).oprBdvouv povdg owtdpdoerg pre [eX<r:prord IRNA, evcb dtr].o 6Xouv 6ro- qopo unoorpoiproto. Kdnoreg rpononotqoetg npouno- 06rouv 6ro6o1u<q 6pdoq noMcov ev(0grov. Aenrop6pereq yro rn 6opurri Bdoq r1g rponono(- nonq rou IRNA on6 5v(upo p6llg dpXroov v' ovo6riov- tor. 'Evo ewunoloror6 nopd6etypro e[vor o prqXovropr6q p6oa; tou ono(ou 11 opXoroo(vr1 (archaeosine), puo rpo- nonorqg6vq G, erodyerot oro Bp61o D ouyr<exptpdvutv IRNA lov oplo[orv. I-ro vo npooeyy[oer q Bdoq nou np6xetror vcl rporonorq0e(, q ono[o rqvovrrd Bp(oxeror ewocptootrr6vrl pr5oo qrov rprroroyrl nuprlvo tou IRNA. to 5v(upo frans-yLuxo(ul.doq tRNA-youovrv4g (tRNA guanine transglycosylase) euodriver pro 6poonrc1 eno- vodtdro[r; rou IRNA, dlpuoupycitwog 6ror gro evoMo- mrrrl rptrorqyi dotrlq yvrrlmrl cog 6o94 tr. Enovo6ro- td(erg rqg 6op4g tou IRNA mo nLq(oto enoy6prevqq npooopproyr;g (induced fit) 51ouv en[or1g noporrlprlOet Boorxeg evuorcg
Kor oe dMo 6v(u;ro rponono(qoqg ror e(vor r<ow6 Xo- porrrl pronr6 orrlv ovoyvd:ptoq. Or rpononot4p5veq Bdoerg npoo6(6ouv ou(r1p5v4 oro0ep6qro oro IRNA ror puO;r((ouv qv ovoyvcilptorj roug on6 npolreiveg ror dMo RNA oro prllovroprd qg petdgpooqg. l-ro nopd6erypo, oe oprerdg neptnrdr- oetg, 61ouv oorpcig npoodroprore( p6i.or yro nq rpoflo- nor4pr6veg Bdoerg mqv ovoyv6proq on6 rrq oprvoo- xuAo-tRNA ouvOerdoeg (Bldne Ev6tryeq 25.9 6o9 25.11).Ie noM6g neprm6oerg, roo16oo, o BroLoyrx6g p6),og tqq rponono(r1onq rou IRNA nopog5ver dyvo:- sroq. H EIK0NA 25.7 nopouord(er opropdveg on6 tq nro xow6q rpononor4;rdveg Bdoerg. Or tpononor1oag rrrlv nuptptdrvcbv (C r<or U) yevrrd e(vor fuy6repo nol,rjn).o- xeg on6 erefueg t<ov noupwcbv (A r<ot G). H ouv406orepq rponono(qon rnq oupr6(v4g Kot rrlq rutoo[vr1g elvor q pre0u)r(rrloq, q ono(o ;rnope( vo ouprBe( oe notrl,5g 6ro<poperu<69 O5oerg rou 6olctu).[ou. H peOu- tr(roor1 or4 06oq 5 r4g oupor<fi.r1q 6qpuoupye( q prBo- 0upu6(vq 0. H Fdon qg 0upu6(vr1g e(vor row5or;prq pre rr1 0upr6ivq oro DNA, oor6oo oro IRNA e(vor npoo6e- 6ep6vr1 oe prB6(rlnopd oe 6eo(uprB6(q. Awri 4 0upu6(- vq undplet qXe66v oe 6l.o ro IRNA arq 05o4 54 tou pp6Xou TqrC. H rfeudooupr6(vq efvor 1.uo ewunootoxr; rponono(r1oq ouprd(vqg nou nporUnrer pe drdonooq rou yLur<o(nrxoU 6eoproU, rrlv onotc c<3:.:-:a -:::. propr6vr; neprorpogrl rou oneLeu0rpc0Us,,,o- 51.-:-, : _ xr enov6voo4 rou dvOpor<o C5 pe rov ovOpci<: l' -: ptF6(rlS.'Etot, q ql,reudooupr6(vr1 6lo0ereL evov I'r-. r.,. r<o(ttu<d 6eopr6. I1e66v 6).o ro IRNA neprdXouv q,er5.- oupr6(vr1 mr1 06oq 55 rou Bp61ou fqJC. H 05o1 56 en(or1g e(vor e[orperrr<d ouwrlprlprdvrl oq xuroo[vqt 1 o).).4Lou1(o TQC mrg 06oetg 54-56 nopSXet rr1 Boorl yto rrlv ovolloro0eo(o ouroU rou rpr4potoq rou tRNA. H drudpoouprd(vq (D) nou 6rlpLoupye(tor p5oco r<o- peoproti tou 6Lnl.oU 5eo;roU nou ouv66er ro C5 xoL C6 qg oupor<fi.r1q e(vot oxe66v ou<oupevrrq oro Bp61o D rotv IRNA. H ogo(peoq rou 6Ln).ori deoproU qg D xoto- orpSqer rnv opopolr<6qto ror ro en(ne6o oXqpro rou 6or<ruL(ou qg oupor<il.qq, ono6(6owoq ULo oouvq0L- orrl 6opq, q ono(o tpononore( e).orpptrtg to o"1r1po tou o<porpu<oU nuprlvo rou IRNA. To vour<Leoo[6ro tvoo(vq (l) Bp(or<eror rovovu<d oro x0rropo og ev6rdpeoo npoi6v r1g Broouv0etu<r1g o6ori rotv nOuprvtr-rv. Om6oo, 6ev evOotl.iotrbve-rOr dpeoo mO RNA. Aw(0ero, 11 6nop[r1 rou efoprdror ond rnv rpo- nono(4o4 qg A. H evocogdtooq qq I orr; 06o11 rou 5'- ov-rrrot6tr<ov(ou ouvero<p6pet no).ri oro ooro06g (euydpopro Bdoe<lv oqv rp(q 05or; tou r<codtxov(ou rou mRNA (B,l.dne Evorrlro 25.6). Ot rpononorqoetq rov A r<oL G oupd nopdyouv rv- Tponrniririuive_t $ciueit 00 H H Prpo0ugtdivrl g1 Atudpooupr0irtr (D),l,.rOoorp16rvn (q.r) 4-0ttooupt0[vrl 0uotohoyrdg pdoeLg 0 ,lt Hl{cffi0H o'ffi' 0uprdivrl NH, t' ,'@; KurtD[vr1 NH, I HN -ctc-Nrt A 11 )cH ,HN'"lt1-u-N' . -'-Y3l ( ntH-ciiocf . -lFl Cli.iir1C:. H^ o :l ;;'"";"r I Lh; I g5 -c-6-N., i*-'tr- H / ,ll )bH ;r.-, . J )H 2HN'u1r],-u-N/ l/x1,'"' -11'zHN' Iouovoofu1 7-ge0utroyouovoo[v1 Krouoorvrl(0) touoioo(vr1 (Y) Kor or reooeprg pooerg rou tRNA pnope( vo rpononorl0ouv. 2S.5 To IRNA lleprdXer TpononoqgSveg Booerg 721 N--:il, xloC-6-t',t. nd=*..8-N)bn UeOutrodevootvn(m6R; trt6
re6q v6eq 6op6q (Ex6vo 25.7). [-to nopddetygo, 6[o notr0nhoreg oerp5g vourtreott6(tov Boo((owqt ornv rponono(r1on rnq G. Ot Bdoetg Q, 6n<oq 11 r<touoo(vt1 (queuosine), nept6louv 6vov np6o0eto donrUAto Tlewe- vutr(ou nou npoor(Oeror p6oco ev6g deopoti NH orq pe- 0utropd6o qg 7-;re0uLoyouovoo(vr1q. O 6ortUltoq newevutr(ou gnope[ vo ;retog6pet notrileg dMeg o;16- 6eg. Ot Bdoetg Y, 6nog 11 ouoioo(vq (wyosine), nep6- louv 6vov np6o0ero dol.c0Lro nou ouwtlretot Ue rov (6ro ro 6qhir0fuo nouplvqq. Aut6g o np6o0eroq dorcrri- Ltog ;rerorp6per Uto puo proxptd ovOpoxtxti otruo(6q, orqv ono(o gnope( vo npooreOorlv 6togopettr6g, t<otd nep(molor1, opd6eq. 25.6 Or lpononornp6v€q Bdoelq Ennped(ouv To Z€uydpopo AvTtKoOtKoviou-Ko6lroviou r Tpononouloerq oTo ovrtrodxtivto ennped(ouv ro npdruno rou ooro0o0g (euyopriUotoq rot ouvendlg e(vot or1povru6g yro rov xo0optopd rnq etdxdrnrog tou IRNA. Tpononorfoerq rou IRNA p6oo t<ot 6(nlo oro owtKo- 6u<6vro enrlpe6(ouv rqv tt<ov6tqtd tou vo (euyopdrvet pe ro rrrl6u<6vlo rou mRNA. Ot neptoo6repeq rpono- notrloerg owo0 rou e(6oug ewon((owor orrg 06oetg 34 t<ot 37 tou Bp6Xou tou qwtKo6txov(ou t<ot yevtxd LetroupyoUv neprop(owoq ro e0pog r4g 6ro06otpqg Krvrlonq oro owu<to6x6vto. Me rq oepd tou, outS 6t- euroLUver r4v oyt<upo6eo(o tou IRNA orq 05o4 A tou prBooopror(ou. Aut6g ot rpononot(oetg enqped(ouv ro (euydpcopo rov xodtrov(ov rt 6tot enrlpe6(ouv dpreoo nc0g 0o ep;tqveUoet to xtittopo to v6r1po tou IRNA, Ot rpoTronotnU6veg Bdoerg enmpdnouv enrnp6- oOeto np6runo (euyoprilprorog erc6g on5 ere(vo nou Boo((owor oro ouvqOtogdvo xot to ooro069 (euyd- popo rov A; C, U xot G. H tvoo(vr1 e(vot t6to(tepo orlprowrri 6tov Bp(oretot orrlv np6tr1 06oq rou owmo6xov(ou (vouxl.eotbto 34 orqv oMqlouX(o), enet6ri e(vot tt<ovtl vo (euyopdroet pe onorod{nore on6 ng pdo'etq U, C rqt A (ElK0l'lA 25.8). O p6Log qq woo(vqg ovo6exv6etot ornv qfloKodxono(- rlon rov r<co6u<ov(tov t4g tooLeur(vqq. E6ri ro AUA t<to- 6rxonote[ r4v too].eur(vq, evr6 to AUG tq trre0etov(vq. l-to vo 6roBdoet rrlv A ornv tprn 06on tou rto6troviou, to IRNA 0o Xpero(6tov 1.uo U orqv np6rn 06oq rou ov- rrxrodxov(ou - coor6oo owrl4 U orrlv ooroOf 06o11 0o (euydptrrve ovoyroortrd Kot Ue rqv G. 'Etot, onoto6rl- note IRNA Ue Uto 5' U oro owtt<to6x6vt6 tou 0' ovo- yvripr(e t5oo to AUG 6oo Kot ro AUA. Aur6 to np6p)qpo enil.[etot pe tr1 o0v0eor1 ev5q IRNA tootreu- rfvqg nou 6ro06rer A34, qv ono[q qKo].ou0e( q rpono- no(r1oq qq A34 oe 134 on6 to 6v(upro 6eoprwdo4 rqg o6evoo(vqg rou IRNA (IRNA adenosine deaminase). 722 XEOA,A|0 25 0 [eveudq Kdtdtxog --.2@1i , :qKxqpo u'nr6ffi* lvos(vn ,ryIdrXopo I6Kxopo Adevivrl IoKXopo flqili,i& I5.& H rvoolvrlpnope(vo (euyoptitoetge rqv U m, , - rrlv A. Irrl ouvdXeto, q 134 prnope( v' ovoyvop(oet Kot rq rpic rco6rr6vto rqg LooLeur(vqg: AUU, AUG t<ot AUA. Ttg neproo6repeg <popdg, 11 U mrlv npcbq 06o11rc- owtxro6u<ov(ou en(oqg petotpdnerot oe Uto rpononc.- 11p6vr; pop<p4 pre 6torpoperu<59 r6t6qteq (euyop6uc- rog. To nopdyo.lyo qg U nou neptdXouv m4 06oq 2 puo 0eurrl optddo ow( rou o[uy6vou eprgov((ouv ou[r- ;r5vr1 ex).ercrtx6t4to mo (euydp<opo Ue rnv A oe o["- Kpron Ue rqv G {;:{lll&. j:I !l Avrrr<to6tr6vLo ga 'Evaq 6esU6q Dav enupxei :oKXopo --@: ,. IdxloPo S 0 I H-5- I H :orxqpo H 2-OerooupoKiln (euyqptivet Udvo Ue A CH" t' HCffi"O.. ,w-,.:x* Adev(vn 'TIlj.1,,'.-* HWH ru**ooo Ell(ONA 25.9 H rpononoinon orrl 2-Oetooupt6ivrl neptop((et ro (euydptopo g6vo otlv A, dt6rt gdvo 5voq deopdq H pnope( vo oXrlpolore[ge qv G, Boorx6g evvoreg lvoorq
ouprdrvo-5-o(uo[rx6 o[ri r<or ouyyeve(g rpononor4oerg orqv npdrq 05o4 51ouv r1v o[roor1;re(crrrrl rdr6rrlro vo enmp6nouv oro povo6rrd IRNA vo 6roBd(er rp(o 11 or<6- Uq Kor ro rdooepo ouvrilvul.ro ro6x6vro NNA, NNC, N- NU xor NNG. Aw5g roOc0g r<or dl}eg tdr6qreq (euyoprirprotog 6e(Xvouv 6rr undpXouv notrl.o[ rp6nor yro rqv Koro- or<euq ptog o;166o9 IRNA u<ovri:v v' ovoyvrop((ouv ro 61 t<codtt<6vto nou owrnpoocone[ouv opuvo[6o. Aev ent- rpore( rdnoro rdro(repo np6runo oe xdnorov opyovr- o;16, nop6tt q onouo(o proq ouyKeKprprivrlg o6o0 rpononofioqg prnope( vo epno6(oer rn Xpion oprqr6- vrov nporUnov ovoyvripror1q. 'Eror, pro ouyrexprpr6vrl orroySvero rcodu<ov(rov droBd(eror on6 IRNA pe Srogo- pertrd owxro6 rrdvro oe 6 ro<popetu<o ri g opyovLopro rj g. IuXvd ro IRNA dro06touv enrrol,umdpeveg rxov6- rqteg ovdyvoonq ouyKexprl.uivcov ro6tr<ov(otv, 6ror riore rivq ouyxerprprdvo xr.o6rr6vro 6ropd(eror on6 6[o 11 neptoo6tepo IRNA. Ie ow5q lg neprmc0oerg (ocrrg undpXouv drorpopdg oq npog rlv onoreleoporrr6rq'ro rov evol,l,omu<rilv owrdpdoeolv ovoyv6proq g (yevrrd, ro rodtx6vro nou XpnorponoroUwqr ouXvd, ouv{Ocog droBd(owor nro onoreLeoprotxd.) Or npoBl,6gerg tou ooro0orig (euyopriporog efvor noL0 ouprBotdg ;re ro nerpol.rotrrd 6e6oprdvo yto 6Lo oXe66v ro IRNA. Oqr6oo, undplouv e[otpSoerg, ong ono(eg ro xro6rr6vto nou ovoyvop((owor on6 6vo IRNA dtoE6pouv on6 ere(vo nou npoBLdnowor on5 rouq Kq- vSveg qg omdOerog. Aur6 [oog npor0nrer on6 rqv en(6poor1 yemovu<cilv Bdoeov Kor/ri rnq 6rog6pgoo4g tou Bp6Xou rou owrxrodrrov(ou orrl ouvo).rx4 rprroro- Wt 6opn rou IRNA. l-'lepottSpo ev6e[[erg yro rn onUo- o(o rqg neprBdl},ouoog 6oprrlg nop6Xowot on6 rrlv onop6vooq onopodxri:v preroMoyprfuov orcl ono(o 11 olr.l.oyri ptog Bdoqg oe xdnorq dMrl neproXrl tou prop[- ou peroBdLLet ulv u<ov6rrlro rou owrrro6rr<ov(ou v' ovoyvro p (er rcr:6 rr6vto. GCU 2 5.7 Yndpxouv Inopo6rr6q lpononorlroerq Tou OrKouu€vtKorj Ku)6rro c Ie optou6vo ei6q 6Xouv ougper otr)royrig oro yevenrd rtirdr- KO, c Aur6q or drtroy6q etvor ouvrl06orepeq or0 pnoXovdprord yo- vtduirporo, 6nou pnopei v0 KoTooKruoore[ 6vo rpu)oyeveux6 d6vrpo yro ilg otr)roy6g. e Iro nupnvuo yovrduirgoro, ot otrtroy6q ouvn0oq enrlpeo(ouv privo ro rodx6vro repponopou. H ou<oupevrr6r4ro rou yoverrorl r<cbdu<q e(vor o(ro- orlge(oq, coor6oo undpXouv oproprdveg e[orp6oerq. Aur6g ouvrl0rrlg en4pe6(ouv ro xrodx5vro nou eprnl,6- Kowor mrlv Svop[q ( rov tepgorropr6. Or o]}.oy6g oro xripro yovr6rilproro (Borctrlprord q nuprlvu<d) ou- voQ(owor or4v EIK0NA 25.10. IXe66v 6Leg or otrl,oy6g oro nuprlvrKd yovt6uil;roro nou enrtp6nouv o' 5vo r<o:6rr6vto v' owrTrpooroneUer 6vo ogtvo[ri enqped(ouv to rro6x6vto reppotrogo[: o Irov nporopurirq Mycoplasma capricolum, ro UGA 6ev Xp4orponorekor orov repporr.opr6, oMd ro6xo- noef r4v rpumoEdvq (Trp). Irrlv npoyporr5r4ro e(vor ro r<up(opXo ro:6u<6vro qg Trp, evcb ro UGG Xpnorponorefuor ondvrq. Yndplouv 611o ei64 TrptR- NA, ro ono(o neprdxouv ro owu<rodrx6vto UCAe (6roBd(er ro UGA xot UGG) ror CCAe (6roBd(er pr6- vo ro UGG). o OproUSvo Blegopr6orp6po (provor<rjrropo npot6- (roo) 6roBd(ouv ro UAA r<or UAG rog ytrouro;r(vr1 ov- r( yro orlproro repUorropori. To Tetrahymena thermophila, 6vo Ble<popr6og6po, nep61a rp[o ei- 6n tRNAot". 'Evo tRNAGrn pe owx<o6rr6vro UUG qvo- UUB 'n, ,IU: "."INil,, 'iy33 ,,, UUt ,,, 'iUt "''" NUtt sT0P-n',,,o iUBt sT0P-Trp'cvsser cuu lccu lcAU p;q HICGU ffi .'' 'issl ".'isii::: :issl ^,' 919 !:1rryt i999 l:19 i:" itl tlgi: AUU I ncu i RRu ^^^ ^, i AGU ^-- AUa 1e r I ncc -u" , I AAC utn * I AGC ue' AUA tte-ry6x1g IRCR ""'lAAA,,,^ ,, lRcR Rrg-N6NE nud ii,r"rt '''"'"M i AaC I nno tv' K ! roc nt[ WCS W 3i3 *' D GAA ^n^ Glu E uAu GUU GUC ,,, GUA JiI: GUG GCC v GCA ^'o ^ : trUU GGU GGC GGA GGG Glv 1 i , r:, -'i.: 0t peroBotr6q rou yeveruou r66txo oro porqproro fl euxopuolro nuprlvrro yovrdrrirporo ouv10otg onodr6ouv ogtvo[50 ,. <oDtrovto Il(nS I geropotrtrouv evo rodtrovto 6rot 6ore vo Unv ro0op(er ntr6ov ronoro olrrvo[u. Mto Ueropotrn oro vorlgo ono evo ,- ,,0[u oe ronoro otrtro elvor oouvl0Loq. 25"? YnopXouv Inopodrreg TpononoLrloerg rou 0rxoupevrxou Krrrdrx: 723 Boorxeq eworeq G
yvop((et ro ouvnen KodK6vto CM r<ot CAG yto 11 yl.outopr(vr1, 6vo 6erlrepo e(6og pre qwtro6tr6vto UUA ovoyvtopi(et t6oo to UAA 6oo t<ot to UAG (oripqrovo Ue rnv un60eoq tqq oord0etog), ev<ir to rpfuo e(6og pe owrrodtt<6vro CUA ovoyv<op((et p6- vo to UAG. O neptoptol.t6q qq et6r6rrltog rou no' pdyowo oneleu06ptoorlg eRF, cborc v' ovoyvop((et pr6vo to ro6x6vto teppotropo0 UGA, en(oqq Xpet- 6(eror c0ore vo Un ouUPe( np6topog repprottopr6q oro veonpoo6topt(6prevo KtoSK6vtq yLourog(vr1g, o Ie rdnoto dIIo BLeqopr6og6po (Euplotes octacari- natus), to t<todu<6vto tepprottol-toti UGA enovonpoo- drop[(etot oe ruorefvq. M6vo ro UAA lprlotgonot- e[tot tog xo6tr6vto tepporropoti, evdl 6ev undpXet UAG. H oIIoVn oro v6rlpro tou UGA prnope[ vo ont- reuX0e( pSoto tponono(r1oqg tou owtt<to6tt<ov(ou rou tRNAcv' pe 134, 6rot coare vo gnope( vo 6toBd(et ro UGA pro(( ge to ouvriOq rro6tr6vto UGU t<ot UGC. To UGA 6Xet 6ml'6 v6rlpro oro Euplotes cras- sus (BL6ne Ev6trya 25.8). o I'5vo (u;roprir<r1to (Candida), to CUG enovonpoo- drop(etot oe oep(vq ow( qq Leuxffiq. Aur6 e(vot 6vo ondvto nop6Setypro enovonpoo6roptopori on6 5vo rodx6vto pe v6qPro o' 5vo d.tro. l-evrxd, yro v' onorcnloet xcodtronorrlurrl Aenoupy(o 6vo t<to6tr6vto repportoUo0 np6net vo oupBotiv 60o r0- nor otr),oycirv: 6vo IRNA np6net vo petoMol0e(cirore v' ovoyvtop(et ro rrodtx6vto rot o nopdyowog onetreu06- poonq rd[4q I np6ner vo ;reroMol0ef 6ore vo Urlv rep- 1.roil(et oe ow6 ro xu:6tr6vto. O dMog ouvriOlg t0nog otrtroycirv e[vor 11 onciLeto tou IRNA nou owtorotXe( o' 6vo t<to6tx6vto, 6tot dlore to rodu<6vto vo Urlv r<oOop[- (et nl,6ov xdnoto opwo(tl. 'Otreq ow5q ot oMoy6g e(vot onopo6u<5q, yeyov6q nou o4go(vet 6rt pr6Mov ouv5B4oov ove(dpqto oe et- 6u<6q e[e].rrcrtr5g oep6g. Iuyxewpdlvowot oro t<trl6u<6- vro repportotrtoU (orog enerdri oe our6g ttg 06oetg 6ev undpXet owtxotdmoorl ev6g oprtvo(dog on6 xdnoto dLlo. M6).tg xo0teprirOrlre o yevettr6g r66trog, oe npcilq-ro ord6ro rrlg e[6].t(qg, onotod{note yevtrrl peto- Bo).4 oro v6qtrro ev6g rodtrov(ou 0o nporol'o[oe owt- rordmoo4 oe 6leg ttg npore(veg nou neptSxouv ro ouyxexptgSvo o;rwo(0. l-lt0ov6v 4 petoBoLq 0o rpov enrBloBrlg yto optop5veg on6 out5g ttg npotelveg, on6te 0o eniley6tov opvrlnrd. Ot onor.tr'(vouoeq Xpn- oerq rov rodtxov(tov teppottopo0 (oog owtnpooto- ne0ouv rn "oUIInqJn" touq pe oron6 t4v rovovtrrl rrodtxonoirloq. Av optopr6vo xto6tr6vto teppottopo0 Xpnotponototiwov ondvto, 11 entorpdteuo{ toug yto xio6rronofqor1, g6oto o]'Loycirv sro IRNA nou enttp6- nouv rov enovonpoo6toptop6, 0o ntov nro nt0ovt1. E[otp6oetg orov otKoupevtr6 yevettr6 rtil6tro en(- oqq ouprBoivouv oro prttoX6v6pto noMrirv et6riv. H El- K0NA 25.11 nopouotd(er prto cputroydveto (e[e].trtrr<6 66vdpo) tcov oMoydrv. H txov6tqro KorooKeunq ptoq guLoydvetog unodqtrcivet 6tt unrlp(e 6vog ott<ougevt- 169 xri6xog nou pretoBtrrl0rlre oe 6td<popo orlpe(o r<o- 0txoupevudg rtiDtroq EIK0NA 25.1"1 0r Ueropotr6q rou yeveuo0 xtit0txo oro ptrol6v- Dpro pnopouv vo ewontototjv oq rputroy6veto. 0 etrdxtmoq opt$ U6q ove[oprntov petopo)rtirv npo5pXerot onri qv undOeorl 6n u Ueropol6q AUA=Met roL AAA=Asn npoyporonototjvrot 1 roOe- pio ove[opqto 6uo Qop6q rot 6tt 11 npdrtpq geropo)rn AUA=Met owtorpdqrlxe mo eXwddePPo. rd qv e(6LrEn tov l.tttoXov6p(trlv. H npotp6teprl oMe V{ ntov n Xpnoqoflohon tou UGA yto u1v rcodmonoi- non rnq rpumorpdv4g, 4 ono(o e(vot t<ow{ oe 6tro ro pr4 <punxd groX6vdpto. Optopr6veg prroXov6ptordg oMoydg rdvouv tov ni6t- Ko qnlo0orepo pe rrlv owxotdoroorl 60o rurl6xov[trw nou e(Xov dtorpoperr6 v6rlpro on6 6vo (e0yog nou dletro (6ro v51pro. Ze0yr1 ro6xov(tov nou owqretoln(oqrov rqd ow6v rov tp6no nepr.ogBdvouv to UGG rq UGA (rot ro d6o yto rnv rpunroqdv4 ow( to 5vo yto rqv tpumog6vrl ror ro dMo yto repUortoU5), 6no4 entonq rot to AUG xa AUA (r<or to 60o yto rr1 geOetovwn ow[ to 5vo Yto rn UE- Oerovfvq rot to dtrtro Ytq rnv toolsur(vq). l-rot( ot oMoy6g tou t<ti6u<q prnopo0oe vo e(etu1- 0otlv eur<ol.6tepo oro prmoX6vdpto oe o0yr<ptor1 Pe rov nuprlvo; To prrol6vdpto ouv06ta l,iyeq npoltetves (-10) xt 6tot to np6BLqtrro rqg onopp00prtoqg ).6yto otrLoytilv mo v6qpo e(vot notr0 l'ry6tepo ooBop6. [lt0ov6v ro rpononoqpr5vo xto6u<6vto 6e lp4otponoqOrlrov eme- vri:g oe 06oetq 6nou ot owtxo-roordoetg oprwo(6rov 0o rpov entBtroBe(q. Etigrprovo Ue rnv un60eo4 qg oordOetoq, 6voq eA6- Xrorog opt0trr6g 31 IRNA (erc6g tou evoprcr{ptou) eivo onopo(rqrog dlore v' ovoYvoJptqro[v rot to 61 r<ro6xo- vro (touldlto,ov dUo IRNA onqroUwot yto rd0e oro- y6veto teoodptlv rodtrov(rov xt 5vo IRNA e(vot ovoyt<o(o ovd (erjyog rcodrov(ou r1 [e1optor6 ro6rG vro). To onodonx6 truolovdptor6 yovtd(trlpro t<ov Oqtro- ottxtirv, toor6oo, rtodxonote( trrcvo 22 IRNA. To IRNA, nou ro6txonotoUwqt oro nuprlvK6 yovt6(olpto, 6ev et- odyowot mo puoX6v6pto rov 0rlLoortr<rilv: ouvenc0g, efdyetot to oupndpoopro 6tt npSnet vo undpla rdnotq rponono(qor1 oroug t<ov6veq rnq oordOetoq yto vo ei- vot 6uvorlq pretd<ppooq oro gttoXovdptor6 p$ootopr6- tto. E(vot ev6totpSpov to yeyov6g 6tt oro prroX6vdpto Uro pn tpononot4pt6vrl oupr6(v4 or4v npriltrl 06sn tou qwrxo6trov(ou e(vqt trov{ vo (euyopdrvet Kot Ue rtq r6ooeptg Bdoetg orqv tpfur1 06oq tou t<to6xov[ou. Mto Inov0utroro 'EwoUo Motrorro EXrvddepgo Nrlgortil6erg 11troru6trptv0eg Murrlteg ZUUeq 0uro AUA 724 KE0A Al0 25 0 levettxdq K66Lr0g
rdroro Url rpononotnpdvn oupt6[vr1 undpXet oro IRNA nou qwmpooonefouv Kot rtq orcril tetpol.teLe(g oxo- ySvereg ro6trov[tov: Pro, Thr, Ala, Ser, Leu, Val, Gly t<ot Arg. Aur6 elorrcbva ro ouvohr6 opt0pt6 ttov IRNA nou onomotwot oro pmoX6vdpto t<otd ormdr. H petorponq t<lv AGA rot AGG oe rol6x6vto tepgortoporj oro ptro- l6vdpro tov 0r1troonr<drv (B).6ne Ex6vo 25.11) ovorpe( rqv ovdyrq yro 5vo entnp6o0ero IRNA, on6te o ouvolt- 169 onomoUprevoq opr0pr6g IRNA e(vot orptBri:q 22. H gerotponrl tou AUA oe preOetovfuq getrivet nepom6p<o rrlv ovdyxq yto rponono(noq rnq rvoo(vr1g orq 05oq 34 rou tRNAIr" (BL5ne Ev6r4ta 25.6). Or 6rogopertro( rov6veg oordOetog yro 14 petdrppo- on oro ptroX6v6pto Kot rov nup(vo nr0ovdrg nporU- nrouv on6 6rogop5q orrl Lentopreprl doprrl to:v ow(morlrov prBoorrlpor(cov nou Uerogpd(ouv to 60o yovtduill.roro. Ito xurroponLool.totrd p$ootoprdno, ot rpononor{oerq orrlv U34 Xprlorpronototiwot rirore vo eneKro0e( q trov6rrlto onor<o6trono(qoqg optop5vtr:v IRNA (BI5ne Ev6trpa 25.6). Aw(Oero, oro 1.ttto1ov6pto- r<d prBooro;rdrro, ot rpononoqoetg orqv U34 Xpnoqo- nororivror yro vo neptop(oouv to (euYdpopo oro xco6u<6vro nou neptdXouv A ( G orqv tp(tr1 05oq, o0pt- govo pe toug ouvqOetg rov6veg qg oord0etog. Tpono- nouloerg orrlv U34 6wcog undpXouv oe Uroxovdptord IRNA nou owrnpooonefouv oprwo[5o yto 6rpre].e[g oprd- 6eg rco6u<ov(cov rilore v' onogeuX0e( to rorodtdBoopto nou 0o ouv6Bowe oe ow(Oerr1nep(mcoor1. 25.8 Ndo Aurvot6o Mnopel vo Evotoyorul0oriv oe Opropdvo Kol6rr6vro Tepprorroporj + H evotouorool rrlg oetrrlvoruotefvr1q oe optop6vo roldtrovto UGA onoue[ rn 6poon evog oouvrl0torou IRNA oe ouvduool.to pe notrtreq nporetveg. " To oouvrl0roro oprvo[0 nupotruolv1 pnopel vo evootportl0el oe opropdvo ro:dtrovto UAG. * Ito gtreQoptdotpopo npo:ro(oo Euplotes crassus ro rtrl6tr6vto UGA ro0opi(et tooo q oetrqvoruorelvl 6oo rot qv ruorelv1. Yndplouv 60o yvocrrd nopo6erypoto oro ono(o 6vo r<ordrr6vto teptrrotrol-toU Xpnotponolefuot yto vo xoOo- ploer dvo oouvrlOtoro oprtvolri 6togopertr6 on6 ro 20 xlooorxd optvo[6o. M6vo ouyxer<ptptdvo t<tl6tr6vto reptrrorrotrroU enovepUnveUowot ror' out6v tov tp6- no. Aw6 6e(yyer 6l ro v6r1;io rrlg rprnL5tog enqped- (eror on6 qv tout6rqro dMtrlv Bdoecov rou mRNA. 'Evo r6roro 6116 v6r1po ev6g ouyrerpt;t5vou xtodtro- v(ou np6ner vo 6Lor<pr0e( on6 tov ove(dptqro on6 ro ou pup po(6 ptevo nlrl pr1 enovonpoo6toptopt6 tr.ov t<tl6t- rov(ov oe optop6vouq opyovtogoUg 11 oro pttox6v- 6pto, 6nog neprypdqetot orrlv Evdr4ru 25.7. H oelrlvoruoteivrl, orqv ono[o to Oe[o rqg Kustd- vqg 61er owrKorooro0e( on6 oetrrivto, evoo4rcnthrcrot oe ouyKeKptg6vo xtodu<6vtq UGA oe yovi6to nou xto6t- ronororiv oeLrlvonpcoretveg rot ortq rpetq enxpdreeg rnq (oflq. Iuvr'10oq out6g ot npo:retveq rorohtlouv ofer6oovoytr:yx5q owtdpdoetg. To t<otd.tr otno oetrrlvo- ruoretvqg runtrd ewon((etot mo evepy6 r6wpo, 6nou Sreurotrfvet dgeoo rn )nUtKIi ow(6poor1. l-ro nopd6ety- 1.ro, ro xtr:6tr6vto UGA r<ooop((et tq oetrqvoruoretvq oe rp(o yov(6to rou E. coli nou rco6tronotoUv toodv(upto qg deu6poyovdoqg rou UUpUnYrtroU: to evooporo- pr6vo oe).r1vro ouvdmet dpreoo 6vo rorol.uttr<6 r6v ptol'u- B6ow(ou mo evepy6 rdwpo. Ot opyovtopro( nou prnoporiv vo xco6tronotrjoouv rrl oelrlvoruore"wq 6to05rouv 5vo oouvrlOtoro IRNA, yvco- o16 og IRNAS"", to ono(o 6Xet prlxog n6vo on6 90 vou- rl.eor(6ro rot nept6Xet ot5LeXog ono66rtq rot or6treXog T noxfi.ou pqroug. Aw( yto emd (eUyr1 Bdoe- ov oro or6treXog tou qnodSKrq rot n5we oro or6e1oq T (pto 6opd 7/5), to Bor,crlptoK6 tRNASu" 6to06tet pto 6oprl 8/5, evc0 ro tRNASec ttov oplo(tov Kot rov euKo- pucorrilv nr0ov6v 6ro05tu prto 6ogrl 9/5. Autd to IRNA neprdXouv en[o4g 5vo owtro6tt<6vto S'-UCA, nou rouq entrp6na vo StoBd(ouv to UGA, Ie 6Loug rouq opYovt- opro0q, ro tRNASec npdrto ol.twoorutrtc0verot Ue oepfu4 on6 rq oepul,otRNA ouvOetdorl (SerRS) nopdyowog oepuLotRNAsec. ftq Borctripto, ro 6v(upro ouvOdoq qg oel4voruorefv4q pretotp6net to Ser-tRNAS"" oneuOe(og oe oetrrlvor<uoreivulo (Sec)tRNAS"c Xpnotponotc0wog o:g 66q oe).4v(ou to oetrqvogtootpopx6. Ito opXo(o Kor rouq europucilreg, to Ser-tRNASec op1x6 Qoogo- puLuliveror on6 rrlv rtvdorl PSTK nopdyowqq Qooqo- oepul.o(Sep)-tRNAsu". I' 6vo 6eritepo Bqtrlo, ro Sep-tRN4s"" perorp6neror oe Sec{RNAsec on6 ro 6v(u- po SepSecS. H e(otpenrr'1 et6u<6qto qg PSTK efuot o[roo4;re(rorq: to 6v(upro e(vot xov6 vo <poorpopufuril- ver onoretreopottrd to Ser-tRNAsec, evd) onoxle(et to rovovtr6 Ser-tRNAS"'. H orot6Ltrr1l4 qroo<popuL(rooq rou Ser-tRNAser on6 qv PSTK 0o prnopotjoe vo npoKo- Moer evool.rdrroorl qg oeLqvoruorelv4g oe rco6u<5vto oepivnq. H enil.oyri rov rrodu<ov(tov UGA nou np6remot vo epprlvewoUv org oetrrlvoruore(vn roOop((etot on6 qv tonrr4 6eureporoyrl 6oprr] tou mRNA. 'Evog Bp6Xoq goupr<6tog r<o0o6u<d tou rodtxov(ou UGA, nou xole(- rot motye[o SEC/S, onomefuot vto rrlv evoropdtrrlorl t4g oeLrlvor<uoreiVrlq Kqt rov onorLetop6 qg np6o6eorlg nopoy6wcov onetreu06pooqg. Eto Borcr{pto, ro fiol.- 1e(o SECIS Bp(oretot oxptBcirg 6(nlo oro rto6tr6vto U- GA, evrir oro oplo[o Kot rouq europucilteq orrlv 3'-oper6gpqorq neproXri tou mRNA. Iro E coli, o e[et- 6rreupr6vog nopdyowog enttrrrptuvorlq SelB, oMrltrent- 6pd onortrermrxd pe ro SectRNASec 6q1 pe rov6vo dLl.o o1-uvoorutrtco gdvo IRNA, ou pneprl.oprBovo;t5vou rou np66po;rou Ser-tRNAS"'. O SelB enloqq npoo66ve- rot dpeoo mo ototle(o SECIS. To onot6leoUo rnq dpdonq rou SelB e(vot 6tt pr6vo to xto6tx6vto UGA nou en(oqq nept6Xouv 6vo r<otdtrtr4l,o tonoOerqpdvo SECIS 25.8 N6o Aprvo[60 Mnope(vo Evoroltotto0otjv oe 0ptop6vo KodrrovroTeppolopou 725 Boorxeq eworeq
npoo6{ivovror nopoyoytKd oro SectRNASec oq prBo- orrrpxq 05o4 A {EiK0NA 25.12}. To oplo(o Kor or euKo- pucbreq 6ro06rouv 5vo og6Loyo rou SelB, toor6oo yro vo enmpdQouv oro prBooro;rdno vo erooydyer q oetrq- voxuorefvr1 onomo[v enrn].6ov rrlv npoteivrl SBP2. 'Evo dMo nopdderypro evoropdrtoorlg ev6g er6u<ori ol.uvof6og e(vor q ronoO5rqoq qg nupoLuolvqg oe ouyrerpr;r5vo rodrr6vro UAG rou y6vouq Methano- sarcina rcov opXo[rov, 6ncog en(oqg Kqr oe ].ryo Borcrr1- pro. Iro Methanosarclna, nupol,uo[v4 undpXer oro evepy6 r6wpo rcov geOulorpovogepoocirv pre0uLopr(- w1g, 6nou nof(er oqprowtr6 p6],o orrl plpur<r1 ow(6po- or1. H evotol.rdrcoorlr4g nupol,uo(vqg npo0no06rer pro e[erdrreup6vr1 o;uvooru].otRNA ouv0erdo4, rrlv nu- poLuoul,otRNA ouv0erdo4 (PylRS), q ono(o oprvoo- r<uLrc0ver 5vo efer6u<eup6vo tRNAPyl pe nupoLuo(v4. To tRNAPvr nepdler ro owrxro6rr6vro S'-CUA, fiou ro Kq0ror6 xov6 vo 6roBd(er to UAG. 'Onorg Bp60qr<e yro ro tRNASec, ro tRNAPvr en[oqg eprgov[(er oouvq0tmo 6oprrd XopqKrnprorrrd nou 6ev undplouv oe dl'l.o IRNA: yro nopdderypro, 6e 6ro06rer ro moOep5 vou- x.l.eor(6ro U8, evc0 o Bp61og D ror o peroBAqr6g Bp6- Xog efuor oouvqOtoro Bpole(q. O prr;lovropr6q Ue rov ono(o ouyrer<pr1.r6vo rodrr6vro UAG droBd(owor oq nupoLuofurl 6ev SXer 6rol,eurov0e(, enet6rl 6ev rot6- orq 6uvor6 vo rouronorqOe( oogrilg 6vo ororXe[o beu- reporoyorlg SoUnq oe 6Lo rq mRNA ro ono(o evoo- porcbvouv ro opuvo{ri. Enrn}.5ov, 6ev 61er ovoyvopr- me( rdnorog er6rx6g nopdyovrog enrprp<uvoqg trre or6Xo ro Pyl-tRNAeu. i1o),6 np6ogorq qvoKo^.rig0qr<e 6n ro rodrx6vro UGA ro0op((er rrlv evoopdrolor; e(re qg ruore(vqg ei re rrlq oei.qvorume'lvqq mo B].egopt6og6po npcor6- (ao Euplotes crassus. Arrn Xprion rou UGA Bp60rp<e 6l ouprBofver or6;rq ror pr5oo oro (6ro yov(6ro, evril q enti.oyrl rou oprwo[rlog nou 0o evocol.rorcoOe( e[optdror on6 rr1 5op{ rqg 3'-oprerdcppoorqg nepropig rou mR- NA. To UGA npoodtop(er qv r<uorefv4 yevu<d cro Eu- p/ofes ror 6e Aenoupye( crlg xrodrr6vro teppoloproti. Iuvenc0g, ow6 uno64Lriver 6rr er6x4 rog npog 14 06o11 6rrq Xprloq prnope[ vo ouprBe( pr6oo oro n].o[oro ev6g rodxov(ou nou 6ev Xcnorpoflorekor erddMrrlg yro rep- prorropr6 oro ouyKeKprp6vo opyovrop6. npood6ver mRNA f lX&N,q :;"1t 0 SelB e(vor evog nopoyovrog enrprlruvorlg, nou npoo66ver etdrrd oro tRNASec 6vo rodrr6vlo UGA, ro onolo oxo- trou0e(ror ono pro 6opr1 oretreXouq-ppoXou oro mRNA. 726 KEOAAAI0 25 0 levelrog K66u<og 25.9 To IRNA Zeuyopdrvouv ErAerrrrd pe AprvoE6q Mdoro AprvooruAo-tRNA Iuv0erootirv * 0r optvoor<utrotRNA ouvOerooeg e(voL 1uo orroy6veto ev--- Uov, r0 ono[o npoo0rlvouv olrrvo[rlo oro IRNA nopoyovi:, o;.uvooru),otRNA oe pro ovr6poo1 6uo pqporov nou Xprl- gonore( evepyero ono ro ATP. * Ko0e ogrvoorutro{RNA ouvOerool optvoorutruiver otro ro :r. NA oe pto loo6exrlrq opoOo nou ovlnpooorneuer dvo oul.,e. xprpdvo oluvo[u u H ovoyvtirprorl rou IRNA ono lg IRNA ouv0erooeg Boor(e:: 0e Uro ouyKeKptpevl ogoDo vourtreol0[rrlv, Trlv (opo00 TC-- rorqrog,, rou tRNA, ro ono(o ouXvo ouyrevrp6vovroL : - ot6treXog rou onod6rrrl ror ro pp6Xo rou ovrlrodxov(ou To optvo[6o etod,plowot orn Uerogpoorrxq o6o ;,: otrl rnq dpoorlg rov oUrvooKUIo-tRNA ouv0erooc, or ono(eg nop6louv to onopoLrrlro onorodrr<onorr..- 16 Bqpo nou perorpriner rrq nlrlporpop(eq rov vo-- r).eivrr<rov o[5crrv mqv no].unenrLdrr<r1 o].q].o u Xi : Otreg or ouvOerdoeg 6pouv pr6oor ev6g Bnxovrofrc- 6[o Bqpdrov, nou nopouord(mor ofllv f i{*iiir r: :.] o To otuvoErj npoto owLdpd pe to ATP on6re oXqu:- t((eroL 5vo ollrvoorul.o-o6evuLrr<d ev6rdpeoo ncc- i6v, evur one).eu Oepoveror n u poqoo<prr-lprr<d. M d,c c : r1g evdpyerog nou oneleu0epurveror orrlv udpoi.-- oq rou ATP noyr6eferot un6 Uopgn 6eopoli ;rrx.:- ovudp(q ut{lnlnq ev6pyetog orrlv o6evu).u<r1 oproE: o Irn ouvdxero, e(re | 2'-OH e(re 11 3'-OH o;ro6o r.c- ewon((owor oro 3'-A76 vour<.treor[6ro rou IRNA c.- rL6pd pe to xopBovu).tr<6 dropro dvOpor<o rou *.- r<roU ovudp(rr1, nopdyowoq oprvooKu).o-tRNA *: row6Xpov4 one).eu06,prooq AMP. OL r5ooeptg IRNA ouv0etdoeg nou e(voL er6rxrig 1',: rq y).outopr(v4, to yLoutol.ur<6, trlv opyrv(v4 rot rq Luc.- v4 onoLroriv rlv nopouo(o IRNA yro vq ouvOdoouv r: ol.rLvoorulo-odevuLtr<6 ev6Ldpeoo. Ze ourd ro dv(uprc 4 IRNA ouvOerdorl oomd 0eope(tor rog 6vo prBovo"- xLeonpoteivrr<6 ootl-rot[dto (RNP), oro ono(o }.error..rc- y(o rou RNA e(vor vo Boq0d tr1v npore'tvr1 vo 6torr1pa. 1-uo r<oroi.urtxd enopr<q 6roprdpgooq. Iro deUrepo Bn- po rrlq oprvoor<uLirr;orlq, ro rUnpo rou o;-rLvo(5og ro- o1-rrvooru).o-o6evu).tr<oU gero<p6peror oto RNA ro" RNP (6't).o6n, oro IRNA). Kd0e IRNA ouvOerdorl e(vor ex).ercrtrr1 pr6vo yLo dvc opttvo(ri pero[ti 6].otv rrrtv oprLvol5orv rnq KUTroprKf c 5e[opevr1g. En(or1q 6tor<p(veL 6].o ro tRNA rou Kurrc- pou. Iuvrl0rog, r<dOe opLvo(0 owrnpoooJneUeror onc neproo6tepo rou ev6g tRNA. I-loLLon).d tRNA (ocr-rc lperd(owot yro v' onowotiv oe ouvrovul.ro r<ro6rrdvLc evri prepu<dq eopdq 6rocpopelr<d IRNA (euyoprbvouv ;.-rs to (6Lo xo6Lx6vto. lloL).qn^.d tRNA nou owrnpoot.rJ- Boorxeg eworeg
'Ev(ugo neproXn ogrvo[6og lleproXli tRNA [ieoLo vn Aip H ouv0eroorl nePLdXet 3 neptoxdg npdodeonq fl "-H-C-NH" Y -u- l O-U.U -o-p=o DU-T=U + AEevocivti To opwo[t rot ro ATP oplgori(ouv opwooruLo-AMP To tRNA npoo06veror To tRNA tDoprriverut Ue ouwo[ri n H-C-NH2 c"t+B:.'0-H r G,f 0 S Aoevootvrl R I H-C-NH I QDtro 'u lCS { EIK0NA 25.13 MLo optvoorutrotRNA ouv0etdorl entqop{(et to IRNA pe 6vo oluvo(ti. neriouv ro (6to optvoqf KoloUwot loodeKrlKd IRNA (isoaccepting IRNA). 'OLo rq IRNA eprgov((ouv u1v rovovtrrl tpttotoyri 6oUn qfriUotog L (Bldne Keqdtroto 24, MetdEpao4), To IRNA 6rnldrvetot t<otd t6toto rp6no c0ote to cr6LeXoq- ono65r,cr1q Kor ro or6^exoq T vo q(4Uor(ouv 1"tto ouvo- (ovrrui orriLrl, evdt to ar6lexoq D rot to or6leXoq tou qwKo6txov(ou vo qXq;roil(ouv pro((tov xdOeto BpoX(o- vo oX4prorog L. O Bp61oq tou qwtrodtxov(ou Kot ro drpo-onob6rcqq CCA tono0etoUwot oe ow(0eto dxpo rou 1-rop(ou rot onSlouv nep[nou 40 A. H otpotptrcl ne- ptoXrl rou oppoti rou IRNA, q ono[o ouv6Set rrq 60o r<d- Oereg orr1l,eg, onorctre(tot on6 to 9p6Xo D, to Bp6Xo T, ro ;retoBtrqt6 Bpo{ovo rt 6vo dtoorrltrrorr6 tptlpto 60o voux.treort6(rov preto[ti rou oreMXoug-ono6drctq Kot rou oretr61oug D. To neptoo6tepo IRNA nepdXouv ;rxp5q geroBLrlrsg nepto16g, nou onote^,o0wot on6 5vo Bp6- Xo 4-5 vouxLeortd(ov, evc0 eldXtoreq too6emtr6g opt6- 6eg e;rqov((ouv 6vo ;reyol0tepo pretoBtr4t5 BpoX(ovo, nou oupnepiloprBdvet 6vo (euyoptogSvo (6(xl'tovo) o16- LeXog, ro ono(o npoe(51et on6 6vo otpotptx6 nupfvo. To xorv6 oxnpo L t<ov IRNA e(vot 0eprel,tdr6eg yto rqv oLl.ql,en(6pooq 6l,tov ttov IRNA Ue rouq nopdyoweg enrprlruvorlg rot to ptBootoptdtto. Iro nLo(oto owr;g rqg r<otvqg 6opnq oXipcroq l- nou evtq4detot on6 rrlv nopouo(o ouvrrlprlpdvtrlv tpr toroycbv oMrlLent6pdoeov pt6oo crro ogotpx6 nuprl- vo, or oLtrrllouX(eq rou IRNA onoxtr[vouv mtg neptoo6tspeg O5oetg Kqt rov teoodptov Bp6ltov tou proplou. Auri 11 notxt),6rr1to drlptoupye[ ].em6g dtogo- p6g orn yov(o peto[ti trov 6Uo BpoXt6vov tou L t<ot, to or11.rowtr6tepo, odqye( oe nopoNoy5g orqv nope(o rou notruvouxtreor6txoU oxel,erorj xotd pr4xoq rou po- p(ou. Aufi orprBdrg q 6opxrl notril.6tqto e(vot 11 Bdorl yro q Srdxptorlrov ;rop(<ov on6 ng IRNA ouv0etdoeq. Ot IRNA ouv0etdoeg 6torp(vouv to IRNA p6oo 6tio yevrxcOv prrlXovto;uilv: dpeo1 @irect) t<ot 61tpeor7 avd- W@o4 @direct readout). Etrlv dpeoq ovdyvtoo4, to 6v- (upro ovoyvtop((er dpreoo Lettoupytt<5q oprddeq et6tr6g yr.o ouyKeKprp6veg Bdoerq-n.X. 6vo entgovetor6 opttvo- [rj puog IRNA ouvOfidoqg pnope( vo 5eX0e[ 6vo 6eopr6 u6poy6vou on6 rrlv e[orux].tr<ri oprwopddo rqg youov(- vnq Gnv N2 qg G), pto oprd6o qg eLdooovog otitrot<og nou 6ev undpXet ortq dLLeg tpetg Bdoerg, Aw(0ero, orrlv 6;rpeorl ovdyvoorl, to 6v(upro npoo66vetot d;reoo oe pq etdu<d rprlporo tou IRNA, ro ooKxopoogooqopt- 16 oreLer6 rot gr1 et6xd tprr]goto ttov vourtreortdxrbv Bdoeu:v. l-to nopd6etypto, oll,rlLouX(eq oro pretoBLrlr6 Bpo(ovo r<ot ro Bpol(ovo D ev6g IRNA prnope( vo 6qgt- oupyrjoouv 1.uo t6to(tepq entrpdveto, ouprnlrlpoprotrr{ pr6vo ge qv ow(orotXrl IRNA ouv0etdorl rot rop[o d],- lq. Kot' our6v tov tp6no, voux.treoil6to to ono(o on6- louv on6 q 6tdcpooq ev(UproutRNA 6rlprtoupyorjv ;tto 6o;ui drdqoonq nou ge r4 oepd r4g npooS6vetot dge- oo. T6oo n dueon 6oo rot q 5prpreo4 ovdyvtoo4 ouvti- 0tog trettoupyotiv cno ntro(oto oprotBo(og enoy6pevqg npooopproyrlg: ot oMoy6q dropr6prptoorlq t6oo oro tR- NA 6oo Kot ro 6v(upro ouptBo(vouv prerd qv opXtrrl np6odeorl oXq;rot((owoq 6vo nopoytoyx6 xototrurtx6 ofgntroro. Kot oroug 66o pqlovtopotig ou;yd en(or1g oullper6Xouv pr6pto vepo6 orrlv entrpdveto enoqfg ge- ro[ti tou IRNA ror rou ev(Ugou. l-to nopd6ety;ro, 5tov q ouvOerdorl tou yl,outol.ttvul'o-tRNA (GlnRS) npoo66- ver ro tRNAGrn, 60o neptol6q tou ev(Uprou neptorp6rpov- ror rl pto oe oX5or1 Ue rnv dlLr1. Tout6Xpovo, to 3'- pov6x).rovo drpo xot o Bp6Xoq tou owtxtodtrov(ou tou IRNA uE[orowot onpowtr6g oMoy6g 6topr6prpoorlg oe oriyxpro'q pe ttg unoOerx5q 6opr6q rouq orn Un ouv6e- 6epr5vr1 r<otdoroorl. Ie noLtr5q neptntc0oetg, ot xo0optor6g tou IRNA nou e(vot ovoyxo(ot yto er6trcrl ovoyvdrptoq ewon(owot oro drpo rou pop(ou, 6r1Lo6n crro or0reXog-ono65rt4 Kor ro Pp5Xo tou owtrodtrov(ou. Yndplouv oor6oo nopo6efuproro qro ono(o to vourtreot6to rou rptroro- yoUg nuprivo nop6louv to orlpoto tout6t4rog. 'Evo dMo xow6 vourtreot(dto toutono(r1or1g e(vot n *pdon 6rdr<ptor19" ("discriminator-base") orrl 05or1 73 tou tR- NA, 11 ono[o ewon((erot oprSoog 5'qq 3'-re].tt<{q qI n- l,ouX(og CCA. E(vil ev6tocp5pov 6tt q otrLrlLouX(o owt- rro6u<ov(ou rou IRNA 6ev e(vot ovoyxoortxd onopofu4- rn yro rnv ovoyvcirpto4 et6trriv IRNA ouvOetoociv. l-evt- r<d, 11 oprdSo tour6ultog rou IRNA e(vot tdtofuep4 yto rdOe IRNA ouvOerdoq. 25.9 To IRNA Zeuyoptirvouv Er),exnrd pe ApLvo[60 M6oro AgwoorutrotRNA luv0eroodtv 727
Or roOopror5g rout6qtog norrilouv toq npoq rn oqproo(o rouq Kor preprx5g gop6q ouwqpoUwor r<ord qv e[5].tlr1. H ouwrlprlorl rov ororxe(ov rout6rqtog ovo6erKv0erqr on5-r4v rxovSrpo noMc0v IRNA ouv0e- roorilv v' otruvooruLrcilvouv IRNA nou npo6pXowor on6 drogoperxoriq opyovroUorig. Yno06oerg oXenxd Ue rrlv opd6o rcov ororle(tov row6ul-rog rou IRNA nou e(vot onopo[r4to yro rnv ex].er,crr4 ovoyvdrprorl on6 puo tR- NA ouvOetdoq npo6pXowot on6 preldreg rpumdMov IRNA ouv0eroo6v ;le nep(O).ooq orcr(vtov X, r<).oorrc] ye- verLxrl ovdl.uon Kor rn vfuo trreroMo[ty5veor1. H oprorxrl on66er(f 6n gro ogd6o tow6r4rog rou IRNA e(vot ro- Ld xo0oproprSv4 ),opBdveror qn6 nerpdproro preropr6- oxeuorlq, oro ono(o 11 nr0ovo).oyoti;revr1 oprddo vou- rl,eol6(ov evocopor6veror o' 5vo IRNA on6 pro 6ro9o- penr4 roodercnrrl oprd6o. I-ro nopd6erypo, q owrrord- orooq 15 vouxLeon6[ov oro or6].e1og-ono66rcrq ror to 9p6Xo rou owrxrodu<ov(ou rou tRNAAsp on6 r' ow(- ororxo voux.eor[6rq rou tRNAGln en6rper+re orq ouv0e- tdorl rou yl,ouroprvu).otRNA (GlnRS) v' optvooKu- Luioet ro rpononorqpSvo tRNAAsp pre yl,ouro;r[v4 pre onoreleogorrx6rrlro rr erLerrx6rrlro ouyr<p(orpeq Ue rtg ow(ororleq ornv rovovu<r1 ow(6poo( qg. l-1oLl,6g IRNA ouv0erdoeg opuvoor<uLuilvouv erdu<d 1.uo "1-uvui),xq" ("minihelix") rou IRNA, q ono(q qnore- Lefuqr on6 ro BpoX(ovo-ono65rcrq xor ro Bpol[ovo TrfC rou pop(ou. Ie opropr5veq neprmriroerg, rog undorpcol.ro pnope( vo l,eroupyrioer Uro purpo6lrro rou IRNA, nou onoretrefuor ;r6vo on6 6vo orrileXog-onod5rcrq xlerqr6 oro dnro drpo rou on6 5vo oro0ep6 retpontr6 Fp6Xo. T6oo or pwdlrreg 6oo ror or pxporiLxeg oprwoor<uLui- vowor fuy6repo onorel,eoportxd qn6 ro or5poro IRNA. Autd rq nerpdproro 51ouv rdnoro orltrroo(o, rosr6oo, yto rnv e[e],trcnxr1 ovdmu[r1 tolv ougn].6r<rov r<ov IRNA ouv0erood:v. I' 5vo np6r;ro e(efur<rx6 ord6ro, ro IRNA (oog onote}orjwov pr6vo on6 ro BpoX(ovo-ono65r:rr1 tou oriyXpovou prop(ou. 25.'l O Or AprvooruAo-tRNA luv0erdoeq KorordooovTor oE Ario Orroy6vereq * 0r oprvoorutro{RNA ouvOerooeq dtorp(vovTor oTrq orroy6ver- eq To[nq I ror ll pe goorl rdroirepo porLBo otrtr1trouXulv ror tdr- olrepeg 6opLreg neproX6g. l-iopd rrlv rorvrl enoupy[o roug, or ouvOerdoeg e(vor opr<etd erepoyevqg oprddo ev(riprcov. Atoxp(vowor oe 6r-io orxoy6vereg. Or IRNA ouv1eraoeq Tar1q / e(vor r<upftog provoprepe(g r<r epr<pov((ouv 6oprr<d nop6proreg neproXSg (nruX6g) Rossmann (Rossmann fold do- mains) cro oUrvoreLrr<6 dxpo 11 xowd oe our6, ot ono[eg oupprer5Xouv mr1 6ropr6pgoon rou evepyori r<5,wpou. H nruXrl Rossmann onoteLe(ror on6 5vo B- 728 KEOAAAI0 25 0 levettroq Kdrdtxog muX<ot6 g0Mo 5-6 nopdMrlLcov muXc0oeov Ue otr vernx5g SLrreg. Aurq q neproX4 e(vor opr6Loyn rnq neptoXrlg rou evepyo0 r<6wpou rrov deudpoyovoorirv ror unefOuvn yro rnv np6odeor; rou ATP, rou oprvo- [6o9 r<ot rou 3'dxpou rou IRNA. 'O].eg or ouv0erdoqg TdEnS I nepr6louv puo neproXrl "np6o6eonq rou ono- 66rcrq", q ono(o evo<rt1lor6veror srnv (6ro 05oq rqq rnuxnq Rossmann oe Uro rotvri 06o4, 6nou en(oqg npooS5veror ro l.rov6xLrovo drpo-onod6rcrqq rou tR- NA. Ie peprrd 6v(upro, ourl 11 06oq en(or1g nepdXer 6vo evepy6 xSwpo ene[epyoo(og (BI6ne Ev6r4ta 25.11). Or xopBo(urelrxSg neprolrlg rotv ouvOerooriv tdlqg I npoo66vowqr ornv eoo:reprx( yrov(o rou tR- NA oXqgorog L xor oro Bpo(ovct rou owrKo6rxov(ou xor q Aenoupy(o roug e(vor vo drorp(vouv ro IRNA. AUo rotvd BpoX6o ;ror(Bo o).Lr1).ou1rc6v qg nrulqg Rossmann egnl6rowor ornv np6o6eoq rou ATP. Ercr6q on6 rdnoro nepropropr6vq oproi,oy[o prero[6 ].i- yrrlv ev([;rov, 6ev undplouv orlprowrx6g opror6qreg orrl 6op( 4 qv oLlql,oul(o rov ev(rigtov rdfirlg l, err6g qg nruxnq Rossmann. Ot fRNA ouv9xdoeq Tdlryq il e(vor en(or1g erepoye- ve(q. H tetoproroyrls 6opn roug yevxd e(vor 6rprepqg, roor6oo prepu<69 qopdq oXrl;.ro{(ouv oprorerpogepq 4 ozBz ereporetpouepn. 'Onog ro 5v(upo td(r1g l, ot tR- NA ouvOerdoeg rd[qg ll 6ro06rouv puo 6o;rxd ouwn- pnUSvn neproXrl evepyori r6wpou - ornv npor<erpdvrl nep(nrororl np6retror yro pro prrrcrl o/B neproXrl nou 6rog6per on6 r1v muXr'1 Rossmann. To evepyd xSwpo rov IRNA ouvOeroocilv rd[rlg ll ewoni(owor npoq ro t<opBo[ure]lx6 dr<po rov no],unemr6(cov. Ie ourrl r1v oroy6vero e(vor ouwrlpqpr6vo rp(o BpoXSo ;ror(Bo ol,- l,4Louluilv orqv neptoXri rou evepyoti r6wpou: ro 5vo on6 outd Letroupye( mov no)r,upepropr6, evdl r' dMo 6Uo 51ouv r<orolunr6 p6).o. YndpXouv 23 <pu).oyevmrrd 6rorptr5g orroydvereq IRNA ouvOmoorilv. 'Ewexq xorordooowor orqv td[q l, ev6 or un6lotneg 12 e(vor dv(upo rdEnq ll (ElK0]lA 25.14).'EXer ev6rog5pov 6r undpXouv 6rio 6togopetu<o( rfnor ev(rjpov LysRS nou ov{rouv oe 6rogoperrxdg rd- (erg. l-1 p6orporo en(or1 g ovor<o).ri g0r1 rov 6 [ o orroydver- eq Url Kovovrrtiv IRNA ouvOetoorilv nou neprop((owor oe oprol-uivo e(6r1. Aurd ro 6v(upro e(vor 11 nupol.uoul,o- IRNA ouv0erdoq (PylRS) rd[r1g ll (neprypdgetor ornv Ev6r4ra 25.8) xor n eooeooepuLotRNA ouv0erdorl (SepRS)td[49 ll. H SepRS neprop((etor oro pre0ovoy6- vo (puo un6ro[r1 rov oplo(rov) ror ro orevd ouyyevx5 Archaeoglobus fulgidus. llpoo6Sver gologooep(vr1 (Sep) oroug ono66meg rou tRNACys nopdyowog 5vo peprrtirq oxuLuop6vo Sep-tRN4cv". 'OLor or opyovtopro( nou 6ro05rouv SepRS en(or1g 6to05rouv to SepCysS, 6vo ove(dprqro rrlq gtrrogopK4q nupr6o[d],qg ouvo66 5v(u;ro ro ono(o perorpdner ro SeptRNAcv" oe Cys-tR- NACys. O 66rqg Oe(ou nou Xpnorponorefror on6 ro SepCysS in vivo e(vor dyvotmog. E(vor o[roorlpe(trrro 6rr opropr6vo pe0ovoy6vo 6ro0drouv qv o66 6Uo B4- ptdrov rolv SepRS/SepCysS rot, nop6M4Lo, ro rovovr- r<6 5v(upro CysRS. l-1op6n undpXouv 23 rpuloyeverrxd 6rorpudg orro- Boolxeg ewocg
Apwooxu),0- tRNA ouvOerdotg Tdqn I Td[n ll EIK0NA 25.14 AroXoptopog rov IRNA ouv0er0o6v oe d0o Td- [erg, nou otoo5rouv onor]retmtxd opotpo(eg oU66eq UoTiBov otr- )rnlouxtdrv rot 6optrd evepytiv neptoXtitv. Irlgetdtvetot 11 reroprotoynq 6oprl rou ev(upou. tlol)ron)to( 6e(xreg unodet- xvoouv ofl 4 retoptoroyrlq doun dtotp6pet oe nox0rouq opYuvt- oUouq. H retoprotoyrlg 6o911 qq PyIRS 6ev 6Xet xo0optore[ oorptirg. ydvereq IRNA ouvOetoooiv, ot neptoo6rcpor opyovtol.to( dto06rouv g6vo 18 6v(upro. H GnIRS Kot rl ooilopoYtvu- tro-tRNA ouv0etdorl (AsnRS) tunrd onouotd(ouv on6 ro penepr6pto rov opYovto;rc0v. l-to vo ouv06oouv Gln- lRfrlfiotn Kqr AsntRNAA"n, ouro( ot opYovtouo( dto06touv [eX<oprmd 6v(upro trr' evepy6rrlto y],ouroputro{RNA ouvOerdoqg (GluRS) Kot oonopruLo-tRNA ouvOetdoqg (AspRS), to ono(o 6ev epgov((ouv xov6rlto dtdrptorlq (nondiscriminating-ND). H GIuRSND ouv06tet 16oo Glu- tRNAGru 5oo rot pepxdrg oxutrttopt6vo Glu-tRNAGrn, evr6 4 AspRSND ouv06ret 16oo AsptRNAA"p 600 rot peptrcig oxuluol.uivo Asp-tRNAA"n. Itrl ouv61eto, to peptxcOg orulrcopdvo IRNA ;retotp6nowot oe GlntRNAGrn 6q1 Asn-tRNAA'n pre q opdorlq Utoq oprt6orpovotpepdoqg nou e[optdtot on6 to IRNA (amidotransferase-AdT). Ot AdTs e(vot o[tooqpre(<oro noLupreprl 6v(upro nou 6ro05- rouv rperq 6ror<pm6g evepy6qteg (ElK0l'lA 25.15)' flprirro nopdyouv opr;rorv(o o' 6vo evepy6 x6wpo pr6oco onog(- 6ooqq ev6g 66q o(tirou, 6noq 11 yLoutopr(vq 4 r1 oonopoy(vq.'Enstto, 11 oprprcov(o pretog6petot 6to;t6oo u ev6g evdoptoptorori 6to[].ou rot ovo6tletot on6 pto deUrcpq 05or1, 6nou npoo66vetot oro 3'-drpo tou trd- 0og or<uLuo1l6vou IRNA. Iro de0repo evepy6 x5wpo, pro rrvdorl y-gtrrotpopuLttilvet rqv r<opBo[u]'oprd6o rrlg ni.eupu<r1q oLuofdog rou ogwo[6oq oro Glu-tRNAGh n ro AsptRNAA'n. T6Log, q oprpov(o ercron(et ro Qooqo- prr6, g4qpror((owoq Gln{RNAGrn ri AsntRNAA'n. Yndp- louv [eXcoptordg oroydveeq AdT nou enevepYoUv rot sro 6Uo Ld0og orultorp6vo IRNA n neptop((owot pt6vo sro oxrlUorropt6 Gl n-tRNAGr". Or ouv0erdoeq rd(nq I r<ot ll 6tor<p[vowot lemoupyt- xd on6 oprerSg on6q.retg. [lptirtov, ro 5v(upo tdfiqq I ol.rlooxullc0vouv ro IRNA or4 2'-OH e6on qg A76, evcir ro 5v(upro rd[qg ll yevtrd opttvoot<utrtcilvouv to IRNA arnv 3'-OH. H 05on qg opXtxrlg ogwooruL(trloqq q(eil- tBNAGrn >-Gru.tRNAcrnKGlu + ATP Gln + ATP tRNAAsn Asn-tRNAAsn y-Asp-tRNAAsnN Asp + ATP Asn+ ATP Asp EIK0NA 25.15 Mryovrogo(yto rn ouvOeon GlntRNAGln xqt Asn- tRNAAsn. Ie xdOe nepinroorl, 11 dvto nope[o unodexvUet qv odti ev6q Bnporoq nou rotqluerot on6 q oupponxn IRNA ouv0etdol. 0t xor6repeg o0o( Dtlo Bnpdttov onovroUv moug neptoodrepoug opyovroporjg. Anoretrouwot ono pto pr1 dteurptvtortxrl IRNA ouv- Oerdor1, nou orotrou0e(rot onri rq 6poor1 ev6g tRNA-e[opr6pevou ev(rjpou opt6orpovorpepoons (AdT). (eror ge tqv roteUOuvo4 np6odeonq rou IRNA ndvo oro 6v(u;ro. Ot ouv0etdoeg t6[r1g I npoo6Svouv IRNA orrlv nl,eupd rqq eldooovog otiLor<oq tou oretr61oug- qno66Krn rot onoro6v to pov6xl,tovo 3'-rel,x6 drpo vo 61et 6opuj <poupr<6tog yto xotdMqtrrl owtnopd0eoq pe to opwo(ri ror to ATP cro evepy6 r<6wpo (EIK0NA 25.16). Aw(0ero, ot ouvOetdoeq td[qg ll npoo66vowot orrlv nleupd t4g pre((ovog orilorog tou oreMXouq-ono66tc4 rou IRNA rqt 6ev onomoUv vo oxnportare( goupr6to peto[ri tou 3'-dxpou Kot rou evepyoti t<6wpou. Yndplet en(onq Uto Unxovtorxrl 6rogopd nou np6ogoto ovoKo- , 0r xpuorotrtrLrdg 6op6q unodetrvuouv ort ot opl- voorutrotRNA ouv0erooeq ro(nq I rot ll npoodtlvovrot orlg ovr[- 0ereq oQetq rorv tRNA unoorptlportov T0uq. To tRNA onerrovl(erot epuOpo rot 1 npotelvrl ruovrl. H tporoypotpto nopo- Xopn0lre ono tov Dino Moras, lnstitute of Genetics and Mo ecular and Cellular Biology (IGBMC). Asn Asp Ser His Lys Thr Pro Phe Ala Glv Sep Pvl Gln (o) Glu (o) Arg (o) Lys (o) Val (o) lle (o) Leu (o) N4et (0,02) Cys (0, o2) Tyr (02) Trp (02) (oz) (q2) (oz) (oz) (oz) (oz) (oz) (0, oz0z) (qz, o+) (oz, oz9z) (0, ) (?) Gln + ATP Asn + ATP J*.1& 0L A;rrvoorutrotRNA luv0erdoeq Kororoooovrotoe Auo OLxoYsveLec 729
).0q0r1re: 11 roXUqro ow(6pooqq tcov ouvOetoocilv rd- (qq I neprop((*ot on6 rrlv onetreuO6pcoon optvoorufuco- pSvou npoi5wog IRNA, ev6 or ouv0etdoeg tdEnq ll neprop((owor on6 npoql6repo Xrlpur<6 Bqgoro Kq/n eu- orr6g ovo6rord(erq oro evepyd xdwpo. 25.1 I Or Euv0erdo€q Xpnorponoro0v Tn Ar6p0oon Aorrpitov yro vo BeArrdoouv rnv ArpiBero r H er6xriuru rou (euyoptirlroroq uero[0 ogLvo[6ov roL IRNA pu0gi(eroL p6orrl ovndpooeov 6r6p0too1q 6orq[ov ot ono(eg u6potrtiouv trd0oq oxnUottop6veg optvooxutro-odevu)txdg ev6oeLg roL oluvooxutrotRNA. Or oprwoor<utrotRNA ouv0etdoeg np6net vo 6toxp[vouv 6vo er6u<6 opuvo[ri on6 rrlv rurroprr( 6e(opevrl tov oprwof6tov Kot rov ouvorpdrv prop(tov, 6nog en(,cr1g rot to ouyyevrl IRNA oe prcl ouyKeKprprdq toodererxrl opd- 6o (runtrd 5vo 6ro9 rp(o) on6 to otlvotro tcr:v IRNA. H Srdrproq rou IRNA prnope[ vo enmeul0e( ge Bdoq ng 6rogop6q ong 6op5g oXrlprotog L (BMne Ev6tqra 25.9). Aur6 ou;rBo(ver t6oo r<otd to opXx6 Bf go np6o6eoqg 6oo xqr mo en(ne6o rrlg enoy6gevnq npoooppoyiq. To prl ouyyevrl IRNA nou npo6pXowor on6 dtrtreg too- 6erctx6g oprd6eg 6e 6ro06touv rqv ntuipq oprd6o vou- r.l.eol6(rov rout6rrltog. 'Eror, 6ev efuor oe 05or1 vo enovodtord[ouv rrl 6opr( roug c0ore vq utoOetjoouv puo 6rop6prpoon ornv ono(o to owt6poortr6 telu<6 drpo CCA eu0uypoprpr[(etor r<otdLtrr1tro Ue rnv xopBo- [u].opd6o tou ogwof6oq Kot ro o-gooeoptK6 tou ATP. Autq 11 on6ppr{.rr1rov Un ouyyevd:v IRNA o' dvo ord6to qg ow[6pooqg nprv on6 rr1 orJv0eoq tou pepmrig ot<u- Luog6vou IRNA prepu<69 ovog5petor og rtv4nx{ 6l6p- 0roo4 6orrp(ov (kinetic proofreading). H o6uvo;t(o rtov Un ou1ryevc0v IRNA v' oxolou0rloouv to Xqprxd Brl- Uoro rnq oploorutr[oo4g npo6plerot qn6 to yeyov6g 6rr ro IRNA dr(ororot on6 to Sv(upo notrU toX[tepo on' 6,rr pnope( v' owt6pdoet (EIK0NA 25.17). Aw(Oero, or IRNA ouvOerdoeg o6uvotoUv vo dror<p(- vouv oproptdvo 6opxd nop6ptoto oprtvo(6o rotd r1v no- pe(o rrlq o[vOeoqg rov oUtvooKu].otRNA nou oup- Bo(ver oe 6rio Brlpoto. E(vor r6tofuepo d0orotro Yto ro 6v(upro vo 6tot<p(vouv 60o opuvo[6o ro onofo 6tog6- pouv pr6vo oq npoq to prrjr<og tou ovOpot<tt<of ordr.e- to0 (6ql,o6ri rord pro opd6o CHr) ti optvo(6o (6tou prey60ouq nou droqdpouv ;t6vo oe Uto 06oq. Fto nopd- 6ery;ro, o 06l,oro9 np5odeo4g optvo(5tov rqg tooleu- xutrotRNA ouv0er6orlg (lleRS) 6e prnope( vo 6tot<p[vet rqv rooLeurfrn1 on6 rr1 Bofuvn opretd rol.d c6me vo no- pepno6(oer 14 o0v0eor1 orlprowxrlg noo6rrltog ValtR- NArr". llopopo(cog, q Bo].utrotRNA ouvOetdorl ffalRS) ouv06rer Thr-tRNAVar oe orlpowx6 Po0U6. H lleRS, 11 VaIRS r<ot toutrdXrorov emd entnp6o0e- reg IRNA ouvOerdoeg (or erdu<69 yro q treurfur1, rrl Ue- 730 KEOMAI0 25 0 [everxdq Kti:Otxoq To ouyyevdq tRNA nupo0oreI otr],oyn rnl dtoutjprpoon Autvoorutr[toon Xop[q qtrtroyn orn droprlprpoorl oumo(ver opyd Aprvooxutr[oor1 oulrBoiver yprlyopo EIK0NA 25.17 H oprvoorutr(roorl Tov ouyyevtirv tRNAs on6 q ouvOetdoq pooi(eror ev g6peL oe peyotrUtepl ouyy6veto yto outd ro ei6q, oe ouvduoo;rd pe ooOevrl ouyydvero yLo p1 ouyyevrl e(6q. Enrntr6ov, ro gr1 ouyyevrl tRNAs oOuvototiv vo unooro[v ntrtlpoq ng induced-fit 0rogoprptorr6g peropotr6q, nou onououvrot Yto T0 6rfrpo rorotrunxd prlporo. Oetov(vr1, qv oLov[r4, qv npo].(vt'1, q gowu],o],ov(vr1, q Opeov[vr1 r<ot q truo(vr1) efvot txov6g vo 6top0cbvouv r' ol.rwooruLo-o6evuLtxd Kot oUtvooKUl,o-tRNA ro ono[o oX4goil(owor mo evepyd rdwpo toug ;16oro entnp6- oOerov evepyorrltov nou efue u6potr0ouv ro ol.ltvooKu- ).o-AMP onod[dowog el,erjOepo oprwo[ti xor AMP efue udpotrtiouv to preptrcbg orul.urrpdvo IRNA ono6(6owog eLeU0epo oprwo(rj ror onoorutruop6vo IRNA. H u6p6tru- orl rou otrrwoot<ul,o-AMP ovo<pdperot oS rpo-peragopt- xr1 6t6p0ao4 (pretransfer editing), evdr q u6p6l'uol1 rou ol.rwoorulo-tRNA tog usra-peraEoptxfl 6Ldp0uo4 @ost- transfer editing) (EIK0NA 25.18). Ir4v nep(mooq'rnq npo- pretogopu<r|g 6t6p0too4q, nt0ov6v 6vo pdpog qn6 ro ).dOog olqpotroprdvo ol.twoorulo-AMP 6(orotot on6 to evepy5 r<6wpo n 6nemo u6potrrietot gq ev(uprtrd oe 6rd- Lupo (o oprwoor<uLrx6g eorepx6g 6eop6g e(vot qXenxd omo0rlg). Aut6g o rUnog ow(6poor1q ene[epyoo(og ;rnope[ en(oqq vo 0ecopr1Oe( oq Uopeli rrvrlnxrlg 6t6p- Orooqg 6orrp(cov. Aw(0eto, 16oo 11 npo-pretogoptxtl udp6uor1 rou Ld0og ol.uvoorulo-o6evutrtro0 6tov npoodeOe[ oro 6v(upro, 6oo t<ot 4 roro].u6prevr1 on6 6v- (upro preto-perogogxri 6r6p0oor1 e(vot yvoor6q oq Xq- prx{ 6rrip0too4 doxrltitov (chemical proofreading). ouvbderqr opyd Boorxeg eworeg
To x6wpo 6rdp0oorlg e(vor prxporepo onc iJ sJ,:3...: IuveeTtK6 r6vrpo .,::,' Ktlvrp,o 0Lo:;-:-: AMP ndnNH' tcooH To q[rvooKUIo-tRNA udpol0erot ouv0erdoeg e(vor duvotri v0 rpoYpoTonote(Tot oTo ord0Lo nptv on6 qv opwoorutr[too1 (npoperoqopxn ene[epYoo(o), oro ono(o uDpotrueror n Un ouyyevnq optLvooxul,txq oDevu)xr1 opoDo. Evotr- trornro 1 enrnpdo0eto, evd6XetoL vo )ropet Xtitpo I udptjtruon tou trov0oop6vo oXtlportop6vou olttvoorutrotRNA pero q oUv0eoq rou (peropetoqoptrn ene{epYooto). Motrov6r or owtdpdoetq npo-Ueroqopu<r1q 6t6p0oorlq prepx6g qop6q ouprBo(vouv onouo(q tou IRNA (6nLo6n npw on6 qv np6o6eoq tou IRNA), 11 nopouo[o tou tR- NA yevrrd Betrruilvet nolri rqv onotel.eoportx6rqto tqg u6p5luoqg. To noto 6r6p0toor1 0o enu<pouloet, TIpo-Ue- rorpoprx4 6vowt trleto-prerocpoptxrlg, notrfi.et ovdloyo pre q ouv0et6oq. H ow(6pooq dt6p0cooqg Boo((etot orov rl.ootr6 pllXovropr5 6rn),o0 qOpoti (double sieve), 6nog nopou- ord(eror yto rrlv lleRS mrlv El(0NA 25.19, xotd tov ono(o rog B6o4 yro q dtdrptoq XpnotUonote(tot to pr6- yeOog rou optvo[6o9. H lleRS 6to06ret 6uo evepyd r5v- rpo: ouv0ert16 (r1 r<5vrpo evepyono[r1oqg), nou ewon((eror arrl ouvq0rl mupl Rossmann td(r1g l, rot x6wpo 6t6p0toor1g (n, qltu6q, u6potrutu<6), nou ewo- nf(etot orqv neptoXrl np6o6eo4g tou ono65rcrrl (B^.6ne Evoqta 25.10). H rpuoroMtrn 6oUn rqg lleRS 6efiyet 6n ro ouv0*x6 r6wpo e(vot notrri trrtr<p6 yto vo enrpS- qrer rnv e(oo6o qg Leur(vqg (r1 n).euptr{ o}.uo(6o qg Leur<(vqg 6ror<Lo6((etot oe dtoEopettr{ 06o4 oe orly- Kpron Ue rqv tootreux(vq). Itrlv npoyl.tonr6tnro, oTIo- rle(erot q evepyono(4or1 6Ltov ttov otrtwo(6cov nou eivot preyoLrirepo on6 qv tootreurfv4, ener6rl 6e gnopotiv vo ero5tr0ouv oro ouv0ettr6 r5wpo. 0o16oo, oprqrdvo pu- rp5repo o;rrvo[6o nou dtotrlpofv enoprrl trov5ulto , NH, RCH tcooH H Leu e(vor nolU peydtrrl yto vo ltrlpdoet mo ouveertKd Kdwpo @-rcu"'" H lle xopd mo ouvOerxo x6vrPo, dxt d[oq rot oro xdwpo Dtop0toonq @-w 9@H Val nepvd ond to ouv0ettrtj r6wpo mo rdvrpo 6trjp0toonq EIK0NA 25.13 H ouv0er6o1 rou tootreuxutro-tRNA 6Xet D6o evepydg neptoX6q. ApLvo[60 UeYdrUrepo qq lle Dev pnopotlv vo evepyonoulOouv, dt6l 6ev eqontovrot orrl ouvOettrrl neptoyt'1. Aprvo[60 ptxp6repo qg lle onoporp6vovrot, xoOtitg pnopotlv vo ero5trOouv onlv neptoXrl ene[epyooiog. np6o6eoqg -n.X. n Boltvn- prnopotiv vo etoSL0ouv oro ouv0errx6 r<5wpo Kot vo npoo6e0o6v mo IRNA. To ouv0ertx6 xdwpo Lettoupye( rog npcitog q0p6g. To r5wpo 6r6p0tr:o4g e(vot ptrp6repo on6 to ouv0ettx6 rqr 6e prnope( vo tpttro[evrioet rn ouYYevq tootreurivrl, comSoo npoo65vet q Bol(vq. 'Etot, ro ValtRNAlle u6potrtieror oro r6wpo 6t6p0ooqg, nou trettoupye[ og Serirepog n0U6q, evdr to lletRNAlle 6ev u6pol'Ueror. To prowSl.ou rou dmLorj rlOprod e(vot 6vo e0xol.o rot yevrrd orpp5g prowSl,o rnq Lero-Uero<poprr{g 6t6p0to- onq. :rnv lleRS, 6nog entoqq xot oe dtreg IRNA ouvOe- rdoeg 6t6p0rrlonq rd(nq I rot ll, to 66o r<6wpo (ouv0erx6 r<or 6r6p0roo4g) on6Xouv opxetd, 10-40 A. l-lpoxeqr6vou vo Trpoypcnonoq0e( 4 Uero-UeroQoprrq udp6Luoq (5t6p- Orr:on), ro drpo-onod6rcrqg rou preptr<cirq oxuLtrop6vou ol.rwoorutro-tRNA petoton((etor xotd pnKoq'rnq entqd- veroq rou ev(0Uou (EIK0NA 25.20) on6 to ouv0err6 xSwpo oro rSwpo 6t6p0coor1g. Aurn n pretot6ruoq nepr.I.oprBdvet puo peroBotrrl orq 6topr6p<poorl rou dr<pou-ono65ffi'l rou IRNA. Itrg IRNA ouv0erdoeg td(4q l, to dt<po-onodSrrqg uroOere( 6rop6prp<ooq goupr<5tog 6tov npooS6verot oro ouv0ertr6 rSwpo (Blene Ev6tryo 25.10) xt er<tetopr5vq 6oprl 6tov npoo66verot oro r6wpo 6t6p0oor1g. Meror6nrorl ev6q LdOoq opuvo(6og ndvro mrlv ent- <pdvero rqg IRNA ouvOetdorlg e(vot ntOovrl oro Unxovt- o;16 qg Utroperqeoptxrlg ene[epyoo(og enet6q to oprwo[ri e(vor ol.rorono].u<d npoode6epdvo oro IRNA. Av- r[Oero, n npoUeroeopxrl 6t6p0rrloq ouprBo(vet npototi oXrlproume( o;totonotx6g 6eo;r6g preto[ri o;rtvo[6og r<or IRNA. [-r' out6 ro ].6yo, e(vot on(Oovo vo npoYporo- noefuor npoperoqopxrj 6t6p0ooq pr6oo orqv neptopl 6r6p0oo1g rou ev(6prou, enetbq 6ev undpXet Unxqvt- o;169 pe tov ono(o vo nopegno6((etor 4 6tdorooq tou :5."i : 0r luv0erooeg Xprlorponorouv U Arop0toon AorLp[t.rv yto vo Be],rLooouv Ti'lv AxoiSsir 731 npdodeon tRNA ADevutr(toon Ydpotr[etor ro o;uvooxutro-odevutrtx6 06pTrorl tRNA Itomd oUrvoEu +
EIK0NA 25.20 H ouyxpumoNLx4 0opr1 qg treuruhotRNA ouvOe- rdqg (LeuRS) npoodedey6vq mo tRNAk, oq dropdprpoorlqg pe- roperoqopx4g ene[epyoo(og (oprorepd) ror q dLopdprpoon Tnq olrLvooru)(oor1g (deErd). H neproXrl ene[epyooiog Ue BooU ruovo XpOUo reprorp6qeror orlgovrLrd ;rero[rj rov 600 dropoprpdroetov, evtir ro drpo-ono66mqg Tou IRNA perorp6nemr pero[u prog eme- rop6vlg etrxoerdoug 6rog6prptoo4q @pmepo) Kor Uroq drogoprpto- o4g d(xqv rpoupr6rog (6eELd). Avoruntir04re rordnv odefog ond J.J. Perona, el al., Biochemistry 46(2007):10419-10432. Copyright 2007, American Chemical Society. 0t rptoroyporpieg nopoXrrlprl0q- rov ond John J. Perona, University of California, Santa Barbara. pr1 ouyyevorig ol.uvoorul,o-o6evul,u<orj r<oOcbq nopeUe- ror on6 ro ouv0errx6 xdwpo mo r6wpo 6r6p0oor1g. Aw(Oero, 6rov dropeooLoBe( rdnoro 6v(u;.ro, q ow6po- on rnq rrpoperoeopu<r1g 6r6p0ooqg pdMov ouprBo(ver pSoo oro 5pro rou ouv0enro6 evepyoO r<6wpou. 25.12 To KorooroArrrd tRNA [leprdxouv MeroAAoyUCvo Avrrro6rr6vro, To Onoio AroBd(ouv N6o Ktrl6tr6vro * 'Evo rorqorotrnro IRNA runuo rp6per gro perotrtro[r1 0T0 ovl- rodrrdvro nou otrtro(er ro rodrxovro oro ono(o onovro. * '0rov ro vdo ovlroDtxovro owlororXel oe rtodrrovto Tspuol- opo0, evotogortilveror 6vo o;uvo[u ror 11 notrunenrLdLq otru- o[do enerreiveror ndpo ono ro roldxdvro reppologori. Auro odqyeL oe oouvOelrl roroorotrq oe pro 06o11 oouv0elxrlg gerotrtro[1q r1 oe ovoyvtool n6po ono evo rovovtxo rodrro- vro reppoTtopou. a Auouv0enrrl roroorotrl ougpo(ver 6rov ro IRNA ovoyvtirpi(er rlvo Drorpoperrxrj rodrr6vro ono ro ouvrlOroprlvo, onore 6vo opLvo[0 ovnro0rororol ono ronoro otrtr0. H onop6vo-ro4 peroMoypr6vorv IRNA unrlp[e 6vo on6 ro roXup6repo epyo).e(o yro rrlv ovd).uor1 r1g rr<ov6- rrlroq rou IRNA v' owrororXe( oro xcr:6rr6vrd rou oro mRNA Kor rov r<o0oprotrr6 rov enL6pdoe<rlv nou 6Xouv drogopetLrd rprrlproro tou IRNA orqv ovoyvroprorl Ko- drr<ov(o u-owrxodLr<ov(ou. To peroMoyprdvo IRNA onopovcovowor 1dpr1 mqv rr<ov6rqtd roug vo [enepvoUv lg enrmo.loerq rov Ue- toMd{eotv oe yov(6ro nou ro.l6rronoro[v no].unem(6to. Iq yevrxq yevertxq opoLoy(o, ;uo pret6Mo(rl nou pno- pe( vo unepvu<qoer lg enLmcooerg ;uoq dM4g prerdMo- lqg ro).ekor xoroorotrrrx{ (suppressor). Ie ouorr;;roto IRNA r<otooroLng, 11 nporoyevrig ;re- 732 KE0MA|0 25 0 leveruog Ktirdrroq rdtrl,o(r1 oMd(er 6vo rrrl6u<6vro oro mRNA, on6re ro no- Lunemrdx6 npoi6v dev e(vor nl*iov Leroupyu<6. H 6eu- repoyevf g roromoLtrx{ per6},}.o[r1 ;reroBd].].er ro owu<to6x6vro evSg IRNA, riore v' ovoyvrop((et ro pretoA- Loyprdvo ro:6x6vro ow( rou ou0ewxo0 rodu<ov(ou-ore Xou (i e[(oou r<o].d). To opuvo[ri flou evooUorc0veror onoroOrord rq Leroupy(o tou nolunemt6(ou. Or xcno- oroLe(g ovorp6powor <rlg oouvOerrxoi (nonsense suF pressors) rl 6uouv0enxof (missense suppressors) ovdloyo Ue rn qUon rnq op4Knq prerdMo[qg. Mro oouvOerrrrl gerdMo[r1 petorp6net 5vo xco6xe vro nou roOop(er 6vo opuvo[ri o' 6vo on6 ro rp(o xor6t- r6vro tepportopoti. I' 6vo rrirropo dyptou rUnou, pro r5roro oouv0erxrl gerdMo[4 ovoyvrop((eror p6vo on6 dvo nopdyowo oneLeu05ptr:oqq, o ono[og reppori(u tr1v npolreivoorivOeorl. Oar6oo, q 6erireprl xoromo.f,n- xli gerdMo[n oro owrKo6rx6vro rou IRNA, 64puoupye( 6vo ol.uvooxuLotRNA u<ov6 v' ovoyvrrlp((et ro ro:6rxo vro repporroproU. Evorotrrorrivowog 6vo o1.rlo[0, q xo- toorolt] on6 rr1 derirepq prerd).Lo[r1 enrrp6ner rr1 ouvSXroq t4g perdgpooqg nSpo ond 14 06oq qg oouvOerrrfg perdLl,o[4q. Awrl 11 v6o 6uvot6qrq rou UeroepofirroU oumrlprorog entrpSner rrl orjv0eorl ev6g nL4poug nolunemt6(ou, 6nog go(veror mnv EF ,(0NA 25.21. Av ro o;uvo([ nou evoroporri:veror pr6oro xo- romoLrig 6rogriper on6 ro ou0ewrrS optvo[r3 orrl ouyrer<ptpr6vq 05or1 rou nol,unemrS[ou dyprou r0nou, r'1 6poorqprdrrlro rou noi,unemr6(ou prnope( v' oM6[a. Or 6uouv0eru<59 perotrLd[erg oMd(ouv 5vq xodrKG vro nou ovrr.flpoooneUer rdnoro oprwo(ri oe rro6rx6uo nou owr.ilpoocrlneUer 6vo dtri.o oprwo(ri ro ono(o o6w+ re( vo Leroupyrloer orn 06on rou ouOewu<orl. C-unxcr, onoto6{note owmordoroorl qrwo[riog ouvrord pro 6r-r ouv0enxf perdMo(r1, oo16oo, oqv npd[4 ovrXyeferor p6vo ov otri.d(et rr; 6poorqpt6rrpo rou nolunemrdiou.) H perdLl.o[q rotoordMeror qn5 rrlv evotoprdrooq eire rou oplxori oprwo[6og efue xdnotou dMou optwo(6og nou onoro0rord rn treroupy(o rou nolunemr6(ou. H Ell(0NA 25.22 6eflyet 6tr q 6uouv0enrry xcrrooroNi pnope( vo enmeul0e( 6nog ror q oouvOerrruj, dqAodli petotrl.dooowoq ro owrKo6x6vto ev6g IRNA nou 1.te- togdpu 5vo ono6err6 ogrvo[ri 6ors v' owrorotle[ oro peroMoypr5vo xtodx6vro. Iuvendrg, 4 6uouv0ertxn xe tooroltj ouvisrotor oe otri.oyrl tou prlv[proroq rou Kq> 6txov(ou on6 6vo o;rwo[0 o' 6vo 6trtro. 25.1 3 Yndpxouv Aouv0errroi KorooroAelq yro KdO€ Ko6rr6vro Tepponopo0 * Ko0e runoq oouvOertrou rodxov(ou rorooretr),eror otto evo tRNA pe perotrtroypevo ovrtrodrr6vro. * 0proU6vo onovro roroorotrtro IRNA nepteXouv perotrtro(erc oe otrtro rprlporo rou pop(ou. Or oouvOerrxo( xorooroLe(g 6roxp(vowqr oe rperq Ko- Boorxcg evvorcg Boorxfg eworcq
Ayproq nlnoq: ro xodxd,vto UUG 6tq0d(erfl qnd rc Leu-tRNA AUG HH8 UM Aouv0errrd [er0uoypo: ro rodtrovto UAG repUo((et _-:- - Ai,picq ruiroq: ro rodtrovto GGA dtopo(eroL ono ro Gri-:i',: ,-.r* ILeu w UAG Arjn 880 i:^i. ,rd IGlv 13 nopoyoutoq W onetreu0epurons I ^radIArg Korumolttxn uetdllo{n: otrtrd(et ro ovrLxtodtxovto tou Tyr-tRNA AUC UAA Ell(0NA 25.21 0t petotrtrd{erg Xop(q vonUo Uropotlv v0 K0T0- orolOorjv ond 5vo IRNA Ue Uer0,tr0Yp6vo ovrtroldtrdvto, ro oro[o erooyer 6vo outvoEr] oto Uer0II0YU6vo xtootrovto II0pdYov- roq uro nporetvl oh6rhnpou p4rouq, onou ro opxtxti xordtroLno Leu owrroromoOqre on6 Tyr. rqyopieg, Uro yto r<d0e t6no x<o6tt<ov(ou repportopo6. H EIK0NA 25.23 neprypd<pet ttg rdt5rlteg optopSvtov on6 ro u g rotrrite po Xopoicn p toU5vo u g r<otooroLe(9. Or rotoorotre(q rou rlLer,cpou (amber suppressors) e(vor or onl.orjorepot. Iro E. col/, toutrdltorov 6[t IRNA 6Xouv ;retotrLoX0e( c0me v' ovoyvtr:p((ouv rco6tr6vto UAG. 'OLq tq xoroorqlru<6 IRNA rlLercpou nepdlouv ro owtrto6tr6vto CUAe, TIou ot rdOe nep(moon npo- 5pXeror on6 tov 6ypto ttino tr-r5oo oMoy4g prtog pr6vo Fdonq. H pretdMo[r1 gnope[ vo ewon((etot oe onot- odrlnote on6 ttg tpetg Bdoetq rou owtK<o6tt<ov(ou, 6ncog go[vetot oro supD, supE rot supF. Kd0e Kqro- orotrnr6 IRNA ovoyvtop((et pr6vo to rcodtx6vto UAG ov- r( yro ro npoqyoUprevo xto6tr6vto ({ ro6tr6vto)' T' oprwo(6o nou evoopor6vowot e(vot oep(w1, yLoutopri- vq 4 rupoo(vn, 6nlo6n tout6orlpo pe r' oprtvo(6o nou petotp6powot on6 t' ow(orotlo IRNA dyptou nlnou. Or rorooroleig qq 6Xpog (ochre suppressors) en(- oqq nponimouv on6 pretoMd[etq oro owtt<to6x6vto. Ifto yvooro( efvot ot supO rot supG, ot ono(ot evoogo- r6vouv rupoo(vr1 rl Luo[vr1 ow(orotXo npoq ro xto6tr6- vrq rnq 6Xpoq (UAA) xot tou r1lertpou (UAG). To Knrooioi.fi: ro AGA 0to9d(etot ond ro perotrtroyu6vo Gly{RNA EIK0NA 25.22 H dtoouv0ertxn Uerdhho[n npoygotonote[tot, orov ro ovnrodxdvto rou IRNA perdrtrdooerot 6ro 6ore vo owt- ororXel oe ]roOoq xodtr6vto. yeyov6g out6 e(vot ouprBot6 Ue rnv np6p^e$n qg un6- Oeonq tr1q ocrd0etog 6tt to UAA 6e prnope( v' ovoYvo- prore( pSvo rou. 'Evog r<otootol6og UGA SXer pro onp6oprevrl t6r6t1- to. flpo5plerot on6 to tRNATrp, oIId n ;rovo6rni pre- [eveuxdg rtinog IRNA Ayptog tunoq Koromol6og supD (su1) sr.ipE (su2) supF (su3) supC (su4) Ko6trdvro/ Avrtrodrxdvto/ owtxrodtrdvto xrodtxdvro Ser UCG Ser CUA UAG UAG AUC AGA UAA UCU A UUA UAG A UUA UAG A UCA UGG Lys Trp c UAU Tyr c UAU Tyr Gln Tyr Tyr CAG Gln CUA UAG CUA UAG A GsupG (suS) Lys supU (su7) Trp ElK0l,lA 25.23 To rorqorotrffio tRNAs xtop(q v6nuq 6nuLoup- youvror on6 perotrtro[etg oto ovrtrodxdvto. Auouv0erLxrl: ro AGA dtopo(etoL ono ro Arg{RNA 25.13 YndpXouv Aouv0euro[ Kotoototre(g yto Kd0e Kodtr6vto Tepuottouou 733 AUG
tdMo[n rou e[vor 11 owu<otdoroo4 A ow( G mq 06o4 24. Autrl rl oLloyrj owu<oOrord 5vo (erJyoq G-U mo m5Lelog D pr' 6vo (eUyog A-U, ou[dvowog dror t4 oro0ep6qto qg 5Lu<og. H oMql,ouXfo rou owu<<odr- rov(ou nopop5ver (6ro 6ncog Kcu. orov dypro rrJno, C- CAe. Iuvencbg, q perdMo[q oro or5],e1og D npdnet pe xdnoro tp6no vo preroBdMer q 6ropr6peool.l rou Bp61ou rou qwKodtxov(ou, ennpSnowog mo CCAe vo (euyoprilver pe ro UGA p' 6vo oouvq0toro ooroOdg (euydpopo qg C pre rrlv A. To r<oroorotrru<6 IRNA e[o- r<oLou0e( v' ovoyvrrlp(er ro oUvq0eg ro6x6vro UGG. Mro nopegqeprlg rordoroon nopqrrlpe[ror or4v ne- p(mtoo4 ev6g ouyrexprpr6vou euKopuoJlKorj IRNA. To rlnop rou Bodtori nepr6ler tivo tRNASer pe ro owtrtodr- r6vro ,CCA+. Or rov6veg ootdOerog npoBl,6nouv 6rr out6 ro IRNA 0o 6npene v' owrororxe( oro rco6u<6vto rnq rpumoedvqg UGG, oMd orrlv npoyponr6rrlro ov- rtororle( oro xor6rr6vro repponoUori UGA. Iuvendlg, oe ourj rlv rordoroo4 ro UGA prnope( vo roroor6Me- ror guou<d. H yevu<q orlpoo(o ourc0v rov noporrlpqoecov 6,yxer- ror orrlv enroqpovo4 6n 11 ovoyvrilproq xo:6xov(ou-ov- rrro6rxov(ou e(re mo IRNA dyprou rUnou e[te mo peroMoyprdvo IRNA 6e pnope( vo npoBLeq0e( nl4prrrg on6 ng oXeuxdg oMqlouX(eg ro:v rprnLerriv, evri oe pepx6g neprmtrloerg prnope( vo enqped(eror on6 dMo XopoKrn proru<d tou prop(ou. 25.14 Or KorooroAeiq Mnopel v' Avroytovi(ovTol Tnv Kovovlri Avdyvroon Tou Kd,6lKo * To roroorotrrtxo IRNA ovroyulvl(ovror ro IRNA oypLou runou nou nepteXouv ro [6ro ovurotdrrrlvro yto qv ovoyvoorl rou ovrlorotXou rcodLrovlou (r1 rodrrovLrrlv). e H onoretreopolrrl roroorotrl ervor enrptroBqg, enerdrl odrlyer oe ovoyvooq nepo ond ro xovovtro xto6rr6vro TeppoTropou. * To rodrrovlo UGA egqovi(er dLoppol ror 0ropo(eror eoqotr- g6vo ono ro TrptRNA pe ouXvrirqro l -3% YndpXer puo evdrocp5pouoo 6rogopd preto[ti qq ouv4- Ooug ovoyvcrtptorlg ev6g xrodLxov(ou on6 ro ow(oror- Xo oUrvooKULotRNA Kor rrlq r<ordoroorlg orrlv ono(o 1-uo perdMo(q enrpdner o' 6vo xoroorotrrrx6 IRNA v, ovoyv<op(oer 5vo v6o rot6rx6vro, Iro xrirropo dyptou r6nou, p6vo 6vo v6rlpro prnope( v' ono6o0e( o' dvo 6e- dop5vo rco6rr<6vLo, ro ono(o owrnpooconeUer efue 6vo ouyrexprprSvo opuvo[ti e(re 5vo o1pro rep;ronopro0. I, 6vo r[rropo nou <pdper Uro Koroorortxr1 prerdMo[r;, ro peroMoypr6vo rro6u<6vto pnope( eke vo 661etor ov- royovrop6 on6 ro roroorotrru<6 IRNA e(te vo 6topd- (eror pe ro oUvq0eg v6qprd rou. 734 KEOAAAI0 25 0 l-evelroq Kcirdtxog 'Evo oouv0etx6 rotosrotrtu<6 IRNA np5ner v' clwo- yr.uvrme( roug nopdyoweg oneLeu06pooqg or onofor ovoyvtop(ouv to xrodrx6vro (-o) repprorroproU. 'Evo 6u- ouv0errr6 roroorotrrrr6 IRNA npdner v' owoyrovrore( to IRNA Trou owrororloriv oro v5o r<ro6tr6vr6 rou. Ie rdOe nep(mcooq, o Bo0pr6g rou owqyovro;rori en4pe6- (er qv onoretreoporrr6rqto rrlq KqrofioLrlg: ouvenrig, 4 ono-releoportx6r4ro ev6g ouyrerprprdvou Korooro- L6o e[oprdror 61r pr6vo on6 rr; ouyySvero prero[( rou owrxco6rr<ov(ou rou Kor rou ro6rrov(ou-m61ou, oMd ror on6 rr1 ouyrSwpoorl rou oro rUnopo, 6ntrlg en(- onq Kor on6-ng nopop5rpoug or ono[eg 66nouv rrq ov- royrovrmrxSg owrdpdoerg repprotroprof 4 evootpdrtrl- onq. H onoreleoporu<6r1to Ue rnv ono(o droBd(etor 6vq r<o6x6vto enrlped(erot on6 rqv ew6nrorl rou. Iuvendlg q 6rcrooq rlg oouv0ettx4g r<otooroLrlg on6 6vo 6e6o- pr6vo IRNA norril.er nolrj ovdLoyq Ue ro ntro(oro rou r<o6u<ov(ou. Or enr6pdoerq rov yerovxriv Bdoeov rou m R NA orrlv ovoyvcil pro4 rardtrov(ou -owrrro6 rr<oviou 6ev efuor xo.trd rorqvoqrSg, orr6oo ro nLo(oro prnope( v' o).}.d(er rq ouyy6rqro pe rnv ono(q 5vo rco6rx6vro StoBd(eror on6 6,vo ouyr<erpq6vo IRNA rord ;to rd!r1 peyS0ouq xor nl.6ov. 'Evog oouv0etrr6g rotoorol6og onotrrovcbveror Xd- pq ornv xovdr4rd rou v' onowd oe 5vo preroMoyprdvo oouv0erix6 r<trr6rr6vro. H (6ro oMrlLoul(o, oo16oo, ou- vtord 6vq on6 ro rovovrrd orlporo tepportollof rou rurrdpou. Oetrrp4ru<d, ro peroLl.oy;r6vo IRNA nou Ko- toor6Mer rqv oouv0etxrl gerdtrio(r1 npSner vo ;rnopef vo r<otoor{i[l.er rov KovovtK6 repproropr6 oro 16}ro9 rd- 0e yovr6(ou ilou Xpnorponoe( ro ouyrerprprdvo r<o6rxo- vto. H EIK0NA 25.24 6efiver 6n ourrj r1 nopoploo4 rou repponoporl roroi,4yer orq oUv0eorl ev6g prorprjrepou no).unenrr6(ou p' enrnp6o0eq xopBo(wel,rr4 oLl.r1- Merdqpool dyprou rrjnou AijG 0 nopoyovroq onetreu06ptoorlg reppor[(er rr1 ouv0eorl oro xrodrxovro repgolopori Kotomo)r1 ntrdrrpou Li;l::, To roroorotrlxo tRNA 6LoBo(er ro rrrl6rxovlo UAG ror 11 nporTeivrl enerre[veror doq ro enopevo xrodrxovro repportogou ,::i i-'r;;' -ii: ,, ,t Q1 qgyy6t.rro( rorooroLe(q enronq dtopo(ouv ne- po ono rovovuo rotDLovro repponopou, ouv0erovrog nporrelveQ. ot onoleg e(voL porpurepeg ono erelveg rou oyptou runou. UAG UAA @ I UAA @llopriyovroq onetreu06prrlorlg UAG AI.JC "$Tvr ITyr loolrcg eworcg
AouX(o. To noLunent(6to 0o rotoffi[et orqv en6pevrl -p*i5.o teppottopori nou 0o Bpe0e( orq qdorl tou nAoto(ou ovdyvooqg' H ruX6v erctetop6vq r<otomotrri rou repUortogoti prnope( v' onoBe( entB)'oBr1q Yto ro xfrropo', nopdyo*og prorp0tepo nolunem6to pe oI- IoyUdvn LettouPY(o. 'b, *o.o*oLe(g ril'er<rpou ouvrlOcog e(vot oXettro onoteheopottro(, yevtrd xotd 10- 50% ovdl'oyo Ue ro oJ*nuo. Awri 11 onotetreogottx6trlto e(vot nt0ovrl enetOrl' ro r<rl6tt<6vto tou riLer<rpou Xpnotponotofwot oXenxd sn6vto Yto repporLoU6 qq petdqpoo4q oro E' "bli. R*(et-o, t<otoorotre(q cbXpoq onopovtirvowot 60' sKo,o. E(vot ndwo no}'ti Lty6tepo onorcLeogottro(' pe 6poorrlpt6trlro x6ro tou 10%. 'Otrq tq rUrropo Ue Ko- .o*oLtiq drXpoq 6Xouv pdtrl'ov cprcoXrl ovdmu(q' ye- yov6g nou uno6rll'tirver 6tt 11 rotootolrl t6oo tou UAA 3oo oo, tou UAG e(vot rotootpemtr<r1 yto to E' coli' ru' 0ov6v enet6rlto rto6u<6vto 6Xpqq UAA lpqot;ronotefuot oqy6tepo oq ro Kovovtr6 orlpro teppottotrlo0' T6Loq' ro UOR e(vor to Lty6tepo onotel'eopotu<6 on6 to rco6t- x6vto teptrtottopo0: oe rUrropo dyptou t6nou, StoBd(e- rot eogol.prdvo qn6 to TrptRNA pre ou;y6trlto 6o9 t<ot 1-3%. ir/oorqUtq, ro UGA Xprloqonote(tot ouly6tepo or,6 tqu tptntr6to rll'errpou UAG yto tov repgonog6 rcov BorcrqPtor<dlv Yovt6(tov. 'Evo duouv0eur6 rotoorotrtrr<6 IRNA nou ovttppo- ne( 5vo preroMoypr6vo rrodr6vto oe pto optop6vrl 06o11 pnopef uo ,,ooydyt, plo ovent0Uptqrrl petdMolrl o' 6vo 6)tlo yov(dro.'Evoq rotoototr6og 6top0drvet gto getdl'- Ao(r1 owu<oOtorciwog 6vo oprwo[rj pr'dvo dMo orn Ue- -oi.LoVUgrn 05or1. Ee dLLeq 06oetg, toor6oo, 11 (6to owrotdmoorl owtxo0ror6 to oprtvo[r3 dyptou ttinou U' e* v6o ogwo[6. H pretoBo].rl Unopgi v' ovoore0ret rqv xovovtrq'treroupy(o tou notrunemtS(ou'- Aw6 06tet ro x0rropo oe 6il.q1-rpo: np6net vo rotooteilet 6vo opt- olrevo petof.loyp6vo t<todx6vto 1op(g 6gto9 v' oMd[et nohti to t<ovovtx6 v6qprd rou ot 6Meq 05oetq' Euve- nOS, n onouo(o roXupcirv 6uouv0ertrc0v roroorotrStov l,t" pti vo e(r1yr1Oei on6 rg rorootpemx6q entd pdoetq nou'0o nporotrorioe Uro YevKeuUdvq r<ot onotetreopott- r{ owtrordorooq oPwo(6tov. Mro pretdLtr'o(q nou 6rlptoupye( dvo- KffiomqtrttK6 IRNA gnope( uo 5Xe, 60o ouv5neteg' l-lprinov, enttpdnet sro IRNA v' ovoyvcop((et 6vo v6o xto6tr6vto' Aerjtepov' lreptx6q Qop6q nopeprno6(er to IRNA v' ovoyvop(oet ro'ro6ir6vto orq ono(o npoqyo0gevrog onowo[oe' E(- ,o, oqpo*,*6 6tr 6tr'or ot onotetreoponxo( xotooroLe(g nie'..pou npo6pXowot on6 petdMo[n eY6S owtyp6- qou p,oq nitou&qorooq opr66oq tnrul :-e ow6g ttg t"p,*Oot,S, -o ^jr.opo nepdlet noMg IRNA xovd v' onownoouu oro rodu<6vro nou onorel'e( tov ou0ewtr6 m6Xo tou IRNA dyptou t[nou' 'Etot, 11 pretdtrtro[r1 6ev rotopye( rqv ovoyvdrptorl rov ndrottitv xto6tt<ov(trlv' to onofo L(onolou0o0v vo xo}'Umowot enoprdrq on6 to tnftrn.ns opd6og. Itrlv oouvdOtotq xotdorootl xotd rn, onoto g6vo 5vo IRNA onowd o' 5vo ouyxer<ptpt6vo xto6tt<6vto, r<dOe pretdl'tro[r1 nou nopeprno6((et rnv on6rrrrloq e(vot 0 ovorlq6 Poq. H rotomotrn ouvn0oq o[ro]'oye(tot oro ntro(oto 1.uog petdMo[r1g nou oMd(et qv ovdyvooq evoq Ktltdt *ouiou. Oo16oo, oe oprop5veg xotoordoeq, 6vo Krru6t- r6vto reppottoprori 6toB6(etot tr:g o1'tloff, nopot Fe Xopu'1l,6tepr1 oqy6tqto. To npd:to yvtr:or6 nop66etypo ovor<oLrl<p0r1Ke oro yov(6to Yto rnv nporotvq rou nepF Blqgotog tou RNA qdyou QB. O oppottopr6g Loqro- VOrw ocol-tott6(ov QB npouno06tet roromolrl tou xo6trov(ou rep;ronopro0 oro drpo owoU tou yovt6(ou Ue XoUnLn orlr6tt'1-o, tilore vq nopoX0e( ;rto prtxpri no- boinio'uopiov nptotetvnq ue uto r<opBofuretrrxq en5- "roon buotoorrrd, out6 to xto6tr6vto tepgottoprori eprpov((et Stoppod, O l6yog e(vot 6tt rc TrptRNA ovo- yvop((et to t<co6x6vto pre Xo;r1trr1 ouly6trlto' ' Au&yurorl n6po on6 to t<co6tr6vto teppottoproU ou;rBoiver Kot orouq euxopuciteg, -6nou Xprlotpronoe(- roiouyy6tepo qn6 RNA to0q. Aw6 (ocog nepil'o;rBdvet oo.o*tM tov UAG/UAA on6 to Tyr-tRNA, GlntRNA 11 leu-tRNn ri rotomotrrl tou UGA on6 to Trp{RNA q ArgtRNA. H 5r<roor1 rqq preptr<r1q rotomolf,q unoYo- pt-ftaol, on6 to *nlo(oro' tou xtrl6xov(ou' 25.15 Io PlBootol..ldrto Ennped(et rnv ArpiBero rnq M€TdQPoonq o H do[rl tou 165 rRNA ottg 06oetg P xot A rou ptpootopor[ou ennped(et Uv orp(Feto tnq UetdQpo0nq' H ouyy6trpo L60oug xord rrlv evoopdtooq rov opt- vofStov oe nolunem(dto np6net vo 6torrlpefuot XoUn- In, ornv xtr(pror<o tou ev5g )rdOouq ovd 10'000 ouwoeeir, c6ote vo 6too<potrtore( 6rt ot lemoupytr6q r6r6rqteg tcov rto6tronototipevtov nolunemt6(tov 6ev oUAEoru r<otd tp6no entploBri yo to rrirropo' Ad0r1 Ut"pti vo ou;rBo6v oe rp(o yevt16 or66tq qq prer6- epqonq (F)rdne Er6vo 24.8 mnv Ev6tqta 24'3): -'. n ,ibd-,on ev6q IRNA pre to r<otdM4o opwo(0 e(- vot oorpciq xp(oq-rr1' Aut6 e(vqt Spyo qq optvooKu- r,o-tRliA ouvOerdoqq nou XopoKrrlp((etor on6 ouXv6ulto L60oug 1:105 6coq 1 :107 o;ttvooruLr6- oetg (BL6ne Ev6qteq 25.9 5trrg 25'11)' . n UeioQopd tou otoototi o1'twoorutrro:prSvou IRNA o-ri, p,B6oiru6tto, 5pyo torv nopoy6wcov 6vop(r1q f t",U,inruonq, nop6let 6vo prqXovto;r6.nou enou[6- vet'rq ouvo)ttrr1 enil'errtr6rqto' Entnl'6ov, ot ouYKe- np,giuo, nopdyovteg Boq0orlv otq dtepyoo(o oyrupo6eofoq rou ogtvoorul'otRNA o"rtq ptBootollt- x5q 06oetq P rot A. . Kpiorun en(oqq e(vot q et6x6t1to qq ovoyvrirptorlq rto6u<ov(o u -owtt<<l6tt<ov(o u. 11 op6tt o r oro0ep6 g np6o6eoqq norr[trouv ovdtroyo pre to (e0yog t<to6t- xov(ou-owrxrodu<ov(ou, 11 ev6oyev4q et6x6tqto no u oXeil(etot pe ro crxnUortopr6 gtoq ouyyevorlg dvowt gr1 ouyyevotq drLrl)rouliog tpttilv (euycirv B6oe<rlv 25.15 To PtBootoudtto Ennped(et qv Arp(peto uq MetdQpoons 735
(nep(nou 101 5<og 102) e(vot nol,ri 1o;r4Lri drore vo e(oo<poL((et ouyy6qro trdOoug < 10-s. An5 xop6 erle 6toruno0e[ r; un60eoq dl o Borrrl- pror6g nopdyowoq enrprqr<uvoqg EF-Tu npoo6dveror oro RNA ove[dpqro on6 qv oM4],ou{o, r<o069 npd_ ner vq Uerogdper oro prBooopdtro 6Lo t, ol.rwooruLo- IRNA (err6q rou evopxrrlprou IRNA). O EF-Tu ovoyvrop((er r5oo ro opwo[u<6 rUnpo rou 6eopo0 opr_ voor<utro{RNA 6oo ror ro o6pro rou IRNA, 6nou rup[<og npoodSveror fio ooKxopogoogoprxd oreler6 tou metr51oug-ono65rrq Kor rou meLdXoug T. MeLdreg orrg ono(eg perp4Ory<e 11 ouyy6vero np6o6eoqg rou EF-Tu oe otom6 xor Ld0og ogrvoorul.uo;r6vo IRNA 66er[ov 6tr 11 roX6g rqg np6o6eo4g oro opuvo[0 e(vor owrorp6<prrlg ovdLoy4 Ue rrlv rq40 np6o6eoqg oro oc0_ Uo rou IBNA. Me dMo L6yro, oo0evrirg npoo6e6eg6vo otruvo{6o eoreponororiwor oroord pre ogtXrd npoo6e- 6ep6vo otil.roro IRNA, evc0 orprXrd npoo6e6e;r6vo opr- vo[5o eoreponoroUwq oroqrd pre ooOevrilg npoo6e6e- prdvo oril.toro IRNA. Iuven6g, rq ooord oxul,uo;-uivo ollvoo<ulo-tRNA npoo66vouv EF-Tu pre opr<erd nop6_ 1.roro ouyydvero. H ex).eru<6tr1ro mr1 ouvoLxrl perd_ qpoon e[oorpo].i(eror ener64 11 ]d0og or<ul,(oor; ev6g oo0evrig npoodedeprdvou ogrvo[6og o, 6vo oo0evc6g npoo6e6ep6vo oc0pro IRNA nopdyer 6vo pr4 ouyyev6q ol.uvoor<ulo-tRNA, ro ono(o oMrll,enrdpd noLrj ovoro_ rel,eopo-nrd Ue rov EF-Tu. En(onq, dvo )td0og orullco- pr6vo ol.uvoor<ul,o-tRNA nou npoo66veror nto orptXrd orov EF-Tu pnope( v' onorLerore(, enerd4 e(vot 6uoxo_ l,6repo v' oneLeu0ep6oer ro oplo[x6 rord].orno rord qv oyxupo6eofo rou mo p$oocrlprdrro. 'E1er Bpe0e( 6n geroMd[elq orov EF-Tu ;rnope( vo roroorerl,ouv Ld0r1 preror6nronq rou nLoro(ou ovdyvcrl- oqg (BL6ne Ev6trlro 25.16). Aw6 unodqLdlver 6rr o EF- Tu 6e <p6per onLcilg ro oUrvooKULo{RNA orq 0dor1 A, oLid en(or1g epnldreror orrlv ronoOdrrlon rou eroepX6- Uevou oprvoor<ul,otRNA oe oX6or1 pe ro nemr6uLo-tR_ NA orrl 05or1 P. l-lopopro(og, preroLld[erq orov nopdyowo 6vop[qg elF2 r4g (ripqg ennp6nouv rqv 6vop[q qg gerdgpoorlq o' 6vq Kodrx6vro dvop(r1g, nou peroMdooeror qn6 AUG oe UUG. Aur6 uno6ql,ri- vet 6rr o elF2 Boq0d firlv oyKupodeo(o rou tRNA,r,r"t or4 06o11 P. H 6r6p0oor1 dorrp(rov (proofreading) ndvco oro pt- Booco;rduo, d:me vo evtoXu0e( n eyyevnq 1o;r4Lr1 er6r- r6rqto nou enrruyxdvetor ;r6vo on6 ro (euydpopo Bdoeov gero(ri xto6xov(ou-owrr<rr6u<ov(ou, npounoOri- rer enrnp6o0ereg o).L4l,enr6pdoeLg nou nop6lowor qn5 to ronrr6 neptBdMov qg 30S unopovddog, Kord rr1 tremoupy[o rou coq Srop0orjg 6orrp(trlv, ro prBoorrr- prdno evroX6er rrlv 4nro eyyevn enil,ercnr6rpo rou (eu- yop<ill.rorog rrrlv rprvouxl,eorr6(cov r<ord 1000 gopdq (EtK0NA 25.25). H enil.oy4 rou ogrvooKuLotRNA on6 ro prBooro;rd_ no ouprBo[ver oe notrtrd orddrq r<o16 pujrog r4g o6orj p5oro rqg ono(og ro rprroroy6g oUprn).or<o EF- Tu:GTP:o;uvoorulo-tRNA nou oxrlpor((eror prerd rqv ol.rwooxul(oo4 pretorpdper ro oprvooKuLotRNA orn pr- 736 |GOA Al0 25 0 l-everrdg Kdr6rrog Kd00 IRNA gnope( vo ero6tr0er oq 06o11 A To oooTo IRNA otr),r1trenr6po pe ro rRNA EIK0NA 25.25 Ko0e oprvooxutrotRNA pnope( vo rono0eqOei oq 06o11 A (p6otrt rou EF-Tu), otrtro p6vo ere(vo nou (euyopdrveL ge ro ovlrodrxrlvto, pnope( vo drlproupyrloer oro0eponoLrlnx6g enog6g pe ro rRNA. Xoprg oureg ng enoq6g, ro ogrvoorulo-tR- NA 0roX6eror 6[o onri rn 06on A. Booropu<{ 05oq A. llpc[rov, oX4pror((eror dvo prd].].ov ooro0Sg oplu<6 orltrrnlor<o np5o6eo4g pre ro prBooro- prdrro. Irq ouv6Xero, undpXet 6vo Bqpo ovoyvc0prorlg rou xrodrrov(ou, xord ro ono(o to op1tr6 o0pnLoro enovo6rqtdooeror enrp6nowoq ro (euydpopo rro6r- rov(ou-owxco6rxov[ou orq 06oq A. Ou;r40e(re dtr 4 6r- nLovrl 06oq P <prLo[eve( ro nenrt6utro{RNA (BA6ne KegdLoro 24, MeruEpoory). T6oo ro opXtx6 Bqpro np6o6eoqg 6oo ro enorSLou0o B4po ovoyv6ptoqg rou ro6rrov[ou e(vor owrarpemd, To ].d0og (euyop<o- ;r6vo ol.rwoor<uLotRNA prnops( v' onoBlq0o[v oe ourd rq 0166ro prSool ev5g ouv6uoollorj ou[qtrr6v4g rolrirrl- toq 6rdmooqg ro/r1 preropr6vrlg roXriqrog oriv6eoqg tov roro(euyoptoprSvtov outrrnl,6rov. Merd qv ovoyvcbprorl xo6txov(ou-owtrro6u<ov(ou, pro np6o0et4 peroBol,4 6ropr6prp<oor1g nupo6ore[ rrlv udp6truoq rou GTP. Arol,ou0e( q onel,eu06poon go- ogopxori on6 tov EF-Tu nou npoo66ver GDp. Aurl 11 oneleu0d,proorl nupo6ore( puo 6M4 ercero;r6w1 oMo- y4 6rotrr6pgoonq Ue rqv ono(o ro oUprn).oro EF- 'Evo trdOog tRNA dtol6erot errog
Tu:GDP 6r(morot on6 ro oripnLor<o opuvoor<ul,o- tR- NA:ptBooogot(ou. M6vo prerd 14 dtdorooq rou EF-Tu outrrBo(vouv ot opronxSg ovo6rord[erg nou oXet((owor Ue rqv oyrupodeo(o rrlq qUrvooKu).u<1g pi(og orrl 06o4 P qg 50S unopovddog, r<o0rr1g ror q enox6l,ou04 owG dpooq peroqopdg rou nemt6(ou. Ercr6g on6 enrLoyrl mo nprirpro orddro np6o6eo4g, onoBoLrj rou LdOoq (euyopro;r6vou ogrvooKULo-tRNA prnopef enfoqg vo npoyporonorqOe( pretd ro B{po qg udp6},uorlq rou GTP. E66 onopoln ouprBo(ver enerdri q ro16111ro tr1g oprorxrlg oMoyrlg 6rop6pgrooqg e(vor noLrj Bpode(o orqv nep(mu:or1 ev6q LdOog or<utrr<oprdvou ougnL5rou. Iuvenc0g, q ouvo).x4 er6x6rrlro enou(dveror enerd4 ro IRNA npdner vo 6r5l0er on6 60o B4proro enrLoyqg npo- toU oXrlportore( o nemr6rr6g 6eqr6g. H orp(Bero rou (euyopdrporoq ro6trov(ou-owu<ro- 6xov(ou m4 06o11A drotqpe(ror pe mevq nopor<o),o0- 0r1oq rov orepeoxrlUrrrriv ror 4l.errpoororrrriv t6tottirrrlv tou rprvourcl.eorr6(ou. Tperq ouwrlprlpr6veg Bdoerq rou 165 prBoor,tprroU RNA (A1492, 41493 ror G530) oMq),enrdporiv orevd pre rrlv el,dooovo oULoxo rqg 6lrroq ro6u<ov(ou-owrrodu<ov(ou oro 6rio nprilro (e[yq Bdoeov ror e(vor u<ov5g v' owr].ogBdvowor pe orp[Bero rrlv nopouo(o r<ovovrr<drv (euyri:v Watson- Crick oe out6g ng 06oetg. Iqv rp(rr1 (ooroOri) 05orl gnope[ vo rpr].o[evr10orlv oprogSvo Url KovovrKd (eUyq, enet6q ro prBoocopux6 RNA 6ev nopor<ol.ou0e( ro (eu- y6popro 16oo mevd. Te).Lrd, o r<0prog L6yog nou ono- BdMeror ro L60og or<ul.urtprdvo IRNA nprv r1 prerd ro Fnpo rnq udp6l.uorlg rou GTP e(vor 6n 6ev owono- xp[veror n].{pog mtg onopo(rrlreg npo6toypoqdg ro- rd t4 l,enrogepr] enr0ec6pqorl on6 ro prBooropdno ornv 6,Lu<o r<o6u<ov[ou-owu<ro6rxov(ou { xot oe dMeq 06oerg. llp6orporo, ovoxol,ri<p0r1re Svog enrnp6o0etog pr4- lovro;r6g nou ouprBdMer qrrlv er6r6r4ro qg prerdrppo- orlq: ro prBoorrrprdno efvor rrov5 vo 6e[dyer 6].ey1o nor6rqrog trrerd rr1 oJv0eoq rou nemrdrxori 6eqroU. Ie otrr6 ro pr11ovtopr6, 11 o0v0eoq ev6g nemr6rro0 6e- opoU nou npoxUmet on6 6vo KoKororprooUSvo ol.rwoo- xulo- IRNA orq 05or1 A,odrlye( oe yevu<6repq onrilero a6x6rrpog orrl 06oq A. Me q oerpd rou, ow6 o6qye( oe npcbtprq InEn rnq prerdgpooqq. To prBootopdtro ovoyvrr:p((er 1.d04 gerd rq oUv0eoq nemrdxd:v 6eopuiv enrOe<opcirwog rqv orpgri ouprnLq- porporrx6trlto qg 6Lrxog xro6rr<ov(ou-owrr<o6rrov(ou orq eSon P. Iuv6nero rqq ).dOog evoopdtoo4g e[vot 11 oulq pdvq rrov6rrlro tov nopoy6wrov oneleu Odprooq g vo npoodrivowor orn 05oq A riore vo npoxoLoriv np6- opo reppouopr6, or5pr1 rr 6rov 6ev undpXer xdnoro xrodu<6vro repprouol.roti. Enrnldov, ou[dver 11 ouXy6q- ro Ad0oq ro6u<ono(4oqg or4 drnLovrl 05oq A. H eno- x6Aou0r; 6rd6oor1 rtov ).o0rilv tefurd o64ye( oe np6opo rlepgmropr6. To r5orog ulg ;rerdgpoorlg, 6nog unoLoy((eror on6 rov oprOg5 6eoprcbv uUnlnq ev6pyerog nou np5- ntt vo u6poi.u0orlv, oogdrg ou[dver on6 trg 6tepyo- oteg 6t6p0o:o4g 6orrp(ov. To np6o0ero evepyeror6 x6mog e[oprdror on6 to orddto mo ono[o onoBd].- etot ro ).dOog or<ulurtp6vo IRNA To xorcq .3, l: i- t((eror Ue rrlv onoBoLrl nptv on6 rrlv udpo,r.r;- ::, GTP orpopd ;r6vo oqv nopoyroyq tro0oq oxuALoo;rc,3, IRNA ond qv IRNA ouv0erdo4. Om6oo, ov ro GTP u6polu0e( nprv on6 q drdcrrooq rou KoKoroLprooge- vou oprvooKuotRNA, ro evepyerqK6 x6moq 0o efvoL preyolUrepo. Aorpoltrrg, to prSyroro r6orog ouvdderoL pe rov np6orpo repprortopr6 qg prerdgpoorlq nou ono- 6(6et prr; Lenoupyrr<6 npoi6v xord tov SLeyXo noLdq- rog perd rn perogopd tou nentr6(ou. Irr;v npoKer- pr6vq nep(moror;, np6ner vo r<oroBLrl0e( oAdr<L4po ro evepyeror<6 ripr4pro nou oXet((eror Ue rrl oUv0eorl rou nolunenrrd(ou drllg ro oqpre[o qg npdotprlg one].eu- 05poo4g. 25.1 6 Merqr6nron Tou nAoroiou Avdyvoonq Mnopei vo IuUBri oe ONo0np6q AAAnAouxieq * To ntroloro ovoyvoorlg enripeo(erot ond qv otrtr1trouXro rou mRNA rol ro pgooogrr6 nepgotrtrov. * 0t otrto0t'jp6q otrtrqtrouXieg enup6nouv o' evo IRNA v0 UrTo- ron[(eror roro pro pool oqorou (euyopooet pe ro ovrtrotdr- x6vro rou, peropotrtrovrog erot'ro ntro(oro ovoyvooqg. * H Uerorppool opropdvrrlv yovLdlrov e[oproror on6 rrlv rorlrrl npoyp0pUoTr0Uevq geroronrol rou n),oro[ou ovoyvtoolg. To ou;rBdwo enovoxo6rxonoi4o4g (recoding) ou- vrlOtog nepr).opBdvouv oL).oy5g oro v6rlpro ev6g ;r6vo xto6u<ov(ou. IXenxd nopoderyproro e(vor -ro gorv6pe- vo rnq IRNA xorooroXrlq @vorrlta 25.12) Kqr n oUor- onolu<4 tponono(4o4 ev6g opuvooxuAo-tRNA (Evrir4- ra 25.8). Oor6oo, rperg dMor rUnot enovoro:6rxono(- qorlg enr<p5pouv nro oLroupevtrdg preroBoLdq oro no- poy6pevo noLunentr6rr6 npoi6v. l-lp6rerroL yro rrl geror6ntoq rou n).oro[ou ovdyv<oor1g (frameshifting) (e[erd(eror oe out4 rqv Ev6r4ro), r4v nopdKopqn (bypassing) xor rr1 Xpqoq 6rjo mRNA yro rrl o0v0eorl ev6g nounemr6(ou (r<or or 6Uo e[erd(owor orrlv Evd- qra 25.17). H ;leror6nroq rou nLoro(ou ovdyvooqg oXer((eror pe erdu<d tRNA oe 6[o nepLordoerg: . OproUSvor preto).).oyprdvor IRNA r<otomoLe[g ovo- yvrop[(ouv 5vo "r<ro6rr6vroD reoodpov Bdoeorv ow( rtov ouvriOrov rprcbv Bdoeotv. o OpLou6veq ,,o).to0rlp6S" oMrlLoul(eg entrp6nouv o' dvo IRNA vo pietortvlOe( ndvo mo mRNA orrl 05or1 A r<ord pLo Bdoq e(re oe 5'e(re oe 3' xotefOuvor1. MeroL).d(erg peror6ntorlg tou n).oLo(ou ovdyvo-l- or;g nporUnrouv on6 ).dOog ovdyvurorl rou r<to6trov[- ou rou mRNA. Avr( vo 6roBd(er pro tprn).5ro, ro ptBoooprdrto 6toBd(er e(re 6vo woudro e(re svo Kou- 3i.iC Meroromorl Tou lltroro[ou Avoyvoorlg Mnopei vo Iupper oe 0trr00lp6q AtrtrqtrouXreg 7A7 Boorxeg 6woreg
oprdro vouKtreorrd(ov. Ie rd0e nep(moto4, q ouv51t- on rnq ovdyvrr:or1q rprnLer6v perd to ouyrerpt;l5vo ouprBdv ono6i6er 6vo notrunent(6to er<r6g droto[ou. Mro peror6nron rou noro(ou ovdyvoorlg pnope( vo xoromotre( on6 5vo IRNA rxqv6 vo 6roBd(er 5vo ro6t- xov(o 6rio 11 reoodprov B6oecov. Ir4v nepintoorl rov r<codu<ov(olv reoodpov Bdoeov, to IRNA 6to05tet 6vo exrerog5vo 0p6Xo owrrodrr<ov(ou, nou onotetre(rqt on6 or,mdl voux.treor(6ro ow( tov rovovtr6v entd. l-to nopdderygo, 1.uo G e(vor 6uvor6 vo etoXoprioet oe Uto oerpd opxerc0v ouvel6prevov Bdoecov G. O Korooro- ).6o9 qq prerot5ntorlg rou nloro(ou ovdyvtooqg e(vot 6vo IRNAGly ro ono(o nepr6let pro np6o0etrl Fdon mo Bp61o rou owrrordrrov(ou xor gerorp6net ro ov- rtr<odrr6vro on5 rq ouvqO4 rprnl,dto CCCe oto r<ouopr6ro CCCC+-. To r<oroorotrrrx6 IRNA ovoyvo- p((er 5vo "Ko6rK6vro" reoodpov Bdoeov. Opropr6vot roroorotre[g perot6nroqg rou n],oro[ou ovdyvoorlg ovoyvcop[(ouv ndvo on6 5vo (KodtK6vtoD reoodpruv Bdoeolv. l-to nopd6etypto, 6vog Bor<tqpto- 169 roromol6oq tRNALv'owrororle[ oe AAAA n AAAU ow( rou ouvq0oug rodu<ov(ou MA. 'Evoq dtr).og r<o- roorol5og droBd(et onotoS4nore "Ko6tK6vto" teood- ptov Bdoecov pe ACC otrq rperq nprirreg 06oetg: 4 en6pevrl Bdo4 e[vor o6rdEoprl. Ie owdg ttg neptntri- oerq, or evotrtror<trr5q Bdoerg nou e(vot onodertdq orrlv r6topr4 0do4 rou 1loxpfrepou (Ko6rKov(ou" 6e oler[(owor Ue rouq ouvrlOetq rov6veg rrlg oordOetog. To xqroorotrrrx6 IRNA gdMov ovoyvcop((et 6vo ro6t- r6vro 3 Bdoeov, oMd yro xdnoto dl).o L6yo -nt0ov6- repo l.6yrrl orepeoxnlJtrrlg nope;rn66ror1q (steric hindrance)- 11 drnhovrl Bdoq ono<ppdooetot. Aur6 npor<oLe( nop6l,etr.l.rr1 prog Bdorlg npototi to en6pevo IRNA Bpet 5vo rodrx6vro. Koroordoerq orrg ono(eg q pretot6ntorl tou nloto(ou ovdyvrrlorlq e(vor <p u orotroyrr6 ou ;rBdv nopor; po tiwot oe tpdyoug xot dLtroug rorig. TSroro oupBdwo (otog enrl- pe6(ouv q ouv6Xtorl r1 rov reppottqr6 qg pretdqpo- onq Kor orpeilowor orrg eyyeve(g r6r6qreg tou mRNA. Itoug petpoio[q, 4 perdgpoorlrou npciltou yovt6(- ou reppror((erqt qn6 dvo oouv0ettr6 rtodtr6vto ornv (6ro <pdoq ge ro nl,o(oro ovdyvoorlg. To derircpo yovi 6ro Bp(or<etor oe 6rorpope'rx6 ntro(oto ovdyvtr:oqg rot (oe opropr6voug ro0q) Uero<ppd(eror on6 gto perot6nt- oq nloro(ou ovdyvtoo4g nou entrp6net pto evotrtroyrl oro 6e[repo nAofoto ovdyvcooqq rt 6ro nopordpmet ro rcr6rx6vto repgortopo0 (FMne tlK0NA 26.28xt enionq lr1v Ev6t4ta 17.12). H onoreLeoporr6tlto qg preror5- nronq rou nl.qro(ou ovdyvcoorlg e(vot XogqLrl, tuntrd -5"/o. H Xopnfui onoteleotrrotx6t4to 51er oqproo(o yto t4 BroLoy[o rou rori, xoOc0g n o0[non rqg onoretreqto- ttx6tlrog prnope( v' onoBe( entBLoBrlq. H EIK0HA 25.27 nopouord(et qv nop6proto rordoroo4 rou orotle(ou Ty rnq (UUnS, orrlv ono(o to t<to6tr6vto teptrrottol.toti rou tya npdner vo TropoKopq0e( on6 pto petor6nto4 rou nloro(ou ovdyvcooqq <iore vo 6toBoore[ to enor6- Lou0o yov(6ro fyb. T5roreg roroordoerg unoypogp(ouv rr1 orlpowtxrl Meroronrorl rou ntrotofou ovoyvoorlg -1 o{o perpoid HIV n/ NNNNUUUUU IJAG GNNNNNNNN Teleuro[o xodrxovro nou 6Lopd(etor mo opXtxo ntroloto ovoyvcrloqq Ilptino xrodrrovro nou 6r090(sr0r oro v6o ntroloro qvoyvoolq Avdyvtoorl Xcoprg peroronrol rou nloto[ou ovoyvooqg NNNNUUUUU UAG GN N N NNNN N Avdyvoon gerd rr1 gerorontoq rou ntroto(ou ovtiyvtooqg NNNNUUUUU GNNNNNNNN 'Evo IRNA, nou otrroOolver roro ;.tlo Bool oro (eu- yopto;.ro pre 6vo rordtrovro, npoxo),e[ ;lerordnrol rou n],oLoio- ovoyvoorlg 11 onolo rorooretrtrer Tn InEl H onoretreoportroq,: e(vot-5%. oppl 6rL ro onovro (oMd npoBL6Qtpo) ouptBdwo "Lov- Ooop5vr1q ovdyvtooqg" e(vot 5vo ovoyt<o[o Bqpo rqc EvotrloxrtroI tp6nor [etdqpqonq ono6i6ouv Tya 11 Tya-Tyb llporeivn Tya 5fufofofufufo) EvopIn An(n tltl - I tvterordnrol rou ] 'Evop[r1 nlqro(ou ovdyvtoolg AnEn Xroprg ArgtRNA, ro To Gly- IRNA lloporoUQ1 ougpotvet orov to ptpootr.tporto pero- ruve(ror roro plrog rou mRNA, 6ro1 6ore ro nenrtdutrotRNA qc Oeor1g P onetreuOeptlverol ono ro (euyoptol.to pe ro raldtxovtc rou xr 6netto (euyoptlver ue ronoro oLtro rodlrovto oe pteyotr0re- p1 onooroorl. Xrpopxrl rpoTelvn Tya-Tyb 738 KE0AM|0 25 0 levertrdg KtitdLxog llotrunenrldro -Leu
quoro}.oyrKriq Uerdepoonq. Aw6 role(tor npoypqppo- nop6vrl peror6ntoq rou ntroloiou ov6yvtooqq (pro- grammed frameshifting). IupBo(ver oe ouyKeKprpSvsg neprol6g pre ouXv6qro 100-1000 eop6q peyoL0teprl on6 rq ouly6rqro Ld0oug oe Un npoypo;rporoprdveg neproX5g (-3 x 10-s ovd r<todrr6vro). Aur6g o rinog peror6nrorlg tou nLoro(ou ovdyvto- orlg epgov((er dtio rowd Xoporcrqptonxd: o Mto "oAro0lpn" otrtrntrouX(o ennp6ner o' rivo o1-u- voorui,otRNA vo (euyopciloer pe ro rto6rr6vr6 rou Kr Snetrq vo UeroKrvrlee[ +1 rl -1 Bdo4 rime vo (euyopdloer Ue Urq enu<olunr6prevq rptnldto, en(- oqq txovrl vo (euyopdloer Ue ro qwtro6rK6vr6 rou. o To prpootrrprdrto xoOuorepe( orrlv nepLoXrl rnq pe- ror6nrorlg rou nLoro(ou ovdyvoo4g oqqvowog Xp6vo oro ogrvoor<ulo-tRNA yro v' ovo6rotd(er ro (euydpogd rou. H om[o qg r<o0uor6pr1oqq ;rnope[ vo e(vor 6vo 6rnLov6 rol6rx6vro nou onotre( 6vo ondvro ol.uvooruLo-tRNA, 5vo ro6rx5vro reppon- opoU nou ovoyvrop((erot opyd on6 rov ow(ororlo nopdyowo oneLeuO6pooqg rl 6vo 6optr6 epn66ro sro mRNA (n.1. 5vog "rpeu6or6pnoq,, Uro r6ro(repr1 dropr6p<pcoorl rou RNA) ilou nopoKol,fer ro prBooto- prdrto. H o).(o0r1o4 ouv(qrqror oe preror<(vqo4 efue npog 14 pro efue npog rrlv dMrl r<ore60uvor1: peror6ntorl -1 npo- rco.l,efuot 6rov ro IRNA perorrve[rot npog ro nioo, ev6 peror5nrorl + 1 6rqv ro IRNA getorrve[ror npoq ro pnpog. Ie rd0e nep(nroor1, to onor5Leo;.ro e(vor vo ercreOe[ gro tprn].6ro ercr6g gdorlg m4 0rio4 A yro to en6pevo o;.rwoor<ulo-tRNA. H gerot6nroq tou nloro(ou ovdyvoo4g npoyUqronore(ror nprv on6 r4 orivOeorl ttov nemt6xc0v 6eopr<irv. Iro ouq06orepo rUno, nou nupo- dorefuor on6 pro o).ro0qpri oMrlLouX(o oe ouv6uoop5 pe [ro ro0o6u<r1 goupr<fto oro mRNA, or neprBdMou- oeg otrtr4).ou1ieg enqped(ouv rrlv onoreleoporu<6r4rd rnq. H petor6nrorl rou nl.oro[ou ovdyv<ooqg orrlv Eu<5vo 25.27 6etpter rrl oupnepupopd gtog runLrrlg oLroO4prlg oMq).ouX(og. H oLLrll,oul(o enrd vourl.eorrd(atv CU- UAGGC ouvq0cog ovoyvop((erqr on6 ro Leu-tRNA oro CUU, nou oxolou0efuor on6 ro ArgtRNA oro AGG. To ArgtRNA e(vot ondvro, r<or 6rov 4 onovr6rqrd rou npo- xoi.e[ r<oOuordpnon, ro Leu-tRNA otrro0o(vet on6 ro ro- 6u<5vro CUU npog trlv enu<ol,um6;revq rprn).6ro UUA. Aw5 nporoLe[ puo petor6nrorl rou nl.qto(ou ovdyvrrl- oqg, ener6rl q en6;revq rprnl,5to orqv (6ro <pdor1 ge ro v6o (euydpopq (GGC) 6roBd(stor on6 ro GlytRNA. H oL(o04or1 ouvrl0cog flpoyporonorefuor orq 06oq P (6rov ro Leu-tRNA ornv npoyUorx6r4to SXer yfver nemrdu- )otRNA nou peroq6per. rrl veoouwe0erprdvrl oluo(6o). H peror6nror; rou nl,oto(ou ovdyvroo4g o' 6vq 106r- r6vro repponopori nporol,e( en6rcroorl r4g nporeivqg. H Bdorl ornv 3' nl,eupd rou r<o6rxov(ou repgottollori enrlped(er q oXenrq ouXv6rrlro rou repUorrotrroU ror rrlg peror6nrorlq Kt 6tot enqped(er r4v onoteLeoport- r6tqro tou o(prorog repprorqrorJ. 25.1 7 AAAo lupBovro EnovoKo6lKonolnonq, MerocppooTtKn nopdKop$n Kot o Mnxovtopoq Tou tmRNA yto Tnv AneAeu0dpoon Tov KoOnAtrrpdvov P rBoooUoTiov a H n0porouUl ouv[ororor oqv uovdrrlro rou prpoorrllror[ou vo repgor[(eL rq perorppoorl, v' one)eu0epdrveror ond ro mRNA ror vo [ovopXi(er q perdqpooq nep(nou 50 vour]reor[6ro ro- 0odLro. e To prBootouono nou efvor roOqtrolg6vo novo mo mRNA pero q peprxrl ouvOeoq pLog nptoreiVrlq pnopei v' onetreu0epo- 0o0v ono rn Dpdon Tou tmRNA, ev6g povodrrof RNA ge Xo- poxrrlprorxo 16oo rou IRNA ooo xoL rou mRNA. H nopdxo;rQ4 (bypassing) neprlo;rBdver Uro perorf- vnorl rou prBooropro{ou nou oltr6(er ro ro6rr6vro ro ono(o e(vor (euyopro;rdvo pe ro nemtdutro-tRNA orrl 05or1 P. H oli.r1l,ou1(o prero[ri rov 6rio rodrrovfrov nopol.e(netor ror 6ev owrnpootoneieror oro no).une- nrt6rr6 npoi6v. 'Onrog nopouord(eror orrlv EIK0NA 25.28, out6 ennp5ner vo ouvexr.orel r'1 perdgpooq n6- po on6 onoro6rlnore ro:6x6vro repgortoprori ruXo(ver vo Bp(or<etor finv nopeprBoM6prevq nepro14. To nro ewunooror6 nopd6erypro nopdroprprlg e(vor ro yov(- 6ro 60 rou rpdyou T4, 6nou to prBooroprdrro perorrve(- tor 50 vour<,eor[6ro r<ord UriKoq rou mRNA. To r),er6( yro ro orjmqpro nopdroprqqg e(vor 6tt undplouv novopordtuno (r1 ouvcbvupro) r<o6rr6vro oe rd0e dxpo qg oLLqLoul(og nou nopoi.e(nerot. Aurd nqpoKopt|n 60 vourtreottd(tov oro yov6ro 60 rou rpdyou T4 Tetreurq[o xo0rxdvro mo oplud ntro[oro ovdyvoorlq [lptirrl xoldrrovLo oro veo ntroroto ovoyvrooqg Avdyvtooq Xorp(q gerordnrorl rou ntrqtofou ovoyvroolg GAUGGA UGAC......,.....AUUGGAUUA Avdyvoorl gero q perordnrorl rou ntroroiou ovdyvrrlorlg GAUGGA:.)".;;.tt; . . ,t.:',-::-::.r,, UUA Mro peroronrorl nLorolou ovoyvrrlolg +1 onoue[- r0r yro Trlv erqpoori rou yovrdlou tyb rou ororXe(ou Ty rotv (upo- pur1rov. H peroronLorl npoyl-roronore(Tor 0e Uro otrtr1trouX[o enro pooeov, orrlv ono[o 6uo rtoDrr<ovto Leu orotrouOouvror ono 6vo onovro xotdrrovro Arg. GAUGGA UGAC...,........AU ' .: : , '[116 Iuppowo Enovorodtxonoqolg: tVleroqpomul tloporo;lfr1 ror MqXovrolrog tmRNA yto Anetreu0epool KoOqtrot;revov PLBoor*rur i-. 739 Buorxeg eworeg
UeprK6q qop5q ovqq5powor oq ileproxSq "onoye(- ooqq" Kor (npooye(o:or1g". l-lprv on6 rnv nopdKouqJn, ro prBootol.rdrro tono0ere('roL pr' 6vo nenrt6utrotRNA (euyoptop6vo ge ro rcodx6vro onoye(rrlor1q or4 05oq P, evd: 4 ddero 05o4 A ovop6ver rlv e(oo6o ev6g o;rwoo- rui.o{RNA. H EIK0NA 25.29 6e(;yer 6n ro prBoocopdrro olro0q(ver rotd pr4rog rou mRNA oe owf r4v rord- orooq Srog 6tou ro nemrdutro-tRNA prnop6oer vo (eu- yopriloer pe ro ro6rxdvro ornv neproXrl npooye(rooqg. H otrl.qLouX(o rou mRNA nupo6ote( rqv nopdr<op- Un. To or11lowu<d XopoKrnprorxd efuor to 6rlo rcodrr6- vto GGA yro rnv onoye(cooq Kor rnv npooye(coor'1, ro nopepBoll6pevo 6rdm4Uq, Uro doprl p(oXou-0ql,tdg nou nepil.o1-rBdver ro r<<o6rrdvro onoye(oor1g ru 5vo rtrl- 6x6vro repprortopro6 6fuLq oro rto6x6vro onoye(ooqg. To md6ro r4g onoye(ooqg npouno0Ster ono65' opeuon tou nemrdutrotRNA on6 ro xrrl6rr6vro pe to ono(o e(vor (euyopo;r5vo. Ar<oLou0e( puo peror<[vqorl rou mRNA nou ro egno6f(er vo [ovo(euyopr6oer. 'Ener- to, ro ptBoocoprdrro oopciver ro mRNA, 6o9 6rou ro ne- nrrdulo-tRNA (euyopciroer Ue ro rro6rr6vro otrlv ow(6poor1 npoo'ye(<oo4g.'Eneno, q nporeivoorJv0eorl [ovopX((er, pr6llg 5vo o1-uvoor<u].o-tRNA eroS].0er orn 06oq A rord to ouvr10r1 rp5no. 'Onog q peror6ntoq rou nloro(ou ovdyvtoorlg, 6ror ror 4 owfdpoon rnq nopdroprQqg Boo(eror oe pto noU- otl rou prBooroprorfou. H nr0ov6rrlro 6rdoroorlq rou ne- mrdu).o{RNA on6 ro rrodx6vr6 rou onl 06o4 P ou(dver on6 ro0umSpnon ornv efoo6o rou oprvooKutro{RNA orn 06on A. H or6p4o4 ev6g oprwo[6og ;rnopei vo nu- fficnucAc........i [lpooye[roor1 To lterTrOutro-tRNA Eovo- (euyopovet ire v5o xtodtxovto 1:'r :ii:,; Ir' .111 Iq geooDo i1g noporo;it11g, 6vo prBootoporto pe roreAlpp6vq Oeon P Unoper vo repporloer q Ueroqpool. Otrr- oOo[ver xoro pqrog rou mRNA oe ;:ro neproXti, onou ro nenri6u- tro{RNA (euyoprlver pe evo rodrrovro 0TI 0dort P 'Ene uo q Uerotppoorl ouvey[(erot. 740 KE0AAAI0 25 0 l-evelro_c Kridrroq po6orrl oet nopdr<opr{.rq oro Borcrrl pror<d yov(6Lo elottfoc rrlg r<o0um6prlolg nou ouprBo(vet 6tov 6ev undpleL 6ro0Sorpro oprLvoor<ul.otRNA yto vo eto6tr0er oq 06o11 A Iro yov(6to 60 tou <pdyou T4, Svog p6).oq qg 6opnc rou mRNA e(voL vo geuilvst rrlv onorel.eotrlorrxdrqro rou repporroprori, npoxolrrlwog Sror qv r<o0uor6pqor nou onotrefuor yto rnv ow(6poo4 onoyefooqg. H 6rdorooq rov ro0qlu:trrdvrov prBootopror(rov mo Borcrrlpro r<or oprop6vo prLtoX6vSpro ennuyldveroi prdcc- ev6g provodLr<o0 uBpt6[ou mRNA{RNA nou xotrefuoL tm- RNA rol nepLdlet 66o LeLroupyu<69 neproXdg. H pro ne- pro1q onoprrpe(tor 6vo rprrlpro rou tRNAAu, ovri r OeUrepq rordrNonoLe( 6vo ppolJ no).unem(61o. To tmR- NA opXrr<d oprvoor<ul.rcoveror ond tqv oLovuAo-tRNA ouvOerdoq (AlaRS). Ir4 ouv6XeLo npoo6dver rov nopo- yowo EF-Tu ru 5ner-ro og rproroydg orjprnl.oxo npoo- dSveror orq 05oq A rolv r<o0r1).rlr;l6vov prBoorl-l;.tot(tov Idvo oro prBooroprdlo ouprBo(veL prerogopd nemL6[or- nou ouvddeL qv olov(vq pie ro ropBo(ureLrxd dr<po rt'1c r<o0r1Lcop6vq g, veoouweOerpr5vrlq npolrelvq g : row6lpo - vo, ro mRNA nou Bp(or<eroL ndvo oro prBoorr.ll-rotto ov- lxoOrordror ond q 6e[repr1 nepLoXrl rou tmRNA. Im ouvdXero, to tmRNA tretroupye( cog erpoye(o yLo tq orjv- 0eor1 66r<o enrnpdoOerov o1-rtvoldorv, preto ro ono[c undpXet 6vo r<rodu<6vro rsppolopori yro rov repUorroUC rqg perdgpoonq Kor qv oneLeuO6pcoorl rrlg npir.lreir,lq l-i np6o0eq r<opBo(ureLu<r1 oh}.q),ouX(o 6po rog ot11-rc yro enordlou0rl ovoyvioprorl ond npr.,:redoeg, or ono(ee ono6oproriv t4v r<o).oBropr5vq npu.lrelvq. 'EroL, ro tmRNA Lenoupye( og 6vog prqXovropr6g el.6y1ou noL6trpog yrc ttlv ovorrix..rr.lo4 r<o0q).rilptdvov ptBootopor(orv Kot rrlv onopdr<puvoq xo).oBropdvrov nporeivov, ot ono(eg er- 6oMrog 0o ouootrlpe[owov. 25.1 I llepiAn(rn H oM4LouX(o rou mRNA, nou dLoBd(eror orrg rprn),e- rsq Ue rote(Ouvoq 5'->3' olsr((eror pSoo rou yevert- r<otj xrrr6rxo Ue rnv oL).r1LouX(o rc'rv oprrvo(5ov rov no).unemt6[tov, nou dtoBd(eror ond ro N- npog to C- rel.rx6 dr<po. And ng e[r1wo r6ooeprg rprn].6req, e[qv- ro Ur.o xco6rronoroiv oprLvo[5o Kor rperq nopexouv or;;roro reppoloprori. To ouvc6vupro r<ttl6rr6vro, nou owrnpooorne[ouv ro i6to opuvo[5o, 6Xouv ouyyeve[g oMrll.ouX(eg, ouyyd pre puo preroBotr4 orriv tpfur; Boon rou xro6rr<ov(ou. Aur4 q er<cprjl.roq rqq tpkqg pdolq oe ouv6uoopd U' 6vo np6runo orjp<povo pe ro onoio 1qpu<6q nop6proro opuvo[do re(vouv vo x<r.lStronototjv- ror on6 ouyyev4 rordtx6vro, eloXtoronoLe( tLg enrdpo- osrq rov pteroltrd[eo.lv. O yevettr6g r<trl6rr<og e[vot orxoupevrx6g r<or npdneL vo xo0tep60r1re no).ri nportl,ro rotd rqv e[6].1[r1. Or oMoy6g oro nupnvrKd yovrdurr- poro e(vot ondvteg, oMd oproprdveg oMoy6g npoypo- ronoLrl04r<ov r<otd rlv e(6Lt(q rov l.utoXovdp(orv. I-loMonLd IRNA prnope( v' onowo[v oro i6ro r<or6L- r<dvLo. H otr-rodo rorv IRNA nou onowoJv mo noLr(i.o xroSu<6vto yro r<dOe opuvo[d e(vor loporrqproru<r1 yto Anoyelo.lon To ptBoocrrgorLo peroxrveltol roro prlrog rou mRNA
Kdee opyovroU6. H ovoyv6pton KodtKov(ou-owtKo6t- rov(ou egntr5r<et oor60eto orqv npdrtrl 05oq tou owt- rodrr<ov[ou (tpir4 06or1 rou ro6tr<ov[ou), 11 ono(o ennpSner os optopdvo IRNA v' ovoyvcop((ouv noMo- nLd r<odr6vto. Otro to IRNA nepdXouv tpononorrlprS- veg Bdoerq, ot ono(eg etodyovtot on6 6v(upo nou ovoyvrop((ouv Bdoerg-m6xouq orn 6opr4 tou IRNA. To (euydpolpo ro6trov(ou-owtrot6trov(ou enqped(etot on6 rpononoqoetg rou (6tou tou owtt<todxov(ou 6nog en(oqg xot on6 ro (ougQpo(5pevo", 6rlLo6n rtq Yero- vrx6g Bdoetq, r6(u:q orrlv 3'nLeupd tou owtt<trl6trov[ou. A[ronotdlwog t4v oor60eto ro:6xov(ou-owro6u<ovfou, to 1.troX6v6pro rov onov6utror6v opt<otiwot oe 22 lR' NA yro rrlv ovoyvdlptoq 6trov ttov rcto6tt<ov(tov oe oX6or1 pre ro ouvr]0q eL6Xtoro opt0pr6 tov 31 IRNA: oe our6 ouprBdMouv ot oll.oy6g oto l.tttoXov6ptor6 rrirdxo. Kd0e oprwo(ti ovoyvcopf(erot qn6 pto ouyrerpt;r6vr1 o1-uvoor<ul.o-tRNA ouvOetdoq, q onoio ovoyvcop[(et en(- oqg 6Lo ro IRNA nou rto6tronoto6v ro ouyrerptpttivo optvo(ri. Optopr5veq ogwoorutro-tRNA ouvOetdoeq 6to- O5touv pto trettoupy(o pretor6ntorlg tou nl'oto(ou qvd- yvoonq, 11 ono(o ent0ecope( ro oUtvooKULo{RNA t<ot u6potrUet ro ).dOog ouvde6etrrSvo ol.twooruLo-tRNA. Or oprwooxul.o{RNA ouvOetdoeg notx(ouv no}.0, oor6oo xototdooowot oe 6Uo yevx6g ogddeg nou eU- qov((ouv oprotBo(o onorlet6pevo got(Bo oM4),ouXrcilv rot np<oreivrdrv 6o;rcbv ottg rototruttxdg neptoX6g toug. Ot 66o otrrd6eg tcr:v ouv0eroodlv 6torp[vowot en[or1g on6 r4v qp1Kn neproXq rnq optvooKuL(toorlg orrlv 3'-te- hxn ptp6(n tou IRNA, t4v roreU0uvon rnq np6odeoqq rrlg Sllrog-onod6rcr4 tou IRNA rot to puOptoneptoptort- r<6 Brjpo rnq optvooKul(ooqg. Mto roOopto;.t5vq opd6o voux.treold(tov tou IRNA, nou xotrefuot o;rd5o tout6rq- roq, ovoyvop[(eror enil.erctrd on6 t1 ouv0erdorl, 4 ono(o lp4otpronoe( 5vo ouvduoop6 dpeoov ru Sppeocov pulXovtoprdrv ovdyvcoorlg. Ie nol'L5g neptnt6oetg, 11 opd6o rout6qtog tonoOetefuot oro dr<po tou owtrcodt- rov(ou Kot rou 3'-ono65rrn tou 1-top(ou. MeroMd[etg Unope( vo ennpS{.rouv o' 6vq IRNA vo 6toBd(er dto<popertrd ro6u<6vto: r'1 ouvrlO6orepn Uop- <pr1 rdrouov pretoMd[ecov ouprBo(vet mo (6to ro owtKo- 6tr6vro. H ueropotrn tou owtrcodtt<ov(ou prnope( vo enmpdrper o' 5vo IRNA vo rotoorc(tret prto petdMo(r1 o' 5vo yov(6to nou r<odtronote[ npote(w1. 'Evo IRNA nou ovoyvop((er 5vo rrr:dtr6vto repporl.opoU nop5Xer 5vov oouv0err6 rotoorol,5o, evdl 5vo IRNA nou ;retoBdMet ro oprwo[0 nou owtorotxe( o' 5vo rco6tx6vto e[vot Svog 6uouv0enr6g r<otooro5o9. Ot r<otooroe(g trrrv rco6t- rov(cov UAG e(vot onoteleo;.totx6tepot on6 touq ow(- ororxouq rov t<to6trov[ov UAA: out6 e(qyefuot on6 to yeyov6g 6tt to UAA e(vot to nto 6tode6opt5vo rro6tx6vto reppottopo0. H onotetreopottr5tqro 6trtov ttov t<oto- orol5tov, oo16oo, Boo[(etor oro (oupqpo(6pevo" rou r<dOe [eXo:ptorof r<o6xov[ou-or6Xou. Meroton(oetg tou ntroto[ou ovdyv<oor1g +1 ri -1 nt- 0ov6v npoxotroUwot on6 olloO4pdg oMrltrouX[eq oro mRNA nou enttp5nouv o' 5vo nemt6ulo-tRNA vo ol't- oOrloet qn6 to rco6tr6vt6 tou t<otd p(o Fdon efue oe 5' efue oe 3' rote0Ouvor1. OptoprSveg flpoYpopporrotrr6veg petoron(oerg tou nLoto(ou ovdyvotolq ::- ':::: -: tor on6 qv oMql.ouX[o tou mRNA onoiro- ':]. '.: 6x<ppooq puoLxtov yovt6[cov. Ilopor<o;rqq o-*p :.' . drov 6vo pLBooto;rdtto mopord rri pretdgpool Kot []e- -rorwe(ror xotd prlrog rou mRNA pe ro nemtdu).o- tRNA orl1 06ori P 6<r-r-c 6rou owd (euyopooeL p' 6vo rotoMq- Lo opuvo(ti, ondre f,ovopXi(et q pretd<ppool' H Xplort rou tmRNA nopSlet 6vo prrlXovto;r6 etr5y1ou not6tqrog yro rrlv ovot<tixlool r<o01trtl;r6vov ptBooropiot(rov xot r1v onoproxpuvoq ovent00prltov r<o}oBtlp6vov npote'[- vLr<.ov npo[dwirlv. BrBAroyporpio 25.1 ELouytoytl "Epeuvo Nirenberg, M W. and Leder, P. (1964). The effect of trinucleotides upon the binding of sRNA to ribosomes. Science 145, 1399- 1407. Nirenberg, M. W. and Matthaei, H. J. (1961). The dependence of cell-free protein synthesis in E coli upon naturally occurring or synthetic polyribonucleotides. Proc. Natl. Acad. Sci. USA 47, 1 588-1 602. 25.3 H Avoyvtoptol Ktrtdtxovlou-AvTtKoDtKov[0u l-lepilogPovet Aoro0eto 'Epeuvo Crick, F. H. C" (1966). Codon-anticodon pairing: the wobble hy- pothesis. J. Mol. Biol. 19, 548-555. 25.4 T0 tRNA IlopoYovrot pe EfleEepYooio Meyotr(repov IlpoDpdptov Mop(ov Avoor6qoq Hopper, A. K. and Phizicky, E. M. (2003). IRNA transfers to the limelight. Genes Dev. 17, 162-180. 25.5 To tRNA ilept6Xet Tpononotlpeveq B00etq Avoouinrlorl Hopper, A. K. and Phizicky, E. M. (2003). IRNA transfers to the limelight. Genes Dev. 17, 162-180. 25.6 0r Tpononoulpeveq B6oetq Enrlpeo(ouv T0 ZeuYopoU0 AvrtroDtxov(ou-Ktodtxov(ou Avoordnnon Agris, P. F. (200S). Bringing 0rder to translation: the contributions of transfer RNA anticodondomain modifications. EMB0 Repofts 9, 629-635. 25.7 Ynopxouv Inopodu6g Tpononotfloetg rou 0troupevtrou Ktl0tro iti:rlilri i: :_rl: 0sawa, S. et al. (1992). Recent evidence for evoiution of the ge- netic code. Microbiol. Fev. 56, 229-264. BrBtrLoypotpio 7 41
Santos, M. A. S., Moura, G,, Massey, S. E., and Tuite, M. F. (2004). Driving change: the evolution of alternative genetic codes. Tends in Genetics 20, 95-1 02. 25.8 Nrlo Aprvo(eo Mnope(vo Evoroporo0ouv 0s 0pr0Udv0 Kto0 rxrjvro Tep gonopou Avcoxonqaq Krzycki, J. (2005). The direct genetic encoding of pyrrolysine. Curr. 1pin. Microbiol. 8, 7 06-7 1 2. Ambrogelly, A., Palioura, S., and Sqll, D. (2007). Natural expansion of the genetic code, ffat Chem. Biol.3, 29-35. 'Fncr rrn Srinivasan, G., James, C. M., and Krzycki, J. A. (2002). Pyrroly- sine encoded by UAG in Archaea: charging of a UAG-decoding specialized IRNA. Science 296, 1 459-1 462. Turanov, A. A., Lobanov, A. V., Fomenko, E. D., Morrison, H. G., Sogin, M. 1., Klobutcher, L, A., Hatfield, D. 1., and Gladyshev, V. N. (2009). Genetic code supporls targeted insertion of two amino acids by one codon. Science 323,259-261. 25,9 To IRNA Zeuyoprirvouv Ertrerlro pe Aptvo[60 Mdorrt Aprvoorutro-tR NA Iuv0aroodlv Avacxrnqoq Giege, R., Sissler, M., and Florentz, C. (1998), Universalrules and idiosyncratic features in tRNA identity. Nucleic Acids Res. 26, 501 7-5035. lbba, M. and Stpll, D. (2000). AminoacyltRNA synthesis. /nnu. Rev. Biochem. 69, 61 7-650. Perona, J. J. and Hou, Y-M. (2007). lndirect readout of IRNA for am inoacylation . B i o ch e mi stry 46, 1 0 41 9-1 0432. 25.10 0r Aprvooxutro-tRNA Iuv0erooeg Kororoooovror oe Auo 0troy6vereg Avroxanqol lbba, M. and Sqll, D. (2004). AminoacyltRNAs: setting the limits of the genetic code. Genes Dev. 18,731-738. 'Ipeuv* Eriani, G., Delarue, M., Poch,0., Gangloff, J., and Moras, D. (1990). Partition of IRNA synthetases into two classes based on mutually exclusive sets of sequence motif s. N atu re 347, 203-206. Rould, M. A. et al. (1989). Structure oI E. coliglutaminyltRNA synthetase complexed with tRNAGln and ATP at 2BE resolution. Science 246, 1 135-11 42. Ruff, M. et al. (1991). Class ll aminoacyltRNA synthetases: crystal structure of yeast aspartyltRNA synthetase complexes with tR- NAAsp. Sclence 252, 1 682-1 689. Sauerwald, A., Zhu, W., Major, T. A., Roy, H., Palioura, S., Jahn, D., Whitman, W. B., Yates, J. R. 3d., lbba, M., and Sqll, D. (2005). RNAdependent cysteine biosynthesis in archaea. Sci- ence 307 ,1 969-1 972. 25.11 0L Iuv0erooeg Xpqorponorouv q Arop0trror'1 Aonp[tov yro vo BstrTrooouv Trlv Arp(pero , a;,a:.ti: Dock-Bregeon, A., Sankaranarayanan, R., Romby, P., Caillet, J., 742 KE0AAAI0 25 0 levelr6g Ktirbtroq Springer, M, Rees, 8., Francklyn, C, S., Ehresmann, C., and Moras, D. (2000) Transfer RNA-mediated editing in threonyl{R- NA synthetase. The class ll solution to the double discrimination problem. Cell 103, 877 -884 Hopfield, J. J. (1974) Kinetic proofreading:a new mechanism for reducing errors in biosynthetic pr0cesses requiring high speci- Iicily. Proc. Natl. Acad. Scr. US,4 71,4135-4135. Silvian L. F., Wang, J., and Steitz, T. A. (1999). lnsights into edit ing from an lle-tRNA synthetase structure with tRNAlle and mupirocin. Science 285, 107 4-1077 . Tukalo, M., Yaremchuk, A., Fukunaga, R,, Yokoyama, S., and Cu- sack, S. (2005). The crystal structure of leucyltRNA synthetase complexed with tRNALeu in the posttransferediting conforma- tion. rVal. struct. Mol. Biol. 12 923-930. lvnnrrlnnm Beier, H. and Grimm, M. (2001). Misreading of termination codons in eukaryotes by natural nonsense suppressor tRNAs. rVuclelc Acids Res. 29, 47 67 -47 82. Eggertsson, G. and Soll, D. (1988). Transfer RNAmediated sup- pression of termination codons in E. coli. Microbiol. Rev. 52, 354-374. Murgola, E. J. (1985). tRNA, suppression, and Ihe code. Annu. Rev. Genet.19, 57-80. 'Ip*uvc Ruan, 8., Palioura, S., Sabina, J., Marvin-Guy, 1., Kochhar, S., LaRossa, R. A., and Sqll, D, (2009). Quality control despite mis- translation caused by an ambiguous genetic code. Proc. Natl. A- cad. Sci. USI 105,16502-16507. 25.15 To Prpooropono Enlpe o(er qv Arp(peto rrlg Meroqpooqq A r:nnp inn.rnn v uu,lsr a ivr I Daviter, T., Gromadski, K. B., and Rodnina, M. V. (2006). The ribo- s0me's resp0nse to codonanticodon mismatches. Biochimie 88, 1001-1011. 0gle, J. M. and Ramakrishnan, V. (2005). Structural insights into translational tidelily . Annu. Rev. Biochem. 7 4, 129-177 . 'f;*ruv* LaRiviere, F. J., Wolfson, A. D., and Uhlenbeck,0. C. (2001). Uni- form binding of aminoacyltRNAs to elongation factor Tu by ther- modynamic compensation. Science 294, 165-168. Ogle, J. M., Brodersen, D. E., Clemons, W. M., Tarry, M. J., Carter, A. P., and Ramakrishnan, V. (2001). Recognition of cognate transfer RNA by the 30S ribosomal subunit. Scr'ence 292,897-902. Zaher, H. S" and Green, R. (2009). 0uality control by the ribosome following peptide bond formation. Nature 457 ,161-166. 25.16 Merordntoq rou fltroro[ou Avoyvtoorlg Mnope[vo Iuppe( Ie 0trroOqpdg AtrtrntrouXreq -' :;:.1::.: t:a :.. Baranov, P. B., Gesteland, R. F., and Atkins, J. F. (2002). Recod- ing: translational bi{urcations in gene expression. Gene 286, 187-202.
Gesteland, R. F. and Atkins, J. F. (1996). Recoding: dynamic re- programming of translation. / n nu. Rev. B i och e m. 65, 7 41 -7 68. 'trnar rrrn Lpouvu Jacks, T., Power, M. D., Masiarz, F. R., Luciw, P. A., Barr, P. J., and Varmus, H. E. (1988). Characterization of ribosomal frameshifting in HIV-1 gag-pol expression. Nature 331, 280-283. 25.17 Atrtro Iugpovro Enovoroldxonotri0r1q: Meroqpoonrrl lloporop{r1 Kot 0 M1xqvtoUdg rou tmRNA yto qv Anetreu06ptoorl rtov Ko0t'1trtrt1t6vtov Ptpootogot[trtv Herr, A. J., Atkins, J. F., and Gesteland, R. F. (2000) Coupling of open reading frames by translattcnar D)!ass -: :--, := Biochem. 69,343-372. Gallant, J. A. and Lindsley, D. (1998). Ribosomes can s c.:,:- and beyond "hungry" codons, resuming protein chain elongatic: many nucleotides downstream . Proc. Natl. Acad. Sci. US/ 95 13771-13776. Huang, W. M., Ao, S. Z, Casjens, S., 0rlandi, R., Zeikus R Weiss, R., Winge, D., and Fang, IVl. (1988). A persistent un- translated sequence within bacteriophage T4 DNA topoiso- merase gene 60. Sclence 239, 1 005-1 01 2. Blpro":::.: 743
Lewin γονίδια χ ( μέρος iii)

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Lewin γονίδια χ ( μέρος iii)

  • 1.
    Meroypoqn Kol q;" r q "4 't{n MeroypoqlKoi Mnxovlopoi 25 3u r? OoToypoQ(o ;le tqv o6eto rov Jie Ren rot Jun Ma, U- niversity of Cincinnati College oI Medicine rot Cincin- nati Children's HosPital. Metoypoqrl otouq [lPoxoPutireq Metoyporpn orouq Europudrreq Iuppoqn rt EneEepYooio rou RNA Iro0eporrlro rL Evtontopdq rou mRNA Kototrurtro RNA Meroqpoorl XprlotgonottitvToq T0 [evertro K66tro KEOAAAIO 19 KEOAAAIO 20 KEOAAAIO 21 KEOAAAIO 22 KEOAAAIO 23 KEOAAAIO 24 KEOAAAIO 25 509 III
  • 2.
    O Phantatomix/Photo Researchers.lnc ffi * K0T0 rIV ertqrlKuvon, I UeToypoqxn Quo0tr[60 UcToKI- veiror roro prlrog rou DNA rr r; otruoOo rou RNA enr- prlruveror oe roreuOuvorl 5'-+3' pe npooOflr4 vourtre- ond[olv oro 3' oxpo. * H Ueroypoqrl oropord ror ro d[r],trrvo rou DNA [ovo- oXrlporl(erot g6trrq q RNA notrupepdorl onoouv6e0e[ ond ro DNA erpoye(o oe Uro 0eon repponopou. H Borqprorrl RNA l-lotrupepooq Anoretre[ror onri l-lotrtr6g Ynopovodeg * 0r nupqveg rtov porqprorov RNA notrugepoorirv e(vor notrupeprl oupntroKo -400 kD pe yevxl Oopn 02pB'o. e H rordtruol enuetrelrot on6 rtg unopovodeg p xor p'. To Otr05v(upo r4g RNA l-lotrupepooqg Anorele(ror ono rov lluprjvo rou Ev(rlgou ror tov llopoyovro lypo * H poxrrlptorq RNA notrupepriorl uno6rorpeiror orov 0z0p'o nupnvo rou ev(upou nou roro),uet Tn UeToypo- qr1 ror tqv unopovodo o nou onoue(ror privo yro r4v evop[r;. ffi ffi @*83 Meroypoefi oTouq flpoKqpudlreq Etooytrlyrt * H Ueroypoqn npoyporonore[ror Ue Qopo 5'npog 3' o' dvo ergoye(o nou 6Xet qopo 3'npog 5'. H Meroypotprl llpoyporonore(rot ge Zeuyopropo Booetov oe gro u0uootr[60, A(euyoporou DNA * H RNA notrupepoor; OroXopr(er toug drlo rl,drvoug rou DNA oe pLo eqlgeprl <quootr[do> rot Xprloqionolei rov 6vo rtrdrvo og erpoye[o yro vo roreuOuver q ouvOeo4 luog oupdrlpolUolrnq otrtr1trouXrog RNA. * To Uflroq rqg EuootrOog ervor 12-14 bp evtir rou uppt- d(ou RNA-DNA p6oo oq rpuootrOo 8-9 bp. H Avr(dpoon qq Meroyporp4g'EXerTpio lro6ro " H RNA notrupepdorl npoo06veror oe pto 06011 unorr- vr1rr1 oro DNA yro vo oXrlpor[oer 6vo rtrerord oupntro- K0. * H RNA notrupepooq [envo q peroyporprl oqou ovor[er ro d(xtrtovo rou DNA yro vo oX1por(oer pro peroyporprrq rpuootri6o. 510 KEOAAAI0 19 Meroyporpq otoug l1poroputirreg ffi 19 r.t I rss Ew tp6)'eta Keqt€vou : Richard Gourse
  • 3.
    tl 0 nopdyovToqo Tpofiororel lg rdulqreg npoodeolq oro DNA r4q RNA notrupepoorlg, 6ror drore n ouyy6- vero qg yto ro yevrro DNA vo ger6verot ror, ovr[Oero, 1 ouyy6vero Trlq yro Touq unouvrlrdg v' ou[over. l16q Bp(oxer H RNA l-iotrugepdorl rLg Atrtr1trouX(eg rov Ynoruv116v; * H np6odeon rrlg RNA notrupepoolq otoug unoxrvlreg e(vor rooo ypnyopl tlore potrtrov dev nporeuor yr' ontrr1 6roXuor1. * H RNA notrupepoorl npoodeveror oe ruXo(eg 06oerg oro DNA ror lg ovro),trooer pe dtrtreg otrtrntrouxieq 6otq drou ppe0ei evog unorrv4qg. To 0tr06v(u;ro Tpononoe[ror Koro rr1 Arepyooio Avoyvdr- pronq Kr Anetreu0epoorlg on6 roug Ynorrvrlr6g s OTqv Il RNA notrupepoorl npoo6everor o' rlvov unoxr- vt1it1, dLoXropt(er roug rtr6voug rou DNA yto vo DrlpL- oupyrloet Uro UsToypoQrKn Quootr(60 xr evotopordrver vourtreoridro oro RNA. * H rvop[n ono rrlv RNA notrupepoo4 dev elvor novrore enuuxqg. * 0 nopoyovrog o(ygo ouvrlOoq onetreu0ep6veror ono qv RNA notrupepoor; orov 1 veoouvre0eqlevrl otruoOo RNA onorrrloet pflrog -1 0 pooeolv. 0 fiopoyowoq Irypo Etr6yXer rrlv l-lpdodeol oro DNA Avoyvop((ovrog IuyxerpLp6veg Atrtrqtrou1ieq rov Ynoxr- vrlrdv s 'EvoS unortvrlrrlg npoodropl(erot onri qv nopouo[o ppoXeuilv ouvorvenxrirv oltrrltrouXuirv oe ouyxerprp6- veg 06oerg. e 0t ouvorvtTrreq otrtrqtrouXieq Tou urroKrvrlrrl ouvrl0oq onore),ouvror on6 gro noupivq oqv oqeqpro, evo e[0Uep5q Ue otrtrr]troux(o nopepqepl Ue rnv TATAAT orq 05or; - -1 0 xr 6vo otrtro e[ogepdq ge gro otrtrr1trou- XLo nopegqeprl pe r1v TTGACA oq 0eor1 --35. * 0t Eexttptorol unoxrvlr6g ouvrlOog drorp6pouv ond rr1 ouvorvelrrl otrtrr;trouX(o oe Uto n neproorirepeg 0e- 0etq. e H onoretreogortx6rrlro rou unortvqq pnope( eniolg vo enlpeo(eror xor on6 enrnprioOero orotXe(0. H Anore),eopotxdn1ro rov Ynorrvqrtlv Mnope( v' Au[o- ver 11 vo EtrLordveror ono Merotrtro[4 e 0r uopvlnr6qD UeToMoqerq nou pertirvouv qv orroTt- leogortxoqro rou unorrvlrrl ouvrlOtrrg pertilvouv tr1 oupg6prprooq npoq nq ouvorverx6g otrtr1trouX(eg, evtrr or uoeflrdq, perotrtro[erq 6Xouv ro ovrf0ero onor6tre- 0u0. * Merotrtro[erq oqv -35 otrtrntroux(o pnope( vo enrqped- oouv rr1v opXrxl npdodeol i1g RNA notrupepoolg. * Merotrtrd[etq onlv -1 0 otrtrr;trouX(o pnopel vo enrlpeo- oouv Trlv np6o6eol 11 n1v ovridpoorl onodroro{qg nou perorp6ner 6vo x),erord ofgntroro oe ovorXrti. l-lotrtreg lleptoXeg qg RNA l-lotrupepoorlg 'EpXovror Ape- oo o' Enogrl pe rov DNA Ynorrvrlq q H do[rl rou o70 ol],0(er orov ouv66er0r Ue Tov nupflvo tou ev(upou, enupenowog ortg neproXtlg yto npoode- o1 oro DNA v' otrtrqtrenr6pooouv Ue rov unorrvlrfl. * l-lotrtr6S neptoXdg rou 070 otrtrrltrenrdpouv Ue rov urroKr- vrlTn. * H unouovodo o enfor1g oul.rpotrler on]v 0rc, , -- -- rou unortvrlq" H Anorunoorl e(vor Mrl0odog YmInS Avotruoqg yra :: Xoporrrlprop6 rtov Atrtrrltrenrdpooetrlv RNA l-lotru pepoorl c - Ynouvrltrl ror, [evxdrepo, D NA- l-l poreivtirv * 0r 0uv0rvtTrK6q otrtrltrouxreq -35 ror -10 nop6Xouv ro neprooorepo onpeio enoqnq yto rrlv RNA notrupepoorl orov unortvrlrfl. 6 T0 qgelo enoqrlg pp(orowor rupro:q or4 pro ntreupo rou DNA. 0r Atrtr4trenrdpooerq Mero[0 rou I-lopoyovro Iiypo rot rou fluprlvo ir;q RNA llotrupepooqq Atrtro(ouv Koro 11 Atotpuyrl rou Ynoxrvlr1 " Mro neproXrl rou otylro rorotroppdver ro 0toutro e[ridou rou RNA rot npener vo perouvr;0ei yto vo dreurotrtivet rr1 ouv0eorl rou RNA. * Avenuuxe(q evdp[erq ouv1Oog ou;rpofvouv nporoU ro 6v(u;ro oXqpor(oet 6vo npoyporrr6 oupntroro entgrl- KUVorlq. & 0 flopoyowoq orypo ouvrl0oq onetreu0epoverot ond qv RNA notrugepooq g6trrg 1 veoouvre0ergevil otruol- 6o RNA vo qrdoer oe Urlxoq -l 0 nt. H Kpuorotrtrrrl Aoprl Ynoderrvuer'Evo Movr6tro yto 11 Meror[vr1o1 rou Ev(ugou e To DNA petouvelrot 6rop6oou evdg droutrou oqv RNA notrupepoorl Kor npoypoTorrote( pto on6roprl orpoqn oro evepyo r6vrpo. * Atrtroyeq orrl 6roprlpgtrloq ouyrerprp6vov eu6trrrrov e[oprnporov p6oo oro dv(upo etreyXouv qv e(oo6o rov vourLeond(orv oro evepyo x6vrpo. Mto Iropoqpdvl RNA llotrupepoon Mnope(vo iovopXioer # Mro oTopoTrlpeq RNA notrupepoorl pnopei vo [ovopXr- oer rn UeToypoErl xripovrog ro RNA peroyporpo yro vo Dqproupy1oer evo rorvoupyro 3' oxpo. H Borrqprorf RNA l-lotrugepool Teppor((er oe Atorpudg 0eoerg € Ynopxouv 6uo ro[erq otrtrr1lou1uirv reppolopou. Erei veq nou ovoyvtop[(ovrot prlvo ono rrlv idro rrlv RNA notrupepoorl Xop(q rr1 oupperoXq rovev6g rurroprrou nopdyovro ouv40rrlg ovorp6povror a "evdoyaveC reppouoreq" (intrinsic terminators). Atrtreq onououv 1-rLo xurroptrrl nporelvrl nou ovopo(eror po (p) xor ovo- tp6povtot oq (TepuoTtoTtig e[opr6gevot onri qv p,. * 0 evdoyevnq reppouopog npoUno06rer ovoyvtirpro4 ptog otrtrqtrouX(og reppouopou oro DNA nou rordrxo- norei pro 6opn Qoupr6rog oro RNA npoi6v. 6 T0 orlpoTo yro repgolop6 pploxovroL rupltrlg gSoo oe otrtrltrouXreg nou 6Xouv n0n UeroypoQe( on6 qv RNA notrupepdoq xr 6rol o repporLoprig ppforeror un6 rov 6treyXo rou erpoye(ou ro/r1 rou RNA npoirivrog nou peroypoqer n notruUepoon. llrirg AeLroupye( o l-lopdyovrog p; € 0 n0p0y0vT0g p etvot pto nptrlrefvq nou npooddveror oro veoouvre0eq:6vo RNA rot perorrve[ror xoro pqrog tou RNA yro v' otrtrrltrentdpooer pe qv RNA notrupepo- orl Kor v0 qv onetreuOepdloer on6 ro o0gntroro enrpl- KUV0rlq. #wa :,!E*i ffi g*ee 1;ia;: .dg€!r14 qGL&&; .-ffi+. Ei,E*" Fg@. 4E&*; l I l 4t."7i4., ouvey[(erut ou1v enopevr1 oelido 511
  • 4.
    ffi H Ynepetriro:oqErvot lqpovrx6 XoporqptorLxo fllq Me- roYporllq " H opvlnrr; unepetr(roo1 ou[dver qv onoretreoporuo- qro ronourlv unortvrlr6v eneroq poqSd qv ovr(6poor1 onodroro{1q. a H peroypotprl dqproupye( gnpooro ono ro 6v(upo Oel- rdg unep6trtreg ror n[otrr rou opvrllr6g unep6],rxeg, or onoleg np6neL v' otpoLpe0oUv onrl rr1 yupootl xot qv ronoioopepool. qlE}i H RNA l-lotrugepool rou 0oyou T7 Elvor 6vo Xplorlio l-lporuno IuoTlUo & H orK0y6vsr0 ro:v RNA notrupepootirv tou T7 e(vor g.rovo notrunenr(Oro trovo v' ovoyvrop[(ouv roug unorrvrlr6g rou qoyou ror vo enuel,ouv notrtreg ond ng dp0orrl- proqreg tov notrupeptirv RNA notrugepootirv. * 0r rpuorotrtrrr6q dog6q rov RNA notrupepoorlv qg oL- roy6verog rou T7 pe ro DNA rouronororjv qv neptoXrl npoodeolq rou DNA, oro evepy6 r6vrpo xor uno0et- rvuouv ;lovre),o yro rri 0rorpuyrl rou unorrvqq. ffi O Avtoytr-rvro;rdg yro rouq l-lopoyoweg liypo Mnoper vo Pu0p(oeL qv'Evop[1 * Io E. coli eXer enrd nopdyovreg o(ypo, xo06vog ono roug onoloug o0ei qv RNA notrugepoorl vo (errvfloer oe pro opodo unoxrvrltrirv nou npoo6top((ovror ono ouyrerpLp6veg otrtrntrouX(eq -35 xot -1 0. * 0t evepyrirnroq ToJV 0rorpopertr6v nopoydvrtov otylto pu0pi(ovror ontl Dtorpopelxou g Lr4Xovropoug. ffi Or flopdyovreg l(ypo Mnope( vo Elvor Opyovrop6vor oe Axotrou0ieg * Mro oKotroue[o nopoyovrov oiypo dnproupye(ror orov dvog nopoyovrog oiygo onoueiror yro rq peroyporprl rou yovr6[ou nou rodrronorel rov enoplevo nopoyov- tu, * To npdrruo yov[dto rou qdyou SP01 peroyporpovtor ono rnv RNA notrupepoon rou [evrorr1. e 'Evo 0rt0 ro nptirLgo yov(6ro roDrronotel 6vov nopoyov- ro o(ypo nou trlOei r;v RNA notrupepoorl vo peroypo- ger ro evdtdgeoo yov(dto. 19.1 Erooyoyfi * H Ueroypoqrl npoyltotonoteirot pe qopo 5'npog 3' o' 5vo er- poye(o nou eXet qopo 3'npog 5'. H pretoypo<pr1 nopdyer pto oluo(6o RNA ge raut6o4- pry aMqAouy[a Ue rov 5vo x].6vo tou DNA, nou Uept- x6g gopSg role(ror x<o6rxonor4rrxrig xArivog (coding strand). Aut6g o r<Ltovog SXer <popd 5'-+3' xor e(vor o u pnlr1papaux6q (co m ple mentary) (6nLodn (euyopcbver) ge ro eKpoye(o (template), nou SXeL <po- pd 3'-+5'. I-t' out6 o nop6protog pe ro RNA x).rilvog * Auo ono ro ev0Lopeoo yov(6ro roduonorouv unopovo- 6eq evoq nopoyovro o(y;lo nou o0e( qv RNA notrupe- poorl vo ueToYpouer ro oQrlro yov(6r0. H Inoproyeveon Etreyxeror ono l-lopoyovreq lypo * H onoproy6veoq 6rope( evo porrrlpro o' evo prlrptro rurropo nou urpiororor truor1 rr dvo onopro ttou oTit- treu0ep6verot. I Koee [eXopror6 6ro;rSprogo npoXoper oro enopevo oro0ro ovdnru(qg ouv06rowog 6vo rorvoupro n0p0- yovro olypo nou erronl(er rov npolyorlpevo, * H rnrKorvovlo Uero[u rorv Duo dropepropdrtov ouvrovi- (er qv eniloyri rou rorotrtrr1trou Xpovou yro Trq ovTrKo- tootooetg ttrlv nopoyovrtov oty|to. 0 Avtrepponopoq Mnopei vo Elvor'Evo Pu0prolxo Iup- tsov * 'Evo o6untroKo owrrep[oTropou enup6ner orrlv RNA notruUepdol vo 6ropooer 6topdoou reppouordtv. * 0 Qoyoq trou0Do xpnotponorc[ ouorrlporo ovTtTtpuoTr- olrou yro vo puOpioer rooo ro npdrgo ooo Kot T0 otIiUC perdyporpo, otrtrd ro duo ouorrlporo treuoupyouv p' ev- retr6g drorpopenro0g plXovrol.roug. * H rpooDeq nopoy6vrotv oro veoouvteOetg6vo RNA ouv66sr rtq nporelveg ovTrTrpgoTroUo0 oq 0do1 iou reppolor1 6rop6oou proq 0qtrroq RNA. * AvnrepUortopdq rnq peroypoqrlg oulrpo[veL en[or1g roL oro oneprivro rou rRNA. 0 Kuxtrog rou Borrqplorou Ayyetrrorprjpou RNA * Iro porrrlpto, n UeroypoQn ror 11 ;leroqpoon npoyU0- ronorouvtor rouroXpovo (ou(euyU6vn Ueroyporpl/Uerc- Qpoon), ro069 ro p$ootoporLo [errvouv q perogpoor evoq mRNA npLv otrortrr1pol0e[ 1 ouv0eofl rou. * To poKTrlproKo mRNA e(vor ooro069 Ue Xpovo rlpr(orqc trQov trenrtiv qq topog. s 'Evo BoKTnproro mRNA pnope( vo e[vot notruotorpovtro ge notrtrrlg rordrreg neproXdg nou ovTrororxouv oe 6ro- qoperLro yovi6ro. nepitrnUn ffi ag.i*,#,* 5! L PJ't ,fiFts{ t:t &:!( l xolefuor xtrrilvog p4-expoyeio (non-template strand), evri: ow6g nou l,enoupye( og er<proye(o yro rrl o[v0e- on rou RNA rq}'ekor rLctrvog exproyeio, 6no:g tpofue- ror ornv EIKONA 19.1. H oUv0eoq tou RNA rotoLUeror on6 ro 6v(upto R- NA notruyep6o4 (RNA polymerase). H peroypocprl [e- rtvd 6rov q RNA noluprepdor; npoo6e0e( orov unoxrvqr{ (promoter), puo'er6rrcl neproXrl, orrlv opXtd rou yovr6(ou. O unorrvqr4g nepil.oprBdver to npd:ro (erlyog Bdoecov nou preroypdgeror oe RNA (ogerqpio - startpoint), roOrig Kor rrq neprBdl.trouoeq Bdoerg. An6 ourj q 06oq, r1 RNA noluprepdo4 perorrve(tor xo- rd pqrog rou erpoye(ou, ouv05rowog RNA prdXpr vo qrdoer oe Uro otr).4},ou1(o repporror{ (terminator), 6nou retrerc0ver ro perdyporpo. 'Erot ;uo peroypogrxri pov66o (transcription unit) erre(verqr qn6 rov unort- 512 KEOA,A|O 19 Meroyporprl oroug l-lpor<oputirreg Boorxeg cworeg
  • 5.
    K),6voq un-eKuoys[o 5' TAC* CG ET,qC G GTCAATG CAICTAC CT 3' .l';':;,:j !:.: i1:t-i:ii:l i.; lt & I'jp.'i-l G jrt +,,$*i * A vrlrri 6oq rov repUorrmrl. To Ko(pro Xoporrqptortx6 rnq Ueroypo<pu<qq provddoq, 6noq <po(verot orqv ElK0- NA 1S.2, e[vor 6rt onotel,e[ 6vo rprrlpro DNA nou Xpnorpo- noe(tor og expoye(o yro rnv nopoycoyrl ev6g provo6trof prop(ou RNA. Mro peroypogrxrl provd6o pnope( vo rto6t- ronoe( neptoo5rcpo on6 5vq yov(6ro. Or oMrll,oufeq nou Bp(orowor npw on6 rnv oee- rqp(o ovogdpowor ovo6rx6g (upstream), evdr outdg nou Bp(or<owor perd qv ogeqp(o (g5oo orq Ueroypo- q6pevq ol,r1LouXio) rog xo0o6rx5g (downstream), Iuyyd 4 oMrlloul(o tou DNA ypdeeror 5rot c0ore vo <po(verot pr6vo o rd.cbvog prrl-erproyeio, o ono(og (6no:g npoovo<p6p0r1re) 51et rrlv (6to oMrlloul(o pe ro RNA. Or O5oerg rov Bdoerrlv optOpo0wot Kot npog rtq 6fo roteu0Uvoerg on6 qv ogerrlp[o, nou ovopd(etot + 1, or de oprOpro( ou[dvouv npoq rnv r<o0o6u<q <popd. H Bdoq Trprv rrlv oqeqp(o oprOprefuot -1 Kor ot opvnrt- r<o[ oprOgo[ oufdvouv nrlyo(vowog ovo6trd. (Kop(o Bd- oq 6ev 61et rov opr0;r6 0). To opXrr6 npoi6v rnq Ueroypoqrlg nepil.o1-tBdvet to ou0ewrr6 5' drpo ror ovoprd(etqt rporoyev6g per6- ypqeo (primary transcript). To npcoroyev4 pretdypogo rou rRNA Kqr rou IRNA utp(orowqr pto 6epyoo(o top(- Uovonq rord rqv ono(o otrLr1trouX(eg'rd:v drpov toug r6Bowor on6 ev6ovourd.edoeg. To cirptpro npoi6wo trrrv -35-10-1 +1 +10 --.+ Avo0rro KoOoDLr<o IVleroyporprrq povodo elvor 1uo otrtr1trouXio DNA nou peroypoqerol o' evo evrolo RNA, [extvtlvrog ono rov unort- vrlq r<or retreulvovrog orov repportor1. wNw{s=Ktrtirvoq erpoyaio METATPAOH RNA getoypotpo ElK0liA 19.1 AeLroupyro qg RNA notrupepoorlg e[vor v' ovlypoqet 6vov r],6v0 rou d(r],orvou DNA oe RNA. H otrtrr1trou1ro rou RNA ervot I oupnlqpupattr4 ge rov xtrtrlvo tergoyeLo xat raur 604 p4 ye rov <otdtronotrlrtxo rrtrovo = 5'UACGCGGUACGGUCAAUGCAUCUACCU onepov(rov rRNA r<or IRNA e(vot moOepd, trre 1p6vo (or1q nop6poro Ue ro Xp6vo yevtdq tolv Borcrqp(ov. Av- r(Oero, ro npcoroyevrl getdypogo rou mRNA 66Xowot oXe66v dpreo4 en(Oeor1 on6 ev6ovouxledoeg r<t e[u:- voux.).edoeg. 'Eror, o prSoog Xp6voq (onq tou Borcrqpto- r<of mRNA e(vor pr6vo 1-3 Lemd. Iroug europutiteg, to pretdyporpo rRNA ror IRNA utp(orowot Korepyoo(o t<ot ro npoi6wo nou npor[mouv e(vot oroOepd, 6nog mo Borrqpro, oLl.d to mRNA e[vqt noLti nto oroOepd on' 6,r oro Borcrrlpro. Ot rpononoqoetq Kot 11 ono66ptqoq rov mRNA 0' ovoLu0oriv ctrlv Ev6trya 19.23. H preroypogq e(vor ro np6to ord6to orq Srgpooq rcov yovrd(rov xor ro Brlpo sro ono(o oorefuot nto ou;yd p(Opuoq. Or puO;umtxo( nopdyoweg ouprd r<o0op((ouv ov 6vo ouyxer<prpr6vo yov(6ro 0o pretoypo<pe( pSoo qg RNA noLuprepdonq. En(oqg elSyXouv rot petoyevSore- po mddro rnq Ueroypogqq r<ot dMo mddto qq 6r<- qpoonq ro:v yovr6(olv. Ario orlprowu<6 epor(proro oletrd Ue rn Ueroypo- qq efuor: o t-lcirS Bp[or<er q RNA no].uprepdorl unortvrlt6g mo DNA; Aw5 eivor pr6poq ev6g yevu<6tepou epotrlpo- rog: l-lig (eXcop((ouv or nporefveg ng ouyrerptpr6- veg 06oerg np6odeoqg mo DNA on6 ttg dl,l,eg oMqLouX(eq; o flcirS oLLrllentdporiv or puOprorrr5g npoteiVeg trle rqv RNA noluprepdo4 (r<or pero(U toug) rbme vo evepyonorrloouv 11 v' qvome(trouv ouyrerptpt5vo Brigoro rotd rrlv Svop(r1, r4v entprlruvorl ri tov rcpprotropr5 qq uerqypoeriq ; Ie our6 ro reqdtroro 0o neprypdtloupe rtq otr4Le- nr6pdoerg qg Borcrqpror{g RNA no}.ugepdoqq Ue ro DNA, on6 qv oplu<ri enogrl rrlq pe tov unoxtwltti, r1 Ueroypoen Kor rnv oneleu06ptoon qS on6 to DNA p6- l,rg oloxl4pto0e( ro pretdypogo. Ito KeEdl.oto 20, 0' ovol0oouge tg ow(motleg owr6pdoetg prero[U rov europutoru<c0v RNA notruprepootirv Kor rov ergoye(<ov roug. To Kerpdl,oro 26 nopouotd(et dtdtpopouq rp5- nouq Ue toug ono(oug or pu0ptmtrSg npolteiVeq Kot ot nopdyoweg prnope( vo po4Orloouv ti vo nopepnod! oouv rn Bonrqpror<q RNA notruprepdorl vo getoypdtlet 6vo ouyrerprpr6vo yov(6ro. Iro Ke<pdloro 27 e[erd(ou- 9.1 Erooytoyq 513
  • 6.
    19.2 H Meroypogri Zeuy6polpoBdoeorv A(euydporou DNA llpoylroronoreiror pe o€ Uro (Ouoo^i6o,) pe nrilq or [elotpror6g pu0;uorrr5g owrdpdoerg gnope( vo ouvdud(owor oe nol,unLor6repo 6(rcruo. To Kegd- Loro 30 ovoL0er np6o0eroug rp6noug prlOpuoqg, pero- [ti dMtov n Xpnorporofiorl gu<pc0v RNA, r<or qvo].Uer nrilg ourdg or oMqlenr6pdoerg prnope[ vo ouvddowor oe peyolUrepo 6(rcuo. roypogrx{ quoo}.[6o", mrlv ono(o ro DNA nopo6rxd 6toXrr:p(eror orouq 6Uo provo0g drc0voug rou Kor o xi.cbvog extrroye[o Xpnorpono€(rot yro vo xqreuO0ver q o0vOeorlrou xl.drvou rou RNA. H oLuoGo rou RNA ouw(0erot on6 to 5, npog ro 3, dxpo pre npoo0qr<q v6ov voux.l.eon6(ov oro 3' drpo rqg ou(ov5;revrlg oi.uo6og. H 3'-OH opd6o tou reLeuro(ou vourlsort6(ou nou npoor60rlre or4v ol,uo.(6o owrbpd pr' 6vo eroepl6;revo 5'rpupoorpopx6 vourLeoo(6ro. To etoepX6prevo voux.l,eoo[6ro Xdver ng 6Uo re].u<6g gco- o<popu<6q oprd6eg (y r<or B), evr6 4 opd6o o Xpnoqonor- efuot oro goorpo6eoreprr6 6eopr6 nou ro ouvdder arnv qluo(6o. H roXUr4ro qg ouvoLx4g ow(6pooqg yro rr1 Borcrqpror<f RNA noLuprepdorl prnope( e(vor dog -40-50 vouxLeor6ro/6eurep6l,emo orouq gToC yro ro nepro- o6repo prerdypogo, nep(nou (oog pe qv to1tirqro pe- rdgpooqg (15 opwo[do/sec), oMd no].ri ;ur<p6repq on6 rrlv rolrir4to owrypoqqg rou DNA (-800 bp/sec). H RNA noLugepdorl 6qpuoupye( rr1 ;reroypogrxrl <puooL6o pr6).rg npoo6e0e( o' dvov unorrwlrn. H ElK0l{A 19.4 nopouord(er qv RNA noLu;repdoq vo perorrvefuor r<ord prqrog rou DNA, pre q guool(6o vo peroxrve(ror po(( qg Kor rnv o).uo6o rou RNA v' ou[dva oe pulrog. H 6repyoo(o rou (euyopcilUoroq rov Bdoerov Kor rrlq npoo0{r<r1g Bdoecov ;r5oo orq <puooA(6o roro.)Jeror xt enr4pefuor on6 r4v (6ro ulv RNA noi,uprepdo4. H 6opr1rrlg quooL(doq 96oo oro oritrrnLoro rnq Ue- royporp4g go(veror ar4 Aemopepn dnorln qg E|K0NAI 19.5. Ko0rig q RNA noLuprepdor; perorrvefuor r<ord prj- Koq rou DNA erpoye(ou, ro 6(r),ovo (erultyetor oro pnpoorrv6 pSpoq qg guool,(6og (onUe(o [erutriyprorog e H RNA notrupepoorl droXropr(er roug duo rtrdrvoug rou DNA oe pro eqrlgepl ,,rpu00L[60" Kor xprlouon0re( rov 6vo r],tirvo org ergoye(o yro vo roreu0uver rr1 ouvOeo4 prog oupntr4prrt- porxrlg otrtr1trouXLoq RNA. * To Unroq qg quootrrdoq e(vor'12-14 bp evdr rou uppr6(ou R- NA-DNA p6oo orq quootrOo 8-9 bp. H peroypocprl Xprlor;ronore( q ouprnL4prollolx6rqro ro-lv Bdoecov, 6ncog xor or dMeg owr6pdoetg noLuge- prol.rori, dqLodq 11 owrypogq xoL 4 ;rerdgpoorl. H .,',, ': ,', . ono6(6er q yevrKr1 opxn rnq ;retoypog4g. H o0v0eo4 tou RNA npoyporonore(ror pr6oo oe Uro (Ue- Kodtrdg x.tirvog Ktr6voq eKuoye(o 0r rtrtirvor rou DNA dtoXrop((ovror yro vo oX4porLoouv Uro UeToypoQrKrl rpuootrido. To RNA ouvrlOeror pe oupntrrlpotporxo (euyoputlro rtov pooeov pe rov evo ono roug r),(rvoug rou DNA. 514 KEOA Al0 1g MeToypoQ1 oroug l"lporoputirreg w;.@,TWrx"= ffifl.M-ffix ''r*er'ffi"x-ffi EIK0NA 19.4 H geroyporpq npoyporonorefror gdoo oe Uro quoq- tri60, orqv onoio RNA ouw(0eror pe (euydptopo rov pdoetov 1r, 6vuv on6 roug rtr6voug rou DNA oqv nprioropo [erutrryp6vrl neproXrl. Ko0tig perouve(ror q quootrbo, ro D(xtrrr:vo DNA [ovo- oXrlp0ri(eror n(oo u1g, exron[(ovrog ro RNA un6 popqrl povrlg nol,uvouxl,eo16xrl g otruo6og. To DNA onodroTdoor.o, I H oUvgeo4 Tou RNA [euvd mr1 quoo],6q Boorxdg evvoreg J + 0uootri6o peroypoqftg +,
  • 7.
    Merorw4oq rou ev(Upou '+ 05oqnpdo6eolg rou RNA EIK0NA 19.5 Kord q Ueroyporprl, I rpuootri0o Drorrlpeiror p6oo oq porqproxrl RNA no)upepdol, rou [erutrfyer xor [ovoru)ryer ro DNA rot ouv06rer RNA. - unwinding point) ror ro DNA enqvefurd)veror oro n(- oro rprlpro (o4pe(o (ovorullyprmoq - rewinding point), To pqrog rrlg <puool,[6og e(vor -12-14 bp, evcir ro ;uixog tou uBpt6(ou RNA-DNA pr6oo on1 rpuooLbo pr6llq 8-9 bp. Ko0rig ro 6v(u;ro petorwe(ror r<ord prrlxog rou eK- poye(ou, [ovoq4qpor((eror ro 6[r],rovo DNA ror ro RNA erron((eror o g eLeri 0ep4 nol,uvo u x.leorr6rnj ol,u o(6o. Ie rdOe 6e6op5vq onypf , ro rel,euro(o 14 prBovouxi.e- or(6to rou ou[ov6pevou RNA Bp(orowor oe oUpntror<o pe to DNA xo/r1 ro 6v(upro. 19.3 H Avri6poon Tnq McToypoefiC'Ex€l Tplo lrd6to " H RNA notrupepoorl npoo06veror oe pro 0eorl unoKrvrlTrl oro DNA yro vo oXqporloer 6vo r],eroro oupntroro. " H RNA notrupepoorl [exrvo q Ueroyporp1 orpou ovo[[et ro di rtrrr:vo rou DNA yro vo oXqporloer Uro UeToypoqrKrl rpuootrido. * Koro r1v enrpfruvor1, I Ueroypoqtrl rpuootrrdo perortvelror roro pr;roq rou DNA ror 1 otruo[do rou RNA entplruveror oe roreuOuvorl 5'+3' pe npoo0rlrrl vourtreortdlrrrv oro 3' orpo. * H Ueroypoql oroporo rot ro dlrtrtovo rou DNA [ovooXlporr- (erot gotrrg 1 RNA notrupepoorl onoouvdeOel ono ro DNA er- poyelo oe Uro eeon repportolrou. H ow(dpoon rnq Ueroypognq drorp(veror oe rp(o orddro nou cpo(vowor ornv i', ' : (1) 6vop[n (initiation), orrlv ono(o ovoyvrop((eror o unoxrvrltrlg, 6q;rroupyekor Uro guoo).[6o r<or (erLvd 4 oJv0eorl rou RNA, (2) enrp{xuvon (elongation), orrlv ono[o 11 guool(6o ;."tetoxrve(ror r<otd pr1<og rou DNA, r<or (3) repponop6g (termination), xord tov ono(o to RNA perdypo<po oneleu0epcbverou<or r<).e(ver q rpu- oo).(6o. H (6to q 5vop[r1 prnope( vo 6Lor<pr0e[oe noMd Bqpro- to. H ovayvdtpon rou expoye[ou lexwd pe u1v npoo6e- ENAP:H Avoyvtirptorl rou expoye(ou: 11 RNA noLugepdol npoo0everoi oro 6[x],rovo DNA To DNA {erutrryerot orov unortvrlq llotrrj ppoXe(q qtruo6eq ouw[0ewqt Kr oflelsueep(tvovror EflIMHKYNIH: 1 H notrupepdorl ouv06ter RNA TEPMATIIMOI: H RNA notrugepdon Kqr ro RNA onetreu0epdrvovror ElKOllA 19.6 H peroyporprj 6xer rp(o md0rq: To 6v(upo npoo66- veTor oTov unouvrlr4, onodroroooer To DNA xor nopop6ver or(vr1- ro xord qv 5vop(1: Merouve(ror rord plrog Tou eKpoye(ou Kqrd r4v enrp4r<uvq rqr 6r(ororor orov reppouogd. oq rnq RNA nohupepdo1g oto Sklavo DNA oe pn aA- l4louy[a DNA nou xaleftcl. unoxlrpr1g @romote!. To 5v(upro oXrlprot((er np6ro 5vq xtreror6 o0prtroKo (closed complex) oro ono(o ro DNA nopopSvet 6(r<Lro- vo. 'Enero, (eruLfuer ronrd ro tprrl;ro DNA'rou unorr- vqrrl nou neptloprBdver rn eSon Svop(r1g r4g peroypogqg yro vo q4qpot(oer ro ovol;6r6 oripnAoxo (open complex). O droXroptog6g rou 6u<Lcirvou DNA ro- Ormd rov r<i.c0vo-er<proye(o 6ro0Sorpro yro (euydpcopo Bdoecov pre eroepX6trrevo prBovoux).eor(6ro xor oriv0eoq -rcov npcbrrov vour.treon6rrc0v 6eoprdlv oro RNA. H qdon ouq prnope( vo nopo'ro0e( qn6 qvenrule[g on6nerpeg Svop[4q, orrg ono(eg ro 6v(upro qndprer BpoX6o pret6- ypoqo, runxd rdro rulv -10 vouxleond(cov (nt), evcir e(- vot or<6;rq npoo6e6epr6vo orov unorrvrlri. To 5v(upo ou;yd r<dver 6ro6oXxorig 66xtroug ovenrulcirv Uerqypo- grilv r<or lovopX((er q o[v0eoq rou RNA. H <pdo4 rqg Svop[r1g re^.er6vet 6rov ro 6v(upro retrrd r<oropOcOoer v' enercrefuer qv ol,uo[6o Kor vo onopor<puv0e( on6 tov unorlqrrl. Krivoq erpoye(o DNA lii .: H AvTiDpooq rrlg Meroyporplq'EyerTpLo Iro0ro 515 Boorxeg eworeg
  • 8.
    H enLp-rrp<uvoq ouv(orororoe npoo6eurrrrl preror<fvq- or'l rou ev(0pou trre 6tordpo(r1 tou (euyopri-rprorog rov Bdoeov oro d(x.tro.rvo DNA, ogrlvowog erreOerpr5vo rov r<Lr.rrvo-expoyeio yro npooOrl<q voux).eorL6(rov, Kor Ue- tot6nroq rnq peroyporpLxqg guooLf6og nro ro0o6rxd. Ko0rrrg perorrve[tor ro 6v(upro, o x],covog-exproye[o qg npoorrrpLvd (etuALyprdvlg neproXrlg (euyoprrtver pre ro veoouwe0eLpdvo RNA crro o.r;pe(o qg o0(r1oqg. Nou- r).eori6ro npom[Oewor ol.rorono].Lr<d oro 3' dr<po rrlg oulovdpevrlg oLuo(6og RNA, o'Xqpor((owog 6vo uBpG 5Lo RNA-DNA ;r5o'o mqv lerulryprdvrl neptoXrl. l-1(oot on6 rrlv [erufuypr6vr1 nepropl, o r<].ovog-er<poye(o rou DNA (euyoprrrveL pe -rov ouOeyrrrd r<),rrivo "ro[pr" rou, yto vo lovooXripor(oer rq 6rn).r1 dLrr<o, evctt o ou(ovdpe- vog N).rrlvog tou RNA [enpoBdtr].eL ond ro dv(upo. H nopo6ooLorr; on6r,l.rr1 6l r1 enr;rrp<uvor1 e(vor ;rovo- rovq 6tepyoo(o, o-rqv ono(o -ro 6v(up_ro npocu0e(ror r<otd pqrog rou DNA pe oroOeprl rolirqro nou owrmotle( ornv rrpooenKn rcuv voux).eorr6(olv, dXeL oMdler ro re- Leuro[o XpdvLo. H RNA no].uprepdorl npoyporonore( nor-ioq 11 ordpq ror oropord oe ouyKeKprpdveg oMrl- trouX(eg. H em6nrorl rou 3' dxpou rou RNA ond ro evepy6 r<6,wpo trrnopei vo nporol6oer "ofiroeox6prlor)" rrlg noLupepdoqg r<oL ogo(peoq ;_reprr<rr.lv vour<LeorL- 6(ov on6 trlv cu(ov6prevq o).uo[6o rou RNA fiprv rr]v enoverr[v4oq. O rep ;.rottopr6g nepiloprBdvet ovoyvcirptor; oMrlLou - Xrorv nou 6(vouv mo 5v(upio ro orlpro vo oropotrloeL r4v nepondpot npooOqr<r1 vour<).eol6iov ornv o).uo(6o rou RNA. H guo'o}.(6o xoroppdel<oOriq ro uBp(6Lo R- NA-DNA onodroroooeror Kor ro DNA lovobq;rtoupye( 6vo E(xLrr:vo, 11 611 puoupy(o goo<podLeoreprotv 6eo pur-rv dLoxdmeror rar ro orjpn).oKo Ueroypogqg 6tolrrip((etor oro ouorolKd on6 ro ono(o onopr[(eror: RNA no].upe- pdoq, DNA r<oi DNA ;reroypo<po. H aAA7Aouyfu rou D- NA nou xareu9lva r ArjEn rnq psraypaErlq ovolto(erat reppau.orq. 'l 9.4 H Borrnplorf RNA lloAuUepdon AnoreAeirot on6 noAAdq Ynopovd6eq ". 0r nuprlveq rov Borqproxtrrv RNA notrupepoorirv oivor notru- pepfl oupntroxo'400 kD pe yevLrrl 00prl 0208'0r. ., H rorotruorl enustrefrot ono rLg unopovodeq p ror B'. OL r<o).rjtepo XopqKrnprotl6veg yevettrd xor BtoXqpru<d RNA no).upepdo'eq npo6pXowor on6 Bor<rr1pro, t6(<i.rg on6 ro Escherichia coli. Oo16oo, oL p6veq BorcrrlpLo- r<59 RNA nolupepdoeq yro rtq ono(eg 51ouv npoo6ro- pLore( uUnlrlS ovd).uo11g rpuoro).Lrr<6g 6op6g, npodplowoL on6 60o Oeppr6gil.o Borcrqptor<d e[d4, ro Thermus aquaticus xat Thermus thermophilus. Ie 6o ro Borrrlpro, p6vo 6vog r0nog RNA noAugepdorlg ei- vor uneU0uvoq yrq rq orivOeoq rrov rRNA, mRNA xor IRNA, qw[0ero on6 toug europucirreg, srouq ono(oug ro rRNA, mRNA rqt IRNA runxd preroypdtpowor on6 6rogoperu<59 RNA noluprepdoeg, ng Pol l, ll xot lll. l-lepfuou 13,000 g6pro RNA noluprepdorlg undplouv oro rUrropo rou E. col/, nop6rr o orprB4g opr0;rdg norrf],er ovd],oyo Ue trq ouv0rlr<eg oU[r1oqg. Irqv npoyllorrr6rnro, oe rd0e 6e6opr5v4 orryprrl 6ev ep- nA6Kowqr 6).eg or RNA noLuprepdoeg orq psroypo- <prl, 6l,eq 6prog e(vor npoodedepdveq oro DNA, e(re et6rrd e[re ;rr1 er6rrd. To n)r1peg 6v(u;ro 11 otro6v(upo (holoenzyme) rou E. coli 51er propror<6 Fdpoq -460 kD. To o).odv(upro (qzpp'oo) pnope( vo droXroptme( oe 6Uo oumotrxd, rov nupr;vo rou ev(riprou (ozpp'to) Kor rov nopdyowo o(ypo (notrunenr(dro o), nou epnL6reror er6rxd orqv ovoyvcilprorl rou unoKrvrlrrl. H oUorooq rou oe unoUo- vd6eg ouvor!((erot orrlv EIK0NA 19.7. Ot uno;rovddeg B ror B' go(( rpoyUoronoroUv r4v r<ordl,uoq rou RNA ror qnore^,o0v to preyol,rirepo rpulpo r4g pd(og rou ev(0- ;rou. Or oprLvo[rrdg o].].r1].ouX(eg rot ot rpro6tdmoreg 6op6q rouq e(vot ouwqpnp6veq, x6tt nou rqxuer yro nq peydleg unopov66eg rtov RNA no).upepoociv r<or on6 rtq rperq enrxpdreeg rqg (or1g: Bor,rrlpto, oplo(o xr eu- t<opuc0reg (B).6ne Ev6rrya 20.2). Auto uno64l,cbver 6rr or Boorr5g t6r6rrpeg rnq Ueroypogrlg e(vor roweg prero- [ti rtov no].upeprirv RNA no].ugepood:v 6].cov rrov opyo- vrogd;v. Or B xor 9' Uo(( oXq;ror((ouv ro evepy6 r5wpo tou ev(6prou, rov r<ripro 6(oulo 6ropr5oou rou ono(ou 6rSpXeror ro DNA xo-rd tov r<0xi.o rqg preroypogrlg, ro 6ewepe6owo 6(ou).o 6ro;r6oou rou ono[ou ro prBo- vouxi.eo{6ro rou unoorpriporog ero6plowor oro 6v(u- [ov[6to llpoiriv Aenoupy(eq rpoB B unopovddo (r51 kD) rpoA 2 o unogovd6eq ftA Iuvopgotrdyqorl Tou ev(ugou ;; ^; i'r ql7 ovoyvopron rou unoKrvrlnl xoOeg(o) npooddver rdnotouq 1|i'-onoqrdq l- Kurotrurtxd x6wpo glrpoC p' unopovddo (1 55 kD) o unopovo6o (1 8-70 kD) Etdtxorrpo rou unorrvrlrrl or unopovd6o (r 0 kD) 'Ev(ugo rou E.coli =460 kD 0t RNA notrupepooeg rov euporqpiov eXouv ne.- re tunoug unopovo0ov: oL o, p, p' ror o eXouv potrtrov oroOepc U6ye0oq oro dlorpopetro erDrl porrrlp(ov, evtir 11 o norrltrer ne - prooorepo. rpoD rpoZ 516 KEOAAAI0 19 Meroyporprl oroug llpoxopurireq Booxdq 6woreg
  • 9.
    uo orrlv nope(orouq npoq ro evepY6 KSwpo Kot ro 6[- ou^o e[66ou 6ropr6oou tou ono(ou to veoouweOetp6vo RNA eyrorotre(net ro 5v(upro. Ie oup<pcov(o pre ro p6Lo ourrilv ttov unopovddcov oe 6l.eq ow5q ng Lettoupy(eg, prerotrtr6(erq oro yov(5to rpoB xot rpoC nou xtodronot- oriv lg unoprovd6eg B ror B' en4ped(ouv 6tro ro ord6to rqg pretoypogqq. To 6tprepdg nou oXrlproil(etot on6 ng 6Uo q unopo- v66eq Xp4orpre0er toq u<p(cr:pro Yto rrl ouvopprotr6yrlo4 rou KewprKo6 ev(0pou. H C-retur{ neplolli (C'termi- nal domain - CTD) rov o unopovd6tov en(oqq 6pXetot dpreoo o' enog( ;re to DNA rou unoKtvrlrrl rt Stot oup- BdMer orrlv ovoyvcilptorl rou unortvqr{ (B}.5ne nopo- rdrco). EntnL6ov, ot unoprovd6eg o Kot o e(vot ot rUpteg enrgdveteg qq RNA nol.uprepdorlg yt' oAtrrlLenrdpdoetq pe nopdyoweq nou pu0pr((ouv qv 6vop[q rrlq Uero- ypoqnq. H unoprovddo to en(or1g no(et p6lo orrl ou- voppol6yrlorl rou ev(6pou, 6nog en(or1q Kot oe ouyr<erpq.rdveg pu0ptorrxSg ettoupy(eg. H unopovdEo o e(vot rotd r0pto tr6yo uneU0uvrl yo rrlv ovoyvdrpton rou unortvrltrl. H xpuorotrtrtr<{ 6opf rou nup4vo rou Bor<rqptororj ev(rlprou 6eiXveL 6tt 51et 5vo oXrlgo oo 6oyr<dvo xoBouptorl, Ue rn Uto 6oyrdvo vo oXq;roil(etor tcup(cog on6 rq B uno;rovd6o Kot rnv dtrl.q rup(rog on5 q B' unoprovd6o, 6nog go(verot ot4v Ell(0NA'1S.8. O npoteUorv Dfoutroq yto to DNA Bp(oretot mqv enrgdveto eno<pqg rov unogovd6tov B t<ot B', ot ono(eg oroOeponoto0v tou g 6toltoptoprdvou g, provoti g r<).cirvouq orrl Ueroypo<prxrl quool(6o, 6nog rpo(vetot orrlv EIK0NA 19.9. H 05or1 rotd).uor1g Bp(oxerot orq Bdorlrqq q4totrrqg nou oXrlptot((etot on6 rq (ooY6vto" roJV 0 rot B'. 'Evo H ovodrrl onogl rou nuprlvo qg RNA notrupepo- o1g, onou qo[verot ro oX11.to ,,doYrovdtv ro$ouptou, Tou ev(u- Uou. 0r 0 (ruovo) rot p' (poQ unopovo0eq i1q RNA notrupepoonq eXouv evov 6[ou],0 yto ro DNA erpoyeto. H o I tpolverot npootvl, q o ll ruprvrl rol 1 o xornvl. tlpooopltoopdvo ono. K. M. Geszvain ror R. Landick (ed. N P Higgins). The Bacterial Chromosome, American Society for Microbiology, 2004 , t, H 6opr1 rou nuprlvo rrlg RNA no),upepoolg ono ro porrlpro Thermus aquaticus, Ue U g unopovoDo pntre rot q $' npootvq. Aopq ono: Protein Data Bank 1HOM L. Minakhin. et ai,, Proc. Natl. Acad. Sci. US/ 98 (2001):892-897. on6 to 6rjo roroluttrd t6wo Mg2* nou e(voL crnopofurl- ro yrcl ro prllovLopr6 rrlg rord).uoqg e(vot ogrXro npoo- 6edepr6vo oto evepy6 Kevrpo rou ev([pou (B].6ne Evoulta 19.18). To dMo L6v Mg2- tptovet crro evepyd x6wpo oe oUpnLot<o pe ro etoepx6prevo tpt<prooqopLxd vour.treori6to (NTP). Onoq npoovotp6pOqr<e, o nupqvog rou ev(rjpou rov tuKopuorov dlet qv (6to pootr<q 6o- prrl pe ro Borcr4ptor<6 dv(upo (Erx6vo 20.2). En(o1g ne- pr51er r<onoteg entnp6o0eteg unoprovo6eg KClt nepLLoprBdver oMrl).ou1(eg nou 6ev undpXouv oro Bo- rcrrlpLor<6 6v(upo. Or r<ripteg 6Lotpop6g ovdi.-ieoo mo Bo- r<rrlproxd Kor ro euKopuortKo dv(upo Bpior<ovtoL oXe66v onor).etorLro mqv nept<pdpeto rou ev(0;rou, uoKpro ono ro evepyo Ktvrpo. 'l 9.5 To OAo6vCuUo AnoreAeiror on6 Tov Tov nopdyovTo o Tnq RNA l-loAuuepdonq Iluprivo Tou Ev(tlpou Kot a H Borrlptorl RNA nolupepooq unodtotpe[toL orov 02BB'o nuprqvo rou ev(upou nou roro),0et rl1 UeroYpotp1 KoL Tl'lv uno- povodo o nou onouoltot ltovo yto rr1v 6vop[q. * 0 nopoyovro-Q o rpononote[ rtq tbtorlreg npoodeorlq oro D- NA qg RNA notrupepoorlg, ersL 6ore q ouyy6veto Tlq rpoq ro yevlr6 DNA vo ptet6verot xot, ovr[0ero, 1 ouyyeveto qg npoq Touq unouvrlrdg v' ou[ovel. O nuprlvog rou ev(tlptou 61et yevtr<q ouyy6veto Yto ro D NA, xu p(r.og ).6ytr-l rllercrpoorotLxrrtv oMqenL6pdoe- oiv 1.rero[[ tqg nporelvng, nou e(voL Bootr<r1, Kot rou DNA nou e(vot 6(tvo. Otov npooddverot mo DNA ror' our6 rov rp6no, to DNA nopopevet oe 6(r<trovq lfr.li To Otrodv(uUo qq RNA tlotruUepoonq Anoretreiror ono rov l-luprlvo rou Ev(upou roirov I-lopoyovro o 5'17 Boorxeg eworeg
  • 10.
    / i r::,1 ,.T -"' I llopoyovrog o 0 nupqvog rou ev(ugou npoodeveTot odtorplrog oe roOe DNA. 0 nopoVovrog o perdlvet rr1 ouyy6veto yto npoode- oq ove[opqrq ono rqv otrtr1trouXio ror npoo6[6et eldtroqro yto roug unorrvlreg, Uopqn. O nup1vaq tou ev(l1tou 6yeLqv xav6trya va ouv06teL RNA an6 6va DNA expayeb, oMd 5e pnopel v' av ayva p [(er uno xwryt 6q. H Uopqri rou ev(ripou nou e(vqr uneri0uvq yro rnv 6vop[q rnq Ueroyporprjg on6 rouq unoKrvqrdq ovoprd- (eror ol.odv(upo (o2BB'roo) (EIK0llA 19.10). Arorp6per on6 rov nup{vo rou sv(rlprou enetdri nepdXer dvov nopd- yowo o. O napayowag o$pa 6yt 1t6vo efaoEah((et 6tt q Baxtqpnxi RNA nolupepdorl evepyonoe[ rn pera- ypaqi an6 optopdveg 0ioerq, aMa en[or1q peuitvet xat qv np1odeory oe pr1 edxdq a&rltouy[eq. Xop(g nopd- yowo o, 11 oro0epd ouyydverog yro rrlv np6o6eor1 tou nupfvo rou ev(Uprou oro DNA per6verot rord -104 rpo- p5q r<or o 1p6voq npr(onq rou ougnl,6rou e(vot <1 6ewep6Lenro. Aw(0ero, ro otro6v(upro npoo66vetot fiouq unoKrvrlrSg noLrj nro orptXrd, pre oroOepd o'uyy6- verog xotd pr6oo 6po -1000 gop6q UeyoUropn Kor Xp6vo ngr(onq 6o9 r<or oprer6q cirpeg. 'Etor o nopd- yowoq o ouorcrorrKd onomo0eponore( q pq etStrrl np6odeoq rou unorwrltj. H roX0rrlro pe rnv ono(o ro oLo6v(u;ro npoo66ve- ror oe 6rorpoperu<59 oMrlLouX(eg unorrvqrc0v nopou- ord(er peydl,eg 6ror<updvoetg ror yr'our6 onore)re[ orlprowrr<4 nopd;rerpo arov ro0optop6 rnq "oy0oq rcu unoxwt1r4", drllodTl rqg onotetreoponx6rqrog ev6g (e- Xoptoro0 unorwrlq oq npoq qv 6vop[4 rnq Usroypo- qnq. H ouXy6rrlro rqg Svop(qg nox(trer on6 -1lsec yo ro yov6ro rou rRNA un6 rSovrx6g ouv0{reg 6o9 < 1/30 min yro rdnoroug dLLoug unoxrqt6g. O nopdyowog o ouv40cog onetreuOepc0verot 6rov 11 otruo(6o rou RNA gtdoet oe ;rr1<og -10 nt, oEtivowog"rov nupfvo tou ev(Uprou une0Ouvo yro rrlv entpr{ruvot1. 0 o onomo0eponore( q gr1 el0rKn npdo000n I 9.6 I'ltilg Bpiorer n RNA lloAuuepdon rtq AAAnAouxiaq Tov Ynomvnrdrv; a H npoodeo! rrlq RNA notrupepoolg oTOUS ulloKtvrlTeg eivc. rooo ypnyopn tiore gotrlov 6ev nporeuot yt ontrl OtoXuo1 ," H RNA notrupepool npooDevetot oe ruXo(eg Odoetg oro DN" xor trq ovrotrtrooer pe otrtreg otrtrr1trouXteg 6oq orou Bpe0s. evog unorrvlrrlg. H RNA nol.uprepdoq np6ner vo Bp(oret unortvqt6g ;.re- oo oro nLo(oto tou yovt6uol-rorog. I-1cog [eXorp((ouv c. unorrvrlrdg on6 trg 4 x 106 bp nou onopt((ouv rc un6Lorno pSpoq tou yovtdrcbporoq rou E. coli', N nopouotd(er ontrd prow6Lo yro to ntrrg q RNA nouprepdorl 0o pnoporioe vo Bp(oret rtq oMqlor-- 1(eg rorv unorrvqrcov prSoo oe 6eq rrg oMq).ou1[e: trg ono(eg pnope( vo npoonel.doer. To otrodv(upo r,': RNA noLupepdonq ewon[(et ro Xptopr6ooLio Ue rL- 1o(o 6rdXuoq ror npoo66veror pr1 erdtrd sro opvllr(c cpoprtop6vo DNA. Ie our6-ro pow6lo, to oo5v(ut-: 6r(ororor noti yprlyopo. H 6tdXuorl 05tet 6vo ovrr-rrc- ro 6pto yro rr1 oroOepd qq roXUtrltog otiv6eorlq r-: rd(r1g rou <108 M'1 sec'l yro or6Xo 75 bp. l-ldwoc - npoyprortxr; moOepd npotrl0rlttrrlq toXUtqroq 1fo'- ward rate constant) yro rdnoroug unortvlt5c, in vtt': rpo(vetot vo e(voL >108 M'1 sec1, oro 6pro dLdxuo1c * xor ndvo qn6 our6, H 6nuroupy(o rot rotompocc- f-tog oerpdg oupnL6rcov 5og 6rou (r<ord tUXrl) rl RN: no).u;repdorl ouvowqoet 6vov unortvqrq xot 6rlptouc- y4oer 5vo ovotXr6 orjpnlor<o u<ov6 vq rpttd[et RNA E: 4rov oXerrrd opVn 6ro6Lroo(q. 'Etot, o Xp6vog no, onoLre[roL yto tuXo(ouq r<rix].oug dto6olrr<qq oJvdi- onq Kor 6rdmooqg oe Xolop6g 06oerq np6o6eoqg e .- vot noAU peydoq ror 6e prnope[ vo epprrlveUoet nc. Bp(or<er r; RNA nouprepdoq rov unoKrvnrq tr1g. Iuvo noq, n RNA no).u;repdoq npSner vo XprlotUonotei ci.- o prSoo yLo v' ovo(qrrloer lg O5oeLg np6o6eo4q rqc H Eu<6vo 19.11 6e(yyer 6rt 11 5to6u<oo(o nt0ov6v en.- rolUverot enerdrl o opltr6g m6Xog qg RNA notru;,a- pdonq e(vor oL6xLnpo ro yovt6(<r-r1.to, 6Xt pt6vo g.: ouyrerptpdvq o).r1Lou1[o unor<tvqq. Au[dvowoq r: pdye0oq rou or6Xou, 11 moOepd rqg toX6tqrog 6toX-- orlq oro DNA ou[dver ow(orotlo xoL 6ev e[vot ntrec, neproprmu<1. l-169 pretor<tve(rot ro 6v(upro on6 gLo t-- Xo(o 06o4 np6o6eorlq mo DNA o' 5vov unoKtvfr- I[prgorvo pe opretd 6e6o;r6vo, toutrdXtorov tpe(g 5Lc- <popertrSg dtepyoo(eg 6topoptptivouv l'lv toltitqr: ovo(qtrloqg unortvqtq on6 tqv RNA noLuprepdor I-1purrov, ro 5v(upo prnope( vo petortvrl0e( oe pto 1.tovc- 6tdoroq tulo(o 6to6poptrl r<otd prlxog rou DNA (,.oI,- o0r1or1 - sliding"). AeUrepov, enet6rl ro Xpropi6otr.rl.tc 6Xet 6or6oLridrl muXcor4 6otrrr1 oro Borrqptox6 nupr- voer66g, ;16l.19 ouv6e0e[ oe pro oMq].ouX[o rou Xpo[o- I ffiy r-"*W L"Ah!** '%ffi92 I To revrpLxri dv(uUo npooddveror oe onoLodrlnore DNA To olo6v(uUo npooDdverot mov unoKtvlrn 518 KEOAAAI0 19 MeToypoQrl orouq flporopudrreg Boorxdg eworeg
  • 11.
    EIK0NA 19.11 llporew6pevorUrlxovrouoi yro ro ntirg n RNA notruuepoon Bpioret 6vov unortvqnl. (o) otr(o0non (p) dLorplportxrl perotpo- pd(y)ev6oneproXrxrlouvdeoqxordrdotoolnovonnOnon.llpooopUoog6voon6:C,Bustamante,etal.,J. Biol.Chem.274(1999):166665- 1 66668. orilgorog, to 6v(upro Bp(orerot nMov nto xowd oe dtr- trsg O5oetg, yeyov6g nou petcilvet to 1p6vo nou onqtte[- ror yrq 6r6mooq rt enovonp6o6eorl oe pto 6IIn 06on ("dtotUrluortrrj getocpopd - intersegment transfer" i (ovonndnon - hopping"). Tp(rov, evril e[vot ouv6ede;r6- vo prq erdrd oe p[o 06o11, to 6v(upro gnope( v' otr].d[et O5oetq qro DNA pr6Xpt vo Bpe0ei o unoxtvqtfg (.dpreor1 prerogopd - direct transfer"). I 9.7 To OAodv(upo TpononoteiTot Kord Tn Arepyoofo Avoyvtipronq Kt An€Aeu8dptrlonq on6 Touq Ynortvnrdq * '0T0v n RNA notrugepool npoo66verot o' 6vov unortvrlrrl, dto- Xop((er iouq rtrtirvouq rou DNA yto vo dlptoupyrloet gto pero- vpoqrrrt rpuootrido rt evootgor6vet vourtreorldto oto RNA. & H evopfi ono qv RNA notruuepdoq 6ev e(vot novrore enttu- xts. * 0 nopoYovroq 0tYU0 ouvflOoq oneLeu0eptlveroL ono rrlv R- NA notrupepoorl otov 1 veoouvre0etp6vrl otruotdo RNA ono- rqoer pflrog -10 pooetrlv. Mnoporipe rdlpo vo neptypdtpoupe ro orddto rnq Ue- roypogrlg oriprgovo pe ttg olLrllent6pdoerg preto[ti ttov 6rorpoperrrrov ptoptpcov tqg RNA nolu;repdoqg Kor rou DNA er<proye(ou. H ow[6pooq 6vop[qg prnope( vo neprypoge( pe Bdoq ttg nopopttitpouq nou ouvoQ(- (owor orrlv : o H ow(6pooq oloev([prou-unoKtvrlrrl [ertvd pe to oXrlprolopr6 ev6q xretoro0 6Lpepoiq oupnA6xou, 6nog <po[vetot offlv Etr6vo 19.12(o). "K].etor6" orl- po(ver dtt to DNA nopop5vet 6ixtrtovo. O oXqgotL- opr6g rou xletoro0 6tprepoUg oupntr5xou e(vot qwt- orperrr6g. 'Etot, ouv4Oroq neprypd<petot on6 pto oro0epd tooppon[og (Ke). YndpXet preydLo e0pog orrg nprdq qg mo0epdg tooppono(og Yto ro cDCq- prottopr6 tou xl,etoroU oupntr6rou. o To xtreror6 orjpnloro petorp6netot oe ovotyr6 o01t- rrloxo pe "oflodtdroqn, ;rtog ptr<priq neptopiq DNA pr6oo orrlv oMqLou(o 5nou npoo6dvetot ro 6v(u- pro, 6nog qo(vetot orqv Etr6vo 19.12(9). l-to roug neproo6repouq unoKtvrlr6g, 11 perorponrl on6 to ovorXr6 oro xtretcn6 oriprnloro e(vor ptq owtorpe- nrj ror owri q ow(6poor1 prnope[ vo neprypoqe( on6 11 oro0epd npool0rlrxrlg toX0r4roq (forward rate constant) (kr). Oo16oo, pepxo( unoxwqrdg (n'1' ol unortqt6g tou rRNA), 6e drlprtoupyo6v oroOepd ovorXrd o0ptntroro rot our6 e(vqt to xLet6( yto 11 priOprori roug. O nopdyowoq o no(et orlprowtx6 p6Lo orqv owidpooq ri€nq GMne Ev6r4ra 19.19). Or preroBdoetg nou ou;tBo(vouv on6 tqv 6vop[4 ;t5- Xpr rnv entprlruvorl en(o4g ouvodeUowot on6 orl- ;rowrr6g oMoy6g orq 6ogrl Kot rn oOmoorl tou ougnl6rou, Ot oMoy6g oro oXrl;ro qq RNA notru;repdorlq ouvo- 6efouv ug t<tv4ttx5g ;retoBdoetg nou neptypdgnKov nporlyouprdvtog, ro0rilg r<ot q ;rerdBoon oro o6pntroro enrpulruvoqg (nopouotd(erot firlv EIK0NA 19.13). Ito xtretor6 oUpnloro, to ol,o6v(upro qg RNA noLupepd- oqg roLrimer nep(nou 55 bp- DNA, errew6pevo on6 -- 55 5og -+ 1. To 6[x].tovo DNA npoo65vetot rup(tog xot6 priKoq rqq puog ntreupdg tou oloev(ripou, 6nou SpXetot o' enoqrl ;re nq C-refur6q neptoX6g rov o uflopovd6tov roOdrq Kor Ue rtq neptoldg 2 xat 4 rnq o unoUovd6og (BL6ne nopor6to). Kord q getdBooq oro ovotXr6 o6pr- nlor<o, q drop6prpoorl t6oo qg RNA noluprepdoqg 6o'o Kqr rou DNA oMd(et. Ot nto dpoonxSq oMoy6g orrl 6o- Uri rou oupnl"6rou qo(vowot crqv Ex6vo 19.12: (o) ptto xdprUn rou DNA -90 potp6v, nou enrpdnet orov xtrtivo "i1l ? To 0tr06v(upo Tpononore[ror Koro rr1 Arepyooro AvoyvtirpLolq rr Anetreu06ptlorig ond rouQ YnoxLvnte c 519 Boorxfg eworeg
  • 12.
    eKpoye(o vq npooeyy[oerro evepy6 x6wpo rou ev(6- Uou' (B) dvorypro tov x.trcirvrov rou DNA rou unoxwrltl pero[6 --11 rqt +3 ovoEopxd pre qv ogerrlp(o rnq Ue- roypocprlg r<or (y) r<tre(orpo rov (ooyovrcbv> tou ev(0prou ciore vo neprruxl,cioer to rpui;ro rou unoKrwlr4 nou Bp(- oKeror r<o0odxd on6 rrlv orperlp(o preroypocprlg. Iuve- ncilg, ro oqge(o enorpriq rou ulroKrvrluj mo ovorXr6 oUpnl,oro erre(vowor on5 --55 oe -+20. To en6prevo Brigo e(vot vo evooUorco0oUv rq 5[o npdrro voux.treot(drq Kor vo oXqpronore( rivog <ptrlorpo- 6eorepx6g 6eopr6q prero[ri roug. Aur6 drlproupye( 5vo rproroy6q o0pnAoxo (ternary complex) nou nepr6ler RNA, DNA xor ro rlv(upro. Iroug neproo6repouq unoKr- vqr6g, qXqpror(eror puo oLuo(6q RNA nou SXer pr4r<og opr<er6g Bdoerg, Xrr:p(g getor<(vqoq rou ev(rjprou xotd prrlrog rou erpoye(ou. M6Ltg npoore0e( puo Bdoq, un6pXer rdnoro nt0ov6- rrlro ro 6v(upo v' onel,euOeprioer trlv oluo[6o rou RNA, rorolqyowog oe npoi6wq overtruxo6q (errptonx{g) 520 KEOMA|019 Metoyporprl mouq flpoxoputirreg EIK0NA 19.12 H RNA notrupepoorl nepvoer on6 op- rerd orddLo fiprv rnv enqrlruvorl. 'Evo r],erotri 6q:e- prig orjpntroro gerorp6neror oe pto ovoLXrrl poptp4 u 6neno o' 6vo rproroy6g oupntroro. l-lpoooplroop6vo on6 S. P. Haugen, W. Ross ror R. L. Gourse, Nat. Rev. Microbiol. 6 (2008): 507-51 I dvop{4q (abortive initiation). Merd rrlv one- Leu05prooq rou ovenrruXorig npoi6woq, ro 6v(upro [ovopf(et rrl orlvOeor1 RNA oq 06orl * 1. EnqvoloprBov6prevor r0xi,or ovenruXd:v ev6p[eov 611 prro u pyo rlv o].ryovo u r].eor(6 ro nou ouvrlOtog 51ouv pqxog tr-r6vo Liyeg Bdoetg (nop6n prnope( vo rptdoouv oe grir<og ro 20 nt), npw ro 6v(upro npoyprorxd roro<pdper vo 6rog0yer on6 rov unorrqrl. Or otrtr4l.enrdpdoerg Ue rrlv RNA noAupre- pdoq te).u<d e[otre(rpowor r<ord q 6rorpuyq rou unoKrvrlq. M6Xprg 6rou 4 oluo[6o rou RNA ensKro0e( oe 15-20 nt, ro 5v(upro yevxd 61et nepdoer on' 6treq lg pretoBdoerq nou Xo- porcrqp(ouv 6vo orjprnLor<o enrprlruvoqg. Or d0o nro govepSg pretoBdoerg e(vor 4 qneLeu- 05poor1 rou nop6yowo o [ono6(6eror mrlv Ex6vo 19.12(y)l Kr o oxqporropr6g ev6g ou;r- nL6t<ou nou rorol,o;rBdver pr6vo -35 bp DNA, ow( ralv -70 bp nou e[vor Xoporrrlptmu<6q uov oupnl5rov rov unortvqrrriv. MoLov6rr 4 oneLeuO6pcoorl rou o ouvqOoq ougBo(ver xo- rd q 6roquyrl rou unorwrlrl, our6 6ev e(vot npoononoripevo yro tq prerdBooq oqv en[- prrlruvoq. Ie optopr6,veg neprmdloerg o o SXer rouronor4Oe( oe oU prnLor<o enq-rr1ruvor1g, oi.- )d q oUvdeo{ rou Ue ro 5v(upro (ocog owrro- tomp((er enovonp6o6eorl orov nup{vo rou ev(0;rou r<otd 14 gdo4 rrlg enqlrp<uvor1g. I 9.8 O l-lopdyovroq o EA6yx€r rnv np6o6€on oTo DNA Avoyvrrrpi(ovToq luyrerprpdveq AAAnAouxieq Tov Ynorrvnr6v * 'EvoS unorLvrprlg npoo6ropl(eror ono rrlv nopouo[o ppoxerotv ouvorvelrtlv otrtrrltrouXrtilv oe ouyrerprpeveg 0eoerg. * 0r ouvorvelx5g otrtr1trouXreq rou unorrvlrrl ouvlOtog onore- Lo0vror ond ;-uo noup[v1 orrlv oqerrlp(0, 6vo e(opepeq pe otr- trritrouXro nopepqepfl pe qv TATAAT oq 0601 --10 rL 6vo otrtro e(opepeg pe gto otrtr1trouxio nopepQept Ue qv TTGA- CA oq 0eo1 --35 * 0t qexoproTo( unouvlr6g ouvrl0rog dtotpepouv ono q ouvol- velrq otrtr1trouX(o oe pro fi neprooorepeq 0eoerg. * H onoretreopolr6rrlro rou unorrvqrrl gnoper enloqg v0 cnl- peo(eror rot ono enrnpooOero orotXe[0. Avodtrd DNA I-lepro1r1 3 tou o Tlsproxn 2 Tou o lleptoXrl 1.1 rou o E(opepdg -1 0 Ktrtivog 91-expoye[o Ko0o0rrd DNAfiepLoXr1 3,2 rou o Ktrrirvog ergoye[o Boorx6g eworeg
  • 13.
    .aciexet 10v 0Kot -5: --3: -30 -20 -10 1 +10 +20 +30 -- ,., ...:;iJ:.i'ttti:,: oxluoT((tTot oTtq 10 Bdoetq, .' -- :-:- .,cKorxov€rrtq0T10Q6q0116ro-356(l)qr0-55 -5: -40 -30 -20 -10 1 +10 +20 +30 . .:.:. :..':. r:.,::t, oxnuoll(tT0t ortq 15-20 -5a -40 -30 -20 -10 '1 +10 +20 +30 H RNA notruUepool opxtKo epXetot o' enoqrl pe -,- ,- :ro -55 eolq +20.'Orov dtfororot o o, o nuplvog , .--,, oloretrtrerot oro -30. MotrLq ro ev(upo perortvloel - -, . - --: ','rvLro o0pntroro enLglruvolg. --. - -:,).qAouXio DNA nou l.ettoupyio qg e(vot v' ovo- -:.leror an6 npare'tvtq, o unoKtvnrnq 6tog6pet --: .: cMq).ouX(eg tcov ono(ov o p6og e(vot vo trle- -: . : loowoL. Ot nLrlpo<pop(eg yro tr1 Lettoupy(o tou ,*:...rlrrl nopdXowor oneu0e(oq on6 rlv oMqLou- -- ::.l DNA: n 6oUn tou eivot to or1trro. Aut6 e(voL . : ., .oorxd nopddetypro ptog cis-evepyoU 05o4q, - --: op[m1Ks ortq Eu<6veg 2.16 rot 2,17. Aw(Oeto, - .,:co(dpreveg neptoXdg ononorjv to v64ptd toug ,-,: lroorou q nLrlpo<pop(o prerorpep0e( orrl uopan . r-l.ou dMou vour<Leivtxoti oE6oS n npottelvqq. -,oq rpdnog vo oledtdoouge Svov unor<tvr1t1 0o --:. Jro ouyr<erpttrt6vq oMrlLoul(o DNA v' ovoyvop((e- -: :lro r1v RNA noLuprepdoq. Kd0e unortvqtrlg 0' --::iorjwov on6 tr1 ouyxercptp6vq otrLr1i.ou1(o, r1 --,:.c1rorov, 0o rrlv nepildprBove. Iro Borcrqptor<6 yo- i.-;-io, ro etrolroro prrl<og nou 0o prnopotioe vo no- : rr(01 dvo enopxdg orlpro e(vor 12 bp. (OnoLo6rlnote :::4r.rrepq oMqtrouX(o e(vot nt0ov6 vo oXqprottore[ op- -..:,q rpop6g -onLcog xord t[p1- n 6rot vo npor<o].5oet -.-5q orlporo. To e).d1Loro prrl<oq nou onottekot yto -: . cbrxrl ovoyvcoptorl ou(dvet pre to pr5yeOoq rou Yovt- - ,.roroq - 6vo np6B).npo oro europu<ortrd yovtdttrt- ..::c), H oMrltrouX(o tov 12 bp de Xpetd(erot vo e(vot ouveX6prevrl. Av 6voq ouyrerptp6vog opt0p6q Bdoeolv 6toXrop(er 6rio oro0ep6q BpoXftepeq otrLfltouX[eq, to ouvotrrr6 pulr<oq touq 0o prnoporioe vq e(vot )try6tepo oTro 12 bp, enetdrj q (6to 11 an6orao4 StoXtoptogori onoretre( 6vo tprrlgo rou otiporoq (6orol rot ov q evdt6- peor'1 oMrlLouX(o e(vot doXerrl). It4v npoyprortr6vlto, 11 RNA noLuprepdoq ovoyvop[(u tLg DNA oMq].ouX(eg rov unoKtvntriv oe preyd).o Fq0U6 ptSoo "dpeoqg ovd- Yvoonq" ouyrerptpSvtov Bdoeov oro DNA on6 ouyre- r<pr;r5vo opuvo(6o tou oloev(tiptou. Ot ewunootordg 6torpop6 g tcr4ioq trr:v 6totp6ptlv Bor,crl ptoxtirv uno Ktvn- rdlv ono6(6owot rqrd noLf or4 6totpopenxrl onotetr'e- o1-rottr6rqro ovdyvoo4g tcrlv otrtrrllouXtrirv toug on6 ng optvo(tx6g oLl.qlou(eq rov unopovddtov o Kot o. Or on6netpeq yto rnv ovoyvtirptorl tov tdtorltov rou DNA nou e(vot onopofurlreg Yto rrlv np6odeoq qg RNA noluprepdonq (er<(vqoov ouyt<pivowoq ttg oMt'1- ),ouX(eg 6torpopettxrilv unortvrltc0v. Kd0e onopofurlrl voux.treot6rrrl oltrr1Lou1(o 0o 6npene vo undplet oe 5oug roug unorwqr6g. Mto t6toto oMrll'oufo Xopo- rcqp((eror ouw4p4p6vq (conserved). Mto ouwrlprlpS- vr1 oM4l,ouX(o 6ev efvot ovoyromxd ouwqpqpSvq oe rd0e (eXoprorrl 05o'r1qq, 6r1}'odri enmpSnerot xdnoto norril.opropg(o. l-kiq ovoltioupe ;rto oMrl).ouX(o DNA yro vo npoodtop(ooupe qv 6Xet ouwqprlOe( opr<etd rilore v' owrnpooonefet 5vo ovoyvtr:pfotpro onpo; Auvqnr6g Odoetg ovoyvrirptorlg tou DNA ;rnopef vo npoo6roprorotiv ovotpopu<d pe pto eft6ovu<eup5vr1 oL- Lql,ou{o 11 ono(o owrnpoooTle0et rq Bdoq nou Bp(oxe- ror nro ouy3r6 oe xdOe 0dor1. Mto ouvolvenx{ oAtrqtrou- ;1fo (consensus sequence) xo0opi(etot on6 t4v eu0u- ypdpptorl 6trtov ttov yvomc0v nopo6etyprdtov 6tot rilore vo preytoronotrl0e( q oproloy(o rouq. l-to vo y[vet ono6erc4 puo oM4trou{o oq ouvotvertr4, rdOe ouYKe- rp4l6vr; Fdon qq np6net vo entxpote( or4v ow(orotXq 06o11 r<ot to neptoo6repo npoYpqrtxd nopode(yproro np6ner vo 6ro<p5pouv pr6vo rotd pto 4 6Uo owtroro- mdoetq on6 rq ouvowenrrl oM4trou1(o. 'Evo ewunootox6 Xoporrqptonr6 trrlv unorwrlrcbv oro E. colie(vot 11 iMeulr1 et<terupivrlq ouwrTp1oqq aA- lqAouyuhv oe 6).11 rrlv 6rcrooq ttov 75 bp nou oler(etor Ue rqv RNA noluprep6oq. 0o16oo, rdnoto BpoXSo tprl- proto pr6oo orov unoKtvrlrl 6Xouv ouwrlpqOe( rot e(vot (onr<r1g oqproo(og yto t4 l,ettoupy(o tou. H ouw1p1ory p6vo noAA Fpaxeuiv ouva:euxtisv aM7ouyuitv e(vatru- nx6 yapaxr4ptwtx6 tuv pu1lttottxtitv 06'oeuv (6nuq ot unoxlrySg) t6oo ota npoKapu@rtKd 6oo xot ora euKa- puanxd yovtdubpara. Apr<er6g oMrll.o u1(eq tov Bor,mq p rorcilv uno rtvltriv ouverogdpouv ornv ovoyvdrprori toug on6 to oLodv(u- Uo rnq RNA nol,u;repdonq. Ario oLl'r1i.ou1(eg 6 bp, yvoord og ororleio -10 G10 element) rot orotlefo'35 (-35 element) (6norg rot to prlxog rnq "6toqrnUotxtlg" oMrltrouX(og nou nopeprBdMetqt ;reto[rj rouq - (spac- er>> sequence) e(vor ouvqOtog oqpowu<6tepeg on6 ou- 169 ro0out6g ttq oll4l,ou1(eg ovoyvrirptoqg. Ot oll,rll,ouX(eg rov unoKtvrlrciv nou Bp(orovtot mqv o<peqp(o r<or oxp$rig 5[nl.o t1g, ot otrtr4]'ou{eq Ydpo) on6 ro ororXe(o -10 [yvotmd oq "6teupup6vo orotXefo - :i.* Ol-lopoyovroqItyUoEtreyXeLUvtlpoobeorloroDNAAvoyvopi(ovrogluyrexprpeveqAtrtrltrouXieqrovYno<1,'- - 521
  • 14.
    'l0D (extended -10element) mqv ovodKn nl,eupd ror "6rqxoprorig" (d iscriminator) mrlv xo0o6u<r1 nLeupdl r<or or 10-20 bp og6ocog qvodrKd rou morXe(ou -35 [yvoor6g oq uororxe(o UP, (UP element)], en(oqg o],- Lqlenrdpotiv rord erdu<6 rp6no Ue rrlv RNA noLu;repd- orl Kor ouveroqSpouv orrlv onorel,eoporrr6rlro rou unorlrprl. . :rouq neptoo6repouq unoKrvqr6g ovoyvrop((eror pro neproXrl 6 bp nep(nou 10 bp qvo6trd rnq oqe- qp(oq (r1 npoygoru<r1 on6orooq on5 r1v oqeqp(o 6rog6per eLogpd on6 unoxwqrrl oe unorwqrrl). Au- r1 q e[opepqg o].].q].ou1(o ouvq0rog ovoprd(eror ororXe[o -10, r<ow( Pribnow (Pribnow box) ri oMuirg xourf TATA (TATA box). H ouvowenrrl oMqLouX(o TATAAT ouvor.l;((eror orn UopQn Tao Ags T+s Aoo A.5o Tso or 6e(rcreg 64Lrirvouv rnv eKorofiro(o ovo),oyfo ulg oulv6tep4g Bdorlg, nou norr0.et on6 45"/" 6tog 96%. (Mro 05or;Xcop(g ep<povr1 npo{gqoq yro xdnoro Bdorl 0o 6rlLcov6rov ge ro oriprBoAo N). H ouXv6qto epr<pdvroqg owrmorle( mr1 o4proo(o our<ilv r<ov (euyc0v Bdoeov yto rrlv np6o6eor1 qg RNA noluprepdonS. 'Etor, ro opXrx6 noLri ourrrn- pqpSvo TA ror ro rellr6 oXe66v on5l,uro ouwn- pqp5vo T orqv oLL4LouX[o -10 oulvd e(vor ro(ptog orlproo(og yro rnv ovoyvriprorl rou unoKr- vqrrl. I{pepo yvrop((oupe 5rr ro ororXe[o -10 flpoyUoronore( er6u<69 yro rrlv oLL4Loul(o enoE6g mrg neprolSq 2.3 Kor 2.4 rov nopdyowo o (BMne nopordro). 0o16oo, ouq q neprofi rou unoKrvn- ri e(vot 6ir<).tovq oro x.l,eriyr6 oriprnLoxo ror ;rov6- r<Lovr1 oro ovorXr6: ouvenc(lg, or oM4Lenr6p6oerq UeroEU tou morXe(ou -10 rcot qg RNA notruUepd- o4g e(vor noLUnl.oxeg ror oMd(ouv oe drogoper- xd o166ro rnq 6repyoo(og 6vop(r1g rnq peroypogrlg. . To ouwnprlp6vo e[opepdg -35 bp ovodrrd rrlg oqerlpfog ovoprd(eror orotleio -35 C35 element). H ouvolerrrrl oMqLouX(o e(vor TTGACA. Ie nro tre- nrotrrep{ Uopen, n ouwnpnprivrl o},r1}.ou{o 61er os eqnq Ta+ Gze Aos Cs+ Aqs Ot Bdoerg oe our6 ro morle(o ollrl).enrdporjv oneuOeiog Ue rnv neproX4 4.2 tou nopdyowo o (BL6ne nopordtro) ;re nop6proto rp6no 16oo oro ovorXr6 6oo ror oro xLeror6 oriprnLoro. r H on6moon ;rero[ri rtov O5oeov -35 ror -10 e(vor 16-18 bp oro -90% rov unoKrvrlrciv. Ing un6lor- neg neprmdroerg e(vot '15 bp ro lly6repo xor 20 bp ro neproo6repo. Molov6tt 4 (6n 4 aMrlLouy[a mo peya)thepo pipog u1q napepBaM6pevrlq nepoyfig e[vot oyenxd ao4pawq, 4 on6otao4 elvat xa[pn enetdr1, t6yu r7q elLxoet6o0q E1oryq rcu DNA, xa1o- pGet 6XL 1t6vo tov xardM4io 6nyapop6 rav 6Ao neprcydtv r4q BNA ndtupepdo1q nou a)rLr1ient- 6poiv, aMa en[oqq Katro ye@petpm6 npooavarcit- o1t6 tuv 50o 06,oeav. o H oqetqp(o ouvrlOog (>90%) e(vor truo noup(vr1, ou- 522 KE0MAI019 Meroyporprtmouq llpoxoputirreg yyd 4 r<ewprrq Pdon qq oMr;),ouX(oq CAT. Oordc: q ouwrlpqol ourrlg rrlg rprn).6rog 6ev e(vor tdoo lr- yd).r1 urore vo Oecope[tot oq unoxpeorxd oqpro. o Opropdvo (e[yq Bdoeov ornv neproX( pero(U r: o<peqp(oq Kor rou morXe(ou -10 ovoyvrrrp((or,.:: on6 qv nepropl 1.2 rou nopdyowo o (B).dne .:. por<drco). l-to nopddetypo, Uro oMr;).en(6poorl o : . r<r1 yro t4v o).).4LouX(o prero[U ev6g roro].oi:r:- youov[vr1g orov x.,rrtvo prrl-expoye(o 6tjo O5oetq .:. 0o6u<d rou orotle(ou -10 e(vor e(orpelr<d of lrc,l . rq yro tov r<o0optopr6 rrlg oro0ep6rqroq r3- ovotXro0 oupnl6r<ou. 'Etor, oL 6ro<pop6g orqv o).1.-- Loul(o rou unoKrvrlrrl oe O6oetg nou 6ev e(vor n:, - ouwrlpqpdveg prnope( vo eu0Uvowor yro rnv ere::- yevq roXU rov 6Lo<p6prr-lv unorrvqrov. o Or pdoerq oro 6eupupr6vo orotXe(o -'10 npooeyyil: ,. ror on6 qv neptoXll 3.0 rou nopdyowo o (B).dne :: poxdtrrr). H oM.r;LouX(o TGN oro ovo6rx6 dr<pc .:- ororXe(ou -10 ouprpret6Xet oe oMq).ent6pdoerg L-.:. repo orlUowu<6,9 yro qv 6vop[q rnq peroypcc*: 6rov 11 oMr;),ou1[o rou ororle[ou -35 orov unoKt,,-:* 6e pord(er noLU pe 14 ouvorvelrcl. Aur6 ovo6err,,,: norg ororleroOerotjwor or oMrlLoul[eq rou uno(.,-. 14: prrpr; oprot6rlto Us rrl ouvorvetLrrl oMqAc-,.: oro 6vo ororXe(o ;inope( v' owroroOpuore( ond ue,:. ).r1 oprot6qro o'6vo dMo orotle(o. o H neptoxn -20 bp ovo6u<d rou ororle(ou -35 cr-:.- Lenr6pd pre ro CTD rotv 6Uo o unopovddcov, OL i . nrcuoerg ourcbv rov ol.Lq).en16pdoerr-rv c:-. evepy6tqro rou unoKrvrlrrl prnope( vo e(vor op(,:: oqprowrr<69, ou(dvowog rn Ueroypoerl rou).:, . orov rord p-uo rd(r1 prey60oug yro no).ri er<gpoi:*i. vouq unoKrvnt6g, n.X. urlv yovr6(rov tou rRNA 'O :. or ouyKeKprpdveg oMrlLouX(eg protd(ouv noAu p. ouvorverrrrl, 11 neptoXrl ourl ovo<p6peror c,Dq c.: . 1e(o UP (UP element). H 6o9( ev6g unoxtvrlrq, pre ro enrpen6pevo eupcl norril.otrropg(oq (on6xlrorlq) on6 ro B6trrroro, nop.-- oro(eror mqv I 9.9 H AnoreAeopoTrK6TnTo Tov YnoKlvnTd,v Mnopel v'Au{dvel 6 vo EAAordveror on6 MerdAAoIn + 0r uopvrllx6qD UsToMoqerq nou per6vouv qv onoretreopc. - rriqro rou unorrvrlrl ouv1Oog pertirvouv q ouppopqrr:c- npoq rtq ouvorvenr6g otrtrqtrouXieg, ev6 or ,,0ertr6g,. perc;. tro(erq 6Xouv ro ovri0ero onor6treopo. * MerotrtroEeLq oqv -35 otrtr1trouXro pnope[ vo enrlpeooo-, rrlv opXrrl npoodeoq qg RNA notrugepooqg. * Merotrtrolerq orqv -10 otrtrr1trouX(o pnope( vo enqpeooc-, qv np6odeorl 11 rrlv ovldpool onodroro[lq rou UrToTpsr: evo rLeroro ougntroro oe ovorXro. Boorxeg evvoreg
  • 15.
    l-leproxn 3.0 tleproXrl2,3-2.4 fleptoXq 1 .2 TOU O TOU O IOU O IroLXeio UP -35 Ext -10 Uls +l -t n0txs[0 Tou DNA Kor T0 ouoToTrKo qq 6oUnq Trlq RNA notru;repoolg nou ouppereXouv oTnv ovoyvtirptorl rou unortvlrrl ----,-, 1: tlpoooppool.t6voono:S.P.Haugen,W.RossrotR.L.Gourse, Nat.Rev.Microbiol.6(2008):507-519. - ,- --::.c getoMd[eotv pnope( vo npooq6pouv - -: - -: :: ryerlKo |re q trettoupy(o rou unoKtvrl- - -. .:'-i.cEelq orouq unoxtvrltdg enqpe6(ouv to - -.: : :-: eKepooqg tou(tov) yovtd(ou(tr.rv) nou -, : -, ,*c[g v' oMd(ouv outd xo0outd ro yovt- ' -,: -::.l.ro. OL nepLoo6tepeg prerotrd[etg ovo- -: -:, :. ;e Borcrqpto tog oloydg tqg oMq).ou- - : - - - : l-','ouv oe onotreto rl Uey6).q ;re(tooq qq . --: ::j-: :tDV yettovtrrirv yovt6(ov. E(vot yvoor5g - , :: , nrxeq peroM6[erg (down mutations)' AMo- - - - : :r-'.-iou1(og euO0vowot Kot Yl'o 0erlx6g pe- -:--: iarc (up mutations) ottg ono(eg undpXet -. -: : ". -3-ioYpoQ( on6 tov unor<tvqrq. -- l- :. ,,8ettrd,g,, doo xot ot "opvr1ttr6g" pretoMd- .. - :: --. .:L oe oXeoq pre q ouvry1to1tiv4 onotetre- - -- .- -:: evdg ouyrer<ptp5vou unoKtvt'1tq nou - - - - I : .:.rL.tret notrU. 'Etot, prto oMoyr; nou ovoYvo- . -.-- -: ::i.1rtr<q petdLl.o[r1 o' 6vov unor<tvr;tr| pno- . : - -- , lrcUovrr-loe( not6 o' 6vov 6Mo unor<tvqtrl (o - - - : r. :. cto rorooroorl 0o pnopo[oe vo e(vot o16- :-r:: cnortleoprottr6g on6 tq petol'LoyprSvrl -,: -- -:- :cLJrou unor<tvr1q). Ae6o;r6vo on6 in vivo - . :-: - : .r.;q ovoyvcop((ouv 11 ouvoLtr<r1 rote[Ouvot1 - - r . :.-: -rou npor<ole[ 11 pretdMo[q. - -. ::,,.oEeiq rou ou(dvouv ttlv opot6tqro rov - . - " " - <ot -35 Ue rtq ouvotvettx6g oMrlLoulieg 11 . :- - : - , :- pretolti toug on6oroorl rdto on6 17 bp - - : - : :- lc. ouv rlv evepy6rrlto tou unortvrltrl. l-lo- . -. -: : *iroMo[etg nou getovouv ttlv ogor6tqro - - -:l-::.i nLeupog Ue rrl ouvotvettr<r1 oMrltrouX(o - - :::::::icuv rr1 preto(0 toug on6orooq (on6 to . - -: :-, '7 bp) o6rlyoriv oe petcol-ttivrl evepy6qto -' ,-.n, Or opvqttr6q getotrtrd(etq ouvq0og - .: -:*,c',/Tot ortq TIto ouwrlprlpr5veg O5oetg tou . - ," -:- :riBeBottrrvowoq rnv t6to(tepr1 oqpoo(o ou- - - . ::ceolv Yto rnv onorel'eopottr6qto rou uno- l;coo, undplouv onopodtrdg elotp6oetg oe - --: -: :ouq rov6veq. -: r,opddetYpo, 6vog unor<tvqtrlq ge ouvotvettxdg - , ..'.cr1[eg oe 6].o to oroLXe[o nou neptypdqrlKov nq- :: -:.:(erot mqv Etr6vo 19.14. Aev undpXouv 6ptolq -:-: r. rpuotxo[ unortvqt6g mo yovt6(trl1-to tou E. coli . : -:, ,rro( unoxLvqt6g nou totptd(ouv on6Luto 5le 11 - - r .::.<rl oMlLoux(o oe 6).eg ttg 06oetg srnv npoY- -:- . ::-:l e(vot oo0evdorepot on6 unortvqtdg pe tou- - l.:, evo ).d0og to(ptoopro oro ouvotvettrd e(opepri -10 r<ot -35. Aw6 ou;rBo(ver enet6rl npoo56- vowor 16oo orptXr6 orqv RNA noLuprepdo4 rilore ornv npoyl.tonx6tnro nopeuno6((ouv tr1 6ro<puyf rou uTloKl'- VNTN, l-ro vo ro0optoroUv ot on6l'uteg entmriloetg rov Ue- roMd[e<ov rov uTloKtvnrcilv, pretpdtot in vitro q ouyy6- verq rnq RNA nouprepdorlg Yto rov dypto rot ro pretotrl.oyprdvo t6no rtov uTloKtvntriv' H etepoy6veto tr1q roxfunroq Ue rnv ono(o q RNA nol'uprepdorl npoo- 66verot in vitro oroug 6totpopetxorig unorwrlrdg o1e{- (etor r<oLd ;re q ouly6rrlro rnq pretoyporprlg 6tov r' ow(orotlo yov(6to errppd(owot in vivo. Enercrefuowog ourl qv ovdl.uorl, ;rnopei vo npoodtoptore[ to orddto oro ono(o prto pret6Mo[r1 enrlped(er r1v ono6onx6tqto ev6g unoxrvntn. ALI'd(et q ouyy6veto rou unorwqtrl yro np6o6eoq rqg RNA nol'uprepdorlg; Arpqvet pev ro 6v(u;ro trqv5 vo npoode0e( oM6 ov(rqvo vo (erwrloet rrl Uerqypoeri; Atogoponoe( trlv enrppoq ev6q enxou- pu<ori nopdyowo; Metpdrwoq ttg t<tvtlttt<69 oro0epdq Yto ro ox!1gort- opr6 ev5g x.tretoto0 oupntr6rou Kot rnq gerotponrig rou oe ovotXr5 orjptnl.ot<o, prnopo6ge vo dtorp(voupe to 6rio ord6to rrlg ow(dpooqg 6vop(r1g: . Ot opvnrtK5g geroMd(etq orrlv ol.i.r1}'ouX(o -35 ou- vr'1Ooq get6vouv r1v toX0rlto oxnUortopoti tou xtretorori ouptntr6rou, oLl'd 6ev ovoqr6Mouv rn Ue- rotponf rou oe ovotXr6 oUPnl'oxo . Ot opvnrtKdg pretoL),d(elq orrlv o).trr1l,ouX(o -10 prnopei vo petc0oouv e(re tov op1tr6 oXqpottopr6 ev6g xtretmori ou;rntr6r<ou e[te r1 petotpon{ tou orrlv ovotXrrl Uopqi efue xot ro 5Uo. H ouvowenrrl oMr1tr'ouX(o orq 06o11 -10 qnotelefuqt onorletonrd on6 (e0yq Bdoeu:v A-T, Stopr6prpoon TIou 9on06 orov opXtr<6 6roXoptopr6 tou DNA oe provo0g xLrivoug. H Lty6tepq ev6pyeto nou onorefuor q 6totd- po[r1 tov (euydrv A-T oe ox6on Ue ro (eUyq G-C orlpo(- ver 611 dvo ttrrripo pe (e0yq A-T onotte( prtrp6teprl noo6rqto evSpyetoq yo to 6toXcoptqr6 tov xi.rirv<ov' Ot otrr1Lou{eq op5oo4 y0po rot r<o0odtrd on6 tqv o<pe- tqp(o en(oqg enqped(ouv qv 5vop[r1' Ax6p4, n np6rn neproX{ nou preroypdqerot (on6 -+1 Udxpt -+20) enq- pedEfl rrlv toXriqro Ue rnv ono(o q RNA noLuprep6oq onogot<p0verot on6 Tov unoKtvnrrl r<ot yt'our6 enr6pd orqv roXti rou unoKtvnr4. 'Etot, 4 ouvotrrr4 toXtig tou unorwqtq 6e ;rnope(ndwore vo npoBtre<p0ei on6 ttg ou- IroLXe[o UP 'i:':.t g 4noat^eoporLrorrlro rolv Ynoxrvlrdlv Mnopei v'Au[overot 1 vo EtrtroroveTot 0[o l,ra :):::- 523
  • 16.
    vorvenKdq oMqAouXieg rou,ox6pr4 r<r ov tr4gOoriv un6_ qrlro enrnp5o0ero ororXe(o nou ovoyvrop((er 4 RNA no_ Aupepdo4 elc6g on6 trg oMqi.ouX(eg -10 ror -35. . Molov6rr 4 opor6rqto Ue rrl ouvorveru<r1 oMrlAou_ {o e(vor lprlorpro epyo},e(o yro rrlv ovoyvcilprorl unoxr- vrlrrirv pr6vo Ue Bdon qv oMql,ouX(o rou DNA, 6nog en[o4g xoL nop6rr ot (runrKo(> unoxtvqrdg nepdlouv eudtdr<pneg oMrltrouX(eg -35 r<or -10, noMoi unoKrvrl- tdg 6e 6ro05rouv eurprvrl morlefo -10 ror/ri -35. Ie notrilig on6 ow6g rtg neprmc0oerq, o unoruvrlrlg 6ev ovoyvrop((etor on6 g6v4 rrlv RNA no).uprepdor; xor qnorre( pto enrKouprKri npcoreivrl (,evepyonor4riq, - activator, BL6ne Kegdioro 26) nou unepBo(ver rrlv ove_ ndprero tov evdoyevcirv oMql,enrbpdoeov pero[ti RNA notrupepdo4q Kor unoKrvrlrl. Eivot oqpowrr6 en(or1g vo_rov[oou1,re 6rr "g6trrrm4 evepy6qroD 6e orlpo,v . ";r5yrorr1 evepy6rqro,,. llol,Lof unorrvqt6g dlouv e(e_ ).t10e[ pre oMqtrouXfeg nou on6louv no],rj on6 to vo e[- vor ouvqrverrr6g oxptBrig enerdrl 6ev efvor ro Bdtrnoro yto ro rUmopo vo grrd;yer 16oo nol,[ npoi6v nou KoJ_ 6monorekot on6 ro RNA pietdypoeo rouq. l9.lO l-loAA6q t-teproxdq Tnq RNA l-loAupepdonq'EpxovTor Apeoo oe Enogfi U€ Tov DNA ynorrvnrfi * H doHl rou 070 otrtro(et orov ouvdeeror ge rov nuplvo rou ev_ (u;rou, enup6n0vr0q oTrq neptoXeg yto npoodeoq oro DNA v, otrtrltrenrdpooouv Ur Tov unorrvr;r1 er llotrtreq neproXeg rou 070 oitrrltrenrdpouv pe rov unorLvrlrl. s H unopovodo o en(orig ouppotrtret orr;v ovoyvtirprol Tou uro_ rrvrlq. Onog npoovorpdp0rlKe orqv Evoulra 79.g, opr<erdg neproX6g orrlv unopovddo tou nopdyowo o Kot rrl CTD r;g unogovddog o Splowor o, enogrl pe ro DNA rou unoKrvrlr4. H rowono(qor1 ptog oerpdg drorpoperr_ t<c6v ouvorverLxov otrl.rlLoulLrrtv or ono(eg ctvoyvop(- (owot on6 o).o6v(upo trie dro<popelxoUg nopdyoweg o (dntog qo(veror o-Trlv ' , ., ,.) uno6r;i.rr;vrr o-i. : [6ro9 o nopdyowog o npdner vo dpleror o, enoef *. ro DNA. En[or1g unodrl].rrrvet 6l ot drogoperLxo( nop:- yoweq o npdneL vo npoo6dvowor Ue nop6pioLo rpo:: mov nuprlvo rou ev(rjprou, dror c6me oL enLgoveLs: ovoyvirtprorlg yro ro DNA ftov 6rorpoperrxrr:v nopoyo,,- roJV vo rono0erotiwoL ire nop6;roro rp6no otore ,,,n npoyporonorotiv ng xpforpreg enogdg Ue rov unoKrvr_ q r<owd ortg oMrlLoul(eg -35 xor -10. Enrnp6o0ereg ev6e([erg drL o nopdyowog o(ypro ec Xeror oe dpreorl eno<p4 Ue rov unor<rvr;r4 16oo orrl _35 6oo roL o-cr1 -10 ouvorvelrq oMrlLouX(o nporlLOov on: unoro-roordoerq orrov nopdyowo o nou KoroordMo", peroALd[erg orrq ouvorvelr<6g oMq].ou1(eg" Otov 1.ric perd).).o[r1 oe Liro ouyxercprpr6,vrl 0dorl rou unoKrvr.lr- nopepno6((er r;v ovoyv<rrpro4 on6 rqv RNA noLuprepc- on Kor puo owroro0prorLrq ;rerdMo{4 orov nopdyowc o enrpdner orrlv noi.upepdorl vo lprloLponorqoer ro f.lr- toMoypr6vo unoxrvqq, q nro nr0ovrl eEnynol eivor or. to olerrxd (e[yog Bdoeorv rou DNA dpleror o, enoer pe ro optvolti nou sxer owrKqtooro0e(, I uyr<p(oerg tlv oMq).ouyurtv opxertrtv Boxrq pto r<u_r,, nopoydwov o un6.6er(ov ouwqpqpr6veg neproXdg o-rc o70 Tou E.coli .,,, , ,, ,.', nou oMqenr6poriv dpreoc pe unorrvqtdg" Aur6g or ev6e((eLg rexprqprot0qxov onc rqv avoyvriprol Uroq xpuoroMLr<qq dopnq rou oLor,,- ([pou qg RNA no].upepdonq oe oUpnl.or<o pr, dvo r<.r.c- ol-lo rou unor<rvqtr1. To Borcrqpro Thermus aquaticus xc. Thermus thermophilus derlvouv notg or neprol6q npoc_ 6eo4g oro DNA rou nopdyowo o mulrr.lvowor or ovr_ {dprqreg neproX6g orrg nptrrreivLr<dg neproleq 1 2 2,3-2.4, 3,0 r<oL 4.1 -4.2. H Eu<dvo 19.'16 nopouord(er ro rprrlproro rou nopc- yowo o nou exouv dgeoo pd),o mrlv ovoyvcilprqn roL unorrvqrrl. Auq q eLrdvo de(pet q 6op4 -rou xrlpLcu nopdyowo o oro n).o(oro rou oLoev(0prou. Atio Bpolec rp4proro qg neproXqg 2 rr evo rprrlpo qg neproplq .1 (2.3, 2.4 KoL 4.2) eplowor o' enog4 ;re Bdoerg oro oror- Xe(o -10 r<or -35, owroro(1org. H neproX4 1.2 rou nopo- yovro o Spleror o' enorpr'1 pe rqv nsproX4 rol unoxrv4rq r<o0odLnd on6 ro ororXe[o -10, evot rl neproxn 3.0 dpXerot Ue qv nepropl rou unoKrvrir4 prd).rg ovo6L- rd rou oroLle(ou -10. Ko0etrr(o ond outdg trg neproldq oX4porf(er Bpoldo rp4poro o-ti].u<og orqv npc,-rteivn Ynogovd6o/ yov0ro M6ye0og (# aa) Kord npoo6yyrorl # tov unorw4rdrv Aliflou1(o rou urtoKvlrn nou ovoyvropi(eror ILypo 70 (rpoD) 613 1000 ILypo 54 (rpoN) 477 5 IryiLo S (rpoS) 330 100 Irypo 32 (rpoH) 284 30 ILypo F (rpoF) 239 4A I(ygo E (rpoE) 202 20 TTGACA-16 bp 6o9 1B bp-TATAAT CTGGNA-6 bp-TTGCA TTGACA-I6 bp eog 1B bp-TATAAT CCCTTGAA-13 erog 15 bp- CCCGATNT CTAAA-1 5 bp-GCCGATAA GAA-1 6 bp-YCTGA Iiypo Fecl (fecl) 173 1-Z ? Elx0NA 19.15 0t nopdyoweg o rou E coli ovoyvcop((ouv unorrvrlr6g pe drorpopelrrlg ouvorvelreg otrtrr1trouxreq. 524 KE0AAAI0 19 Meroyporprl oroug ilpoxopudrreg Boorreg eworeg
  • 17.
    H 6opq rounopoyovro o oro yevrrorepo ntroioro rou o),oev(upou: otr).qtrenrdpooetq -1 0 rot -35 0 nopoYovrog o enerrel- vrT6r Kgr or neptoXrlg rou ouvfleovror p' euetrrrroug ouvdereg. To oXrlpo 6XeL npoooplioorel ono: D. G. Vassylev, elal., Nature 417 (2002)'. 712-719. AoUrt on6 Protein Data Bank 1lW7 D, G Vassylev, elal.,Nature 417 2002).712-719 Mro r<puotoMu<q 6o;11 tou oLoev(0prou oe oripnlot<o 1.t' 5vo xldopro tou unoxtvrlni, oe ouv6uoopr6 pre nsrpdpto- ro Ue unoKrvqt6g oro ono[o ot r<].ri:vot tou DNA roro- oreudmqxov ri:me vo nept5Xouv oto(ptomo (eUyr1 (,,etepo6(xLrovo)), 66erqov 6rr o o70 6pXetot Kotd r<rjpro L6yo o' enogq ;re Bdoerg orov xtrcbvo-prr1 expoyeio tou ororXe(ou -10, oro 6reupup6vo orotXe(o -10 t<ot orrlv ile- ptoXrl tou 6tolcoptorrl xot ouve{(et vo 6rotqpe( ow5g rg eno<p6g t<ot getd to (etritrtypro rou DNA outtlg qg neproXqq. Aur6 entBeBouilvet 6tt o nopdyowog o efuot or1trrowx6g orrlv ow(6pooq onodtdtoEns FnEnS) H Xprio'n o-ehroetdriv pro{Brov e(vot t<owr1 oe npo- retveg nou ovoyvop((ouv oMql.ou{eg 6(r}'orvou DNA (BI6ne Ev5qto 28.6). Apwo[5o nou on5louv pero{U roug 3-4 06oetg Bpiot<owot ornv (6to nLeupd qg o-51't- Koq Kor yt'out6 e(vot oe 05or1 vo 6p0ouv o' enoq4 pe yenovrrd (e0yq Bdoeov. H ElKOl'lA't3'17 de(Xvet 6n r' oprwo[5o rord prlxoq ptog n].eupdg tqg neptoplg 2.4 trlq o-6lrro9 5pXowot o' enogrl pe rg Bdoerg ortg 06- oerg -12 5og -10 rnq -10 otrtrr1).ou{og tou unoxtvrprl. H neptoXrl 2.3 potd(et pre npo:teiVeg nou npoo66- vouv 1.rov6rtrrovo vouxleivtxd o(5o rt epnl,5retoL or4v ow(6pooq rn[nq. Ot neptoXSg 2.1 ru 2.2 (ro nto ouv- flpnudvo rpqgo tou nopdyowo o) egnl'6rowot oqv otrLqtren(6poon Ue rov nuprlvo tou ev(ripou. Eu<d(erot 611 6tror or nopdyoweg o npoo66vowot onq (6eg ne- ptoX6g rou nuprlvo trlg RNA no).uprepdoqg, on6re "ov- royrr:v((owotD yto neptoptopr6vo Udpoq tou nup{vo rnq. I-1op6l o nopdyowog o 6XeL neptoldg nou ovoYvo- pi(ouv ouyr<enpt;tdveg Bdoetg oro DNA tou unortv4trl, q N-teLrr<q neptoXrl tou eerJOepou nopdyowo o (ne- proXrl 1.1), evepyolwoq Log ouroovoorotrttrrl neptoXtl, onorpUnret qv noptoXrl np6odeoqg oro DNA. M6vo 6rov q 6Lopr6p<ptr:or1 rou rpononotq0e( on6 11 orjvdeorl rou ue rov nuprlvo tou ev([p-rou, prnope( o nopdyowog o vo npoo6e0e( erdLr<d ortg oMqLouX(eg tou unortvqtrl H oduvopr(o tou etrerjOepou nopdyowo o v' ovoyvolp[oer oMqtroul(eg tou unortvqt4 dleL orlpo- o(o: ov npoodevdrov oe unoKtvrlr6g u.tg e).e00epq uno- provo6o 0o pnopoUoe v' ovome[].et rqv 6vop[q tqg 06on -13-12-11*10-9-8-7 ElK0$iA 19.17 T' oprvo[60 ot2.4 o-5trtro tou o70 6pXovrot o' enoqn Ue ouyrsrprpdveg pooetq orov rodxonoulrtrd rtrtivo qg -1 0 otrtrqtrouXiog rou unortvrlrfl. 19.tr0 [lo),treg l-leptoX6q qg RNA llotruUepoonq'EpXovrot Apeoo oe Enoqrl pe rov DNA YnoxLulnl 525
  • 18.
    neproxeg flp6o0eonq oroDNA tlH N-rdwr1 neptoXrl npoo6Sver nepLoX6g npdobeorlg DNA mov ele00epo o To DNA erronl(et qv N-re)x1 neproXrl rirov oXqgor[(eror ro ouurroKo pre ro DNA EIK0NA 19.18 To N-retrxd orpo rou o epno6(er Trq neprox6q rp6o6s0nq DNA vo ouvoeeofv Ue To oTdxo Touq. Mdtrrq oxrlUoTr- ore( 6v0 ovorXro otjuntroKo, ro N-rs,xd orpo ourlpeuor 20 A po- rpud ror or 6Uo neproX6q rpdodeonq Tou DNA 0r6xouv 11 1uo ond qv dtrr1 roro -15 A. Ueroypoqnq on6 ro ol,o6v(ugo. H EK6vq 19,18 ono6(- der g4qporrd qv oMoyri dropr6pgooqg rou nopdyov- ro o rotd ro oxnporropr6 rou ovorXroU ouprnL6rou. 'Orov o nopdyowog o npoo66verqr orov nupr]vo qg noLuprepdonq, n N-rellrq neproXrl ourlpe(ror -20 A por<pud on6 rtg neproX5g np6o6eorlg rou DNA ror or neproX6g np6o6eo1g rou DNA on5louv preto(ti roug -15 A, nr0ovr6g rilore v' ononrloouv nro enlgrp<uoprdvrl 6topr6pgcoorl, rotd).Lr1l,q yro rnv enog4 pre ro DNA. MeroLLd(erq e(re oqv oLL4LouX(o -10 efue mrlv -35 ovoor6tr.ouv rrlv np6o6eor1 oro DNA puog xotroBro;r5- vrlg erdoXqq rou o70 Xcop(g rqv N-rel,u<ri neproXrj, yeyo- v6g nou uno64l,6ver 6tr o o70 6pXeror rour6lpovo o' eno<p4 Kor Ue trq 6rio d.)l.r1lou1(eg. Aur6 rorprd(er pre rg nlqpotpop(eg on6 rqv rpuoroLllxrl 6oprl rou olo- ev([pou (Etr6vo 19.16), orrlv ono(o e[vor oorpdg 6n o nopdyowog o 61er gdMov enr;rrlxuoprdvrl Uopen, eKrer- v5prevog oe -68 A rov 6Uo orpoec0v rou DNA. MoLov5t 11 neproXq 1.1 rou nopdyowo o 6ev SXet ovoLuOe[ pre r<puoro].troyporp(o, Brorpuorx6g prerp4oerg qq ew6nrorlq rnq oro oLo5v(upro Kor oro ovotXr6 oUpr- nloro unodql.<irvouv 6r oro eAe00epo olo6v(upo 11 N- retur<{ neproX( (neproXri 1.1) Bp(oretor orov rewpr6 6(ou).o DNA rou ev(0prou, puproriprevq ouoroorrrd q 06- orl nou 0o xoroldBer o unoKrvnr{q 6rov q4qprorrme( ro oUpnloro rnq Ueroyporp{q (ElX0ilA 19.19}. Orov ro oLo6v(upro oXqproiloer 6vo ovorXr6 oripntroxo oro DNA, q N-reLtrrl nepropi rou nopdyowo o eKroTr((eror on6 rov r6pro 6(oulo. Iuvendlg, q ew6nroq qg oe q46or1 Ue rnv un6trotnr1 nporelvr1 e(vor notr0 euprerdB],4rr1. A].- trd(er 6rov o nopdyowog o npoodeOe( oro rewptr6 6v- (upo ror (ovd 6rov to oLoSv(upro npoo6e0e( oro DNA. H 5fur<o rou DNA np6na vo trretorwq0e( nepfnou xo'rd 16 A on6 t4v opXxrl qg 05or1 nporeql5vou vu ero6),0er mov xripro 6[ou].o tou DNA rr 6nemo vo perorrvrl0e( (ovd 6ore vo enmpdr.fer oT o DNA vo ero6l0er oro 6(ou- l,o t<otd ro oxrlporropr6 rou ovorXro6 ou;rnL6rou. H El- (01{A 19.20 nopouord(er owrl q perorfur1or1, B}.5nowog 526 KE0A Al019 Meroypogrt mouq l-lporoputlreq EIKONA 19.19 0 nopdyovtog o 6Xet gro enqrlxuop6vrl 0opr1 nou exre(veror rord p1rog qg enrqoverog rtrlv unogov66rov Tou ttu- prlvo 6rov oXrl;iouore( ro otro6v(ugo. oe eyxdporo roprrl rotd pr4rog rou dlovo t4g 6).u<og rou DNA. l-loLot6repo enu<pcnoiloe 11 dnogq 6rr o nopdyowog o efuor q povo6rrq unogovd6o rrlq RNA nolupepdorlg nou npoo66veror orrlv nepropi rou unorwrlrq, I(pepo yvrop(oupe 5rt or C-refurdq neptoyiq rov 6Uo unopov6- 6rov o pmope( entorlq vo no((ouv orlprowx6 p6lo orqv enogf pre ro DNA rou unorwrlrq ;r5oco rqg np6o6eor;g rouq oro ororle(o UP (BL6ne Ev6rqra 79.8). Enerdq or oCTd ouv66owor XoLopd Ue rnv un6l,orn4 RNA nolupre- pdon (FMne Eu<6vo 19.14), ro 5v(upro prnope( vo rprdoer oe opxerd onopoKpuol,l5veg neproX6g evril e(vor ox6pr1 npoo6e6epr6vo qro ororye(q -10 Kot -35. 'Etor or oCTDs nop6Xouv rwqr5q neprol6g yr' oM$.en(6ppooq pe psro- ypogu<orig nopdyoweq npoodedepr5vouq oe 6r6gopeq onoordoerg ovodxd rqq oger4pioq rnq Ueroyporp{q 616- gopov unorwrpcilv (BMne lGgdloro 26,7o Onep6vto). EIK0NA 19.20 To DNA opXtrd 6pXeror o' enorprl pe rov nopoyov- ro o (po0 ror rov nuprlvo rou ev(Upou (yxpr). Merouve[rqr nro po0rd p6oo orov nuprlvo rou ev(Upou yro v0 npoyporonouloer enoq6g pe rrlv otrtr4tr0uX(o -10. M6)rg onetreuOeptoOe[ o o, ro etj- poq Tou nepdoporog nou nept6Xer DNA ou(dver. Avor0ntooq pe r4v d6eLo rov Macmillan Publishers Ltd: Nature, D. G. Vassylev, et al., uol 417, pp. 712-719, copyright 2002. 0toroyporpfo pe qv odero rou Shigeyuki Yokoyama, The University of Tokyo. N-TetrrKn neproxn U-
  • 19.
    19.1 1 HAnorrjntoon Eivor Mi6o6oq YrlnAfq AvdAuonq Vro To Xoporrnprop6 rurv AAAnAenr6pdoeov RNA lloAupepdonq- Y no xw nr i't xaq I €,v rx6repo, DNA-I'I p an ei' v thv * 0r ouvorvenreq otrtrntrouXreq -35 rot -1 0 nopSXouv T0 nepro- oorepo orlge[o enoqrlq yro qv RNA notrugepoorl 0T0v uIIoKt- vnTrl. = To orluelo enoqnq 9p(orovror rupfog oq pto ntreupo rou DNA. H txov6r4ro qq RNA nouprepdo4q (ornv npoyport- xdrqro, r<d0e nporeivqg) v' ovoyvorp((er to DNA pno- ce[ vo XqpoKrnprore( pe onorUnooq (footprinting). Vro oMqloul(o DNA npoo6e6epr5vrl oe Uto nptotelVrl r6meror peputitq on5 pro evdovoux.tredoq nou enrG 0erqt oe (eltrrptoroIg grrrogodreoteptroUg 6eopotig ldoo mo voux.treivrr6 o[6. Yn6 r<otdtr].r1].eg ouvOr1- <eg, rdoe (e1oprm6g grj:ogodremeptr6g 6eopr6q .rnope( vo ondoer oe rdnoro prdpto DNA. Ot O5oetg :ou ndnrowor ovoyvtr;p((owot Xpqotpronoui:wog D- A o4pooprSvo orov 5vo x).rivo, p6vo oro :vo dr<po. [lp6retror yro rrlv (6to oppl nou iraoppd(eror ornv oMqLoriXqoq rou DNA: - peprr<r1 ndr,Uq ev6g telrrd oqpoopr6vou *op(ou oe Uro eunoOrl 06o11 dqpuoupye( 5vo doopro povo6u<oU prqxouq. H 6e(Xvet 6tt petd rrlv Korep- .'ooio pre voux.tredoq to onoop6vo r<i.doproro )NA pnope( vo 6toltoproroUv ovdloyo pe to *r'lKoq rouq pie rlLer<rpog6pnon oe nr1rr1, (o0e x).oopro nou 6Lotr;pe( 6vo orll.rooprdvo :r<po nopdyet pto po6revepVn (rr:vn. H 06ot1 :1q (rrrvqg owtororle( mov opt0pr6 torv Bd- trocrJv oro xi.dopo. To Bpoxurepo r^doporo *erorrvo0wot roXUrepo, 6ror q on6moorl :16 ro orlproop5vo drpo unoloy[(etor on6 :c xdrro pr5pog qg nnKqq, Iro e).eU0epo DNA, r<d0e eunoOnq 6e- :-rog ondet roul.dltorov oe rdnoto p6pto. H =L<dvo 19.20 6e(Xver 61 6rov to DNA Bpi l(rror or oripntror<o Ue Uro npore[vq, ot 05- leiq nou roLUnrovror on6 tqv npotelvrl .cccpuldooowor ond qv n6Qr1, 'Etot, 6tov 5.ro owLdpdoerg rp6(ouv nopdl,).q).o -dvo H onorunool ovoyvtrtpi(eL rtg 0e- -.r-q onou npood6vovrot npoTelvsq oro DNA ono -,' npooroo(o nou nop6Xouv rlvovrL qg 6qgtoup- , .rc evropcirv. DNA prdprupog 1op(g ropr(o np<oreTwl ru 6vo nerpogqn- 16 prfupro pre pr6pro DNA npoo6e6e;rdvo mqv nporreivr; 0o 64;uoupyrl0e( dvo XopoKrnprorrr6 npdruno. 'Oruv gro npoodedepdvrl nptoretvq ovoore(Aer rqv np6oBoo4 qg vouri,edonq oro DNA, or 6eoiro( orryv npoo6e6e;r6- vr1 oMqAoulfu 6e ondve oro rretpapaut<6 1t$pa Ket ctu- t6 to t1t4pa rqq ftnKrrlq 6ev nepdya o4paopiva xid- oparu DNA. Iro prdptupo, onder r<dOe 6eog6g dqpnoupytitwog pro rl,(poro (ovrirv, orrlv ono[o x60e Bdoq owtnpooo- neJerot on6 gro (c0vq. Irqv ex6vo, undpXouv 31 Bd- oerq. Iro npoororeuUdvo xtrdopro, 6eopro( 6e ondve orrlv neproXri 6nou e[vot npoo6e6etrr6w1 11 npcoteivq, on6re 6e dr1;uoupyoriwor (c0veg nou owtnpoooneUouv rl6oproro tcov ow[morlrov preye0c0v. H onouo(o turv (ovrrlv 9-18 orrlv eu<6vo rou'ronoe( ptto 06oq np6o'6e- o4g q ono(o roltjmet rrlv neproXtj nou Bp[oretot 9-18 Bdoerg on6 ro or1;roopdvo drpo tou DNA. Iuyrpfuov- roq rrl Lop(6o tou ;rdprupo Kor rnv netpogotu<r1 op(- 6o pe pro ovt[6poor1 o].].q].orixnong nou rpdlet nopdtrtrr1tro, prnope( vo "6tqpoore6, dpeoo q owforotXq oMrlloul[o, ovoyvrop((owog 6ror tr1 vou t<].eottdu<rj o],- Lql,oufo qg 06o119 np6o6eo4g. 'Oncog npoovo<p6p0r1r<e (BL6ne Ex6vo 19.13), q R- NA nol.uprepdo4 npoo6Sverot srrlv neptoXti rou unoKt- .AJffinffihnffih I*stu@htr@ + /V/Va/1. + /-,/,/VV. }a b + .ra/./vv1. *ry+ /L / HAEKTPOOOPHIH DNA olgoogdvo oro / Ilto gorPtd on6 dxpo rou evrjg rtrtivou m ffi- ro olpoopdvo ry ffi drpo 1ETpAMATTKH ffi ffi 1HKTH NHKTH ffi ffi MAPWPA: ffi To DNAnou 6ev To orlpntroro RNA rduUepdonq- unoKrvnrn ddXerot peprxq en(Oeoq on6 qv DNAdoq I 0r (tiveg nou onouord(ouv ovoyvrop(ouv 06oerg npdoDeonq Anopovrrlorl rou DNA xor onoDrorolrl oe lrovoug rtr6voug ffi eivoL npoodedepdvo ffi sqynougepoorl ffi eXer (tiveq oe Oeoerq ffi fiou owrororxouv oro :::: onoorpo Kooe deol]ou llro rovro oro orll-roouevo oxpo ' ; :: H Anorunorq e[vor M50o6og Ytpllrlg Avoluqg yro Xoporqpto;.ro Atrtrqtrentdpooeov RNA llotruuepoolq-Ynoxtvrlq xct Dl',]i-': *..., * , 527 Boorxeg evvorcg
  • 20.
    v4q on6 -55Scog +20. To oqpre(o oro ono[o q RNA no- Lupepdorl.6pXeror oe dpeorl enoqrj Ue rov unoxl4rrl rquronororiwor Ue rponono(qor1 rrlg reyyu<rjg rnq ono- t0n<oor1 g, 1p4 orprono uirwog owr6 poonl pro no u rpono- notoUv ouyrerptpr6veg Bdoerg oro orjprnl,or<o RNA noLuprepdo4g-unor<rvqri. Me ouq qv npooriyyron, ro ne(po;ro ;rnope( vo ercrel,eore( ge 6tio rp6noug: o To DNA pnope( vo rpononotrlOef nprv npoodeOe( 4 RNA nol,upepdorl. Ie ourrl qv nep(nrooq, ov 11 rponono(qo4 egnod(oer qv np6odeoq r4g RNA no- ).upepdoqg, rouronomfuor trlo Bdo4 6nou q enoErj efuor onopofurlq. o To o0unLor<o RNA notru;repdoqg-DNA pnope( vo rpononor4Oe(, on6te ro np6runo rov npoororeu- pr6vrov (alvcbv ouyr<p(verot ge ro ovr(morxo rou eLerlOepou DNA r<ot rou 1-trl rpononorqpr6vou oup- nl6xou. Kdnoreg (6veg e(ogov((owor, rouronor6v- rog 6tor 06cerg ottg ono(eg ro 6v(upro rlXer npoororeUoer rov unoKtvrlfi on6 tponono(r1or1. iL- Leq (6veg 6Xouv ou[r1pr6vq 6woor1, towonorciwog O5oerg orrg ono(eg to DNA np6na vo 6roqpr10e( oe 1.rro orepeo6op4 mqv ono(o e(vor neproo6repo erre0erp6vo orov nopdyowo n5Qqg. Aurdg ot otri,oy5q or4v euoroOqo[o onord].utfov r1 yeolprerp(o rou oupnL6rou, 6ncog ouvotp((eror orqv El- K0l,lA 19.22 yro 5vo runx6 unorrv4rri. Or neprol5g oro - 35 ror oro -10 nepr6Xouv to neproo6tepo on5 -ro o4pe(o enorpfg yro ro 6v(upo. MSoo oe our6g trg ne- proXdg, or (6teg oprd6eg O5oeov re[vouv'16oo v' ono- rp5nouv rrlv np5o6eon qv 61ouv nporlyoug6vrog tpononot4Oe( 6oo xor ov eprrpov((ouv ou[qpdvrl ri prer- cr:pdvq euoro04o[o orrlv rpoTronoirlorl trrerd rqv np6o- 6eoq. To orlpefo enogqg 6e oupn(nrouv n],ripog pre O5oerg prero,trtrd(eov: ouprBo(vouv ndwcog orrlv (6ro ne- proproprdv4 neproX4. A[r(er vo orlpreuoOe( 6rt or (6reg 06oeLq oe 6rorpopen- roUg unoxlqr5g nopdXouv noMd on6 ro orlpre(o eno- rpr'19, 6oro Kr qv rl Bdoq nou Bpioreror oe Uro ouyKe- xprpr6vr1 06oq 6rog5per. Aur6 unodrlAdrvet 6n undpler rivog r<otv6g ;rr1Xovtotrr6g yro rnv np6o6eo4 qg RNA noLuprepdoqg, nop6rr 11 ow(6pooq 6ev e(optdror on6 r1v nopouo(o ouyrerprprSvrov Bdoeov oe optopr6vo on5 ro orlpre[o enoqrjg. Aw6 ro ;row5].o e[4ye( yror( xd- noro on6 ro o4pe(o eno<pr1g 6ev e(vor 06oerg peroMd- [errlv. Enrn].6ov, 6e Bp(or<erot r<dOe prerd].],o(r1 oe oqpe(o enorp{g: or preroMd(erg ;rnope( vo enrlped(ouv q yettovu<q neproXrl 1o:p(g mqv npoygolr6rnro vq Splowor o' eno<pri pre ro 5v(upro. E[vor r6ro[repo orlpowx6 6n ro nepdproro nou oKo- LouOo0v q orporrlyrxrl rng npo-rpononoqorlg rouro- nororiv Odoetg or4v (6to nepropl Trou npoorore[eror on6 ro 5v(upro Svowt enqK6Lou0qg rponono(qor1g. Au- rd to 60o nepdgoro perporiv 6ro<poperu<d npdyproro. H npo-ponono(r1oq ovoyvop((er dleg ttg O5oerg nou npdner v' ovoyvcop(oer ro 6v(upro yro vo npoodeOe( oro DNA. To netpdgoro npooroo(og ovoyvrop((ouv 6Leg rg O5oerq nou npdygort Splowor o' eno<pq oro 6rprep5g otipnloro. Ot npooroteu;r5veg 06oerg nepil.oprBdvouv 6leg ug 06oerg ovoyvc0proqq r<or peprrdg enrnpdoOereg 05oerg, yeyov6g nou uno6rlLtiver 6rr ro 5v(upro npcbto ovoyvcop[(et puo oprd6o Bdoecov onopo(r4tov yro rnv "npooye(oon) Tou Kr Snemo enercrefuet ro oqpe(o eno- grlg rou o' enrnp6oOereg Bdoetg. H neproXrlrou DNA nou e(vor (eruLrypdvq oro 6rpe- p6g oUpn).oxo prnope( vo rouronorlOe( d;reoo pe Bdon rrq Xqpu<5q oMoy6g orq 6ro0eo4-r6tr1rd qg. 'Orov 6ro- lcoprororiv or x.trri:vor rou DNA, or o(euydpcoreq Bdoerg yfuowor eud.troreg oe owtdpoorrlpro nou 6e prnopotiv vq rrq npooeyy(oouv or4 6rntr4 5Lu<o. T6roto neLpd;roro nepiloprBdvouv 06oerg pero[ri -9 xoL +3 orrlv opXu<r] ov- ildpooq rri[nq. I-r'qur6 11 neproXrl nou [eru]lyeror rord rqv 6vop[4 neprAopBdver ro 6elr6 dxpo rrlg oMq].ou- X(oq -10 ru ercre(veror pr6Xpt pr6).rg prerd r4v oEerlp(o. Av r<orrd[oupre rpro6tdmoro ro orlpre(o enocprlq qvodrKd rqg oLLr'1louX(oq -10 dronrmrirvoupe 6tr Bpi- oKowor 6Lo orq puo nLeupd rou DNA. Aur6 <pofveror oro xdrcrr Srdypogpo qg Eu<6vog 19.22, oto ono(o ro EIKSNA 19.U2 Mto ntreupo rou unoxtvlrrl nepL6Xer to orlpe(o enoqlg yro ro RNA. Atrtrr1trouX(o -35 t ALtrnlouxio -1 0 J+ t,I, ,1. l. I lt^,t+Iitl, I K06rKoIornrtK{iq t Ktrtivoq lxxxxllu.IffiKtrdlvog erpoyeio To neprooorepo orlge(o enoqlq pploxovror oq gto ntreupo rou DNA Tpononouloerq nou egno0[(ouv r1v npdodeorl qg RNA notrugepoorlg 06oerg rinou 1 RNA notrugepoorl npoororeuet ond qv rpononoiqorl l4ero),tro(erg nou xoropyouv 11 perdlvouv qv evepyriqro rou un0Krvlrr1 J + 0 ti ililil1 528 KEOAAAI0 19 Meroyporprl oroug l-lpoxoputilreg
  • 21.
    : ^Joro enoglqonpercovowor oe pro drnArq dtrrro nou j 1.,,'rror ond q prLo nLeupd. To neproodtepo Bp(or<ov- :trr orov r<ovo prq-er<pioye(o. Aur6g oL Bdoetg (or,rg ovo- .,ropi(owor r<otd tov opXrr6 olqpotropr6 evdg xLer- :.or-i 6Lpepo[g oupnLdr<ou. Aw6 0o enSrpene orqv R- ', - .rouprepooq vo n).rloLdoer to DNA on6 14 pro ntreu- - r (or v' ovcryviop(oer ourr; rqv dQrl tou DNA. (o0rig [errvd ro (erritrLypro tou DNA, ;rnopef v' ovo- .*pLoro[v r<or dtr].eg O5oetg nou opXrr<d Bp(orowot --',, oMrl nl.eupd tou DNA. | 9.1 2 Or AAAnAenr6pdoerq Mero(U rou llopdyovro o Kor rou llupfivo Tnq RNA IloAupepdonq AA deouv KoTd Tn Aroguyfi TOU YnOKIVNTfi n ','.0 n00t0Xl rou o rorotroppovet ro dtoutro e(o6ou rou RNA .r. lrprnrr vo perouvl0el ylo vo dteurotruver q o[v0eo1 rou t.i . - ,.nTUXe[q evop[erg ouvq0tog ouppo[vouv nporou ro ev(upo ,,i-porlorr evo npoygortro o0gntroro enLprlruvolg. " I .opoyovroq o ouvnOoq onetreu0epdrveror ono qv RNA :,,tpepoo1 potrtq q veoouvre0elpdvq otruo(60 RNA tptooet :: plxog -'10 nt. - rA noLupepdoq np6neL vo ouprBrBdoer rrg ovdy- ..: !'ro dvop[q pe rtg ovdyxeq yro enrprnruvon. l-1pco- -: , o 6(ouLoq Sroguyqg rou RNA ornv --:iuolx6rqto e(vot r<oreArlprpr6vog on6 rprlpo tou -r:ryowo o, ro ouv66rq (linker) nou ouv6Set rtq ne- - r ,.3Q 3 r<or 4. 'Eror, 11 6to<puyr1 rou unoKtvrlrl np5net : .oorloprBdver ovodLo-rdro(q rou nopdyowo o, .- cri(owdq rov on6 ro d(outro e[66ou tou RNA - -t vo prnope( vo ouvexrore( 4 otlv0eoq tou RNA. ' : - .3oov, q 6vop[r1 onone[ orevq np6o6eor1 p6vo oe - -. <a<prpdveg oMq).ouX(eg (unor<tvr1r5g), evcb q enr- --,-;ol onotre( np6o6eor; oe dreg Ttq oMrlLouXieg - - - :o ouvowrloer to 5v(upo Kord tq preroypogtl. H 6e(Xvet nurg enil.Uetot out6 to 6il.qprpro -.r* rlq owtorpemrlq np6o6eor1 tou nopdyowo o - :, rupqvo rou ev(Upou. Oncog npoovogSp0lKe, ro . --ic ouvrl0og u<p(ororor r<rirLoug oventtuXtov - ::ieov xord q 6Lepyoo[o r4g 6to<puy4g on6 tov --:..'rlll. Ev6oto oupBo(vouv outo( ot oventruXe(g . -- ' :., ro dv(upro 6e perorrvekor ndv<o oro expoye(o, : -' : -oMov rpoBd ro npdlto ).tyo vour<i,eor(6ro tou . :::5Lr<oU DNA, npoBdl).owog ouroUg roug provo6g . -- ,o,Jq novco orqv enrgdverd rou, oe pto dtepyoo(o -:rrl orq <paoqoq tou DNA - DNA scrunching". Ztn r- ,. '.aro. p' 6vov prrlXovtopr6 nou 6ev SXer onooorp4vr- , . .r.r1pcog, to 6v(upo dro<peUyer on6 ouq q <pdor1 --, r','enrruxov r<rjr<.trrrlv r<ot nepvd orrl rpdorl Tnq ent- _ -'- , rplyopo I f novo ww@w ",:1lk!'> / --*ru I i U rpnvopo * : ^m.:@E i ,*oypnyopo I i ^d!%' :*-Ef*eB ! n*rypnyopo I i#@r....... I To revrprxri 6v(ugo repgorl(er rr oneheu0ep6veror EIK0NA 19.23 0 nopdyowoq o ror o nuprlvoq rou ev(ilgou ovo- ruxtrtirvovror oe dtorpopenro oqpeio qg Ueruypoqnq. Molov6rt q oneLeu0Spoorl rou nopdyowo o on6 to o0pmtroro 6ev e(vor onopokrlrrl yto q 6roguyi on6 tov unorwqr], 11 drdoroorl tou nopdyowo o on6 tov nuprlvo rou ev(titrrou ouvrlOoq ouprBo(ver rout5lpovo 11 Lryo perd rq 6roquyrl. O nopdyowog o e(vor oe nep(o- oero or o16o4 pre qv RNA nolupepdoq, ouvencig 11 oneLeuO6pcoon rou on6 ro oLo6v(u;ro 6e y(verot onL6g yro vo ro xdver 6ro06otpro yrq Xp{on o' enrnp6- o0ero owlypoqo rou otroev(riprou. Irqv npoy;rorr6q- ro, ot nopdyoweq o owoyrov((owol yro neproproptdvo owfuporpo rou nupfvo qg RNA noLuprepdo4q, yeyo- v6g nou 6(ver q 6uvot6rqro v' oMd[er ro npSruno t4g peroyporp4g (BI6ne Ev1trTra 1 9.1 9). O nuprlvoq tou ev(6;rou (<,xewpx6 tiv(upto", core enzyme) cro rprprep6g oriprntroxo (nepd.o;tBdvet DNA, veoouweOery6vo RNA r<or RNA noLuprepdor;) ouorool- rd e(vor "x.l.et6ofr5vog" pr6Xpr vo o).oxlr1po0e( 11 enrprl- KUVon. 'On<oq 0o 6oripre, outl 11 e[6].t(q npor0met ev ;rdper on6 rov rp6no Ue rov ono(o ro 6v(upro neprKu- xl6ver to DNA ru ev p6per on6'rqv of[non qg ouyyd- verdg rou yro ro orl;rntroro Ue ro veoouwe0ettrr6vo RNA. KevTprKo 6v(uUo ono0lreupevo oro DNA 0 nopdyovroq o ouv06eroL pe ro revrprxd 6v(upo To otrodv(upo perorrve[ror npog Touq unorLvlr6g To revrptxrl ev(u;ro ouv06ret RNA 3.rtr,rltrenrdpooergMero[0rou[lopoyovrooKorToutluprlvorrlqRNAtlotruUepoonqAtrtro(ouvKoroqAroquyl:0i-'.].,.,-.- 529 Booxfg eworeg a
  • 22.
    To gdppor<o prgopnu<(v4(UdLoq rrlg orroy6verog rou owrBrorrroU prgol.rur<(vq) epno6((er q preroypo<pq on6 q Bor,m4pror<r1 RNA noluprepdoq. E(vor ro rUpto owrBtoru<6 evow(ov qq gupror(coonq. H rpucroMrr{ 6opr1 qg RNA noluprepdonq oe o0gnl,or<o Ue rn pt- <poprnx[vr1 e[4ye( q 6pdor1 rou cpopprdrou: n prqognr- r[v4 npoo6Sveror o' 5vo Ori],or<o rqg B unopovddog, >12 A prorpud on6 ro evepy6 r6wpo, oe ;uo 05o4 6nou nopoxrr:trrJer q 6to6popuj rou enrgnKuv6gevou RNA. Epno6((owoq r1v endrrooq qg oLuo(6og tou RNA ndpo on6 2-3 vouxl,eor(6to, 6tor5nter rrl Uero- Ypoen. 19..I3 H KpuoroAArrrl Aopri Yno6errv0er 'Evo MovrdAo yro Tn Merorivnon Tou Ev(0pou e To DNA perorrve[rol dropeoou evog droutrou orrlv RNA notru- gepooq ror npoygoronore( pto onoropq orpotprl oro evepy6 r6vrpo. x Atrtroydq oq dropoptptrlorl ouyxerprpevov eudtrrrrtov e[oprr1- porov p6oo oro ev(upo etreyXouv qv e[oodo rrov vourtrrol- d[ov oro evepyo xevrpo. Me q droLeUrovoq r1g xpuoro).).rx4g 6ogrjg rov ev- (tipuov on6 Borcrr;pro r<or (tipre oe oUpnloro pre NTP ror/n to DNA, onorcr4oope noMdg nnpogop(eq oxe- nrd ;.re rq 6o;14 r<or 14 l.enoupy(o rrlg RNA nolupe- pdonq rord rrlv enrprrlruvoq. H Boxrqpror<r1 RNA noLugepdorl 6Xer 6romdoerg -90 x 95 x 160 A, evo-l or s0A RNA noLupepdoeg rov opXo(rov Kot rov europu<orrirv e(vor eloEpcbq preyol0repeq, rup(rog ).6yo np6o0e- rov rgqgdrcov o1.uvo(6cov r<o/r1 unopovd6ov rou ev- ton((owor orqv neptg6pero rou ev(ri;rou. MoLorowo, ot nuprlveg rov ev(U;uov prorpd(owor 61r pr6vo r<owr1 6opr{, mrlv ono(o undpler 6voq "6[outroq" eripoug -25 A nou meyd(er ro DNA, oMd r<or r<ow6 pr;Xovt- qr6 yto npooOrjrq voux.l.eou6[ov. 'Evo gowriLo owo( rou 6roUl.ou orq Bor,irrlprorcg RNA nolupepdo4 ono6(6eror orrlv EIK0HA 19.24. H oG Loro ouyr<pore( -17 bp DNA. Ie ouv6uoopr6 pre to -13 nt DNA nou Bp(orowor orrlv neproXq rou evepyof x6v- rpou rou ev(0pou, owtorotloriv ornv npoororeup6v4 neproXrl tru'p<oug -30-35 nt rou noporqpe(ror oe on(> tunc0oerg tou oupnL6r<ou enrprlruvoqg. H o0Aoro enevdUeror pre Oeru<d gopr(o, Xdprl oro ono(o oM4Ae- nrdpd pe rtg n^erupopvrlnr6g qrr:orpoprxdg opd6eg tou DNA. To rorolurrx6 r6wpo (catalytic site) oX4pro- r((etor on6 pro o1rotrrr1 ovdgeoo mrg 6rio ;reydleg unopovd6eg rt otXl.roLor((er oro (oqy6vro" rou to D NA t<oOcbq ero6pleror mqv RNA noi,uprepdo4. H RNA noLupepdo4 neprBdLLer ro DNA Kor oro evepy6 rdv- rpo rnq nepdXer 5vo 6vo rorolurx6 r6v Mg2+. To D|.lA ouyrporefuor orq 05oq tou on6 rov r<o0odrrd ouvde- rrjpo (clamp), evoMorcrtrq ovol.roo[o yro 6vo on6 to ooy6vro. H EIK0HA 19.25 onerrov((er q orporp{ 90o nou ovoyr<d(eror vo npqyporonorfoer ro DNA orrlv e(oo6o rou evepyo0 r<5wpou elon(og ev6g 6rnlovoU npcoreivr- ro0 re[Xoug. To pr{r<og tou uBprdu<o[ RNA neprop[G- ror on5 6vo 6).Lo npo:reivu<5 eprn66ro, yvom6 og rdlu;r;ro (lid). To vourctreor(dro ntoreUerot 6u ero6p Xowor oro evepy6 r<5wpo on6 xdtco, prSoo rou 6eure- poyevorig 6torlLou (mqv RNA nol,uprepdoq rnq E0pnS ovoprd(eror "n6poS", pore). H rpuool(6o nepiloprB6va 8-9 bp uBpr6(ou preroyporptxrl DNA-RNA. To xdLuppro Movr6tro nou 6e[Xver q Dogrl qg RNA no- truUepoolq 6top6oou rou rup(ou 6roul,ou. 0r uno;-rovodec rordrronorouvror Xpopolro coq e[lq: B po(, 0, ruovo, o npootvo, oll, r[rprvo, o, 16rrrvo. Ipoooplroogrlvo onc K.M. Geszvain rorR. Landick (ed. N. P. Higgins). The bacte- rial Chromosome. American Society for Microbiology. 2004 FcTp,,l 530 KEOAAAIO 19 Meroyporprl oroug llpoxopudrreg Boorxeg ewoleg
  • 23.
    Avo6rxrl DNA 'Evo nporeivtro re(Xoqe[ovoyro(et To DNA v0 :,poqr1 oro evtpYo K6vTpo. Nouxtreor[dto pnopel vo oro evepy6 rSvrpo 0toireoou evog nopou oqv npto- :.-ir rLq BdoeLg DNA rot RNA oro 5vo drpo tou : :- tpl6ne Etr6vo '19,25) r<ot n Fdon tou DNA l:.cxeroL mo dMo dt<po rou uBpt6(ou ttvd(etot llpoor(0erot rowo0pu Bdoq xot ondve 0eogo( 'R11i-: lrt;ii,l lltr;111. To 6v(uuo gerortvefiot rot ovodrlptoupye( touq 6eoU00q il ;, r. ll ll:t,i, , -:- - "-..... --, ;:.r.i;il+1li.it:; ', Noudeot(6tq KtuIon tou ev(Upou ir I !1,_! 2To DNA Ero6pXoTot oro ooydvto : .: e[o yto vo enttpSqet to (euydptol-to Ue ro et- : -o'vo NTP. ^N r A . ,-^-^i -'(E* h -^avia rr,lr Ftwr,lnt- :1..q ro DNA unoore(qEn, n rpox(q tcov [elcopt- , - . ,.i.!vcDv U6oo crro 5v(upo dev neptop((erot nlSov :-: :- 5uoKoUqr(o 'rnq 6tnI(q 5trxog, enrp6nowog oto - ,: . : rpoYuoronotrloet rq orpoqn 90o oro evepy6 r6v- -: : I:.c1q, ouprBo(vet ptto peyoLr1 oMoyrl dtopt6ptpoorlq -- l : ro dv(upto nou nepAoptBdvet to "ouvderrlpo" o EIK0NA 19.26 H Ueroxivnon rrlq notrupepdorlg evdg vourhei- vrKorj oE6oq rpoUn0e6Tet Oporion Kot 0v06rlUtoupY(0 6eop6v pe vourtreotldto oe oT00ep6q e6oetq oXettrd Ue Tn Oogn rou ev(rlgou. To vourleoridto oe our6g rtg 06oetg d0d(ouv Kdee Qopd rou To 6v(uUo UeToKtvs(Tot KoTd Uto pdoq ndvto oT0 eK' uoYe(0. vouKleivtKov o[6ov nou Bp(or<owot prto 06oq nto rdrco Kord ilnKoq tou t<.trtilvou. To ;reoo[o n].o(oto 6e(1vet 6tt ow6 np6net vo onpo(vfl 6tt pr6tr'tg npooreOef n pdon Kot nprv ro 6v(uUo petorwqoe[ oe oxSon pe to vour]'ei- vrx6 o(ti, ot enogdq nou 61ouv 6r1prtoupy40e( pe er6x6g 06oetq np6net vo ondoouv 6tot cilore vo Unope( vo Eo- vodrlpuoupyl0oriv pe rtq Bdoetg nou Kqto^,ogBdvouv ng O5oetq our6g prerd q ;retor<[vqor1' Optop6veg rpuoroLLtt<69 6opdq rrlg RNA nolupe' p6onq npooq6pouv oqpowtx6g ev6e([erq yto to nc0q ro 6v(upro 6rotqpe( rtq enog5g pe ro un6orpopd tou ro0drg ondet Kot [ovo6qproupye( 6eoprofg qrn 6tepYo- ofq rou r<[x].ou npoo0rixqq vour],eott6(ov rt evcir uq(- orsrqt petot6ntorl. 'Evo eudNrro t;r(trro nou rol'efuqt 0qtud nupo66qonq (trigger loop) qo(verot 6tt 6ev efuot mulo;t6vo Tlptv rl'lv npoo0(xr1 tov vourl,eort6(tov, dr- d muXrivetot pr6l,tq to NTP etodLOet oro evepy6 x6v- rpo. 'Orov olorlqpo0e( o oXrl;ronopr6g tou deoproU rot r1 pretor6ntoq tou ev(rigou orrlv en6prevq 06o11, 11 Or1Ltd nupo66tr1or1g (erutr'iyerot rot n6Lt, 6roq-r1 yto tov en6- prevo r0xl.o, 'Etot, 1.tto Sopu<r1 oM.oyrl orrl OqLtd nupo- 66rr1oqg ouwov((et Tr1v orotrouo(o tcrrv oupB6wov rqq rot6).uor1q. :: cXqprot((et 6vo on6 to ooy6vto tou ev(fpou nou -ouv ro xo0odtr6 DNA orn 05or1 tou. " : :nd to 6tl.qpr;roto roOe notruprepdonq voux.e'i- :ieoq e[vot 6tt to 6v(upo np6net gev vo 6qpt- -:r,, oerxr5q eno<pdg pe to un6otpopo Kqt Ue ro : . lMo en(or1g xot vo 6tor6r.l.tet out6g ttg eno<p5g , : :.Q [ovooltlpot(oet oe rdOe r<0ro npooO4r<qg -'r::16(ou. Ag ovoloytmo[pre trlv rotdorooq nou . s3Qerot oTnv Mro notrupepdorl *::onote( pto oerpd et6trrov enorpov pre ttq Bd- : :,.l Bp(orowot oe ouyxerptp6veq O5oetq. l-tq no- :...*c, q enogrl "1" dtlutouPYe(tot pe q Bdorl mo :: :ou ou(ov6pevou x.l.<ovou, evco 11 enocptl "2" pt - ::]r orov rLdlvo er<proye[o nou e(vot ougn].qptl1'to- - . :: -. qv en6pevq B6oq nou 0o npomeOe(' Entoq-- . :: -. qv en6pevrl B6oq nou 0o npomeOe(' Entoq- -: ..:lr, ocn6oo, 6rt ot pdoetq nou rotol'oprBdvouvl. el, wv .: :rq 06oetg arouq r<Ltovoug ttov vout<LeivLxriv - i: --, oMd(ouv r<dOe <popd nou npoor[0etot 6vo vou- -: rovo.r ror to xdttrl nl,o(oto r1g etr<6vog 6e(lyouv -. l.c rordoroorl: prto Bdo'r1 np6rettot vo npomeOe( , :. :r(ov6;tevo xtrtrlvo. H dtotpopd e(vot 6tt mo rdttr; - r l.c o oufov6pevog x}'tovog SXet ener<roOe( rord pto : --- -l yeo-lpretp[o Kot rov 6rio ouprnL6rolv e(vot oxpt- :-: - i6Lo, qMd oro xdto nLo(oto ot enorp6g "1" Kot : :ooyUoronoro6wor pe Bdoetq tolv oLuo(6<ov tcov l* il H Kpuorotrtrrrl Aoprl Ynooltrdrver'Evo Movr6tro yto q Meroxtvlon lou Et'[-*:- 531
  • 24.
    19.1 4 Mlolropornp6vn RNA l-loAupepdon Mnopei vo lovopxioel r Mr0 or0U0Tlp6vq RNA notrugepool pnopei vo {ovopXioer ng pet0ypogrl xdpovrog ro RNA peroyporpo yLo vo Drlgroupyloer 6vo roLvorjpro 3' oxpo. H RNA noAutrrepdoq npdner vo e[vor oe 05or1 vo Xerpr- ore( rotomdosrq orrq ono(eg n Ueroypogq ovoor6).- Leror. I-ro nopdderypro, qur6 prnope( vo ouprBe( 6rov ro DNA SXer unoorei BIdBn. 'Evo np6runo oriorqpro yro r6roeg neprordoerg nporUnrer ge 6ror<onri qg enrpr{ruvoqg in vitro e(opc0wog 6vq on6 ro onopo(- rqro np66popro vouxleor(6ro, lepropr6g nou enmpd- ner rn gOopd rou dxpou rou RNA. 0o16oo, to [6ro onorSLeopo 61er xou<dOe ouprBdv nou nporoLef ].d- 0og euOuypdppton rou 3' dr<pou rou RNA Fre ro evep- y6 r<6wpo: r<dr lperd(eror yro vo lovoronoOerfoet ro 3'-OH rou veoouweOer;rdvou RNA oro evepy6 r<6wpo 6ror c6ore vo prnope( vo 5e10e[ en[Oeor1 on6 ro en6pre- vo NTP Kor vo oxrlpor(oer rpologo6eoreprx6 6eog6. H enoveu0uypdprgtorl ennuyldvetor pe ronq rou RNA to dxpo d:qre vo rono0erqOe( orq ocoarrl 06oq yro npoo0qr<r1 enrnp6oOerrr;v Bdoeolv. Molov6rr 11 11 xonrl e(vot Spyo rqg (6ro9 rrlg RNA noLupepdorlg, 6eye(peror nolri on6 enrroupxo0g no- pdyoweg, noL0 6to6e6opr5voug Kor oro rp(o BroLoyrxd Boofi.ero. Iro E coll undpXouv 6Uo'r6rotor nopdyov- req, or GreA xor GreB. H RNA nol,upepdoq ll rov euKo- purorrriv Xpno$onote( yro rov [6ro or<on5 rov TFllS. O TFIIS 51er pu<pri opor6rrlro oq npoq rnv oLL4l,ou1(o ror rq 6opr4 Ue rouq nopdyoweq Gre, otrld npoo66ve- ror oro 6ro rprl;ro rou ev(rjprou, ro 6eurepoyevrl 6(ou- ).o (n6po) qg RNA no).uprepdo4g. Or nopdyoweg Gre/TFllS enrp6nouv or4v noLupe- pdoq vo x6rler prepu<d prBovouxLeot(6ro on6 ro 3'rcLt- rd drpo rou RNA npoi6wog, ennp6nowog ritor mo roro.l.urtr6 rdwpo qg RNA nol,upepdoqq vo eiloveu- Ouypogrore( pre ro 3'-OH. Ko0Svog on6 roug nopdyov- reg erodyer gto orev4 nporeiVrrrl neproXrl [orov TFIIS e[vor gto r<op66],o geu6opyripou (zinc ribbon), oro Bo- rrqproxd riv(upro 6vo onerpoer66g one(popro (coiled- coll)l Bo0rd prdoo or4v RNA noi,u;repdor1, qrdvowog noL6 rowd oro rorolurrr6 r<6wpo. A(o 6(wo oUrvo- [5o or4v ropugrlrou nopdyowo nl,qord(ouv fio npo- -reUov Korqluttx6 r6v ;royv4o(ou rou evepyori xdwpou ennp6nowog rrlv e[oo6o ev6g 6ew6pou r6woq Uoyvq- o(ou ror gerorp6nowog ftor q 06o4 rordluoqg oe pr- Bovourtredoq. Iuvomxd, 4 enrpqxuv6prev4 RNA noLuprepdoq riXer rqv xov6trlto vo [eru],(yer Kor vo [ovoru].(yer ro DNA, vo ouyKpors( roug 6toXroproprdvoug r<).c0voug tou DNA ro0oig ror ro RNA (npoi6v), vo rorol,fer rqv npoo0rlxrl prBovouxi.eon6(rov orqv ou[ov6gevq otruo[6o RNA, vo enmrlpe( r4v np6o6o out4g qg ow(6poo4g Kor -Ue rrl 532 KE0A Al0 19 Metoyporprl oroug l-lpoxoputirreg ouv6popr4 ev6q r1 60o enrxouprxcbv nopoy6wov- vo enilUer npoBLriproro nou dqproupyo0wot on6 qv rorq Ltycov nt on6 ro npoiSv RNA rot u1v enovdvop[n rnq otivOeorlq. 19..l5 H BorrnploKn RNA t-toAupepdon Teppori(er o€ AtoKprrdq @6oerq * YnopXouv d[o ro[eLq otrtrr1trouXuirv reppologori. Exelveq nou ovoyvopf(ovror povo ono qv [6ro rrlv RNA notrupepoorl Xo- p[g rr1 oupperoXq rovevoq rurroprrorj nopoyovro, ouvrl0otc ovotptlpovroL oq "evdoyevelg rep;louoreg,, Atrtreq 0n0rT0u, pro rurroplrrl nporelvr1 nou ovopo(eror po (p) xor ovorpepov- Tot trq <<TepUoTroreg e[oprc0pevor ono rrlv p". * 0 evdoyevrlg repponopdg npouno06reL ovoyvtrlprorl ;rrog otr- trqtrou1rog repportopou oro DNA nou roduonorer pto dogl qouprerog oro RNA npoiov. * To o1poro yro reppolopo pplorovror rupirrrg p6oo oe aAAq- Aouyieg nou eyouv 1764 peroypagei ono qv RNA notruUepo- o1 rr dror o repporropog pp[oreror un6 rov 5treyyo iou erpoye(ou ro/r1 rou RNA npoidvtog nou geroypotper I notru- uepoolt. M6Lrg 11 RNA no).uprepdorl [er<rvrloer rrl pretoypoqq t<ord pujr<og rou erpoye(ou, ouv06rer RNA, pr51pL vo ouvowrloet prto o).).qLouX(o repporrorq (terminator) . Ie out6 to orlpe(o, ro 5v(upo oro;lord vo npoo0drer vour<).eor(6to orov ou{ov6pevo r}.rrlvo RNA, one}.eu- Oepcbver ro oloxr1pcopr5,vo npoi6v xor 6rfororor ond to erproye(o tou DNA. O reppouopr6g npouno0dreL 6rL SLor or 6eogo( udpoy6vou Trou ouyKporoJv ro uBp(6ro RNA-DNA np5ner vo ondoouv ror getd vo [o- vooxrlUorrore( ro 6(x).rrlvo DNA. Meprrdg eop6q e[vor 6Uor<o].o vo npoo6rop(ooupe q 05o4 tepprorLoprorj yro 6vo RNA nou dlet ouwe0e( oe (orwov6 rlirropo, ener6q ro 3' dr<po rou prop(ou Unoper vo 6toonoore( on6 pro 3' efovouxi.edorl r1 vo rone[ ond pLo ev6ovouxedor1, lcop(g v' ogrloer rov6vo (Xvoq qq 05or1q dnou npoyprolr<d reppdttoe 11 RNA no).u;iepdorl: out6 e(vor rdro(tepo rorvd oroug eur<opurrtreg. l-r'ourd ro ).6yo, ot O5oeLg -reppolop.roU oulvd loporcrrlp((owoL xo).0repo in vitro. C)016oo, 11 rrov6rr;ro rou ev(Uprou vo reppot((er in vitro enrlped(eroL noLU ond nopog5rpoug 6norg q rowrxrl roltig xot q 0eppror<poo(o mqv ono(o npoyporonorefuor r; ow(6poor1. 'Eror, o tepporLop6g oe prro opLoprdvq 06,o1 in vitro 6ev onoreLe( on66er(q drr 11 o6v0eoq 0o reppolore( orrlv (6ro 06o11 Kor oro x0ropo (in vivo). 0o16oo, 6rov ro (6ro 3' dr<po ovrXveUeror 16oo in vivo ooo xor pe roOopd ouororLrd in vitro, our6 yevr- rd Oeorpeftor utg Lcrluprl Svdetlq 6tr npdrenor yro rnv ouOewrxrl 05or; tepprorLoproU, Ot i.li,,..::. ., " Ko[ r'jir-:,:... i'. 'rrr oUVoqJ((oUvro 60o r<rjpro lopor<rrlptmrr<d rcov evdoyevdrv repporror6v Boomeq eworeg Boorxeg eworeq
  • 25.
    Ixe6ov otreq otonopo(rrlreq lrliltrouXieq yro tov repgotlopo Ppiorovror otr1v fleptoxn nou usroYpdQnKe H rpoupr5ro oTo RNA Unopei vo onotTtiTot 0t otrtrtltrouxieg DNA nou onotTolJvTot Yto Tov -.:-lrLolto $plorovrot ovootro Uq otrtrntrouxtog rou repportorrl, , ,,-lorrouoq uroq QoupKer0q oro RNA unope( v0 elvot 0n0p0i --tt ntri nsic terminators). l-l ptrttov, ot ev6oYeve(q repUort- : .q, or ono(ot e(vot ove[dprlrot on6 rov entrouptr6 nop6yovro po (rho factor), Yto vo oXqpot(oouv trl 1.-repotoyq doprrl rou RNA nou preroypd<petot onot- :JV llo goupx6ro (hairpin) n).oUoto oe G+C. 'Ero4 11 -.,aypagn eoptatat an6 rc RNA npoi6v xat 5ev xa)o- .*#.w E*G ffiiG b#& L&th U}4F $r$4 "6RqeAg. h* wtrg6lE h&r& ilfl-E ffi ffi neproxrl iloriotq oe G-C oro gtoxo ffiq [Jels il&T TMovoxtrovn oend U EIKONA 19.28 0r evdoyeve(g repponor6g nepiloppdvouv notr[v- dpogeq neptox6g nou oXl;roti(ouv goupx6req Kot notKfiouv 0s g4rog on6 7 etrrq 20 bp. H 6opr1 Utoxou-Ontrtdq nepiloUFdvet Uto neproxn ntroUoto oe G-C nou oxo)rou0eltot on6 pto ostpd K0T0- tro(nov U. p(etu anLriq pe lleyyo n1q a&rlLouy{ag tou DNA xa- td q peraypacpfl. 1o 6e0repo XopoKrnptsrr6 efuot pro oetpd xotdtrotnov U (T oro DNA) nou orol'ou0o0v to pr(o1o qg qoupr<6tog oMd nporlyo0wot qg npqYpm- rrlq 05oqg repUortopoti. Ito yovtd(topto 'cou E. coli undpXouv -1100 otrtrr1LouX(eg nou totptd(ouv pre owd ro r<pttripto, yeyov6q nou unodqcilvet 6tt ndvo on6 to gto6 pretdypqqo rou xurrdpou tepprot(owot o' ev6o- yeve[g reppottm6g. Ot repportor6g nou efopr6wol qn6 rov p (rho-dependent terminators) xoOop(owot on6 t4v ovdyr<r1 npooOrixqg tou nop6yowo p in vitro. En(oqg, pretoMd(etq 6e[lyouv 6rt o nopdyowog epnl'd- Kerol fiov reppronop6 in vivo. Ot tepgoror6g nox[trouv no]li og npoq rnv onotetre- opcntr6rrpo. Og per6ypogo nopopiooqg (readthrough transcripts) ovog6powot to pretdypogo nou 6e oropo- roUv orovtepgonon(. (O 6poq nopoB(ooq lpqoqlonoef rot ornv Evot7ta 25.14ytovo neptypdtpet rrlv xotooroNi rodu<ov(cov tepporropori on6 dvo ptBooropdtto). En(oqq, o rcpponop6g prnope( v' onotpone( on6 et6u<orig entKou- pu<oUq nopdyoweg nou oMrltrenrdpo0v pre to RNA ro/r1 rqv RNA noluprepdorl, rotdoroorl Yvoorn oq qwlrep' ponopr5g (antitermination). 'Eror, 6nog rot orqv nep(- rrroorl qq 6vop[r1q r1 qg entptir<uvonq, o tepporrop6g prnope( vo pu0ptore( oq Unxovtotl6g yro tov 6treyXo rqg 6rgpoorlg rrov yovt6(tov, 6noq 0o 6ori;re oro Ke<pdtroto 27, Zrpaqyxiq tav @dyuv. YndpXouv Kqt dMot nopoM4Lrqro( preto[U 6vop[qg Kor repportopro0. Kot or 6Uo 6epyoo(eg qnomo0v 0rt6- orpro deoprdlv u6poy6vou (opXu<r1 ui[r1 tou DNA r<otd qv 6vop[r1 rot 6tdorooq rou uBpt6(ou RNA-DNA orov tepgottotrl6) r<ot pnope( vq XprlotUonoto6v entnp6o0e- reg nproreiveq (nopdyoweq o, evepYonotqt6g, roro- orol.e(g rot nopdyoweq p) nou otrq}'ent6po(v pe rov nuprjvo tou ev(0;tou. Qor6oo, ev<ir 11 6vop(q Boo(etot pr6vo orrlv oMr[ren(6poor1 prero[U qq RNA notru;repd- onq Kor tou dtxtrtivou DNA, o reppottopr6g nepilopBd- ver Kot ovoyvtilptorl onpdtov oro ;retdypotpo on6 qv RNA nol.ugep6or1. Ot orlpretor<6g pretoMd(etg nou trretcbvouv rllv onore- treopour6qro rou tepgottogoti ouvrlOoq ouprBofuouv p6oo oro p(oxo rqq rpoupr5rog ri ornv nLo0otq oe U oMrllouX(o, 6v6et(4 Yto rrl onUoo(o outriv ttr:v oMq- trouXtc0v oro prlxovtopt6 tou reppottopro0' To uBp(6to RNA-DNA 6Xet preydtrrl ouvetogopd ortg duvdpretq nou ouyrpototiv to otigntroro entpfruvoqg' Euvendrg, to ondotpto tou uBpr6(ou 0o prnopoUoe v' qnooro0eponot- qoet to o(prnLot<o en4.uiruvorlg, o6rlydrwog oe repUo- rrotrt6. Atrtrntrenrdpdoetq rrlq <pouprdtog pe rrlv RNA noluprepdoq ri 6uv6pretq nou oot<o0wot on6 ro crxnpo- nopr6 qq goupr6tog ro0tirq ro RNA (enpoBdMer on6 ro 6(qutro e(6dou tou RNA, prnoporjv np6oxotpo vo eu- Ouypoprpr(oouv pre trd0og rp6no to 3' dt<po tou RNA pe ro evepy6 x5wpo tou ev(Uprou. Aurl q I'ov0oopr5u1 eu- Ouypdprpuoq, oe ouv6uoo1.t6 pe to oouvti0toro oo0ev6g uBp(6to RNA-DNA to ono(o oXrlpro{(etot on6 to (eUyr1 Fdoecov rU-dA RNA-DNA nou npox0mouv on6 ro 4tt1- Uo Ue ro rqtdlotnq U, onosroOeponote[ to otipntroxo enq.rlruvorlg. 19.15 H Bonrlptorrl RNA l-lotrupepdon Tepuqr(et oe Aroxptt6q 06oerq 5:B lGUu UA GG AA I H RNA notrupepdoq ror ro RNA onetreu0ep6vovrot ..'}..I. , [ }'^
  • 26.
    H onoteLeoporrr6qro rourspUorrol,-tou in vitro not- x0,er nol.ri, n.X. on6 2% 6toq 90%. H onoreLeoponr5q- ro rou repUorropori 6ev e[oprdror pr6vo on6 trg oMqLoul(eg on1 goupx6ro KoL rov oprOpr5 ror ng 06- oerq rov roroLo(nov U xo0o6rxd qg goupr<5rog, oMd ror on6 otr).r1LouX(eg 16oo ovodrxd 6oo ror ro0o6u<d rr;g 06oqg repponopori. Aw( vo reppor(oer, ro 5v(u;ro gnope( onl6q vo rdver puo no6or1 npororj (ovop1(oer qv enr;u1r<uvo4. Aut6q or 06oerg no0orlg prnope( on6 p6vegroug vo ennetroUv puOpuoru<o0g oronoUg (Bl,6ne Ev6trya 26.13). To ov 11 RNA noLu;repdorl 0o oroprorl- oer Kor 0' one).euOepciloer rov r<Lrrlvo rou RNA r1 ov on).c09 0o oroporrloer Trprv [ovop1(oer rq ;reroypogri (5rll,odn q brdpr<ero qg noUorlg ror 11 onoreLeogour6- qro drotpuyrjg on6 rrlv noriol) r<o0op(owor on6 6vo no),[n].or<o orivo],o r<tvrpxoiv r<or Oep gobuvotrru<r6v no- poy6wcov nou drlproupyorlwot on6 ro lopor,rqprorrxd qg goupr6roq Kor rou nLo0otou oe U t;rq;rotoq oro RNA, 6no4 enioqg xot on6 rrg ovodrrdq rot ro0o6trdq oMqLoul[eg rou DNA. l-ro nopd6erypo, no0or1 prnope( vo oupBe[ oe 0doerq nou ;rord(ouv Uev Ue repprorrordg, lopor<rqpi(ovror 6puog on6 on6oroorl prero[ri rqg rpoupr6rog Kor rnq oerpdg U preyoLUreprl on6 q Bdtrrr- orrl yro rov reppotropr6. l-loLri Lry5repo e(vor yvroord oXerrxd pre ro orlproro Kor rouq enrroupu<orjg nopdyoweg nou egn),6Kowor srov repportopr6 rcov europurorrxc0v RNA noLupepo- ocilv. Kd0e rd[r1 no].uprepdonq go(veru vo Xpnoqonor- e[ (e1opror6 prlXovropr6 (BMne KeEd],oto 21). 19.16 l-l6q Aelroupyei o llopdyovroq p; * 0 nopoyovrog p e(vor pro npotreivrl nou npoo66ver0r oT0 vr- oouwe0eLp6vo RNA roL perouvelror xoro gqrog rou RNA yro v' otrtr4trenrdpooer U0 Tlv RNA notru;repool Kor v0 Tlv ors- LeuOeptiroer ond ro oupn),oro enrplruvorlg. O nopdyowog p e[vot onopo(rr1r; nporelvq oro E. coli nou npor<ol.e[ tepironopr6 rnq Lleroypognq. H ouyrdwpotoq rou prnope( vo grdver 6otg xor -10% qg ouyr<5wp<oonq rnq RNA no).upepdonq. Tepprorrop6g ove(dpqrog on6 rov nopdyowo p unooy((eror 6rr ouprBo(ver oXe66v oroug l.rrooriq repponordq rou E. coli. H - nopouord(er 5vo prow6Lo yro rq Ler- roupy(o rou p. l-lptito npoo6Sveror oe pro oM4louXio prdoo mo prerdyporpo ovodrrd rrlq 05or1g tepporLollori. Aurq 4 oLLq).ouX(o ovoprd(eror 06oq rut (or<provrj;uo yro: rho utilizalion - XpnorUono(non p). Iq ouv61eto, o nopdyowog p peror<rve(ror rord piqr<og rou RNA drog 6rou npo).dBet -rr1v RNA nolu;repdoq. Orov 4 RNA no- Lupepdoq qrdoer orl 05oq teppotrollori, o p e[ovoy- rd(er ro 6v(upo v' one),eu0epc6oeL to RNA. H noUoq 534 KEOAAAI0 19 Meroyporprl oroug t-lporopudreg {p ra EIK0NA 19.29 0 nopoyovrog p npoo66veror oro RNA oe pro 06- q rut Kor peroron[(eror xord prlrog rou RNA gdXpr vo rprdoeL ro uppiDro RNA-DNA mlv RNA notrupepdorl, 6nou one),eu0eptlver ro RNA on6 ro DNA. on6 r4v nol,uprepdorl orrl 06oq repporropro0 6(ver 1p6- vo orov nopdyowo p vo Uerorontore( oro uBp(6ro xor e(vor o4 1.rowx6 Xopor,r4 prorrr6 ro u tep porrollo 0. E6ci BL6noupe Uro oqpowu<rj yevrxri opXr1. 'Orov yvrop((oupre q 06oq mo DNA mqv ono(o enevepye( pto npore(vq, 6e pnoporipre vo ouUflepdvoupre 6rr owr] oupn(mer Ue rnv oL.4Lou{o rou DNA nou oplrrd ovo- yvop((er. Or 6rio O5oerg prnope( vo drorpSpouv xor 6e Xperd(eror vo undpXer pro ro0opro1.r6vn oX6on prerolri roug. llpdyporr, or 06oerg ruf oe 6rogoperrr6g prero- ypogrx6g ;rovd6eg Bp(or<owot oe 6rdqopeg onomd- oerq nprv on6 lg O5oerg repprorrol.ro6. Mro nopdproro 6tdrproq y(veror on6 roug nopdyoweg owrrepUorr- o1-rori (BL5ne Ev6qta 19.22). E(vor dyvcrlmo rr ouvrord orrlv npoypqrrx6rqro puo 06,o1 rut. Korv6 Xoporrrlproru<5 tov 06oeov rut e(vor H RNA noLupepdon Ueroypdqsr DNA 0 p peroroni(eror rord prlrog rou RNA H RNA notrugepdorl rdver gro norjorl orrl rpouprdro roropqvnpotropo(ver 0 p [etutr(yer to uppGro DNA-RNA tTepgortog6q: dtro ro oumortxd one)eu0epdrvowot Boorxeg eworeq
  • 27.
    Bdoetg C 41'/o <-H onotrotQr1 onorp6net rov TepuortoUo '+ A 25lo u 2oo/, $ 14Yo H e60n ru1 6Xer pro gtrtrltroux(q iltrguoro 0e C Kor QToXn 0e G nou npolyetTot TIq rpoYUoTtKnq(ov) 0eo1q(eu:v) TepUoTt- ouou. H otrtrl^ouxio ovTtoTotxel olo 3' oKpo Tou RNA. 6n q oMrlloux(q rouq e(vot nl.o0oto oe Kordtrotno C Kor Qr@xn oe Kord^otno G Kor 6ev 5xet 6ewepotoytl Eopui. 'Evo flopddetyUo 6(vetot ornv ElK0l'lA 13'30. H C e(vor Ue dtogopd rlnto Kotvrl Bdorl (41%), evcb n G n It- y6repo Kotvn Bdon (14%). Ot 06oetg rut nox(trouv oe pnKoq. Qg yevu<6g rov6vog, rl onoretreogottx6tqto Uroq e6onq rut ouqdva pe to prirog rnq neptoxiq nou e[vor ntro0oro oe C/<ptoxn oe G. O nopdyowog p e(vot pr6l,og ttlg otKoY6vetoq rcuv e[qreptirv eturoocilv nou e[oprc0wot on6 to ATP. Kd0e unoprovd6o 61et pto neproXrl np6odeoqg RNA rot ptto neproXrl u6p6Luo4g ATP. To e[oprep6g treroupye( nep- vdrwog vour.treivtr6 o[ri 6ropr6oou qg onrlg oro p6oo qg 6o;ujq nou oxnUor(erot on6 trg neproX6g np6o6e- oqg RNA ttrlv unogovd6tr:v {EIK0NA 1$.31}. H 6o;tt1 tou p 6(vet xdnoeg evde((erg yto ro n6q 0o prnoporlos vo tret- toupye(. Oq[vetot 6rt ru],(yet RNA on6 ro 3' dr<po yUprrl on6 ro e[coteptr5 tov N-retrtt<citv neptoXcbv rot onpril- )yer ro 5' drpo rqg npoo'6e6epr5vr1g neproXqq oro eoo- rep116, npoq Uto 6euteperiouoo neptol{ nou npooS6vet rtg C-rerr6q neptol6g rou RNA. H op1trci popei rou p eivot 6vog dtoreroprprdvoq SorcrrjLtog, oI'- Ld r1 np6o6eon rou RNA tov petorp6net oe xtretor6 6o- rrr0trro. Merd rqv np6o6eoq orq 06o11 rut, o p Xprlotponoe( qv evepy6qro rng elrrdorlg nou 6to06tet Yto vo pero- ronrme( xotd prrlxog tou RNA, to0o6prevog on6 tqv u6p5l,uor1 rou ATP, ;r5Xpt vo Etdou t4v RNA nol'upe- pdoq. Iq ouv6Xeto, Xdprl crrrlv evepy6t4to efuxdoqg, [eru],iyer q 6(r].ovr1 6oUn xo/ri oMrl]'entdpd Ue rnv RNA noLuprepdorl oupBdMowoq qrnv one).euO5pocn1 rou RNA. O nop6yowog p npSner vo Uerorontsre[ r<otd gti- Koq rou RNA on6 14 06o11 rut oro npoyl.tottt<6 o4pe(o repportoporl. Aw6 npounoOSret 6n 0o petortvefuot ro- XUrepo on6 rqv RNA notrupepdorl. To 5v(upo xdvet pto norioq 6rov <ptdoet o' Svov tepgononl Kot, ov rrlv npo- grdoet o nopdyowoq p, eq ouprBe( tepprottopt6g. Iuve- ncirq, rl 4 norioq 6Xet oqproo(o Yto rov teppottog6 nou e[optdtot on6 tov p, 6nrrlg Kot orov ev6oyev4 repUort- opr6, ener6{ 6(vet to Xp6vo yt' dltro onopo(tr1to ou1.t- Bdwo. H o6(eu[r1 1.rero[U ;retoypoqriq r<ot pretdgpooqq 61er orlprowu<5g npoercrdoetq Yto rn 6p6on tou p' O p np6net npc0to v' onorcrnoet np6oBoorl sro RNA ovodt- rd on6 ro otipilr,oxo preroypogriq rt d'nemo vo UeroKt- vq0e( r<ot6 prrp<og tou RNA yro vo npogtdoet qv RNA no)ruprepdoq. 'Erot, n Spdon tou eprno6((etot 6rov to prBooroprdno preto<ppd(ouv 6vo mRNA. To ouyrexptptd- vo pow6Lo el4ye( 6vo rpotv6gevo nou npoBtrrlpfutoe roug npriroug Bor,mrlptoro0g yevenor5g. Ie opropr6veg neprm6oerg, 1.uo oouv0ettr4 petdMo[r1 (nonsense mu- tation) o' 5vo yov(6to ploq UeroYpogu<r1q provddoq Bp6- 0r1xe 6rr onotpdnet trlv dr<rppoorl rov endpevtov yovrS(rov orrl provddo o16;ro Kt ov Kq06vo on6 to 60o yov(6ro e(Xe tn 6u<rl roug t6to(tepq 06o11 np6o6eoqg mo prBoocol.tdtto, qorv6ptevo yvtrlor6 og notrlx6rqro (polarity). M6oo oe Uto Ueroypogrrci povd6o, ot 06oetg reppo- rropro6 nou e[optc0wot on6 tov nopdyowo p ouvrlOog rotrUmowot on6 ptBootoprdno nou Ueto<ppd(ouv (ElK0' NA 19.32), on6re o p 6e prnope( vo enevepy{oet oe ro0o' 6rrdg RNA notrupepdoeg. Oor6oo, oouv0ertr6g pretotrl.d(erq oneLeu Oep rirvo uv p tBooco;rdno prSoo oro RNA ev6g notruyovt6tot<of onepov(ou, enttp6nowog orov p vo repgoiloet rrl UeroYpogrl np6olpo rt eprno6(- (ouv tr1v 5r<ppoorl onpror<puopr6vrov yovt6(rov rnq pero- neprogdq RNA 0 eloUepqq 0om[trtoq rposddver RNA 3', 0 p exet pto N-relLrq neptoXrl npooDeorlg RNA ror gro C-re),rrn noptoxrl ATPoonq 'Evo e[oUepeq uno Uopqn evog dtoreroppevou borrutr[ou npood6vet RNA roro Urlroq rr1q eloreprrnq enupovetog ttrtv N-retrtr6v neptoXtilv. To 5' orpo rou RNA ervor npoodede;rdvo oe pto deurepoyewl 06on oro tooTept- 16 rou e[opepo0g. I I f i"i i n6q Aeuoupyel o llopoyovroc p 535 AU CGEUACCUCAUAU COSCA*{U CEU C}AqA C{i CIJA C'UCGACiAGAMS$CffUOU CUU
  • 28.
    il: i il:,li-,ri:i:,1: i,5ill[:Hl wrpd(l)p0 ^@ H dpoott rou nopoyovro p pnopei vo ouvbeoer q peroyporpri ror rr1 peroqpool orov evoq p-e(oprtlpevoq TsppqTroTll( pp[oreror rovro oe pro oouv0elrq perotrtroi1. ypogrrlg provd6og nopdrt r' ovorXrd nLq(oro ovdyvro- oqg roug nepeiXov oM4louX(eg dyprou r0nou. l-ror( ro oroOepd RNA (rRNA r<or IRNA) 6ev unoBdL- l,owor oe notrrx5r4to; To IRNA e(vot Bpox6o xr dXouv ercretop6veg deurepoyeve(g 6op6g nou ntOovdv ono- tp6nouv qv np6odeoq rou p. Ttrrrjproro rtov rRNs en(- o4g 61ouv errerop6veg 6o;159, ol,Ld ro rRNA e(vor nol,0 prorprjrepo on6 rq IRNA, ogrlvowog oprer6g eu- rop[eg yro rn Spdon rou p. To r<6rropo 6prcog 61ouv ovom[[er 6vov dMo prrlXovropr6 yro v' onorpd,nouv rov np5opo reppotro;r6 rov rRNA geroypdqov: 6ro0rirouv npolteiveg nou npood5vowor ortg ovol.ro(6preveg 0d- oeq nut (nut sites) mrq neproX6g-o6rlyorig 16oo rcov 165 6oo Kot rov 23S rRNA preroypdqov, oXrlpori(ov- rog oripntroxo owrepponopori (antitermination com- plexes) nou qvoor6l ouvrrl Spdoq rou p. Ot peroMd(er.q rou p nopouord(ouv preydl,rl notrr- Lo;ropg(o oq npoq rtg enrmcioerq rouq orov reppol.- opr6, To Boou<6 onot6l,eopo ourc6v trov pretoMd{ecov e[vor onorul(o rou repprorropori. Om6oo, o Bo0p6q rqg onorul(og, 6nog errrpdror on6 ro nooosr6 ralv nopoBrdoeov in vivo, e[oprdtor on6 ro ouyxexpqdvo r6no nou onorel,e( or6Xo yro repporrqr6. l-1opo;ro(og, 4 e(dpqoq on6 tov nopdyowo p in vitro nou<il.er. Me- prxo( (p-e[opr6prevoQ repporror6g qnonorjv olenrd ut|lnMq ouyrewprioerq rou p, ev6 dMor treroupyo0v e((oou xoLd oe 1op4l6repo en(ne6o. Aur6 unodrll,c0vet 6rt 6rogopenro( repporrm6g ononotiv drogoperu<d en(ne6q rou nopdyowo p yro rep;ronog6 ror yr'our6 owonorp(vowor drogopenrd qro unol,erptrronrd en(- ne6o rou nopdyowo p nou onop6vouv orct preroMoy- 536 KEOA Al0 19 MeToypoQn orouq tlporoputilreg :;- :a: 4..:t.:)!.,, .. To prpootopdrro ouoreud(ouv mRNA n(oo on6 qv RNA no)tupepdoq To prpoorrrpolo egnod((ouv qv npooxritrqoq rou p ro/rl rt1 UeToK[vnon 0 p npoo- rotrtroroL otrtrd to prpootrtpdlo egno6[(ouv 11 petox(vr1oq rou IuveX((eror 1 peroypogrl To ptpootrlpdrro drimowor oq 06011 qq [erdMoqr]q -0 p onoxrd npdopoorl orrlv RNA notrugep6orl pr6vo oretrSXq (ouvr1Oog 1oporcr4p((owor. oq preroMdy- proro droppoqg, leaky mutants). Kdnoog preroMd(etg rou p Urops( vo xorooroLo0v on5 prerotrld[erg oe dMo yov(6ro. Aurl 11 npoo6yyroq npoo<p6per 6vov e(orpenr<6 tp6no yro rrlv ovoyvcbproq npcoreivc0v nou ol.tr,4Lenr6poriv Ue rov p. H B unoprovd- 6o r4g RNA nolu;repdonq epnLdxeror oe 66o r0nouq peroMd[eo:v. llprirov, or preroMdlerg oro yov(6ro rpo B pnope( vo perdtoouv rov reppolopr6 oe puo p-e[op- rriprevq 0dor'1. AeUrepov, or peroMd(erg oro yov(6ro r- po B pnope[ v' onororoorrjoouv rqv rrov6rqro teppottopoti rnq perqypoqTiq orrq p-e[opt6peveg 06- oerq oe Borcr4pro trre ;reroMd[erq rou p. Ae yvotp((oupre oor6oo noro Lemoupyrx6qro 61er q oM4).en6poor1. 19.1 7 H YnepeAiroon Eivot Inpovrrr6 XoporrnproTtK6 Tnq Meroypogfiq e H opvnlrn unepetrlrtrloq ou{over rqv onoretreopoTrKoTlTo ronoulv unouvrptiv enerDl poq0o rr;v ovr[6poo1 onodtdro- Ers. 6 H Ueroypoq1 dqproupye( pnpooro on6 ro ev(u;ro Oertxec unepetrrxeg xor n(oo rou opvqrrx6g unep6trrreg, or onoleq np6net v' orporpe0ouv ono q yupooq xor rlv ronoiooUepoort T6oo ot npoKopuonr<5q 5oo Kor or euKopuco-rLr<6g R- Boorxeg evuoleg
  • 29.
    ',: roluUspooeg Qo(vetot6rt ln vifro evepyoilotouv -- JoroypoQq onoteLeoprottx6tepo 6tov ro expoye(o . ,:i unepetrolp5vo. Meptr<6q Qop6q, q onotetreopo- - ^,irnro rou unoKtvnrrl evtol0etot ewuntootoKd on6 -- . opvrlttr<rq unepe[rtrlotl' I-tor[ optol-ttivot unoKtvn- -:l enrlped(owot neptoo6tepo on6 to Bo0p6 tqg - .aoel(xtooqq on6 dMoug; H nto ntOovrl e[r1yr1oq e(- ,l. ort 11 e[dpqorl ev6g unortvqtrl on6 tr1v unepe]'[- --oq r<o0op((e-rot on6 qv etreUOepq ev6pyeLo nou : .cLre(rot Yto rqv q(r1 tou DNA cn'o otipnor<o tivop- :-l Me q oetpd rr1g, 11 eleU0eprl evSpyero e[optdtot r 3 rllv oMrlLoul(o tou DNA rou unoKtvnrrl: 6oo nto -'.c..loro oe G+i eivot 11 oLLqoux[o tou unortvqtrl -: - owtorotXe[ orrl 05or1 qg pretoypoqtr<r1q quoo]'G l:l rooo nto orlpowtKrl yto tov unortvrlrl 0o e(vot 11 - ,ce).[root1, q ono(o 0o tov ouvSpdpret crrqv r1(r1 -:- DNA. ',1ooto[ro, ot ent6pdoerq rnq unepe]'(xtr-ro4g orrlv . . ':vorlro ev6g optopr6vou unoKtvntrl e(vot nol0 nto -:'.-n,or<o (ntnpo. H e[dpqorl dto<popettxt'tlv unoKt- - ;v on6 ro Bo0pt6 qg unepeL(Koonq enqped(etot ,-.rrq on6 oMrlLouXieq DNA 6[tl on6 rr1 rpuooL(6o, .-..5q q unepel(xcr-ror1 otrI.d(et q yeopretp(o rou oup- - :(ou enrlped(owoq ttg yc^lv(eg Kot rlq onomdoetq -.::[Lt rr.ov Bdoeolv mo 1c,-lpo. 'EroL, 6to<pop6q mo :::,o rqg unepel'lrolorlg prnope( v' oMd(ouv ttg o]'trr1- . 5pooetq preto[ti ttrlv Bdoetov rou unoKtvnrrl roL rcov --.. .c(er-,.rv tqq RNA nolutrtepdorlg. Entn}'5ov, enet6rl - ::opertxd pr5pq tou Xpclprootrlprotog 6Xouv 6to<pope- - - l-q Bo0proIg unepetr(r<.rlonq, ro onot6treogo trlq - .:eit<rr.lot1q onlv evepy6tqto ev6q unortvqtrl prnope( . : rrqped(eroL on6 rq 06o11 rou unoKtvllrq oro Xpirl- - - l!f-lo. (oOoq q RNA no)'uprepdorl ouveXtrlq (etutrtyet rot ::,:ru,Lyet to DNA evul npoXtr.lpel mo ex;royelo (no- ::-:lc(etor orqv ELt<ovo 19.4), e(te o)'6x}r1po to otipr- - :{o rlq pretoypoqqg npdnet vo neprorpo<pe( yUptrl r-: :c DNA, eite ro (6to to DNA vo neptorpoqe( yfpro r-: rcv o(ovo trlg 6trtr<og. l'ltme0etot 6tt nLo xowd - - , itpoYUortr6tqto e[vot to deritepo: -ro DNA nepvd- : ..oco on6 to riv(upo dniog 4 B(6o pr6oo on6 6vo --: -tovt, ',lLo ouv5neto rnq rleptorpo<pqq tou DNA nopouotd- -. :i orlv ,:r,l:l:lrii' it: :':. >To prow6Lo ttov 6[Supcov ne- : - ':i'/ (Nvin domain nodel) Yto rn peroYpotp(, xoOr'trg -. :rrverot q RNA no).upepooq oe crtr5or1 pre q 6tnLq ' ..rc. dqptoupye( 0ettr<6q unepSltreg (nto orptXro tu- . *avo DNA) prnpoord rnq Kot orpr-1vet opvqttr6g une- :':..req (pepir<d [eru].typi6vo DNA) nioo qg l-to r<dOe :-:ccpq iqq d).tr<og nou 6totp6XeL r1 RNA notrupiepdoq' :--.cupye(tot +1 orpotpq pinpoord r<ot -1 orpoqq nG - - l-t out6 11 petoypo<pr1 61r prdvo enrlped(etor p6vo r o r'lv rontr<r1 6oprl tou DNA, oMd en(or1q enrlped(et . r. :rlv npoyl.lottr<r1 6opr1 tou DNA' To 6v(upro DNA yu- : r:I. nou etodyet opvqttr<6g unep6tr'tr<eq oro DNA' t<oL l',,A tonotoopepdorl l, nou otpotpe( opvrlttr<69 unep6trt- . .: ond to DNA, e(vot onopo[trlro Yto v' onotp5qouv .-. ouooopeuoq tonotroytt<cilv nr5oecov roto r1v nope(o - e ileroYpo<pr1g r<or rnq owtYpoQrlg 'Etot, ov ovooro- (Apvnw6g unep6trtreq) MeroyporpdUevo YnePetri*u:- DNA 1Oet.^ec.,e;e;.':- H yupdon etodyet 0etu6q unep6trxeg 19.18 H RNA l-loAupepdon Tou Odyou T7 Eivor Evo Xprioruo l'lp6runo l0ornpo EIK0NA 1g.33 H getoypotpf drluroupYei nto orptlro ru]rtyp6vo (Oerxd unepetrxop6vo) DNA Unpootd on6 qv RNA nohugepdorl' ivdr io DNA n(otr: on6 ro 6v(uUo y(verot trtyotepo orptxtd rultyp6- vo (opvnrtro unePelrrtrlP6vo). Le( 11 evepy6rnro rnq Yupdorlq Kot rrlq tonoioopepd- ons bt npoxtrqOo0v 1-reydLeq oMoy6g orqv unepeL(xto- on'rou DNA, enrlped(owoq rrl UeroYpo<pr1 r<ot tqv owtypog4. Aut6 ovoL[OnKe nponYoupr6vrog mo ntro(- oro rnq owtyporprlg (pMne Ev6rrya 14'2)' * H otroY6veto rotv RNA notrupepootirv rou T7 e(vot povo noLu- nenr[Oto rrovo v' ovoyvtrtpi(ouv roug unortvrlr6q Tou QoYou ror vo entreLo0v notrtreq ono rtg Opoorrlptoqieg rov notru- peptirv RNA notrugePootitv. * bL rpuorotrhtrSq 6ou6q rov RNA nolupepootilv Tnq otKoYe- veroq rou T7 ge ro DNA routonoto0v qv neptoXl npoobeolq rou DNA oro evepy6 r6vrpo xot unoDrltrtfvouv povretro yto 11 DtotpuYll rou unortvrlq. Optopr6vot Borcrqpto<pdyot (n.X. f3,T4, N4) gttdXvouv rtq 6ireq touq RNA nou;repdoeq, nou onoretroUwot on6 provdg nolunenttdtxsq oLuo(6eq' Aut6q ot RNA nou ;.repdoeq ovoyvtr:p((ouv p6vo Ltyouq unortvrltSg oro DNA tou <pdyou, oMd entte}'ofv noM6q on6 ttg evepy6tqteg tolv notru;rep<i:v RNA notru;repoocov' 'Etot,'onotetro[v np6tuno ouorrlproro Yto rrl ;re6tq ouy re r<p tpSvtr:v e tto u py rc6v tq g pietoy p o<pr1 q' 'l-to nopddetypro, 11 RNA noLuprepdorl rou T7 e(vot puo ;rovrl notrunen-rt6tr<r1 oLuo(6o <100- kD Iuv06ret hNn Ut tol0tqto -300 vour<treot(6to/Seutep6Lento orouq 37"i,'notrrj u(.rrlLdtepq on6 t1v toX0rqto t6oo rr1g nolupreporiq RNA noluprepdoqg tou Borcrrlptoror-i [evtorrl 6oo xot ttrlv ptBoocol.tor(ov nou petotppd(ouv ro mRNA rou. 'Etot, n UeroYpoqq nou roteu0rjvetot on6 tov T7 0o unoBoM6tov oe UeroYpoptr<r1 notrtxorn- to, e[5].t[q nou onotp6netot on6 to yeyovdg ort Il us- H ronoioopePciorl I Xotroptivetttg.. J opvnTtKeq unepe^tKeq I A(rtrtovo DNA (10,4 bp/orpoan) .10..!8 H RNA flotrupepoorl rou odyou T7 Etvot Evo Xprlotlio tlpotuno ldorqFo 537 Boorx6g 6woleq
  • 30.
    roypogn on6 rrlvRNA noluprepdorl rou T7 ouprBo(vet oe peroyev6mepo ord6to rqq Lo(;rco[4q, 6rov 11 Srrppo- orlrou p efuor nepropropr6v4. H RNA nolupepdo4 rou T7 e(vor opr5Loyr'1 pe rrg DNA r<or RNA no).upep6oeg enerdri or roroLwu<o( nupriveg rot ttov rprcOv ev(Upuov 6Xouv nop6proteg 6opr5g. To DNA Bpi oKerot oe Uto unotrdun (palm)" nou nepgdMerqr qn6 "Sdrcrul.o (fingers), rt Svov (ow(Xerpo (thumb), (FMne Er5vo 14.7) ru ro 5v(upro Xprloq-ronoe( novo;.ror6runo xotol,ulr6 gr11ovrog6. Irlgepo 6loupre oprer6g rpu- oroMr5g 6o;169 rcov RNA noLuprepooc0v rrov T7 ror N4. H RNA noLupepdoq rou T7 ovoyvcop(et ng oMq- LouX(eg-or61ouq rnq sro DNA pre np6o6eo4 orrg Bd- otrq rnq pe[(ovog oriLoxoq, 5nog qo(veror orrlv Ell(0NA 1 9.34, 1pr1or1.rono rciwog pro grTir<i etdm6rr7t aq (s p e c ifi c i - ty loop)nou q4q;ror(eror on6 puo B rop66Lo. Aur6 ro XopoKrnprorrr6 efuor provodrr6 yro rrq povopepe(g RNA noLuprepdoeg (5e ouvowdror orrg DNA no).uprepdoeq). 'Onog ong noLuprepe[g RNA no].uprepdoeg, o unoKr.vrl- rfg onoreLe(tor on6 ouyxerprpr6veg Bdoerg oro DNA ovo6rrd rqg 06oqg peroypoqrlg, ;rol,ov6n or unoKrwl- 169 rou T7 onore).orjwor on6 Lry6tepeg Bdoerq ouyKpr- trrd pe unortvqrrig nou runrxd ovoyvrop((owot on6 noLu pepe(g RNA no).u prepdoeg. H perdBooq on6 ro oritrmLoro rou unoKrvl1r1 oro oUpntroro r4g enrp{ruvoqg enmuyldverqr on6 6rio oq- gowxSg oMoySq dropr6pgoorlg oro 6v(upro. llptinov, 6noq xor pe rrg notruprepe(g RNA notrugepdoeq, oup- Bo[ver "pdoqol, tou erpoye[ou oro evepyd x6wpo xor ro 6v(u;ro nopoprdvet npoo6edeprdvo orov unoxrvqri r<oOrig 4 notruprepdoq npoyUoronols;( avenruyetq, ouv- 06oeq (abortive synfhesrs), drlproupyriwog BpoX6o pre- rdyporpo pr{roug 2-12 nl. H nepropl np6o6eorlg rou unortvqt{ 0' onoretrotioe eprn66ro mo olqpronopr6 ovenmulrriv npoi6wrov, coor6oo onoporpUveror Ue ne- ptorpogti nep(nou 45 gorp<irv, enmp6nowoq orrlv no- Lupepdo4 vo dtorqprioer enog6g pe rov unorwqrj evcb ouw(0eror ro op1rx5 RNA perdyporpo. Aur6 e(vor ovd- Loyo pre rrlv erc6nroq rou ouv66rq rqq neproplg 3-ne- ptolnq 4 rou nopdyowo o on6 ro 6(oul,o e[66ou rou RNA r<ord r' opXrxd srddro oriv0eorlg rou RNA mqv noLugepq Borcrrlprorrl RNA noLuprepdorl. To RNA ovo- 6Ueror orqv enrqdveto rou ev(riprou 6rov 61ouv ouwe- Merox[vr1or1 rou ev(U gou..-...] H RNA notrupepdol rou T7 eXer pro Oqtrro erdtr6- TIToq rou npood6ver rtg 06oerg -7 6rrlg -11 rou unouvrlrrl, evdr or 06oerq -1 6tog -4 eLodpXovror oro evepyo r6vrpo. 538 (EOAAA|0 19 Meroyporp1 oroug flporopuc0reg 0e( 12-14 vouxleor(6ro. 'Eneno oupBo(ver puo or6prrl peyoL6tepq oMoyrl 6top6pgooqg, xord r4v ono(o puo unopovd6o yvoorrl og neproXq H geroxrve(ror ndvo on6 70 A on6 q 06o11qg oro o6pntroxo 6vop[r1g. Au- r{ 11 tepdorro 6ogu<rj ovo6ropydvoorl rou oupn}.6r<ou enrpr{ruvo4g 6r1;uoupye[ puo oripoyyo p6oro rqg ono[og pnope( vo e(5l0er to RNA prerdyporpo, r<oOrilg Kor pro 05oq np6o6eonq yro ro 1.rov6r<Lrovo Url-eKpoye(o DNA rnq peroypogtr{q quootr(6o9. I9.19 O Avroyulvrop6q yto Touq Ilopdyovreq o Mnop€i vo Pu0pio€t Tnv 'Evop{n * fo E. coli SXer enro nopoyovreq o, ro06vog ono roug onoi oug o0ei rrlv RNA notrupepool vo [errvqoet oe gro ol.rodo unoxrvrlrdv nou npoodrop((ovror on6 ouyrerprpdveg 0trtr1- trouXieg -35 ror -1 0. * 0r evepyoqreg rrrlv drorpopeurdrv nopoy6vrorv o puO;rl(ovror onr! 6rorpopelxouq UIXovroUoLtq. Irrg en6peveg ev6rqteg 0o nopo06ooupe pepu<d no- podety;roro yro rn pU0proq rrlg Svop[qg, rqg enr;rq- KUVonq Kcu rou tep;,rologoU. 'AMo nopodery;roro 0o 6o0oriv oro Kegdloto 26 r<ot 27. O roropepropr6g opl.ro6rotrltcov prero(ti tou nupr;vo tou ev(0ptou, nou e(vor uneU0uvog yro rnv enrpul<uvor1 rrlg o).uo(6oq, Kor rou nopdyowo o, nou e(vor une00u- voq yro rqv entLoyq rou unoKrvl'lrr1, unorfvr1oe to epot- rrlpo ov 0o unrlpXe ndvo on6 6vog tUnog nopdyowo o, o r<oO5vog er6rr6g yro 6rogopelr<q o;rd5o unoKrvn- t6v. H i r :, 6e(1ver rqv opXrl ev6g ouorrlprotog To o,lo6v(upo Ue To 070 ovoyvtop(er gro opddo unorwltriv H owrxordmool rou nopdyovro o nporole( qv 0v0yv6pron 6torpopetrrlg ogridog unoxlqrtiv ond ro 6v(ugo 0 nopoyovroq o nou ouv66er0r Ue rov nuprlvo rou ev(upou ro0opi(er rrlv o;rodo rov unouvlr(rv or0uq ono[oug opXi(er 1 UeroypoQn. To p tptitr)ro erdrrdqrog npooDdveror orn
  • 31.
    oro ono(o UtoowrKqrdsroorl rou nopdYowo o oLI'6Eet rnv enil.oY{ rou unoKtvnri. To E coll oulyd lptlotponoe( evoLtr'otcttrofq nopd- yoweq o Yto v' owonorptOe( oe nep$oMowr6q 11 6to- tpoqir<6q oLloy6q. Ot evotrtrot<ttro( nopdyovrsq nopouotd(owot offlv IlK0t'lA 19.36 (ot nopdyoweg o ovo- prdiowot on6 to proptor6 gdpoq tou npoi6wog { on6 q iettoupy(o ttov yovtS(tov TIou UeroYpdEouv)' O nto 6q0ovognop6yowoq o, un6 xovovu<6g ouvOrlreg une[- 0uvog yto rn UeroYpoqq tov neptooot6pt':v yovt6[tov' eivqr o oTo flou rtodtronoefuqt on6 to yov(6to rpoD' O evoMorrtr6g nopdyowoq os (ot8) Xpqotponoe(tot yto rrlv nopqYoY( noMrirv npoi6wtov oXen(6pevov pe ro stress. Or oH (o32) xor oE (o2a) onoffoUwqt Yto rnv Tlopo- yoyrl npoi6wtov onopo(tqtov yf ondwnon oe ouvOr1- itg nou (e6tnLcivouv npcotetveg oro 1urr996ntro4to *oi *o nep(ntroopto, owtoro(1coq' O oN (os') ouv06tet npoi6wo rup(tog onopofu4to Yto rrlv evotrrg6rroorl rou oiir-or. 6 qi*r (ors; nopdyet Aiyo npoi6wo-onopokqto yto rn peroqop& -o, ot6{pou, evci o oF (o'z8) er<qp6(et npoi6wo onopokqto yto tt1 otivOeoq trov-poorty(tov' H ondwrlon rov ononrultopr6vtov ((edrnl'opr6vov) nproreivrilv efuot pto on6 ug TIto ouwnprlpr6veg pu0pt- orrr<6q onowrioerg oe ol'6xl'r1pr1 q Bto)roy(o' ApXtr6 *o^oL,l.peqre u:g ondwnon crrrlv ou[4oq qg Oeppo- xpoo(oq ir.or yt'our6 ovopdorrlre ondwrlo4 Oepylroti s'hock, heat shock response). Xoporcr4p((erot on6 r1 oriv0eoq ev6g ouv6Lou npoteivrilv, nop6potrov Kol' oro rpio Broioytt<d Booileto, nou npoorore0ouv to r0rro- po ot O neprBoMowu<5 stress. l-1oM6g on6 out6g nq npcotetveg Oepprtxoti shock e(vot oanep6veq (chaper' ons) nou'1r",,irrou, to en(ne6o rov oTloTlruX<opr6vu:v npoteivdrv nporoLtilwoq rnv EllovomUxoon 11 onodt6- told toug. /ro E. coti,4 enoytoyrltov npoteivriv Oep- prxori shock npoYgorollote(tot mo en(ne6o rnq Ue- rcrypoQnq. To yovbto rpoH e(vot 6vog puOprtorqg nou cnoretroi Yto rrlv evepyono(qor1 qg ondrrrrloqq 0eppt- xo6 shock. l-1poi6v tou e(vot o d2, e(vot 6vog evotr"tr'o- rnx6q nopdyowoq o nou ovoYvop((et touq unortult6g trrlv Yovtd(tov oePPltr<o6 shock, H ondrrrqorl Oepgxoti shock (r<up(trtg oonep6veg rot npolrcdoeq) eL6yXetot Ue ndr(v6poUn pU0ptorl' To rtret- 6( yto tov 6)reyXo tou d2 efuot 6tt 11 6to0eoqr6rr6o cr rrilv ttov rurropontrooptottt<cbv nporcoodrv Kor firEPG vri:v eqopt6rot on6 ro ov rtthonototiwot on6 onontuXtop6veg npotetveg' 'Etot, ov etrqrro)Oofv ro en(ne6o rov onortuxop6vov npoteivriv (efue enet6ri ot npotetveg Oepprroi shock ttq enovontulti:vouv eke ener6q rtg 6toono0v, e(te enet6rl 1ogr1tr'drvet 11 Oeppo- rpoo(o), 6e 0' onopoxp6vouv nepott6pto ttq npote6oeg nou 6tqono0v rov o32, o ono(og 6tot 0o enqv6tr0et oe quorotroytrd en(ne6o, Enet6rl ot o7o rot cr32 owoyov(ov- io, y,o to 6to06otpro nupdvo tou ev(tipou, rl.pretoypo<pq on6 touq unoxwqtdg tu:v yovtS(tov Oepprtxo0 shock ent- orpdqet oe Bootrd en(ne6o roOcilq ro en(ne6o rov o24 ^oi .p' "nor6p1o*ot oro cpuotoLoytt<6"Etot, ro otlvotro ttov yovt6torrirv npoi6wrov nou nopdX0rp<ov t<otd tq 6rdpxeto tou 0eppxo6 shock e(oprotot on6 rqv toop- pon(o preto(0 o7o rot d2. Iu;rBot6 Ue rn onpoo(o rou o*oyru,o1.,oti ttov nopoy6wtov o e(vot t<ot to YeYov6q 6rt 11'ouyr6wpoorl rou o7o e(vot ;reyotrtkepq on6 tqv ow(orotXrltou nupqvo qg RNA notrupepdorlg un6 ouv- Orir<eq oris ono(eg 6e ouprBo(vet enoyu:y1tou d2' O o32 6ev e(vot o povodtr6g nopdyowoq o nou eMyXer qv on6wrlorl rov onornuXo:pr6vtov npolrcivdlv' o oi'endvetor on6 q ouoodrpeuon oTlofiruxop6vtov npoteivdlv otov neptnl'oopnr6 ldlpo Kot rnv e(oteprrtl pepppdvn. 'Onog r<or pe rov o32, q nptote6Luorl e(vot to nLiof y,o qv enoyoyri rnq ueroYpoqqq ro:v oE-e[op- rrirpevov unorw4tcilv. To nep(nLot<o r0rl'ogo nou eu- 0Uvetot Yto rn pU0prorl tqq evepy6rnroq tou oE ouvorf.r((etot mrlv Ell(ONA 19'37. O oE npoo6Svetot oe uro nbcoretvrl (RseA) nou Bp(oretot ornv eooteptr<ri Uepppdvn. H RseA e(vot nopdSetypro evSg nopdyowo ili-; (anti'sigma factor). 'Otqv e(vot npoo6e6epr6vrl NEPINAATMA c Krirlrpo ond qv OegS + K6rlruo onri n1v YaeL ( 0o onetreu0ePtivetot oE I v i:il'l 2V a.l r:.':i : iilll r :;l iii:II r i:i.]ill';.., H RseA ouvt(Oerot orrlv eoolreptrrl UeUppovl H rurropontrooportxfl neptoXfl rrlq npoo66vet rov nopoyovro oE' H RseA ro$erot 6tu6o1wo, np6ro orov neptnloogtro ltilpo rot ptero oro ruuopontroopto, To ruuopontrooporLro ro$tpo oneeu0epdlvei tov oE. RseA /-(ot i 6ro no06Yowoq Xpnoit '::-- 070 0t nepLoodrepeg onopotrnTeq ettoupYteq .: --S ss Iionrn rpdon/Ueptr6q onovqoetg stress . - 032 OePutr<o shock '--:-: sE l-leptntrooutr6q/eEorunopteg flporelveq '::', 054 Evor:-rgdtoorl rou o(titrou ._:= sF luvoeorl poorryitov/Xnuetoroqio .:: srecl MeroBotrroUdq/Uerorpopd rou otorlpou Enoq ono Tov 070, to E. co/i 6Xet oprero[q nopo- , Meq 0 nou endyovrot ono ouyxexptp6veg neptpolJrovrtr6q ,,,r01reg. (0 optOpoq oro ovo;to rou nopoyovro 6r1trdrvet U Uo(o :"), NTIAPONAA'MA it.l* 0 Avtoytrlvtol-tog Yto Touq tlopdYovreq o Mnopei vo Pu0liiE :'.' 539
  • 32.
    oro oE, nRseA eUno6[(er rrlv np6odeorl rou oE orov rupnvo rnq RNA nolupepdorlq Kor rrlv evepyonofrlon Tov uTroKrvrlrd)v rou oE. Auro[ ot unoruvr1"r6q preroypd- rpouv npoi5wo onopofurpq yro rrlv enovomdlcrlorl rov onodroroypr6vov neptnloopu<tilv nputreivcbv ri yro 11 6rdoftoon roug. Iuvendrg, 11 neprn].ooptr<ri ondwrlorl 0epprro0 shock e[vor Uro npoooprvrl ondwrloq noA(v- 6poUnq prj0pto4q nou eL6yXerot on6 ng ouyrewpdl- oerq rov yovrdtorc0v npoi6worv rnq. H pri0puoq rou oE owonorpfueror oro enfnedo rov ononruxcotrlSvrov xot ono6 roroypSvov neptnAoopru<riv n pcoreivc0v nopd tr:v owiororXrov xurroponl.ooportrrilv npcoreivcirv. l-1c69 yvcop((er 11 RseA n6re vo oneLeu0epciroer rov oE; O prllovrqr6g neptLogBdvet pu0;u(6pevr1, 6ro6o1r- rrl npolre6luon rnq RseA. H ouoo6peuorl ononruxo- pr6vov npr.ureiv6v evepyonoe( pro nportedoa (DegS) orov neprnloopx6 Xcipo nou r<6Ber ro C-reLu<6 dxpo qg nporcivqg rseA. Aur6 to r6qtpro evepyonoe( pto dMq npo:redoq, q RseP, ourlq gopd or4v Kurropo- nloollotrrrl n).eupd rnq eooreprx4g pepBpdvnq. H RseP r6Ber qv N-relu<q neproXrl qg RseA, oneLeu0e- pc6vowog relrrd rov oE, o ono(og prnope( nL6ov vo npoode0ef orov nup4vo rqg RNA no].uprepdorlq Kqr vo evepyonor4oer rn Llerqypoqrl. 'Eror, 11 ouoocipeuorl onomulcopr6vov npoteivdlv mrlv neprg6pero tou Bo- mqp(ou evepyonore[ rqv oprd6o trov yovr6[tov nou e].6y- Xowor on6 rov nopdyowo o. | 9.2O Or l-lopdyovTsq o Mnopei vo Elvol Opyovtrrpdvot o€ AroAou0ieg s Mro oro),0u0[o nopoyovrov o 6qgroupyeuor orov 6vog nopo- yovToq 0 onorre(ror yro Tn UeToyporprl iou yovrDrou nou rotdr- ronoreI rov enoUevo nopoyovTo. e To npdruo yovfdro rou tpoyou SP01 peroyporpovror ono qv RNA notrupepoorl rou [evror1. * 'Evo ono ro np6rpo yov[6ro rodrronore( evov nopoyovro o nou o0e( qv RNA notrupepoori vo peroypo(.rer ro evdtopeoo yovrDro. c Auo ono ro ev6ro;.ieoo yovidro rodrronorouv unogovo6eg evog nopoyovro o nou o0e( rrlv RNA notrupepoorl v0 UETo- ypoQeL ro oQrl.ro yovr0ro. Ono.lg oro E. coli, nopdyoweg o Xprlorponoto0wot eu- p6og yro rr1 p60pror1 qq 6vop(r1g rnq Ueroypoenq crro Borcrrlpto Bacillus subfilrs. To yovLd(rrrpo rou B. subt/r,s ro-l6Lronore( rouldXrorov 1 8 6ro<popeur<oUg nopdyov- req o, ouyKpw6gevo pe rouq emd rou E. coli. Aev e(vor oouvrlOtoro vo undpXouv neptoo6repor nopdyoweg o on' 6,1 orc E. coli.Irrlv npoypro-rLr6rqro, ro yovtd(topro tou Strepfomycetes coelicolor xo6rronoe( >60! Iro B. subtilis, rdnoror on6 roug nopdyoweg o e[- vor nop6weg oro B).omu<d x[rropo, evut dMor nopd- yowoL pr6vo ong et6rr6g ouvOrp<eg rrlg ).oipro(r1g on6 540 KE@MA|0 19 Meroyporprl oroug l-lporopuoreg qdVo n rord r1v oMoy4 on6 Bl,omrrrl ovdmu(r1 oe oroptoy{iveo4. H rtipro RNA no},uprepdoq nou eprnM- Keror orn rpuorotroyx4 Bl,ooru<tj ovdmu(4 nepdXer tq 6reg unopovd6eg ru 51er r4v [6to ouvo],rr1 6o;ui pre rqv oeBB'crlo rou E coli, oMd enrnL6ov 6rq06rer rot dtrr1 pro unopovddo nou ovopd(eror 6. O r0prog nopdyov- roq o (oA) rou Borcrqp(ou ovoyvop((er unorrvrltdg pre rg 6reg ouvorvenr6g oL,r1).ouX(eq nou Xpnorponororlv- rot on6 ro 5v(upro tou E. coli un5 rg o6r1y(eg rou o70. Or evoMorcrtrdg RNA notrugepdoeg nou neptSlouv 6ro- goperroUg nopdyoweq o Bp(or<owor oe no).ri pu<p6rc- peg noo6rqreq Kor ovoyvrr:p((ouv unorrvrlr6g pe drogopertrdg ouvorveru<rig oMrl).ou1(eg ong neprolSg - 35 ror -10. Meromriloerg on6 qv 6r<Epooq ;uoq opddog yovr- d(rov oe gro dMrl oprddo e(vot 5vo Xopohrnproru<6 t4g ).oipuo[qg on6 Borcrrlprogdyo. Aur6 ouprBo(ver r<ord rrl Lo(Uo:[n rou B. sutilrs on6 ro qdyo SPO1 r<or rou E coli on6 <pdyoug 6nrrrg or T7, N4 11 o L. Ie 6leg rrg neprmri- oetg er,m6g on6 rrg onLoriorepeg, q ovdmu[r1 rou 96- you neprloprBdvet evoLl,oydg oro np6runo Uero- ypogns rord tr1 drdprero rou l,or1.roy6vou rUrLou, or ono(eg prnope( vo en6pXowor on6 q oriv0eorl Uroq no- Lugepdorlg nou rto6rxonorefuor on6 ro qdVo n on6 rq 6pdor1 enxoupxriv nopoy6wrov nou xtr:6rronotorjwor on6 ro Edyo rc eA6yXouv t4 Borrcrlproxri RNA noLupe- pdoq. Kord rr1 Lo(grrr[4 rou B. subtl/ls on6 ro gdyo SPO1, rq 6torpoperu<d orddro qg ),o(pro[r1g etr5y1owor ;t6oo qg nopoyoyrig vSov nopoy6w0rv o. O Lorlloy6vog r<Ur<Log rou SPO1 51er tpfo ord6ro 6rgpooqg rrov yovr6(rov. Apreoo rord 14 Lo(purl[r1, prero- ypdrpowor to nptilpo yovidlo (early genes) rou cpd- you. Metd on6 4-5 Lemd, ro npcirtpro yov(5ro nqUouv vo peroypdrpowot, on6te ntr6ov preroypdgowot ro ev- drdpeoo yovidro (middle genes). Iro 8-12 Lemd on6 q ).o(pro[q, 4 petoyporprl rcov evdrdpeoov yovr6(<ov ov- rtKo0(ororqt ond q geroyporp4 trov 6lpl;trov yovrdiov (late genes). To nprilq-ro yovbro preroypdrpowor on6 ro o).o6v(u- Uo rou Borcrqpiou (evrorrl. Ouoroonrd 6e 6torp(vowor on6 rq yovbro tou (evrorrl, rcov onol<ov or unoruvqr6g 6Xouv r4v ev6oyevrl rrov6rqro v' ovoyvcop((owqr on6 rqv RNA noi.upepdoq ozBB'tooA. H 5r<qpoorl rov yovt6(rrlv rov rpdyrov onore(tor yrq rrq evotrLoy6q rnq Ueroypog{q on6 ro ev6tdpeoo mo 6{.rtpro yov(6ro. Tp(o puOprmrrd yov(6to, ro 28, 33 ror 34, eL6ylouv t4v nope(o rnq perqypo<p4q. Or Ler- toupy(eg rouq ouvorl((owor or4v EIK0NA 19.38. To np6runo qg pU0ptorlg gord(er UE Uro ol,uorbcorq ov- r(6pooq rl xoropp6xrq (cascade) mnv ono(o ro riv- (upro rou (evrmq preroypdqer dvo npcilrpro yov(6ro rou ono(ou ro npoi6v e(vor onopo('rrlro yro q geroyporprl rcov evdrdgeoorv yovr6(cov. Merd on6 ouq rrl pero- ypoqn, 6Uo on6 to evbtdpeoo yov(6ro xrodrronoro0v npoi6wo onopo(rqro yrq rrl pretoyporpri rov 5r.!r1.uov yovr6(tov. Odyor ;re preroMd(erg oro npd:rpro yov[6ro 28 odu- voroUv vo ;reroypdqrouv ro evdrdpreoo yov[6ro. To npoi6v rou yovr6(ou 28 (9p28) e[vor 1.uo npotetvrl 26 Boorrfg ewoeq
  • 33.
    i):.:,.?t;,tt;;,. unoKrvrlT6q rou gdyou ovoyvrrlp[(ovrot onoTo [JoKrnptoKo otro6v(ugo To np6rpo yov(6to 28 r<odrronore( dvo veo nopoyovro o nou onoUoKp0vet ro Boxrrlproro o 0 nuprlvoq tou evordueoou ev(tuou gp2B geroypoqer ro peoo(o yov(6to rou qoyou To evbrdgeoo yov(0to 33 xot 34 xtodtronoto0v yro npoltelveq nou ovnroOtorouv u1v gP28 0 nupnvoq Tou ijiilii:l ev([pou gp33-9p34 uer0YpoQer r0 oqIu0 yov(0ro rou goyou 19.21 H InoploYdveon EAdYxeTol on6 Touq llopdyovreq o * H oroptoy6veol btotpet evo porrrlpto 0' 6v0 UrllptKo rurropo nou utplororot truor1 rL 6vo onopto nou 0netreu0ep6v6T0t, * Koee qsxoptoTo 5t0p6pt0u0 rpoxopet oro enopevo orodto ovonTuqnq ouv06rovrog evo rotvoupto nopoYOVTo 0 fi0u erron[(et Tov ltp0!Y0uUev0 nopoYovTo o. & H rnrKorvrovlo UsToq[ Tov 600 dtopeptoporolv ouvrov[(et qv enLtroyq TOU KoTotrtrntrou Xpovou Yto Ttq ovTtKoTooToostq Tov nopoyovrorv o. 'Evo r<o16 nopddetypro Yto rrl Xp4otpono(non rnq evoMoyrlq tov oLoev(ripov npoq dLeyXo rov oI o- ycrrv o'rr1v 6r<tppooq ttov yovLd(cov d(verot on6 r1 ono- proy6veoq (sporulation), 6vov evotrXot<ttx6 rp6no (tor1g oe noMd Bor<tqptor<d e(6q. Av oro5lorloet 11 Lo- yoprOprrxrl o[[4o1 enetdrl petdlvourot to Opemtxd oro ;.16oo ovdmu[r1q, 11 Bloorrxq 96oq (vegative phase) oU[r1o4g outtrtv ttov Borcrrlpitlv 0o 6toxone(. Autd nu- podote( tq onopLoySveorl, 5vo ovonru(tor5 orddto rord to ono(o to xUrropo e(vot ov0ercrtr6 oe noMd e(6r1 neptBoMowtt<<.6v t<ot 6to-rpo<ptt<ov ntdoecov (no- pouotd(owot orqv ::i"' .l::l ;rl r:r). Kotd q onoproy6ve- on oro B. subtitis, dvo on6 to Ouyotptxd yovtdttol-toto nou npox[ntouv on6 rlv owl.Ypoqrl tou DNA onopo- vriverot orn Uto dr<pq rou rurrdpou, npooxoMrlpr6vo orov Kurroptr<6 n6Lo. IXrlpor((etot 6vo 6tdcppoyl-to nou 6r'lprtoupye( 6rjo oveldprrlto dtopep(opioro: to prrlrprr<6 r<frropo KoL ro npoon6pto (forespore) . To ou[,ov6prevo dLdrppoyl-io noyrdeUet rpnpo ev6g Xptr-r- proocol-iotog oro npoon6pto Kot pretd prto tpovoAoxd- oq (translocase - SpollE) owle[ to un6]'otno Udpoq rou Xpopooto;rorog ;.16oo oro npoon6pto. TetrLxd, to npoondpto ;re to eyxol.no;r6vo 1p<opr6oopd rou nept- BdLetot on6 6vo or<Lqp6 nep(BLqpro xot our6 to on6pto e(vot oro0ep6 o1e66v en' o6ptorov. H onoptoydveoq 6topxe( nep(nou orcro <rlpeq. Mno- pe[ vo 0eopr10e( tlg nprot6yovn UopQn 6totpopono(- qoqg, r<otd trlv ono(o 6vo prqtptr6 r<urropo (ro B).o- onr<6 Borcrrlpto) 6qptoupye( 6Uo dtorpopettr<d 0uyorpt- xo r<rjrropo pre [eltlptor6 nenpo:ptdvo: to prqtptr<6 16r- ropo nou teLtr<d trfetoL xot to on6pto nou one]'eu0e- purvetot, to ono(o 6Xet ewetrolg 6torpopetrr<q 6oprl on6 to ou0ewtr6 BonrrlPto. H onopLoy6veol nepil.ol.tBdvet pto 6poorrr<r1 oLl'oyq orrg Btoouv0ettr<6q evepy6qteq tou Borcrqp(ou, crrnv ono(o epn).5Kowot noMo yov(6ro. Ot oMoydg orrlv 5r<- epoon trov yovt6(cov nou odrlyotiv retrtrd oro oxrlport- opr6 tou onop[ou nporo].o0wot r<up(tog on6 oMoy5q orrlv dvop[q rlq UeroYpo<pr1g, Kdnoto on6 to yov(6to nou tretroupyo0v orrl Btroortr<r1 gdoq odpovonoto[wot r<oro tr1 onoptoyeveorl, oMd to neptoo6tepo ouveX(- (ouv vo er<qpd(owot. Qm6oo, nol').d yov(6to etdtrd yro rrl onoptoydveorl er<<ppd(owot ;t6vo r<otd tq 6tdp- i,.I: H Inoptoy6veorl Etrdyxerot ono roug llopoyovrec o 541 Ell(OilA 19.38 H UeroYpoQn tolv yovtd[rrlv Tou QdYou SP01 etrey- XeroL onrl d6o 6to6oxtx6q ovruoromdoetg tou nopoyovro 0 nou 0trtr6(ouv qv etdLrdrrlto rnq 6vopfiq. kD nou owtt<o0tord tov nopdyowo o rou (evtorri crrov nup4vo rou ev(riprou. Awfi r7 awuardotaorl e[vat rc pova6x6 oultBdv nou ana:reftat Yn rn peruBao4 an6 t7v 1xEpaory ruv nptitryav mryv 1xEpaorl rav evddpe' oav yovt6fuv. Arlprtoupye( 6vo oloriv(upro nou o6uvqte( vo peroypdUet to yov[5to tou [evtorrl rot ow(yt'owd preroypdqer etdtrd to ev6td;reoo yov(5to. Aev yvtop(- (oupre ndlg to gp28 onoporp6vet ro cr43 o6te n ouprBo(- va oro o noLunem(dto tou (evtorri. A0o on6 to evdtdpeoo yov(6to epntr6rowot mrlv en5prevrl evotr).oyq. MetoMd(erq mo yov(6to 33 n 34 nopeprnod((ouv rrl UerqYporpri tolv 6tltprtov yovtS(<ov. To npoi6wo outtilv ttrlv yovt6(rov oXrlprot((ouv 6vo 6t- prepSg nou owtro0tord q gp28 orov nuprivo rrlq R- NA noLuprepdorlq. En(or1g 6ev yvcr:p((oupre nc0g ot gp33 xor gp34 erron((ouv tr1 gp28 (f onoto6rlnote un6L).etppro oA tou levrmri), aMa p6ttq npoode)olv orcv nuprlva rcu ev(i1tou, e[vu Lxavd va evepYonou fioouv rq peraypaEn p6vo orou;, unoKtvqriq tuv 6tltt- pav yovt6[av. Ot 6to6oXrr5q qwtKqroordoetg tou nopdyowo o SXouv 66o entmc0oetg. Kd0e <pop6 nou oLd(et n uno- provd6o, q RNA notruprepdoq ;rnope( v' ovoyvop(oet gto v6o t6(q yovr6[tov evdr tout6Xpovo no0et v' ovoyvtop(- oer rrlv nporlyoripevrl td[4. Euvendrq, owo( ot 6tot<6- meg entgripouv 6pomtr6q oMoydg mrlv evepy6qto t4g RNA nol.uUepdonq. I Boorr6g evvoleg
  • 34.
    Ippoti(erot 6Ldrppoygo rUnopo trtjeror To onopLo onetreu0eptiverot ElK0ilA19.39 H onoproydveon repDoppovsr Trl Oroqopono(norl evdq p)oorKou poxqpfou o' 6v0 UnTpKd xunopo rou trUeTor Kr 6vo on6pro rou qfletreueep6veror. Kerq ourrlq rqq nept66ou. Iro r6loq rnq onoproy6ve- or1g, -40olo rou BoKrrlproKoU mRNA e(vor erdrx5 yr' ourn rn Qdon. N6eg pop<pdq RNA nolupepdonq evepyonoro6wor oe rUnopo nou npoyporonoro0v ofloproySveon. ne- pr6xouv rov i6ro nup{vo rou ev(ripou 6ncoq ror ro B}.o- mtxd x0rropo, oLl,d 61ouv dtogoperu<69 npcoretveg orn 06on rou Bl.oonroU nopdyowo o (oA), Or oMoy5g orn peroypogxf er6x6r4ro ouvo(r((owor qrrlv Ell(BllA 19.40. H Boou<r1 op14 e(vor 6n o nopdyowog o r60e 6roprep(oprorog 6ro6oXu<d erron[(etor on5 Svov vdo no- pdyowo o nou nporole( q pretoypogn Uroq 6rogope- ttxrlg optddog yovr6(ov, Mero[U rrov 6ropepropdrov undpXer enxowrov(o ciore vo ouyxpovroro6v or oMq- ydg oro npoon6pro Kqr oro ;r4rptx6 xrirropo. H or<oLou0(o rrlq oroproydveorlg opx((er 6rov or ne- prBotrtrowx6g ouvOr1reg nupodororiv 6vo r<0xl,ropro gto- ogo-ovoFer66oo4g (phosphor-relay), orov ono(o puo gororpropu<ri oprd6o prerorp6petor rord prirog proq oer- pdq npolrcivrirv prSXpr vo grdoer aro Usroypo<px6 puO- 542 KEOA Al0 19 Metoyporpn mouq llpoxopurilreg gror{ SpoOA. Ie ouq q 6epyoo(o epn},6rowor notr- Ld yovr6rord npoi6wo, rov ono(rov r'1 nolunLor6rrp6 clwrKoronrp((er q Xpfon o4pre(alv el6yXou, 6r1no611 orryptor0nov Kord rq ono(o ro Borcr4pro enrBeBordloei 6rr entOupe( vo ouvel(oer orrlv o66 qq 6togopono[- r1oqg. Autrl 11 puOgronr{ nope(o 6ev np6ner vo },oprp& vetor odu<oroL6y4to, roOrig q rel,rr<q on6qoo4 eivor oprerdrlqq. H evepyono(qorlrou SpoOA Ue qooqopul(oor1 orp ;roro6ore( qv 5vop(q rnq onoproy6veorlg. Ee qoorpo puLuopSvq Uopen, o SpoOA evepyonoe( rn peroypo- gr1 6rio onepov(rov, r<oO5vq on6 to ono(o getoypdrperor on6 pto dro<popenrri Uopen r4g RNA nolupepdorlg tou [evrorrl. To 5v(upo rou fevrorq, Xprlorponord:wog ro yevrr6 nopdyowo oA, pretoypdqer ro yov(6to nou ro6tronop[ rov oF rqr, un6 qv ro0o6r1yr1or1 ev6g 6.]t- l,ou nopdyowcr o, oH, getoypdrper en(or1g ro yov(6ro nou ro6rronote( 6vqv np66popro tou nopdyowo oEs nou ovoE6peror rr:g npo-oE. T6oo o oE6oo ror o npo-d nopdyowor nprv ro olqpolo;r6 tou 6rogpdyUqroq, otrl.d evepyonoro(wor opy6repo. H peroypo<pr1 nou roteuO0veror on5 rov oF ovo- qrdMeror ener64 6vog nopdyowog owroryl.ro (SpollAB) npoo6dveror oe qur6v, eprno6((owdq rov vo g4rlpor(oer 6vo oLoriv(upro. Iro npoon6pro, com6oo, 6vog nop6- yowoq owr-owto(ypo (SpollM) r<oroor5Mer rov Kqro- oroMo. H o6povono(4oq tou owr-owro(ypo etr6yXeror on6 pro oetpd ouprBdwov googopul(coo4g/ono<pro- oEopuL(rooqq, ornv ono(o to npriro Brlpo e(vor n ono- goorpopuL(roor1 on6 Uro eoogcrrdoq nou ovoprd(eror SpollE. H SpollE e(vor 1.uo nproteiV4 qg eooreptx4g pepppdvnq nou ouooopeUerqr orov Kurroprr6 ndLo, pe onot5treopo q evepy6r4to groogordonq nou 6ro0era vo oupmurvc6veror neplso6repo oro npoon6pro. Iuvo mrrd, 11 onoeoogopul(roor1 evepyonoe( rov SpollAA" nou Ue rq oerpd rou ercron((er rov SpollAB on6 ro oF. H onel,eu0Sproorl rou oF rov evepyonoe(. H evepyono(qor1 rou oF orlproro6ore( qv dvop[r1 rnq KuTropoer6u<r1g 6r<Epoorlg rtov yovr6(<ov. Yn6 rrlv xo0o6tiyqoq rou oF, 11 RNA no).uprepdo4 pretoypdger qv npdrrq opd6o yovr6(orv rnq onoproy6veoqg, O Ed efuot unefOuvoq yro 6trr1 q preroypogrl oro npoon5- pro. O oA 6ev rotoorp6geror xord 14 onoproydveorl t<ot, ouvencig, to Bloorrr6 oAodv(upro, EoA, nopop6va oro rUropo nou npoyporonororlv onoptoy6veoq, H or<oLouO(o ouveX(erot xoOdrq oro npoon6pro no- pdyowor npoi6wo ro ono(o npo6pXowor on6 unorrul- 169 nou ovoyvop(owor on6 ro EoF (BI6ne ElK0l*A 13.41). l-ro nopdderypo, ro EoF etr6yXer q orlv0eo4 ev6q prero- ypd<pou nou rcodrxonoe( rov oc, o ono(og ;re q oerp6 tou oXqpro{(er to o},o6v(upro ilou peroypdger ro riore- po yovbro rnq onoproydveoqq. To EoF ovoyvrop((er eni oqg 6vov unonvrlrq nou eLdyler t4v 6rrppoorl ev6g npoi6wog une0Ouvou yrq rnv enrrowov(o pre to 6ropr6pr- opo rou prqrpu<oti rurdpou. To ouyrerptprdvo npoi6v, yvoor6 og SpollR, erxpfueror on6 ro npoon6pto prSoo orrl pepBpdvrl nou 6ro1op((er rq 6Uo 6roprep[opoto. In1 UepBpdvn, o SpollR evepyonote[ rqv nporeivq SpollGA 11 ono(o r<5Ber rov ovevepy6 np66popro npo-oE, pretorpS- Btroortrri poxqpro AvTrypoqo DNA Cq c(q ct^r-i ffi ffi' 1 .>
  • 35.
    MHTPIKO RTIAP0 TooLodv(ugo nepr6Xer nP0:n0Pl0 tov gH n rov oao I /,---f,- "r' 0 oK evepyonoreftol rr exron[(et rov oE I Y 0 evepyonorq96vog oK unoorrlp[(er rn usToYpoQn TOV orlruov yovt6urrv H onoproy6veorl nepLtroppover 6ro6oXrr6g otrtroy6g oroug nopoyovTeq o nou etreyXouv qv erdrrorqro rrlq RNA notrupepo- --( yto TIV evop[1. 0t orotrouO(eg oro pqrprro rurropo (aporepa) ror ro npoonopo (6e(La) oXer((ovrot pe orlporo nou 0tonepvouv ro : orppoyuo (entorl;.ro[vovrot Ue opt(ovflo B6trl). I ,fuJI I 0 evepyonotlg6vog oG unooqp[(et q UsroypoQn rov ri{.rrprov yovt6[tov ffiw 0 oE evepyonore(ror n erron((er rov 043 ITo yovOto oK geroypdrperor on6 rov oE I I I #t nowdg rov finv evepy6 Uopgri, oE, oro prppu<6 r<(rrro- po. (O oE nou nopdyetor oro npoor6pro onodoprefuo$. H oxoLouO(q ouveX((eror 6rqv o oE fio ;rqrpu<6 rftropo owtxo0(ororor on6 rov oK. (H nopoyoyrj rou ox eivfl oprerd nep(nLox4, ener6{ ro yov[6r6 rou 6qpr- oupye(rot on6 6vo ouprBdv ronoerdrro0 ovoouv6uo- opori!). 'Onog ror o oE, o oK ouw(0eror og ovevepy6 np66popo pr6pto, ro npo-oK, 'Eror o oK np6ner vo evep- yonotqOe( pre r6rprpro qg np66poUnq UopAnq nporo0 pnop6oet v' owr.Koroorrloer rov oE Kor vo pretoypdget ro 6r.trrl,ro yov[6ro oro prrlrpu<6 r0rropo. O ouyXpovr- opr6q owcbv rov 60o ou;rBdwcov qro 60o 6toprep(opro- to ouwov(eror on6 brogopetxd or1;roto, Iuvomrxd, r1 evepy6qro rou oE oro truppx6 rUrropo e(vot onopo(- rnrn yro rnv evepyonofioq rou oG oro npoon6pro rot q evepy6qro rou oc ononefuor yro rq 6rlpuoupy(o ev6g o{porog nou peroS6eror 6top6oou rou drogpdyporog yro vo evepyono(oa rov oK. To rUrtrtogo n1q rptoorpo- ovogeroDdoqg evepyonore( q oUv0eoq tou sF 661 169 snPo-E 0 oF owrKoerord To 043 mov nupflvo rou ev(Ugou I V To rewpud odpntroxo rou oF geroypdqer ro np6tgo yov(Dto qg onopoy6veoqg 0 oG e[vor npor6v rtov np6qrov yovr6(rov n1 g onoproy6veorl g 0 oG evepyonorefuor ror erron((et rov oF nP0x0Pt0 &.ry flK0NA 1S.41 0 oF nupodorei U orjv0eon rou en6pevou nopo- yovTo o oro npoondpro (oG) rr evepyonoLe( ro SpollR, nou e[o0e[ qv SpollGA vo K6qrer rov npo-oE. 19.21 H Inoproy6veol EtrdyXeror ond roug flopoyorrec o 5tE +@SpollR l@p
  • 36.
    Y [lporedluorl gK EIK0NA 't9.42 Hdromoupou;rcvn pu0gron Tnq ofloproy6veorlq ouvTov((er To ouyxpovrouo rtov yeyovdrov oro urlrprKd KUTTopo KOr ro rrpoofidpro. Iuvenrilg, n onoproy5veon el6yxeror on6 Uro oKo- LouO(o orrlv ono(eq or nopdyoweq o Kdee 6toprep(opro- roq evepyonoro0wor 6ro6oXxd on6 toug ofupo F, E, G r<or K, r<o06vog on6 rouq ono[oug xmeuOUver rrl Uero- ypoAn ev6g ouyrerptprSvou oUddqg yovr6(rov. H oro- Lou0(q pnope( v' ovqfiopqoro0e[ on6 5vo oroupcor6 np6runo o4trrdrcov nou 6ronepvo0v ro 6rd<ppoy;ro, ouv- 66owoq qv 6rEpoo4 trov yovr6[rov oro dtogopelrd drogepioproro, 6ncoq eo(veror orrlv EIK0NA'l 9.42. Ko0drq evepyonororiwor vrior nopdyoweg o, or noLro( nopd- yovreq ercron((owor, evepyonordlwoq Kr onevepyonor- rbvrog drogopetrr6q opd6eg yovr6(cov oro 6Uo 6roprep[oproto. | 9.22 O AvrrreppoTrou6q Mnopei vo Eivor Pu0prorr16 EupBov * 'Evo ouUntroro ovTrT€pUoTrouotj enup6ner ornv RNA notrupe- poorl vo Dtopooer 6roprloou repportor6v. " 0 tpoyoq IoUB6o Xpr;orgonorel ouor1poro ovruepporLopo0 yro vo pu0pioer 16oo ro nptiulro ooo xor ro 6QLpo peroypotpo, otrtro ro 0uo ouorrlporo trenoupyo0v pe evietr6q Drorpopel- roug l.u1xovro1roug. e H npoodeon nopoyovrov oro veoouvre0erp6vo RNA ouvdtier lg nporelveg ovTrTeppoTropou oq 0eor1 rou repponorrl Dro- Ueoou Uroq 0r1trroq RNA. * Avnrep;loloUoq TIq peroypoqrlq ouppolver enlo4g ror oro onepovro rou rRNA. O owLrepprotto;r6g Xpnorponorr(tor yro rov SAeyXo rnq Ueroypoenq oro onep6vro 16oo tov gdy<ov 6oo Kor. rcov Borrqp(tov. 'Onog qo(veror ornv fifftii :i i.lj, o owrrepporropr6g ovogdparor oe rponono(4or1 rou ev(riprou, to ono[o prnope[ ntr6ov vo nopoBrdoer Svov repprororrl ror vo nepdoer oe yov(6ro nou Bp(or<owot roOodrxd. Ito nopdderygo rnq ex6vog, evcb 11 nporo- 0opro;r5vq o66q yro qv RNA no).upep6or1 e(vqt vo reppo{oer oro r5l.og qg neproXrig 1, o owrepport- opr6g rorolrlyer oe ouveXr(6prevn Ueroypoe4 6topt6- oou rnq neproplg 2. Iuorrlporo owrrepUorro;ro[ e(vot r<otvd orouq LoprB6oerSe(g Borrqprogdyouq (<pdyor nop6prorot pe ro qdyo l,dpB6o nou neprypdeowor mo KegdAoto 27). Aw(Oero on6 roug tpdyoug nou prord(ouv mov T7 tou E. coli Kor rouq rpdyoug SPOl rou B. subfl/is nou neprypdgqxov nporlyougdvog, o LdgBdo 6ev rcodrxo- nore( ofre 6trr1 rou RNA no).upepdo4 ofre roug 6r- roUg rou nopdyoweg o. MdLLov lpqorpronore[ qv nolupepq RNA noLuprepdorl rou [evrorrl yro 6].q rq peroypoqrlrou. Ie o0wopro 6rdorrlpo perd q ).o(pr<l- (r; on6 ro rpdyo, (er<rvd n Ueroypoen oe 6rio npcir- pouq unoKlvnr6q, Pn rot Pr-. 0o16oo, oe ro06vo on6 ourd to onep6vro undpXouv repprortm6g prerd rrg orpeqp(eg rnq Ueroyporprlg ror nprv ro neptoo6tepo yov(6ro nou ro6rronororiv rtg nprilrpreg Lenoupy(eg. O tepprono;r6g rnq peroypoenq oe our6g lg 06oerg po- rord:ver 14 ).o(puo[q. Av 6prcog 11 RNA no],upep6o4 no- poBtdoer rouq reppqrrm5g xor UeroypdQer to npcirrpo yov(6ro nou e(vor uneU0uvo yto rnv owrypo<pr1 rou yo- vrdrdrprotog rou gdyou, 4 ovdnru[q rou ].d;rB6o 0o ouveXrme(. H npcirtq on6qooq yro reppotopr6 el,Syleror on6 qv npcoreiV4 qwrrepuorrol.rori "N", qv nprilq npr.,:ret- ul nou nopdyeror un6 rov dLey1o tou Pr-. H N oXqpo- r((er o0trrnl,oKo Ue rrg npcoreiveq rou [evrmq nou xoloUwqr nopdyoweg Nus (N utilizing substances - ouo(eg lp4orprono(r1onq rnq N) rpononorcilwog qv R- NA noLuprepdoq rord r6roro rp5no riore vo Urlv owo- noxp(veror nL6ov orouq reppotm5q. Irqv npoyUo- rrx6r4ro, ro oriprnLoxo rou owrrepUotrotrroU oXqprorl- (etot ndvro oro veoouwe0erpSvo RNA oe pro oMqlr.ou- X[o nut (N utilization site - 06oq Xprlorprono(4onq rnq N). Or O5oerg nut onorel.o0wqr rup(cog on6 otrl,qlou- 1(eg RNA nou ovo;rd(owor boxA t<ot boxB,6nou ouo- otr:pe0owor or nopdyoweq rou [evrmrl NusA, NusB, NusE (prBoorotrrorrr4 npcoreivrl S10) ror NusG. Or nporefveg owrreppqtogoU nopopdvouv npoo6edepS- veq oe outriq ng 06oetq tou RNA oq dyo 1.t6vt1to oult- nltoxo avrlrepparLolJoti r<oodrg n RNA nolu;repdorl ouv05rer ro 6rjo perdypogo npoq ro 6e[rd xor r' opr- orepd. 'Eror, to veoouwe0erpr5vo RNA ouv66er rpuotrcd ng nporeiveq owrrepporroporj nou e(vor npoodedeprS- veq orn 06o1 nut pe rnv RNA noluprepdoq nou ni.q- ord(er rouq repUortor6g. Mol,ov6rr o or<prBqg 1.r4Xovropr6g Ue rov ono(o ro oripnLoro owrrepporr- opro( e;rno6[(et tov rep;rorro;r6 6ev e(vqr or<6pr4 yvro- m6g, ro 65or;ro ratv npo:teivdlv repporrol.roU mqv RNA no).upepdo4 6rop5oou rou veoouwe0erp6vou R- NA e[qye(rr1v u<ov6r4rd rou v' owreppol((er oe 6ro- 6o1u<orig repprotrmdg nou Bp(oxowor oe on6moo4 erorow66cov q ordpq r<or Xil.r66cov Bdoerov r<o0o6rxd. 0"ffi 544 KE0MAI0 19 Meroyporpn mouq tlporopudrreg nP0rn0Pr0 Boorxeq eworcq m
  • 37.
    lleptoXq 1 Ynouqqg +{-l-leptoXfl2 W HRNA notru[epdon rep[or((et To RNA Bp(orerot grlvo oqv neptoXrl 1 ANTITEPMATI:MOT Ttloo I neproXrl 1 rioo xot 11 neptoXrl 2 getoypdtpovrot 0 owueppoloUoq Unoper v0 etrdyqer Tl UeToypoq1 K00op((0vT0q 0v I RNA notruUepool 0o repporloet r] 0o nopoptooet , -, :pLopevo repportorl rot 0o npoXoprlott orrlv enopevn neptoXrl. - ieuro(o npolrefvq nou nopdyetot on6 ro N-owt- -.:-crrop6vo petdypogo ond rov dtro np<otpo unor<t- -:'-l PR, ovopd(trot <Q,. Onotg n N, n Q e(vot -: *:srvl ovrrrepporrouof. Awtreppror((et rn uero- ::cr1 on6 rov drl.rtpro unoxtvqrq PR, o ono(og nopd- . ivo prerdypogo nou rordtrconote( ttg npotetveg qg .'::"qq Kor rnq oupdg tou tpdyou. 'EroL, 1 6t<<ppootl ---, yovr6(ov tou LdprB6o npoyporonote(rot oe 60o :-:51o, xoOdvo on6 to ono(o etr6y1erot Ue owtreppo- -:-c (BL5ne Ev6rrlta 27.7 Kot - . ). H Q enL- -:.:a o-rqv RNA notrugepdoq vo nopoBtdoet rouq -1:Jorroreq mqv 6q.rrpr1 pretoyporpLr<r1 provd6o, oMd pt', :. 3^r0g 6Lo<popetu<d prqXovtopr6 on6 qv N. Aw(Oetq - : rrlv N, n Q npoo66vet DNA (orr1 0doq qut, Q uti- :a: cn site - 06oq lpqotprono(nonq rnq Q), o).Ld : -q Kor 11 N ouprnope0etot Ut rrlv RNA no).upepoorl . :. le rdnoro tp6no nopeprBo(vet mq 6pdo1 rov rep- -::.;rov oe 6Lo to 6q.rtpro onep6vto. Oofverot 6rt q : ::r1 rrlg Q nepil.opBdvet enttdXuvorl qg RNA noLu- -.:corlq ovdpreoo oe 06oetg no0or1g. (H ouvoLtt<t1 :-3.ro1 trlg ovdnru(rlq rou ).d;rB5o 0o nopouotome( - : Kegd).oro 27). -o onep6vto rou rRNA 0' ovoprev6tov vo 6e(1vouv --'..i<orqto, enet6q e(vot ptoxptd oMd 6e preto<ppd(ov- -: Oordoo, r<o06vo on6 to onep6vto rRNA rou E. coli nepdXer otrLr;l.ouX(eq rinou boxA xqt boxB flou ouvclp- poloyoUv oriprnAoro owrepgortopoU ildvo oro petd- ypqeo, mo ono(q opropr6vot ilopdyoweq Nus e[vqt rowo( pe ro <pdyo LdpB6o. Awd to otiprntroro 6ev ne- pr5louv 6vov nopdyowq {nou N i O, TIou Ko6tKonot- oUwot pr6vo on6 ro yovt6uiporo roJV qdyov, oLLd enopxoriv yro v' oTlorp5{.rouv tov np6opo tepgonop6 ortg oM4trouX(eg qq qoupr6rog Kot orouq ooOeve(g p-e[optc0;revouq repUortor6g nou tuXo(vet vo undp- Xouv prSoo oro 6oprxd yov(6to tou rRNA. O owLreppo- tropr6g e(vqr ndvro onopo(tqtoq yto rnv enopr<r1 nopoyoyrl rRNA, 61r p6vo 6tov o LdptB6o ptotr0vet to rrirropo. Euvenrilg, t<ot6 rrlv eE6fuEn rov Borcrqp(rov, ot nopdyoweg Nus 6ev enrl.6X0qr<ov yto vo 6eurol,0vouv qv Srgpooq rrov yovt6(trlv-rou LdpBdo oLLd prdMov yro v' qnorp6rllouv qv nofur<6qro oro onep6vto tou r- RNA. Or neprol6g-odrlyo( ttov onepov(rov tou rRNA ne- pr6Xouv oMqLouX(eg boxA nou ouvopl-totroyorjv roug nopdyoweg Nus r<oOcirg ot otrtrqtrouXbq boxA (enpo- Bdtr.ouv on6 ro 6(qu^.o e[66ou tou RNA. Onog rot pe rov qwreppotro;r6 cno <pdyo LdgBdo, outrl 11 6epyo- o(o otr)td(er trg r6r6rqrcg qg RNA noLu;repdorlg rotd t6roro rp6no rime vo prnope( nL6ov vo nopoBtd(et rouq repLlorrorSg. Qor6oo, o Urlxqvtop6g 6ev 6Xet o16- prq onooorp4vrore(. To RNA ovTrnpoooneuer lqneproxdgl +2 iti.ii: 0 Awuepgortopog Mnope(vo E(vot Pu0pLorLrc I"-3 ' 545
  • 38.
    iiifiiii -,ovrspus],olt.,.. .. &Ynortvrlqqt @r"liilrw*.wRNA q,rX-, ty*fyyf Meroypogrr4 povrido Ynomvrlqg l-lptotetuq owueppottolrori fL rR1 tR, Mto npolrelvrl ovTtTspu0Ttouou pnope( vo npoodtooet orqv RNA notrugepoorl rrlv rxovoqro vo nopoprooer 6vov erdrxo rs:- poiloq. PL Pnr D ,R) pN pN p0 Ageoo nptirpo Ageoo npdrrpo '0rlrt1ro 19.23 O KtirAoq rou BorrnptoKo0 Ayyetuoqdpou RNA * Iro porr1pro, n Ueroypoqn rot 11 perorppoorl npoypoTonor- ouvror rourriXpovo (ou(euyprlvr1 Ueroyporp1/perotppoo4), ro- 0drq ro prpooropolo (euvouv q perorppoorl evog mRNA nprv otrortrr1po0er 11 ouv0eo[ rou. * To Borrrlproro mRNA e[vor ooro069 pe Xprivo rlpt(au1g triyruv Lenr6v rqg opoq. e'Evo Borrnprord mRNA pnope[ vo e[vor no],uororpovrro pe notrtr6g rtrldrxdq neptoxeq nou ovTroTorxouv oe Ororpopenro yov[6r0. To oyyelrog6po RNA SXer n1v (6Lo erroupy(o oe 6Lo ro r<Urropo, undpXouv 6pro:g orlprowrxdg 6ro<po- pdq oq npoq rrq ).emoprdpereg qg oUv0eorlq Kor rrlq doUriq rtrlv npoKopurorrxd;v Kor roJV euxopuortrotv mRNA. Mro orlpowrxq 6rogopd onlv nopoyoyrl rou mRNA e(oprdrot on6 trg O5oerg 6nou ouprBo(ver 11 preroyporprl ror q perdgpoorl: o Iro BqKrriprq, ro mRNA preroypdgeror Kqr Uero- gpd(eror oro povo6rr6 rurropu<6 6roprdpropro. Or 6fo 6repyoo(eg e(vor 16oo mevd ouv6edepdveg nou ouprBo[vouv rour6Xpovo. To prBooroprdrro npe oroLlc0wor oro Bor,cqpror6 mRNA npw o16;r; ole rl,qpoOe( 11 peroypoqri rou, ouvendrg ro ndtuoapdrto (polysome) ;rnope( vo e[vqr oxdprl npooxoLlq;.uivo mo DNA. To Bomqprox6 mRNA ouv4Otog e(vot omq06g xor yr'ow6 prerorppd(etor oe noLunem[bro pr6vo en( ],(yo ].emd. Aurl q Drep yooio roAefuor ou(euypdvn Feroypqgrj/perdgpo- o4 (coupled transcription/translation). o I' 6vo euxopurotrx6 r<6rropo, q oUv0eorl r<or 4 rrlpr(- Uovorl rou m R NA npoyporonororiwor onoxl,ermx6 orov nup{vo. To mRNA e(dyeror oro rurrop6dt+ o1-ro, pr6vo ogori ol,or).4pro0oUv ourd ro ou;rBdwo. 'Evo tunx6 europurrlrrx6 mRNA e(vor oXeurd oro- 0ep6 r<or ouvel((er vo petorppd(eror en( opxedg cbpeg, poLov6l undpXer peydln preroBLqt6rqro oq npoq rn oro0ep6r4ro erdu<rirv mRNA. H ElK0l'lA 19.45 6e(Xver 6n oro Bor,r4pro n UeroypG grl ror q perdgpooq qXer(owor orevd. H petoypogri [er<rvd 6tov 11 RNA no].u1-repdo4 npoo6e0e( oro DNA ror op(oer vo nopeUeror qrtdXyowog 6vo owiypogo rou ev6g xtrrilvou. l1ol,[ o0wopro, prBooroprdrto npo- llporetvq ovlrepporrogorj Tepporoqg 546 KEOA Al019 MeToypqqq moug l-lpoxoputirreq Boorxeg eworeg H nporeivn ovnrepponoporj 6[ver rq duvordqro oqv RNA notrugepdorl vo nupoptdoet rov teppottoq
  • 39.
    S mi* rri"r'rl,.iI :r lil'rlr ji,;'ir,.)l': j.J :rtitr li rllli*lrtririiijrj lll:l,lir.i..r iir'i1.'r lt: :ti rliitl To 5' oKpo elvor rprQooQoprKd ^urd2,0 nrin l-{ ENA noiupepdrq reppot[(er mc 3' axpo , _ ;'- 6rr' rff$1, - - 1:a::::_-3: / I Y:.+a-:--r/ M -:"t::11)A i.i1:-z 3 C min H ono0rlpqon ouveXi(eror. ru prpoor*uorto olor<trnotirvolrv rq [€,ToQpoun ,a 'n /zrr=-,"{-l- /,r'" I , :,. ; Iuvotprl: oT0 BoKT1pro To mRNA UeToypoQsTot, - : rorppo(eror rot ono6opelror rour6Xpovo. ;<oMrr;wor mo 5' dr<po tou mRNA t<ot (er<tvotiv tr1 pte- rl:!pooq, nprv or<6pq ouwe0e( ro un6lorno prdpog tou * - v u 1.roro g. l-l oMontrd prBooropdtLo rtvo tjwou<otd prrl - . :q rou mRNA ev6o<o or<6;ro ouw(0etot. To 3' dr<po ::r mRNA 6r;;uoupye(rot 6tov repprotrme( q Ueroypo- :- To mRNA 6oo (er ouveX((er vo pretogpd(eror on6 -: crBooco;rdlo, oltrd opr<erd ypiyopo ono6oge(tor :-:- ouvo).trq r<ore0Ouvoq 5'-+3'. To mRNA ouw(0etoL, ...::gpd(eroL on6 to prBoooprdrro r<or ono6ope(tot, :'.: no).r-i ypnyopo, 'Evo [e1opror6 pr6pto mRNA ent- : -'",er ro nolri yro prepu<d emd. Iro Borcrrlpro, n Ueroypoerl not pretd<ppoon npoy- -:.cnoLo0wot pre nop6proto rol0qro - oroug 37oC, - r, - 5 0 vo u x..eot(6ro/6eutep6).emo. Aut6 npooeyy((er -:r.r rrlv roXritrlto qg npurreivoo[vOeor1g, nou o6pd ..:, 15 oprvo[doi6eutep6].ento, Iuvencbg, -1 ],ent6 ,.:.ii(eror yro vo petoypocpei Kor vo Uerogpoore( 6vo -:A 2500 bp, nou owrororXe( o' 6vo noLunem(dro 90 . I Acpdrou [exrvr;oer q dr<gpootl ev6g yovrd(ou, ro m- RNA rou runrKd 0o epqovtote( crro Kt nopo $m oe -1.5l,em6. To ow(ororXo notrunem[dto 0o epgonore( nep(nou 0.5 Lem6 opy6tepo. H Borcrrlprorcl petdgpooq e(vqt no^.U ono6orx4 rot ro neproo6repo mRNA ;rerogpd(owot on6 peydlo opr0p6 ogrXrd ouoreuootrr6vov ptBooroprot(cov. I' 6vo ouyrexprpr6vo nopd6erypro (trp mRNA), xdOe Lent6 npoyporonoro6wor nep(nou 15 evdp(etg rnq Ueroypo- qrjg r<or xo0dvq on6 ro 15 mRNA prdMov pretogpd(etor on6 -30 prBooro;rdrro mo 6rdleq.r;ro prero(0 Uoroypo- gqq rot ono66prqo4g. H oord0ero rov Tleptooot6p<ov Borcrrlptorc0v mRNA e(vor ewunootor<r1. H ono66pr4orltou mRNA opXt(et l,i- yo prerd on6 rr1 prerdgpoorl rou Kot, nt0ov6v, ptSoo o' 6vo l,em6 on6 qv 6vop(4 rnq Ueroypoq(S, To 5' drpo tou mRNA op1((er v' onodope(tor npoto0 ouwe0e( f Ueroqpoore( to 3' 6xpo. H ono66pqo4 qo(verot 6rt oxoLou0e[ to reLewo(o prBootoprdtro rqg noprnrig r<otd prrlxoq rou mRNA. Qsr6oo, npolcope( Bpo6ritepo, nep[- nou Ue rn Urori tolUrrpo rnq Ueroypogrlg r<ot rnq Ue- rdgpooqg. H oro0ep6r4ro rou mRNA 61er peydlr; en[6poor1 mqv noo6rrlrcl rov notrunenl6(ov nou nopdyowot. Iuvq0cog erqpd(etor oe oX5o4 pre to Xp6vo npt(otiq. To mRNA rdOe ouyrerptpr6vou Borcrrlprorof yovt6(ou dXer Xoporcrqprortx6 Xp6vo r1;u(oriq, Ue pr6oo 6po -2 Lemd. Aorpolrig, ourl q oepd ougBdwcov e(vor equcrrl 95- vo ener6{ 4 petoypoqrl, 4 ;retd<ppoo4 ror q ono66pr1- on npoyporonororiwor npoq rnv (6ro r<ote0Ouvor1. H 6uvogu<r1 r4g dxrppoorlq rov yovt6(tov otrttKonote(rot orqv rllercrpovu<r1 gu<poyporp(o qg EIKONAI 19.46. Ittq ouyrerpqldveg (dyvcooreg) peroypogxdq govd6eg, op- retd mRNA ouw(0ewo row6Xpovo r<or ro r<o06vo rp6- per noLLd prBoocoprdrro Trou onooXotrotlvrot pe rrl prerdrppoorl. (Aur6 owrororle( oro orddto nou nopou- ord(etor oro 6e0repo nlq(oro r4g Ex6vog 19.45). 'Evo RNA rou ono(ou 1 oOv0eoq 6ev 6Xet or6pq otrortr4pto- 0e( ouyyd ovopd(eror yew6pevo RNA (nascent RNA). To Bor,cqprord mRNA noxfi,ouv notr[ cog npoq rov opr0g6 r<ov nproreivci;v nou ro6xonotoUv. Mepu<d mR- NA g6pouv ;r6vo 6vo prov6 ORF, 6qLo6r1 e(vot govoor- orpovrx6 (monocistronic). A).Lo (q ntrerov6tqto) rpSpouv otrqtrouX(eg nou ro:6rronotoUv notrtrd nolune- m(6ro, 6nl,o6n e(vor notruororpovrx6 (polycistronic), Ie ow6q rrg neprmcboerg, 6vo ptovo6tx6 mRNA preto- ypdgerot on6 pro ogd6o yenovu<tirv otorpov(olv. Mto rStoro oumorl(o ormpov(rov onopr((et 5vo onep6vto nou eLdylerot rrlg evro(o yevetx{ provd6o (B}.5ne Keqd- Loro 26). 'OLo ro mRNA nept6Xouv rperg neptoX6g. H ro6tro- notrlrtrri neproXrj (ovorXr6 nLo(oro ovdyvcoo4g, open reading frame, ORF) onoretrekor on6 puo oerpd xo6txo- v(cov nou owrTrpoooneUouv rrlv oprwo[u<{ oMrl}.oufo rou nol.unemr6(ou. ApX(er (ouvrlOrog) pe AUG Kor re- Lercbver pr' 5vo on6 ro rp(o ro6rr6vto tepgottol-toti. To mRNA oor6oo e(vor ndwore prorpUrepo on6 t1v t<to6t- xonorrlurrl neproX(, roOrilg nepdler enrnp6oOeteg ne- -lil 'il 0 KUrtrog rou Borqproxo0 Ayyetrrorpopou RNA 547
  • 40.
    t-a To yevvdpevo mRNAnp0e!6x0uv ono ro DNA xor eivor xotruppevo pe prpoorogono Mnopotlpe v0 flopornpnoouUe Ttq UeT0yp0qx6q Uovddeq oT0 PoKrnpro. EuyevLr4 nopoxd)pnon rou 0scar Miller. at proX6g Kcr. oro 6Uo dr<po. Mro enrnp6oOerrl oMrlLoul(o mo 5' drpo, ovo6u<d r4g ro6txonorrltrrrlg neproXrlg, ovogdpetor coq odqy6g (leader) n 5' UTR (5' untranslat ed region, 5' operd<ppom4 neproXri -). enrnp6o0erl oMqLoul(o r<o0o6rxd on6 ro o,j;ro repgotroproU, q4r1- pror((er ro 3' dr<po ror ovoprd(eror oupd (trailer) ri 3' UTR. llop6rt 6ev x@drronotoUv notrunem(6to, ourrlg or oMql,ouX(eg pnope( vo nepr6Xouv orlgowrx6g puOpu- oux6g o6qy(eg, rdicog oro europuronrd mRNA. 'Evo nol,uororpovrx6 mRNA en(o4q nepr5ler ev6o- ororpoux6g neprol6g (intercistronic regions), 6nog go(veror or4v Ell(ONA 19.47. Aur6g ot neproX6g nou<fi,ouv notr6 oe prdyeOog, on6 trr6l,tg 1-2 vouxLeor(6ro ovdpeoo oro rro6rr6vro repporroUoU ev6g no).unel6(ou rqt ro ro:6rr5vro 6vopEnq rou en6pevou nolunenn6(ou 6oq 30 voux.l,eorbro qro Borcrqprord mRNA (xor or<5prr1 ne- ptoo5repo oro RNA trov gdyrov). Ie pto or<po(o nepi rroon, 6Uo yov(6ro orrlv npoytlotrr5rrlro enrKo- trUmowor, rftor riore rlteAeuro(o Bdon qS prog ral6r- ronotrlru<qg neproXrlg vo e(vot xor 11 np6q Bdo4 tqg en6prevrlg. O opr0pr6g rrr:v prBoo<rt1.ror[ov nou eprn].6xowot orrl petdrppoorl ev6g ouyrerprpr6vou ororpov(ou e[oprdror on5 trlv ono6our6rrlro rnq 06o119 6vop(qg orrlv 5' UTR rou. H 05o4 6vop(49 yto ro nptirro ororp6vto y[ve- tor 6ro0Sorprl U6fuq ouwe0e( ro 5' drpo rou mRNA. E' | ..1r: -r-C rT1..: -+ ! il H 5' UTR nporlye(tot H 3' UTR oxotrouoei ro rou rotOtxovlou tlvop{1q xo.r6rx6vro repportopou I H dloototpovrxrl onooToon I I norx[trer ono -1 eulq -40 pooerq I l-lilg prerogpd(owot ro en6pevo ormp6vro; Apoye, or 6rd<popeg ro:6tronor4lr6g neprol6g ev6g nol,uotorpo- vtxori mRNA;rerogpd(owor ove[dpqro rl pujnog q 6r<- gpoon roug ouv65ero6 E(vor rorv6g o ;rqXovropr6g rrlg 6vop{qg yro 6},o ro ormp6vro, ri Unnoq dtorpdper yro ro npcilro ororp6vro Kcu yro ro eooteprxd ororp6vro; H perdgpooq ev6g Bor,rqprorori mRNA npoXope( 6ro6oXu<6 6ro;r6oou rov ororpov(<ov. Orov to prBoorr:- prdtro npoo6e0oriv orrlv npc0rrl ro6rronorqnxrl nepto- Xt1, or en6preveg ow(ororXeg neproldg 6ev 51ouv r<ov getoyporpef. M6]lq y(va 6ro05orprrl n 06on yto ro 6eri- repo ptBooropdno, q perdgpooq 6Xer r16q npolcop4oer yro ro roLd pr6oo oro np6to ororp6vro. Tunrrd to pr- Booropdrto reppror(ouv q prerdgpoorl qro drpo rou npcilrou ororpov(ou (rot 6roXrop((owor oe unoprovddeg) t<t tivo v6o prBooroprdrto ouvoppoLoyekor ove[dptrlro mqv op14 qg en5prevrlg ru:Stronorqttxriq neproX4g. (Or 6repyoo(eq rnq 6vop(49 Kcu rou repprorropoti 0o nqpouoroororiv de(o6txd oro Kerpd).oro 24). 19.24 I'lepiAn$n MLo peroypogrrq povddo onore}.e(ror on6 ro DNP peto[[ Tou unoKrvnrq xor rou repprolorrl, dnou oplL- J ,-.KiL'0ii0f l "iiKll + 'Evop[rl Tepgortopdg 'Evop(rl TspUqTroUdq To porqptoro mRNA nepiloltpovet operogpooTeq Kor geroqpo(opeveq neprox6q. Ko0e rodrronoullrrl neproXrl 6XeL 0 . : qq o1;to ev0pElq rot olpo repportopo0.'Evo tunrro mRNA pnopei vo eyer oprertlg rodrronorllreg neproXeq, 548 KEOMA|0 19 MeToypoqrl oroug l-lporoputrtreg
  • 41.
    (er rot teletc0vet,owtmo[1tog, n UeroYporprl. 'Evoq x).rilvoq tou DNA ournq tr1q neptoXqg Lemoupye( o:g erproye(o ytq rrl o6v0eo4 ev6g ougnLrlp<oprotrr<o0 rl,ri:vou RNA. H uBprdtr<r1 neptoXrl RNA-DNA e(vot ppoxeio Kqt npooopw(, r<oOtig n (UeroYpoqtrrl <pu- ootr[6q" ruvekot rotd prjxog tou DNA. To o]iodv(u;to qg RNA nol,upep6o1g nou ouv06tet Bolmrlptor<6 R- NA drorp(verot oe 60o ouorottr6. O nuprivog rou ev- (ri;rou e(vot 6vo noLupep6g nou nepr6Xtt rtq unopovddeg ozgg'o ot ono(eg enoproUv Yto rnv eflt- prr6uvorl qg oluo(6og tou RNA. O nopdyowog o e(- vor Urq povtl unogovdEo onopoitry4 pr6vo oro qr66to rrlg Svop(qq yto rrlv ovoyvc0pr;or1 rou unoxtvqtf . O nuprlvoq rou ev(0pou SXet prto yevtni ouyydveto yro ro DNA. H npooOrir<r1 tou nopdyowo o petci:vet tr1 ouyy6veto tou ev(rjpou Yto pn etdtxrl np6o6eor1 oro D- NA evrir ou[dver rq ouyy6vetd rou Ylo touq unorwrlt6g' H roXrlqto pe rnv ono(o 11 RNA noLupepdorl Bp(oxet rouq unoKtwltsq qg e(vot t6oo nol'ri ;reydl'r1 dlore 6e gnope( v' ono6o0e( onLcirg oe tuXo(eg ouvowrloetg pe ro DNA ptSoo onLrlg btdXuorlg. Ot ;retoypo<ptro(nopd- yoweq nou npooetrrUouv tr1v RNA nol'uprepdorl oro DNA xot xoteu0Uvouv qv 6preoq owoMoyrl rou ev(ri- Uou UetqEU rnq Utoq n rnq 6Mnq oMrlLouX(og DNA nt- 0ov6v no((ouv p6Lo orrlv ovo(r1r4orl rou unor<wr1ti' I-1oLLo( Bor<tr1ptoro( unortvqtdq touronototivtot on6 6Uo otrLrll,ouX(eg 6 bp ortg O5oetg -35 t<ot -10 oe oxSorl Us r4v oqerrlp(o rnq perqYpoqriq. lqv ovoyvc0- prorl rou unorwqt4, coot6oo, ougBdMouv en(o4q rot 6Mq entKoupxd orotXe(o rou unoKtvnttl ovodxd on6 ro orotle(o -35 (ro orotle(o UP) r<or yUpo on6 ro orot- Xeio -10 (ro 6teupupr6vo -10 rot ot neptoX5g tcov 5to1to- protcirv) . H on6otqorl ;rero(0 rov ouvqtvettrriv oi.Lrll.ouXrc0v oXe66v ndwore e(vqr 16-18 bp. To 6v(u- pro prnope( vo rqtrUntet 5og -75 bp DNA. To opltr<6 *xletor6, 6tgep6g o0gnLoro petotpdneror oe (ovot- Xr6, 6rprep6q o6gnloro pe 6to6oXtr<rl ono6toro[q ptog oMqlouX(og -14 bp nou [er<tvd on6 qv neptoX( -10 rt eme[vetu 6ro9 nep(nou 3 bp r<o0odtrd on6 to oqpre(o Svop[qg. H ou(qgdvq nepterctx6tlro rnq oMrll'oufoq -10 oe (eUVn gdoeov A-T ouprBdLtret mrlv ow(6poor1 ono6tdro[qg. To dtpep6g o0prntroro petotp5nerot oe rpq.rep6g pe npooOri r<r1 n p66 poprov ptBovo u rLeort6(tov. Euvt10 oq oupBo(vouv noLl.oilro( t<0x).ot oventtulriv evdp(ecov, xord toug ono(oug q RNA notr'uprepdoq ouv06tet rt oneLeu0epdrvet notr0 Bpole(g xtrc0vouq RNA Xop(q vo drorpe6yet on6 tov unortvqnl. Ito t5Log outoU tou orod(ou, o o ouvriOog onetreuOepc0verot Kot o nuprlvoq rou ev(fprou nou onop6vet ro],rimet p6vo -35 bp DNA 6vowr rov -70 bp nou rol'6met oro o6ptnf,oro 6vop- (r1g. 'Enemo, o nuprivoq tou ev(0prou petorlefuot rotd prqroq rou erpoye(ou, [etuL(yowoq to DNA xqt ouv06- rowoq to RNA pretdyPogo. O nuprlvog tou ev(6ptou prnope( v' ovoyvop(oet unoruvrlt6g pe 6 totpo petu<5g ouvowetrr6 g oMrllo u1(eg un6 rqv r<o0o6riyqor1 evqtrtroKrtr6v nopoy6wtov o. Eto E. coli, ouro( ot nopdyoweq o evepYonoto0wot un6 ow((oeg ouv0{reg, n.1' 0epprx6 shock 11 6MEnl o(cinou. H yeopretp(o tou oupnh6rou RNA nofuppti' onq-unoKtvtlrri e(vot oXettrd nop6proto yto 5)to ro olo riv(u;ro. 'OLot ot nopdyoweq o er<t6g on6 tov d' ovoyvop((ouv ouvotvetu<d orotle(o nou Bp(orowot ne- p(nou 35 r<ot 10 ovodtrd on6 rr1 06oq 6vop(1q rnq Ue- toypoqrig, npqYporonotri:vtog dpreoeq enoq6q ue Bdoetq owc0v ttov orotle[tov. O nopdyowog o70 rou E' coti 6yet gro N-reLrr<ri ouroKoroorotrtu<q neptopl nou onotp6net qv ovoyvcbptorl rou DNA on6 nq neptoX6q np6o6eonq DNA. H ouroKoroorotrttx4 neptoX4 erron(- (etor on6 DNA 6rov to oLo6v(upro oXrlproiloet 6vq o- vorxr6 orjprntroxo. H *roxrls" ev6q unortvrlrl neptypd<per q ou;y6qro pe rrlv ono(o q RNA noLuprepdorl (er<wd 11 pretoypoqf . Elet((etot pre to Bo0pr6 opror6rrltog ("ouuu6p9oorl") tcov orotle(u:v -35 t<ot -10 rou unortvqt( rot 6l,Lcov ent- rouptrcbv orotle(rov pe lg t6ovtr6g ouvotvelx5g oMrl- l,ou1(eq. H opvrlttr<ri unepel,(rcooq ou[dvet tqv toX0 oprop5vtov unorwrlttitv. H pretoypotprl 6qptoupye( Oen- r<5q unep6trtreq prnpoord on6 rrlv RNA noLuprepdorl rot oqrivet opvrprr<5g unep6trtreg n(o<o qn6 to 6v(upto. To B. subtilrs nept6Xet 6vo govo6tr6 pre((ovo nopd- yowq o Ue rl'lv (6to etdtt<6tt'1rq Ue ro pre[(ovo nopdyov- ro o rou E. coli, ol'l,d en(or1g nepdXet rot notrilouq etrdoooveg nopdyoweg o, rdnotot on6 toug ono(oug evepyonotorjwot 6to6o1trd r<ord rq dtepyoo(o rrlq oro- proy6veorlq. H onoproy6veor; pu0p((mqt on6 Utq oKo- trou0(o nopoy6wtov o, ornv ono(o owtxotoordoetg rcov nopoy6wov o npoYuoronototiwot oro npoon6pto Kqr ro Unrptr6 r0rropo. Arotrou0(eg nou nepil'opBd- vouv 6to6oXtr<n Xpnon 6to<popettxtilv RNA noLupepo- ori:v prnope( en(oqg vo puO;t(oouv rn UeroYpoqrl rord rq Lo(go[q on6 BorcrrlPtorPdYo. H Borcrrlptoni RNA nol'u;repdorlteppror((et t1 peto- ypoei oe 6rio trjnouq 06oecov. Ot evdoyeve(q repport- m6q nepr6louv Uto nl.oUoto oe G-C rpoupr6to nou or<ol.ou0e(tot on6 gto ntro0oto oe U neptoXq. ln vitro ovoyvcop((owot p6vo on6 tov nuprivo tou ev(riprou. Ot p-e[optcirprevot repportor6g onottotiv tov nopdyowo p 16oo rh vitro 6oo rot in vivo. O p npoo66vetot ortg 05- oerq rut nou e[vqr n].o0oteg oe rotdtrotnq C xot <ptcoX6q or 6 oo, npolyorlwot rnq npqYuorrxrig 06or1q repuort- opoti. O p e[vot pto e[opreprlq e].rxdor1 e[optcOprevq on6 to ATP nou UeroKtvefuqt Kotd ;r1xoq rou RNA pr51pt vo grdoat orrlv RNA notru;repdor1, 6nou nporotre( 6t6mo- on rnq RNA notruprepdonq on6 to DNA. Kqt moug 60o t0noug tepponopoti, q no0oq qq RNA nol'uprepdoqq nr0ov6v ouprBdMet orov teppottop6' O owtteppotrop6g lp4otpronote(tot on6 toug tropt- Bdoerde(g qdyouq yto q pri0pton rnq npo66ou'on6 to 6vo srd6to 5r<rppooqg to.lv yovt6(tov oro dMo. Tq no- Lunpolteivtt<d orjprnLor<o nou nept6louv rrlv npotetvrl N 11 Q tou gdyou l'dpB6o 6nrr:g en(or1g rot touq nopd- yoweq Nus prnope( vo ouv6e0ofv Ue rnv RNA notruge- pdorl 6topr6oou RNA rot (oog DNA 0r1Ltdrv, qwtmo[- Xoq, Kot onorpdnouv rov repUortog6 rnq petoypogdg. To oriprnlor<o owtrepUortol.to( nou nepdlet N enttp6- reoLr-,- 549
  • 42.
    ner orrlv RNAnoLutrlepdo4 vo nopoBrdoer rouq reppo- rtm6g or ono(or Bpiorowor mo drpo rrov dtrleoov npciuprov yovr6(rov, evri rq oripnLoro nou nepr6louv Q Xperd(owot oe preroyev5orepo md6ro rqg l,o(;ro[r1g on6 gdyoug. BrBAroypogio 19.2 H Meroypoqrl flpoygoronore(ror Ue Zeuyopo1l0 Booeov oe plo u0uootrrdo, A(euydprrlTou DNA Avoordnq*q Losick, R, and Chamberlin, M. (1976), RNA polymerase, Cold Spring Harbor Monograph Series 6. 'Ipeuva Revyakin, A., Liu, C., Ebright, R. H,, and Strick T. R. (2006), Aborlive initiation and productive initiation by RNA polymerase volve DNA scrunching. Science, 31 4, 1 1 39-1 1 43. 19.3 H AvriDpooq Tr1q MeToypuQflq'EXer Tp(o Irddro 'Ipeuva Rice, G. A., Kane, C. M., and Chamberlin, M, (1991). Footprinting analysis of mammalian RNA polymerase ll along its transcript: an alternative view of transcription elongation. Proc. Natl. Acad. Scr. USI 88,4245-4281. Wang, D. et al. (1995). Discontinuous movements of DNA and R- NA in RNA polymerase accompany formation of a paused tran- scription complex. Cell 81 ,341-350. 19.4 H Borqprorrl RNA llotrupepool Anoretrelror ond l-loii6q Ynopovodeg !.i ::'.': t. {.:, t.:: :. : i ; Helmann, J. D and Chamberlin, IVl. (1988). Structure and function of bacterial sigma factors. Annu. Rev. Biochem. 57 ,839-872. Shilatifard A., Conway, R C., and Conway, J. W (2003). The RNA polymerase ll elongation complex. Annu. Rev. Biochem. 72, ovJ-/ I 3. Campbell, E, A., Korzheva, N., Mustaev, A., Murakami, K., Nair, S., Goldfarb A., and Darst, S. A. (2001). Structural mechanism for rifampicin inhibition of bacterial RNA polymerase. Cell 104,901- 912. Geszvain, K. and Landick, R. (2005). The structure of bacterial R- NA polymerase, ln The Bacterial Chromosome Higgins, N. P (ed.). Washington, DC American Society for Microbiology Press, pp. 283-296. Korzheva, N., Mustaev, A,, Kozlov, M., Malhotra, A. Nikiforov, V., Goldfarb, A., and Darst, S. A. (2000). A structural model of tran- scription elongation Science 289, 61 9-625. Vassylyev, D. G., Vassylyeva, M. N., Perederina, A., Tahirov, T. H , and Artsimovitch l. (2007). Structural basis for transcription e- longation by bacterial RNA polymerase. Nature 448,157-162. Zhang G., Campbell, E. A., Zhang, E. A., lVlinakhin, 1., Richter, C., Severinov, K,, and Darst, S. A (1999). Crystal structure of Ther- mus aquaticus core RNA polymerase at 3,3 E resolution. Cef 98,811-824. 19.5 To 0tr06v(ugo Tl1q RNA l-lotrugepoonq Anorele[ior 0n0 Tov I'luprlvo rou Ev(upou Kor rov n0p0y0vT0 Iry1to 'Ipeuvo Travers, A. A. and Burgess, R. R. (1969). Cyclic reuse ofthe RM polymerase sigma factor. Nature 222,537-540, 19.6 l-ltig Bpiorer H RNA llotrupepoon rtq Atrtrr1trouXieg rov Ynorrvrlrdrv; ,Avoordnqoq Bustamante, C,, Guthold, M., Zhu, X., and Yang G. (1999). Facili- tated target location on DNA by individual Escherichia coli RNA polymerase molecules observed with the scanning force mi- cr0scope operating in liquid. J. Bio. Chem. 274, 16665- 1 6669. 19.7 To Otroriv(upo Tpononorelrot Koro q Arepyoo[o Avoyvroprolg rr Anetreu06proonq 0rr0 Touq Ynorrvqr6q 'Ipeuva Bar-Nahum, G. and Nudler, E. (2001). lsolation and characteriza- tion of sigma(70)-retaining transcription elongation complexes trom E. coli. Cell 106, 443-451 . Kapanidis, A. N., Margeat, E., Ho, S. 0., Kolckhonjia, E., Weiss, S and Ebright, R. H. (2006). lnitial transcription by RNA poly- merase proceeds through a DNA-scrunching mechanism. Sc" ence 314,1144-1147 . Krummel, B. and Chamberlin, M. J. (1989). RNA chain initiation by f. coli RNA polymerase, Structural transitions of the enzyme ir early ternary complexes. B i o ch e m istry 28, 7 B2S-7 842. Mukhopadhyay, J., Kapanidis, A. N., Mekler, V., Kortkhonjia, E.. E- bright, Y. W., and Ebright, R. H. (2001). Translocation of sig- ma(70) with RNA polymerase during transcription. Fluorescence resonance energy transfer assay for movement relative to DNA. Cell 106,453-463. Wang, 0., Tullius, T. D., and Levin, J. R. (2007). Effects of discon- tinuities in the DNA template on abortive initiation and promoter escape by E. coll RNA polymerase. J. Biol. Chem 282,26917- 26927. 19 B 0 ilopoyovrog l(ypo EtrrlyXer qv ilpocdeol 0T0 DNA Avoyvop((ovroq IuyKeKprp6veq Atrtrr'1trouXieq rov Ynonvqrtlv Haugen, S. P., Ross, W., and Gourse R. L (2008). Advances rn bacterial pr0moter recognition and its control by factors that d0 not bind ONA. ffafure Rev. Micro.6, 507-520 McClure, W. R (f 985). Mechanism and control of transcription initiation in prokaryotes. Annu. Rev. Biochem. 54,171-204. Bar-Nahum, G. and Nudler, E. (2001). lsolation and characteriza- 550 KEOAAAI0 19 Meroyporprl oroug l-lpoxopudrreq
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    complex containing thelambda N protein. Genes Dev.9, 2831- 2845 0 son, E. R., Flamm, E. 1., and Friedman, D. l. (1982). Anatysis of nutR: a region of phage lambda required for antitermination of transcription. Cell 31, 61-7 0. 19.23 0 KUrIoq Tou BoKTnploKori Ayyetrrogdpou RM Brenner, S., Jacob, F. and Meselson, M. (1961). An unstable nr::- mediate carrying information from genes to ribosomes for pro- tein synthesis. Nature 190, 576-581. BLptrroyporpLo 553
  • 46.
    @ Carol &lVlike Werner/Visuals Un - :: - H Meroypogt1 oTouq EuKopudrreq ffi Erooyoyrl * lto vq npoode0e( orov unorrvlrrl 11 RNA notrupepool npenet v' ovo(er 1 XpoUouvrl. 0r Europuorxeg RNA l-lotrupepdoeg Ervor flotrupepr1'Ev(upo * H RNA notrupepool I ouvO5rer rRNA orov nuprlv(oro. e H RNA notrupepoorl ll ouv0erer mRNA oro nuprlv6ntro- ou0. s H RNA notrugepool lll ouv06rer ptrpo RNA 0To nupn- vtlntroopo. * 'OtreS or europurrltrreg RNA notru;repooeg 6Xouv -12 unopovddeg rol e(vor oupntroro rtov -500 kD. * Meptx6q unopovodeg eivor rorvrlg Kor onq rperg RNA notrupepooeg. s H Ueyotr[rep1 unoprovodo qg RNA notrugepoorlq ll 6XeL pro CTD nou onore),e(ror ono notrtrontr6g enovo- trrlgerq evoq enro;le pouq. H RNA l'lotrupepdor; 'EXer evo ALpepq Ynouvlr4 c 0 unoxrvlrrls iqg RNA notrupepooqg I onoretre(rot onrj 6vov xevrptrd unoxtvrlrr; rot 6vo ovodtro orotXe(o rou unouvlrrl (UPE - upstream promoter element). * 0 nopdyovrog UBF1 rutrryet DNA yupto onri pto npo- teivrrrl dopr; yro vo rp6pet rovro rov revrprro urroKrvrl- rr1 ror ro UPE. s 0 SL1 nepiloppovet rov nopoyovro TBP nou egntrexe- r0r oTrlv 6vop(q ror onrl lg rperg RNA notrupepdoeq * H RNA notrupepoorl I npoodeveror oro oupntro<: UBFl -SL1 orov revrpmrl unorrvrlrr;. ilFry5. *:, F,F ffi H RNA notrupepool lll Xprlorponorel T6oo Ko0o6rroLr '0oo ror Avodxoug Ynorrvrlr6g * H RNA notrupepoorl lll 6Xer 6uo runoug unouvlrolv. e 0r eou;reptro( unorrvrlr6g eXouv ppoXeieq ouvorvsT.- xeg otrtrqtrouX(eg nou pp[orowor luioo oTn peToypoq.- rq povodo ror nporoLouv rqv 6vop[q 0e Urc ouyrerpq6vrl ondoroorl ovodxd. e 0r ovoduo[ unorrvlr6g nept6Xouv rperg ppoye[eq ou- vorvertreg otrtrrltrouxLeq ovodrro rrlg orperrlpiog oric ono(eg npoo66v0vT0r UsToyp0qxoI nopoyovreg. * 0t TFrrrA ror TFrrrC npoodevovror ong ouvorverx6g otr- trqtrouXLeg rt enupenouv qv npdodeorl rou TFlllB o-qr oqeqp(0. * 0 TFrrrB rp5per cog unopovo0o rrlv TBP ru enupdnel qr npoodeol rrlq RNA nolupepoolg. H Aqeqp(o yro rqv RNA l-lotrugepoorl ll * H RNA notrupepoorl ll yro v' opX[oer Tn UeroypoQr onoue( yevlrouq UeToypoqrKoUg nopoyovreq (ovolro- (ovrotTFlfi). ryE*3 554 KEOAAA|0 16 Iuoqporo EnLdtop0uloqg 20 ji ,$- .:$.'j t ,.1 {.!.: 5i .* .*: ..!' l-
  • 47.
    ffi H TBPervor0txoupevxdg l-lopriyovrog ffi ffi H Bootrrl luoxeurl luvoppotroye(ror orov ynorrvqrrl * 0r unoxrvrlrdg qg RNA notru;repooqg Il oqv oqeqp(o ouXvo 6Xouv pro ppoXeio ouvrrlpq;rrivrl otrtrr1trouX(o PyzCAPys. e To xour[ TATA e(vor rotvo ouorolrd rov unorrvrlrov rqg RNA notrupepooqq ll rur onoretre(ror ond evo ntrouoro oe A-T orropep6q nou pptoxeror -25 bp ovo- 6xo r4g orpeqp(og. * To DPE elvor rorv6 ouorortrd rov unouvqrov r4g RNA notrupepoqg ll nou dev nepr6Xouv rour(TATA. * 'EvoS revrpxog unouvqrrlg yro qv RNA notrupepoo4 ll nepilolrpover ro lnr rol, ouvr1Oog, efue 6vo rour( TATA e(re 6vo DPE. Mnope( enro1g vo nepteyer roL dtrtro eLdooovo ororXe[0. * To orroUepn ru.rv torovdrv rpononoro0vroL np6oxorpo rord q perorivlon Tnq RNA notrupepdoriq. * H CTD ouvrov[(er r4v enetepyoo(o rou RNA Ue q Ue- r0yporpq. a Av ouUpouv ptroBeq oro DNA, 0o enr6top0rrl0orjv rord nporiprlol ro peroyporpopevo yov6to. * 0 TFIIH nop6Xer ro ouvdeopo yro evo oupntroro ev(u- Uov enrDt6p0oonq. ffi O,EvroXur6g llepreXouv Aggi6popo lrorle(o nou ynopoq- Ootjv r4v'Evop[1 * 'EvoQ evLoXuqg evepyonorei rov unorrv4rq nou pp(_ oKeror rr0 xovro rou rot pnopef vo ppfoxerot 0e olror_ od(nore onoorool e(re ovodrro e(re roOo0uo ono rov unoxrvrlrrl. * Mro AUS (ovodurl evepyonorlnx4 otrtrritrouXro) rou (u- popUrrlro oupnepup6peror oov evtoXurrlq, otrtro trer- roupyel p6vo ovodrrd rou unoxrv4r1. o 0r evroxureq oXrl;ror((ouv oupntroro on6 evepyonot- qr6g nou otrtrqtrenLdpor1v opeoo 11 6ppeoo Ue Tov uno- xrvr1rr.1. ffi O, EvroXurdg AeLroupyorlv Au[dvovrog 14 luyr6vrpororl rov Evepyonorrlrov Kovrd orov ynoxrvqrrl * 0r evroxur6q ouv4Otrlg treuoupyouv prlvo oe cis 6Lo- UdpQoo1 oe oX6oq Ue Tov unoKrvt1rr1-oroXo. * Ko0e evtoXurqg unoXpetirveror vo treuoupye( pdvo 6rov ppe0ei xovrd orov unouvrlrrl. ffil H 'Erqpoor'1 rov lovrd(tov lXer((eror pe qv Anope8utrito- on o H onope0utr[too1 oro 5' dxpo rou yovr6(ou e(vot qno- po(qrr1 yro t1 peroyporprl. ffi O, Nrlo6eg CpG E(vor Pu0ptorrr<o[Ir6Xor * 0r vrloOeq CpG nou ppforovroL oroug unortvrlrdg rov r6rootjororo ertppo(dpevov yovrd[tov e[vor onope0u- truop5veg. * Nqo(6eq CpG pp(orovror enfo4g Kor oTouq unouvrlreg op toluivtov yovtd frrrv pe roroe16 rrrl rixrppoori. * To ovOpdlnrvo yovt0(rrllro nepr6Xer -29,000 vrlo(6eq cpG. o H Ue0utrioon prog vrlo0og CpG onorpener Trlv evspyo- no(qo1 rou oXert(dpevou unoxrvrlq, a Koroorotrr1 nporotre(ror ond nporeiveg nou npood6- vovTor or pe0utrrop6vo (euyn CpG" ffi nepiinrpn e H TBP e(vol ou0rolx6 rou nopdyovro rono06qoqg nou onoue(ror yro qv npdodeoq roOe RNA notrupepd- orlq oTov unoxrvnrq qg. o 0 nopoyovroq yro Tnv RNA notrugepoon lll e(voL o TF1D, nou onoretre(ror ond TBP ror -14 TAFs, ;re ouvotrurl Uo(o -800 kD. * H TBP npoo66veror oro rour( TATA oqv etroooovo ou- troro rou DNA. * H TBP oX4pol(er pro o6tro y6po ond ro DNA ror ro ropnrer roro -800. * To ovo6trd ororXe(o xor ot nopdyovTeq nou npoode- vovror 0r ouro oulovouv 14 ouXv6qro r4g evop(r1g. * H np6odeon rou TFrrD 0ro rour( TATA 11 oro lnr e(vot ro npdro p4;ro rrlg evop[r1q. a Atrtrot peroyporprro[ nopoyovreg npoodtivovror oro oupntroro ge xoOoproprlvn oerpd, enexre[vovrog rqv npoororeupdvq neproXrl oro DNA. * H RNA notrupepoorl ll opX((et rq peroyporp4 pdtrrg npoode0e( oto oupntroro. H'Evop[1 Arotrou0efrot on6 Ko0oporl rou ynortvqrrl rt Enq1xuvorl * 0r TFrrE rot TFrrH onououvror yro rrlv r4ir1 rou DNA ror rr1 peroxfvqol qq notrupepdonq. * [t0 vo [errvrloeL 4 ro0oporl rou unorrvrlrrl ror 11 enqrrl- ruvorl npener vo rpootpoputrtro0e(11 CTD. * Ie pepuoUg unorrvrlr6g, yro vo repporiorr rl ovenrTu- Xrlg 6vop[1 lpero(erot enrnpooOer4 gootpoputrioorl qq CTD. 2O,l Erooyrrlyri e lro v0 npoode0er orov unoxrvqq q RNA notrupepool np6ner v' ovol[er 1 Xpoporivrl. H 6vop[q rnq UeroypoAnq o' dvo er<proye[o Xprr;prorG vrlq nou e(voL 11611 ovotXr6 onotre( np6o6eoq rrlg RNA no).upepdoqq orov unoxrvrlll Kor Ueroyporpxu_lv no- poy6wotv orouq evroxurdg. /n vitro, preroypo<pq o, 5vo er<proye(o DNA onore( pro 6roqoperu<4 unooprd6o geroyporprx<ilv nopoy6wov qn6 exe(voug Trou oTror- roUwor yro rn peroypogri rou erpoye[ou rnq Xpopo- rfur1g (oro KeqdLoro 28 0o e[erdoou;re nc6g ovoryer q Xpropror(v4). KdOe nprrlteivq onopo[rr1r4 yto qv 6vop- En qq petoypogrlg, q ono[o 5prog 6ev e(vor 11 (6ro rpnpo tqg RNA noLuprepdoqg, op((eror oq Ueroypo- gtr6g nopdyowog (transcription factor). l'1oMoi prero- ypogrxo( nopdyovteg 6pouv ovoyvrop[(ovrog cis-evepyorlg 06oerg oro DNA. H np6o6eoq qro DNA, to016o,0, 6ev e(vor o provodu<6g rp6nog 6pdorlg ev6g UerqypogtKo0 nopdyowo, o ono(og prnope( v, ovo- yvopr(er 6vov dMo nopdyowo ryrqv RNA noluprepd- l:i.1 Erooyoyri 555 Boorxeg eworeg
  • 48.
    on, 6noq en(onqKor vo evoopor6verqr USoo o'6vo oUpnLor<o 6vop[r1q p6vo nopouo(o oprercitv 6Mov npcoreivcbv. H r<p(or;rq 6orrproo(o yro rn ouprpretopl qrn Ueroypogrrrl ouoreurl e(vor Lemoupyrrrl: yro vo Bp(oreror puo npore(w1 o' 5vo ouyr<erprprSvo unoKrvrl- fi 11 o;rd6o unorwqrriv np5ner vo e(vor onopo(rqt4 yroq peroypogrj. Mro o4gowu<{ 6ro<popd peto[ri tqg pretoypoqrlg Tov euKopuoru<rilv ror rov rrpoKopuorxrilv RNA e(vot 6tt oto prev Bor<rrlpro n Uerqypoerl npoyproronore(rot o' 6vo er<proye(o DNA, evrir orouq euKopurilreg o' 5vo erpoye(o Xpro;ro{vr1g. H Xptoprodvq oMd(er ro ndwo ror np6net vo LoprBdveror un6rlq oe r<d0e FriUo. H Xpro- porrvn npdner vo Bp(oretor oe ovoffi 6opu] xor, or6prrl xot t6re, to or,nopreprl rolv voux.Leoocopoilcov npdner vo 51ouv onopor<puvOe( on6 rov unorwrlrl 6ore vo gno- p6oet vo npoo6e0e[ 11 RNA no].uprepdo4. Aur5 preprrdg gop6q npouno06ter preroypoqri on6 6vo ouon4pd r'1 rpuq6 unorwrlrrl, efue orov (6ro e(te srov qwrouv0etr- 15 x).rirvo. Mro 6e0repq o4prowrrf 5ro<popd eivor 6rr n BoKrn- proxrl RNA nolupepdo4, Xdpn orov nopdyowo o nou 6ro06rer, pnope( vo dtoBdoet qv oMrlLouX(o rou DNA ytcr vo Bpe( rov unor<tvrpf Kqr. vo npoo6e0e(, evc0 pro europuamr{ RNA no},uprepdorl 6e ;rnope[ vo droBdoer to DNA. l-r'out6 11 6vop(q o' euKopuoru<or3g unoruvrl- 169 nepil,oprBdver peyd).o oprOpr6 nopoy6wov nou np6ner vo 61ouv npoo6e0e( oe pro nou<tl(o cis-evepy6v ororle(ov npoeroq.rd(owoq ro 66o9o9 yro rnv np6ode- on rnq RNA nolupepdonq. Awo( or nopdyoweg ro- LoUwor poorxoi UeroypoglKo[ nop6yovreq (basal transcription factors). M6).rg oupBe( ow6, q RNA no- ).ugepdo4 npoo66vetor oro oUpnLor<o Boorr6v peto- yporprrcbv nopoy6wov/DNA. H ouyrer<pr;r5vq neproXrl np6o6eoqg op((etor og xewprx6g unoruv4r{q (ri nu- pnvoq rou unoKrvrlrrl, core promoter) xor nept6ler 6leg trg onopofuqteg 06oerg yrq rrlv np6o6eor1 ror rq Lemoupy(o r1g RNA nolu;repdoqg. H (6ro q RNA nol,u- pepdorl npoo6Sveror y6pco on6 qv orper4pio (start- point) rqg peroypogrlg, qI d 6ev 6plerot oe dpreorl eno<pr1 Ue rnv er<retoprdw1 ovo6xrl neproXrl rou uTroKt- vrprl. Aw(O*o, or Borcrlproro( unoxwqrdg nou ovoLrl- 04rov oro KerpdLoro 19 ro0op[(owor r<up(rog oe oXSoq pe q 05o4 np6odeoqq qg RNA noluprepdorlq ornv neptoXq rowd mrlv o<peqp(o. Evcil to Borcnjpro 6Xouv pr6vo puo RNA noLuprepdorl nou preroypdger. Kor lq rperq r<ripreg td[erq yovr6(rov, oro euKqpuorxd rUrropo q preroyporp4 unodropekor oe rperq xoulyop(eg. Kd0e r<oqyop(o peroypdgetor on6 pro drorpopeur<q RNA nol,uprepdoq: . H RNA nolupepdoq I geroypd<per ro 18S/28S r- RNA. . H RNA noLuprepdoq ll preroypdqet mRNA ror Lryo prrpd RNA. . H RNA nolupepdoq lll peroypdger ro IRNA, to 55 ptBootopu<6 RNA r<or dMo ptxpd RNA. Aurr1 r4v err6vo SXoupe yro lq xripeg rd[etg rrrlv yovt6(orv. Onog 0o doripre oro KeqdLoro 3C :::::: reg ovoxoL0qlerq ond Bo0ptdcordq ouorolliec :' :. - pou rou yovr6rrlrporog (whole genome t I 'c ,-". . onor<d).ur.l.rov 6vov v6o rdopo owrouv0errxr.,.lv *:r r , - . tpcov (antisense transcripts), preooyovLdror<cov -:.r, : : rporv (intergenic transcripts) ror peroypdcpc, :-:: :- Xpropror(vqg (heterochromatin transcripts). Ae ,,' . - - -: - pe or6pq t(noro oXerLro Ur rouq unoKrvfro( l--- rorv rdl,eov 11 yro rr1 priOprorl roug, yvcop[(or*, ---: 6t notrl.d (Ud).).ov ro neproo6repo) ond ro c, , .:.: trr6vo petdypo<po nopdyowor on6 qv RNA t:c: -*.:: oq ll. Or Boorr<o( UrroypoqrKo( nopdyoweg e[vcL :-:: - rrlror yrq qv Svop[r1, oMd or neproo6repoL 5: ,.:: d(owor orrl ouv51ero. l-ro rLg rperq euKoputrtrL<.: :',,,- no).upepdoeq, or Ueroypoqtro( nopdyoweq. r:::- -: (6to to 6v(upro, e(vor unerjOuvor yro rrlv ovoyvog.l- -: oM1l.ou1(og DNA rou unoxrvrlrrl. I-ro 6).eg r.c : -. : purrrflr<5q RNA no).uprepdoeg, oL BooLro( pero.:- .: nopdyoweg 6qpuoupyoriv pro 6oprr; fiov unoKt,,-:- -: pdlowoq ro 0161o nou ovoyvop((etot ono :- . = " - no).upepdo4, l-ro rLg RNA no).u;repdoeg I roi :--: or nopdyoweg e[voL oletrr<d on].o(, evol yro :- , :', - no).upepdorl ll ouyxpotoriv 1,.uo opr<erd peyo.I.r :*::: Or Boou<o[ nopdyoweg evcovowot pe rnv RNA r:' - -. pdoq ll yro vo oX4por(oouv 5vo orjprnLor<o nc" -:: Bd).).eL rnv qeernp(o xor ro0op((ouv rq 0ec- -- 6vop(qq. Or Boorxo( nopdyoweg po(( ;ie qv R: -- Lupepdoq onoteLorjv q Boorxl ;reroypoqLr<1 ouci. -- Or unorrvqrdq yro rrq RNA noLuprepdoeg I xc. i: oKovror (r<up(orq) ovodrrd rnq oeernpiog. Ar,tL:. - noMo( unorrvrltSg qg RNA no).u;,repdorlg lll Bc.;. : roL Kq0odrxd tqg otpeqp(og (p6oo orr1 Ueroypc- . povd6o) . Kd0e unoxrv4rrlg neptdler XopoKrrlpr:...: oprd6eg BpoXeLo.lv ouvtrlplpr6vcr.lv oLLqLouxro . -: ovoyvrr;p((owoL on6 rrlv KotdMrl^n rd(n Boorr<i", -." toypogu<rov nopoy6wov. Ot RNA nol,upepdoeg . : ovoyvrr; p ((o uv puo crlettr<d neproproprdvrl o ;rd6 o r- : : . vq-rctrv r<or Boo((owot oe puKp6 opL0p6 enlKouptKr, -: poy6worv. Or unorrvqt5q ilou XpnortrlonoLorjwor ond rqv' :',: noLupepdoq ll eg<pov((ouv preyoLUtepq noLrt).opro:: - orqv oM4).ou1(o r<or opyovrr.rvowor on6 6rdgopc 3:::- -rrlproro (modular organization). 'O).oL or uiloKrvrlrr. :- - RNA noupiepooqq ll nepL).opBdvouv oMrl).ouxrec . : rd orqv ogetqp(o mrg ono(eg npoodSveror q pc, .- ouor<euq rot q noLuprepdorl r<o0op((ovrog rq ii-- 6vop[r19. A].].eg nro onogoKpuoprdveg otrtrq,o1,, : : ovodLrd (q ro0odxd) roo0wot evrolur6g (enhancer r<or r<oOop((ouv ov 0o er<<ppoore[ o unortv4qq Kor, /.-- p[rog, ov q 6x<ppooq rou 0o e(vor oLxoupevu<r1 q r<urr:- po-et6txrl (cell type-specific). O evLoXutrlg e(vot 1-l: L6rofrep4 05or; nou epnl.dxeror crq Ueroypogq xor rcr- ronorekot on6 qv nopouo(o otrtrr1trou1tiov or onorri dLeye(pouv rqv 6vop{r1 oMo ewon[(owor oe 6Ldgope I onoorooetg ond rov rewpu<6 unorrvqrrl" Irorle(o ra. evroXurcov oulvd e(vor mdloL yro roroerdrxn n XpovLxr pti0;uo4. Meprxo( evroXurdg npoo6Svouv peroypogt- 556 I(EOA Al0 20 H Meroypog4 oroug Europudlreg
  • 49.
    EvLoXurnq Ynouvn"rnq -1 00 bp,nEPtdxet oPKtTeq notrri r<ovrw6q 06oetg npdo6eorlq Yto UeToYPoQtKoIq ono rov xevrptro unoKtvlrll Unopo( vo xuPoiveruL onri 1 006eq bP PdXPt kb H onootoorl rtrlv evloxUT{iv +-1 00 bP, nePtdXer dtdonoPreq ohtrr1)rouXieq nou nPoodevouv . rn Booun ueTqYpqQtKn ouoKturl 2O.2 Ot Europuortr6q RNA lloAupepdoeq Eivol floAuUEPfi 'Evquuo nopoyovreq , : . , .Evo runrxd yovioro nou peroypoqeror ono rrlv RNA nou;repooq ll exer 6vov unortvrlrrl r10u eK1e[veTot ovobtro rnq oeolc ln. onou (errvo 11 Ueroyporpn. 0 unorLvrlrflg nepulXet op*rii BpoXeo (-iO np) o,o,Xiiu otrtrntroux(oq.nou npooo6vouv preroypoqtro0g no- j0y0w0q, oroonopro oe novrrr ono -100 bp..Evoq evroxurflg nou nepr6xer p,o n,o'or1.,r,oyt1 ouorotx(o orotxe[ulv nou en[olq npoootlvouv -eroypoqrrouq nopoyovreq, pnopei vo ppiorerot o, onioioJn,.i*r,,i,u i*o,ouro6orv Lp eolq opxerd'v kb (To DNA unope( vo elvot etrtrul- Jsv. | 0v06r0Toyp6vo pe otrtro rpono eror 6ore or usT.yp.QrKoi nopoyoweq orov unorrvqq K.r T.v rvr'xuTn v otrtrrrtrenrbpo.v titore vo :xf Uorloouv 6vo peyotro npolreivtr6 o[pntroro)' <orjg nopdyoweq nou treLtoupyofv pre o]']'r1]'enL6pd- otrq'U,*ptlq'eprBetreLoq r<ot Bp(ot<owot xowd orov uno- .,ul.h ,u, dLLo, prnope( vo Bp[or<owot oe on6oroot'1 ,<lL,iolu (euyrr-rv B&oeolv. H r' ''r ",,' '" ' nopouotd(et ttg :iru,o6q rdt6trlteq rr,ov unoKtvrltotv xot trov evtoxuttilv' r"ll.o pugU,*tr<r1 Oeoq nou npoo65vet nepLoo6tepouq "o"ni*o,is nopd Oertxo0q pu0gtor'59 Ylo..to' dI'eYXo iq !,t oypoQnq r<qtrekot onootonrp4g (silencer)' 'OL unortvrlr6q nou er<ppd(owot t6tooUototo t<ot e[- '.'ot onopoittltot oe 6tro to t<trropo (ro yov(5to rouq Ue- .,^X ,p"pei ovocptlpowot tr;g 6lolerptcrtx6 yovi6to' ntrt'"r"irping genes) tixouv ovo6trd orotxe(o ol'trq- r.cuxloq nou ovoYvcDpiqo*ot on6 otroupevtrotiq evep- . onotrlt6g (u biq u itous activato rs)' Kovtlvoq ouv6uoopr6g -or'lriorTt opdyowo 6ev e[vot onopo(tqto ouorotLr6 1., uno*,rn-n, yeyov6q nou uno6qtrtovet 6rt q evop[r1 :no tqv nNl no).upepoorl ll prnope[ vo pu0prtore( pe .oMolq dtotpopettro0g tp6nouq Ot unortvrlt6g nou t.,ppa(o*o, un6 etdLr6g ouv0r1<eq dlouv orotle[o o]'- i."1ou1(og nou onotto0v evepYonotqr6q 6ro06otpouq -ouo or,q ouyr<er<ptp5veq ouv0rlreq' -- , "pVarroq ev6q evtolurl 11 onootonqrq protd(et -t -nu !*t*otXq ev6g unot<tv1tr1' 6qodq.onotetre(tot l:ro 1.uo notrA(o e(opqpotcov 6nou prnope( vo npoo6e- 3oiv 0ettxo( q opvqtir<o( pu0;rtot6q oe oqtXtd ou- ,otuooUgu.l ouorotl[o Ot evtcrlut6g 6e Xpero(etoL vo .o(orowot rowd orov unortvrltrl' Mnope[ vo Bp(ot<ov- .or ovo6tr6, p6oo o'5vo yov(6to 11 nepo on6 to 6pto avoq yovt6[ou Kot o npooovoto]'topr6q toug o'letLrd pe -o yov[6to 6ev 51et,o1proo(o Ot npulte[veq flou npoo- 5evowot oe orotxeto iou evtoXurl oMrlent6po0v pe .pr-"iv"s npoodiaepteveq oe orotxe[o tou unortvlrq' ro0 ouXvd ptSoto ev6to;reo0lv nou ovo;'to(owot ouve- vcpyonorrlr6g (coactivators)' ' H pretoypotpq tov europurrltcilv un6rettot TIto ouxvo ;e Oettxll p,igU,o.t. 'Evoq petoypoqtr6q nopdyowoq -opeXt-o, un6 toroerbtxd 5}'ey1o Yto vo evepyonotrloet ,uou unoo,untrl q prto ogd6o unorrvqtolv nou nept6Xouv .rto r<otvr1 otrq}'ou1[o or6Xo Aurl ri 6tepyoo(o nept- Lo;rBdvet opretd Briproro: (erw6 pre,dvotytrro rnq Xpo- ,.ii*., ro'ono(o orol'ou0ef 11 np6o6eor1 tov Booxriv ;;;l;;v nopov6wtov xor ^qr6nw qq nol'uprep6- [[;:'[Eeu,on r-,t et6txrl rorooro]'ri ev6g unor<rvr1t(- or6Xou e(vot trtY6tePo r<owt1' Mro euxopucottKn UeroYpoqrrci govd6o levtrd ne- or6xet 5vq uovo6tr6 yov(6ro Kot o repportoUoq flpoYUo- i;,i;;;,';epo qnb to t5trog rqg rtodtronotqtrr<r1g neptoXfq. O rep;rort4t6q 6ev 6Xer 16oo UeY6In puO;tt- *l^n'iihu*(o'6oo oro TlpoKoputotu<d ouorrl goto' ot nrun'r,oLugepdoeg I r<ot ll teppot((ouv oe 6tor<ptt6q otr- iniouxfts'oe xo0optop6veq owr6p6o.ets' op6 o tp6- ;;;;AuL-,opro6 rrlg'RNA notrugepdorlq ll 6ev e(vot oo.inq. lldwoq to <n1powtr6 ougBdv orrl dlptoupy(o ior-ilaopou ev6q mnruR 6ev e(vot ow6q ro0out6g o tepproro;r6q oM6 prto ow6poorl nS$nq rou rporoYe- uo,iq UetoVpdqou (BL6ne KeQdtroto 21)' " H RNA notrupepoorl I ouv0eret rRNA orov nuplvtoKo' * H RNA notrupepoorl ll ouv06tet mRNA oro nuprlvontroo;t0' s H RNA notruuepoon lll ouv06ret uLrpo RNA oro nuprlvontroolto' * 'OeS ot europuortr6q RNA notrupepooeg 6xouv -1 2 unopo- vodeq rot e(vot ouptntroro rcov -500 kD' - 1y1a,*ei unopovodeg e(vor rotvrlg Kot oTtq rpetg RNA notru- Uepdoeq. * it ueyor[repr1 unopovodo qg RNA notruuepoonq Il 61eL pto CTD nou onoreLelrot ono notrtroneg enovotrlQetg evoq enroUePoUg. Ot tpetg europutottxSq RNA noLu;repdoeg Bp[oxowoL ot oio,popt trSg 06oetg orov nuprlvo' ot ono[eq owt- 20.2 0r Europuortr6q RNA tlohuuepdoeg Eivot llotruuepn'Ev(uuq 55'',
  • 50.
    ororxofv oro droeoporrKdyov(5ro nou Ueroypd- 90UV. H evepy6rrlto nou (eXop(et neproo6repo e(vor qg RNA nol,uprepdonq l, 4 ono(o ewon((eror orov nuprlv[- oro (nucleolus) r<or ovoLoprBdver yro rrl preroypogri rtov yovr6frov nou rto6rronoroUv ro 18S ror 28S rRNA. E(vqr uneU0uvn yro 14 orivOeol qg geyoUrepnq no- o6qroq RNA rou rurr6pou. To dMo rUpro dv(upro e(vor q RNA noLuprepdorl ll, nou Bp(oretor oro nupqv6ntroopro (nucleoplasm) (ro rpnpo rou nuprlvo ercr6q rou nuprlv(oxou). Awrnpo- oone6et ro geyotrUrepo prdpog tqg un6Lornrlq Kurro- pu<{g evepy6rnroq Kor e(vqr uneUOuvn yro 14 oUv0eo4 rov neptoo6reprov on6 ro erepoyev{ nuprlvrx6 RNA (heterogenous nuclear RNA - hnRNA), nou e(vor np66popro yto noMd mRNA ror 6Xr pr6vo. O rLoou<69 opropr5g rltov 5rr ro hnRNA nepil,oprBdver ro ndwo mov nup{vo ercr6q on6 ro rRNA ror ro IRNA (en(oqg, oritrrgovo Ue rov r.troorr6 opto;r6, mRNA Bp(orerot pt6- vo oro r<urtop6ntroopo). H e[5toor1 rou hnRNA pe oriylpovo ;ropror<d epyoLe(o onorol.0mer noLl,d RNA nou 51ouv prev purprl ouyx6wp<ooq oMd peydL4 oq- proo(o, 6nog en(oqg ror noltrd dLLo RNA rrov onoftov q l,enoupy(o pr6lrg op1(ooge vo rorovooUpe. To mR- NA e(vor to Lry5repo dgOovo on6 ro rp(o rUpro RNA Kor owrnpooone(er p5tg 2/"-5% rou rwtoponl,o- olloru<o0 RNA. H RNA notrupepdoq lll e(vor 6troooov 6v(upro on6 qv dnor,Lrq qg evepy6qroq, oMd nopdyer 5vo orlvoLo oro0epc0v, onopo(rqrov RNAs. Iuv06rer ro 55 rRNA, to IRNA ror dMo puxpd RNA nou onoretrofv ndvo on6 6vo r6topto rov Kurropontrooprorxcirv RNA. 'Otreg or europurotrr6g RNA noLupepdoeg efuor pre- ydtreg nprrlreiveq nou Letroupyoriv og o0prnlor<o -500 kD. Tunxd 51ouv -12 uno;rovd6eg. To r<o0op6 5v(u;to prnope( v' ovo).dBer e(oprtir;revq on6 ro er<proye(o geto- ypoqn rou RNA, 6e prnope[ 6puog vo npor<o].6oet 5vop- [r1 ent].ercttxd orouq unoKrvnrdg. H RNA no].upepdoq ll tou Saccharomyces cerevisiae e(vot lopoxtrlptmtr6 nopd6erypro yro rq yevx4 oUoroorl tov euxoputottxc0v ev((;rov Ue rrl ouyKexptpr6vq evepySrlro Kcu ilopou- ord(eror orqv E|(0NA 20.2. Or 6rio ;reyotrrirepeq unoUo- vd6eg e[vor o;r6],oyeg pre rtg B r<or B' unopovd6eg qg Borcrqprox{g RNA notruprepdonq. Tperg on6 trg un6trot- neg unoprovd6eg efuor rowdg oe 6leg lg RNA noLuge- pdoeg, 64).o611 e(vor ouotottrd rot ttov RNA noLuprepoorilv I r<or lll. Enrorlpro(verqr 611 6ev undpler rdnoto unopovd6o oXenr<r1 pre ro Borcr4pror6 nopdyov- to o. H Lemoupy(o rou nepdXetor orouq Boou<oUg ge- roypogrro[q nopdyoweg. H preyoLritep4 unoprovd6o rrlg RNA nol.upep6orlg ll SXer puo xoppofu-retrrx{ neprol{ (carbory-terminal domain - CTD) nou onorel,efuot on5 noMontr6g eno- vol,4rferg Uroq ouvorveru<4q oMqtroul(og emd optvo- (6ov. H oll.qLouX(o e(vor l.rovodrrrl orrlv RNA noluprepdoq ll. YndpXouv -26 enovol,rll.rerg oroug (u- gog6xrpeg rqr -50 mo 0r1tromu<6. O opr0pr6g ttov enovol4r.peov e(vor oqprowu<69 ener6{ or ono}.orgdg IXertxrl ge rn Borrnptorn unogovodo p l-lpood6ver DNA 'EXer CTD : (/SPTSPS)n [(uun n=26, novrlrr n=52] Ixenr<r1 ge rn Borrr1prorn unopovodo I tlpoo66veL vourtreor[0ro IxerLrrl pe rrl Boxrnprorn unogovddo o Kotvrl ror olg rpeiq notrupepooeq Korvrl ror oug rpe[q notrugepooeq Korvrl ror olg rpe[g notrugepooeq Konoreg unopovodeg elvor rorveg oe otreq rLc .-- [erg rorv europutolr6v RNA notrugepootirv. 0ptop6veq en,.-- . oXer[(ovror ge rr1 porqproul RNA notrupepool. Auro ro o'1--- e[vor ;-uo npooo;-ro[roor1 rou ltrerrpoqopllrou npor0nou Tl( ..:. Oopnq RNA notrugepoolg ll Uq (uUnq oe pro n1rrrl SDS tlmE - unogovoDeg vo droyroproro0v roro p6ye0og. rrou oTropoKp0vouv (runrxd) ndvo on6 ng ptodg eno- vo},r1r,Uerg e(vor 0ovorlq6peq. Or CTD ;rnope( vo e[vot nolri groogopu).urlprdveq oe rordLotno oepfurlg rl epe- ov(vr1g. E;rnL6r<ovror otrl pU0puon rnq ovr(6poo4g 6vop[qg (B].6ne Evotrlta 20.8), arrlv enrprfxuvoq qg preroypo<pr1g ror oe 6Leg ng mu15g qg ene[epyooiog tou mRNA, or6;rq Kor srrlv e[oyroyri rou mRNA oro rurrop6nl.ool-ro. Or RNf noLuprepdoeq rov Urroxovdp(cov Kor rs)v XLcoponl,oordlv e(vor pur<p6repeq ror prord(ouv Ue rn Po- rcrrlprorrl RNA nol,uprepdorl nopd pe rdnoro on6 to 5v- (upro rou nup4vo (enet6ti e[e].(10qrov on6 to euBor<trlpro). Aogdrcbg, ro yovr6uiproro roJV opyovr- 6[ov e(vor no].ti gxp6tepo, on6re q rontrrl no].uprepd- o4 np6ner vo pretoypdQet olerrxd L[yo yov(6ro ror o SLeyXoq rnq Ueroyporp4g pdllov e(vor ndpo noAti onLoUorepog. Aurd ro 6v(upro protd(ouv neptoo6tepo oro 6v(upro trov Borrqprorpdyorv nou 6e Xperd(etot t' onoxp(vowor o' 6vo noLunlor<6repo neprBdMov. Mro rUpro npornr4 6rdr<ptorl pero[U rov euKo- purorxriv ev(6prov ouvdyerot on6 t4v ondwqo{ toug oro 6u<ur<Lu<6 omonem(dro o-o1.rovn(vq (ro rofu<5 ou- mortr<6 mo e(6r1 rou UovrroptotS Amanita). Ouotoon- r<d oe 6Lq rq euKopuorrrd rrjrropo, q evepy5qro rqq RNA nolutrrepdonq ll ovoor5Merot yprlyopo on6 Xo- prrlLdg ouyxewprrloetg o-o1lovn(v4q. H RNA noLuprep6- oq I 6ev ovqor5trtreror. H ondvrqoq tqg RNA notrugepdorlg lll 6ev e(vor t6oo r<oLd ouwqpnp6vn: oro (rou<d rUrropo ovomSMeror on6 utp4ld en(ne6o, oLld otoug (ugoprixqreg ror ro 6wopro 6ev ovoor6l- Leror. 558 KEOAAAI0 20 H Meroypoqrl orouq Europu6reg
  • 51.
    : C 3H RNA l"loAupepdon 'Exet ivo Atueplt '-crrvnrfr . I - .orLvlrtlS qg RNA notrupepoorlg I onoretreftot ond 6vov .!, .31K0 unorrvlrrl rr 6vo ovo6rr6 ororxe(o TOu u[IoKtvrlTrl , r! - upstream promoter element). . I apoyovroq UBF1 rutriyet DNA ytlptrt ono gto nporeivtrtl : : -- yLo vo rp6per rovro rov revtptro unorLvrlrfl xoL to UPE. . - SLl nepttroppovet rov nopoyovto TBP nou egntr6rerot - -, evop[1 ror on6 ug rpetq RNA notrugepooeg. r - r,[ notrupepoorl I npoo66verot oto ouplntroro UBF1-SL1 . ., rtvTprKd unoKtvnTrl. - =iA no).uprepdorl I preroypd<pet p6vo to yov(dto : :c prBooco;rtr<6 RNA, on6 5vo pr6vo tUno unoKt- :. -'-i.cu1(eg t6oo rou ;reydLou 28S 6oo Kot rou pt- . ::, I8S rRNAs, nou opy6repo u<pforowot ene(ep- r l : Jr rotortrlrloerg Kor rpononotrloetg. YndpXouv -: ''.1 ovr(ypogo rnq Ueroypogrxrlg ptovd6og. =, :'-'.cooowor Ue pn peroypog6pevo 6toor4port- .: r;,lri poro (non-transcribed spacers) r<ot e(vor Avo0rx6 ororle(o unortvlq r60 -150 -140 -130 -120 -1 10 opyovtopr6vo oe uto oumotl(o, 6n<og neprypoqerol o-trlv Ev6t4ta 7.3. H opydvooq tou unoxwrfr( ror ro ou;rBdwo qg 6vop(nq nopouotd(owot ornv El- K0NA 20.3. H RNA notrugep6orl I u<p[motot tog o]to 5v(u;ro nou neptdXet entnp6o0erouq nopdyoweg onopo(trlrouq yro tqv 6vop[q Kt eluorporefetot on6 roug ueroypoqlKorig nopdyoweg oneu0e(og orov urroKrvqrrl un6 Uopgn ev6g ytyowto(ou ou1.t- ntr6rou. O unorrvqrrlg onoretre(ror on6 dUo [eXorptmSg ne- ptoX6g. O r<ewpx6g unor<rvqtrlg neptBdMer rrlv oge- qp(o, on6 -45 Srog +20, rt enopre( yto rrlv 6vop[r1 rrlq peroypogqq. l-evu<d e(vot nl.otiotog oe G-C (oou- vrlOroro yro Svov unorwrlui), er<r6g on6 ro p6vo ouv- rrlpqpr6vo otorle(o olilrlLoul[oq, pto BpoXe(o oLLql,oul(o nLo0oro oe A-T yripo on6 qv o<peqp(o. H onoretreopotrr6rlto rou KewptKo6 unortvrlti ou- [dvet no].6 on5 ro ovodtr6 motle(o tou unortvqtl (UPE, preprx6q gop6q ovopd(erot rot ovo6u<6 pu0pu- onr6 morXe(o, r1 UCE - upstream control element). To UPE e(vor dLl.q pro ntrotjotq oe G-C oMqlouX[o nou olet((eror pe rnv oM4trouX(o rou KewptKoU unortv4- tr1 rr ercre(veror on6 -180 6o:q -107. Aw6g o trinog op- ydvoor'1q e(vor r<otv6g orouq uTloKtvqr6g qg pol I oe noLld e(6r1, pol,ov6rt or (6reg or oLlql,oulieg 6tog5- pouv nol0. fltrorioro oe G-C n)oIoro oe A-T Kevrprx6q unortvlrrlg -40 -30 ,20 -10 +10 +20 0 UBF npooddverqr oro ovo6rr6 motXe(o rou unoKtvnrn To otro6v(ugo qg RNA no)tugepdorlq I nepfiouBdvet tov nopdyowo npdodeonq mov revrptr6 unorrqq (SL1) nou npoo66vetot mov xewptrd unorwlq 0r peroypotprrdq Uovo6eq rrlq RNA notrugepooqq I 6Xouv rivov revrprro unortvlrrl nou dtoXtopi(erot ono'70 bp ono ro , ., .: morXe[o Tou unoKrvr]Tr1. H npooDeorl rou UBF oro UPE npooyer qv npoodeoq tou nopoyovro CBF (core-binding factor) oTov Kov- '..- -.oxLvlrl.0nopoyovrogCBF(SLl)rono0ere[tr1vRNAnotrupepoollorrlvoqeqp[0. 28.3 H RNA l-lotrupepoorl 'EXer 6vo ALpepq Ynoxtqrfl 569 eworeq
  • 52.
    H RNA nolupepdonI onore( 6Uo enrrouprro0g peroypoqrKoUq nopdyovreg. O nopdyovroq nou npoo66vetor orov rewprr6 unoxtvqr{ e(vor o SL1 (n TIF-1B rqr Ribl oe 6rorpoperrrd e(6r1), o ono(og ono- rele(rot on6 r5ooeprg npcoteivu<69 unogovddeq. 'Evo on6 ro ouorqrrxd rou SL'l e(vor q nptore'iv4 nou npoo66ver r4v otrtrqtrouyio TATA (TATA-binding protein - TBP), en(o4g onopo(rr1q yro rnv 6vop[r1 on5 rrg RNA no),upepdoeg ll roL lll (BL6ne Ev6rqra 20.6). H TBP 6ev npoo66veror oneuOe(og oe DNA nl,orioro oe G-C r<or 11 np6odeorlrou DNA e(vor oppro- 6r6rqro dlLtov oumorrxtirv rou SL1. flt0ov6v, r1 TBP otrl,qlenrdpd Ue rnv RNA noLuprep6o4, (oog Ue Uro r<orvf unopov66o 11 6vo Xoporcrqptmrx6 nou 61et ouv- rqprl0e( mrg noLupepdoeg. O SLI ennpSner or4v R- NA nol,upepdorl I vo [errvrioet on6 rov unorrvqtrl oe XoUnIn Boorrrl oulv6rqro. Boorr6 ro0qxovrou SL1 e(vqr vo e(oogol(oer 6rr q RNA nol,ugepdorl SXer ronoOeqOe( ooord ornv ogern- p(o. 'Ontog 0o 6oripre opy6repo, pro nqpepeep4g Let- roupy(o nop6Xeror orrg RNA noAuprepdoeg ll ror lll on6 5vov nopdyowo nou onotel,e(ror on6 tr1v TBP ror 6A- treg npolretveg. 'Erot, rivo rorv6 XopoKrrlprorrr6 qg Svop[qg ror on6 nq rpsrq no],upepdoeg e[vot 6tr orqpi- (owot o' 5vov "nopdyowo rono06r4oqq" (positioninQ factor) nou onorelefuqt qn6 TBP ouv6e6e;r6vq pre npo- retveg erdu<5q yro rov rd0e r0no unorwrlrrl. O or<prB4q rp6nog 6pdonq r<rlv e[oprc0prevov on6 qv TBP nopo- y6w<ov tono06rqo49 6roq5per r<ord nep(moorl: orov unorrqr4 yro rrlv RNA noluprepdorl I 6ev npoo6Sver DNA, ev6 fiov unoKrvrl{ yro qv RNA no},uprepdorl ll e(vor o r<riprog rp6nog yro rnv tono0Srrloq rou ev(tiprou oro DNA. H 6vop[r1 Ue uqrnln ouyy6rrlto onome(ro Ueroypq- 9116 nopdyowo UBF. llp5r<enot yro 6vo govodr6 no- Lunem(6ro nou npoo66veror o' 6vo morXe(o nLorioro oe G-C oro UPE. O UBF 61er 6Uo Leroupy(eg: 6eye(- per rrlv one).euO5poorl rou unorwqrq on6 r1v RNA no- Lupepdoq r<ot, 6nog 0o 6ori;re opy6repo, dreye(per ro SL 1. Mto 5v6er(r1 yro ro ndrg oLlrlLenr6pd o UBF pre tov SL1 6(verot on6 rr1 oqproo(o t4g on6oroorlq Uero- Ef UBF ror rewpu<ori unonvqt{. H ouyrerprpr5vq oMq- l,en(6poor1 prnope( vq rpoflonorq0e( pr6vo ov q on6moorl oLld[er rord 6vo dprro opr0p6 mporpc0v tou DNA 61t 6prrrg ov or nerpquoru<o( lerpropro( nepr- LoprB6vouv etooy<oyrl ror prori:v orpogriv. O UBF npooS6verot orqv eLdooovo o0l,or<o rou DNA xor rul(- yer ro DNA oe Uro 0qlld nou oXqpor(et orpo<p{ nep(,- nou 3600 orqv enrgdvero rnq nporetvrlg, Ue onor5treogo o xewpx6g unor<wrlrjg xor o UBE vo oup- nLqord(ouv, enmpdnowoq srov UBF vo 6eye(per qv np5o6eoq tou SL'l orov unor<rqq. H Etr6vo 20.3 6efiyer qv 5vop(q og oerpd 6ro6oXr- rcirv oMq).enr6pdoeov. H RNA noLuprepdo4 l, ocr5oo, ug(ororor tog oLoSv(upro ro ono[o nepdler rouq nepto- o6repoug r] 6).oug roug nopdyoweq nou onomofwor yrq rnv 5vopEn ror prdLl,ov enrorpore0eror oneuOe(og orov unoxw4trl. 2O.4 H RNA noAup€pdon lll Xpnorponorei T6oo Ko0o6rro0q'Ooo ror Avo6lro0q Ynorrvnrdq * H RNA notrupepoorl lll 6Xet 6[o runoug unorrvr[drv. * 0r eoolrepuol unorrvqr6g 6Xouv ppoXeteq ouvorverx6g otrtrn- trouXteg nou pplorovror p6oo oq peroypoqxrl ;rovo6o ror npc- ro),ouv n1v evop(q oe pro ouyrerptg6vl onomooq ovodtxo, a 0t ovodtro[ unonvrlr6g nepreXouv rperq ppoXereq ouvorvrTL- rdq otrtrltrouxieg ovodrrd qg oqerrlprog olg ono(eg npoobe- v0vTor UtToyp0rpuoI nopoyovreg. * 0r TFlllA ror TFlllC npooOevovror oTrq 0uv0rvrTrK6q otrtrltrou- X[eq rr enup6nouv Trlv npoo0eol rou TFlllB oqv ogerqp[0. q 0 TFlllB rpdper oq unoprovodo qv TBP Kr EnrTpener rrlv npoo- deon rrlq RNA notruUepooqq. H ovoyvrrlpror; rov unoKrvnrcllv on6 qv RNA no).upe- pdorl lll ovo6etrvUer ewunootord rouq oxetKoUq pc- Loug rtrrv peroyporptxrrtv nopoy6w0lv Kor r1g notrl- Uepdonq. Or unorrwlr6q rorordooowor oe 6[o x0prec roqyop(eg nou ovoyvop((owot pe 6rogopelrorjg rpo- noug on6 6rogoperu<59 opd6eg nopoy6w<ov. OL unox.- vqtdg yro to yov(6ro rou 55 RNA xor rou IRNA eivc. eourepLxo( (internal), 6r'1Lo6r1 Bp(or<owot ro0odLrd onc qv oqeqpio. Or unorrvqr6g yro ro yov[6ro tou snRNA (pur<p6 nupqvu<6 RNA, small nuclear RNA) Bp(r<owc. ovo6rxd rnq oeernp(og r<ord to ouprBoru<6 np6runc tcov dMov unoxrvrltulv. Kor ong 6Uo neprmo.loerq, rc [eXoprord ororle(o nou e(vor onopoirrlro yro r; ].e:- roupy(o rou unoKrvnr4 onoreLoriwor onor).erortxd onc oMrlLoul[eg or ono(eg ovoyvop((owor on6 Ueroypoei- roUg nopdyoweq, nou pre rr1 oerpd roug r<orcu0Uvou,. tqv np6o6eoq rqg RNA nol.u;repdorlg. Ot 6op5g rprciv r[nrllv unorrvqtutv yro rrlv RNA no Lupepdol lll ouvotU((owor ornv' : "ir --' ,. Yndplou, 6Uo t0nor eocoteprroU unor<rvr1rq. Ko06vog nepr6leL ;-rrc dtpeprl 6oprq, orqv ono(o 6Uo BpoXdo ororle(o oMr- A.ou1(og 6tolcop((owor ond pro preroBLqrrl oMrlLoulic Oct PSE TATA I 0L unouvrlreg yro qv RNA notrupepoorl lll pnopei , onoretro0wor ono 6rpepe(g otrqtrouXreq ro0odxo rr1q oQeqpioq T: boxA vo OtoXtrlpr(erot efre ono rc boxC elre ono rc boxB, eue o .: [eXorprmeq oiitroux(eq ovo6xo qq orpeqp(oq (Oct PSE, TATA) 560 KEOAM|0 20 H Meroypoqrl oroug Europudrreg Boorx6g ewomg
  • 53.
    - - :<Lvnrqqtinou .1 Yto ro ptBoottlptr<6 yov(6to tou :: ::ctelenoL on6 prLo oL}qtrouX,rr boxA nou dtoXtlpG -. :. ond 6vo ev6tdpreoo orotle(o (indermiediate ele- --.=-l-lE) ond prto boxC otrtrr1ouX[o 0]'6r]'qpr1 q -.:.o1q boxA-lE-boxC ouXvd ovorptipsrot olq 5S eorrlre- : ,- cuOprortr<r1 neptold (5S internal control region - l- -: :rl (0pr1, prdvo ro motle(o boxC e(vot onopo(-rqto r rn peroYpoAn O r0nou 2 unor<tv4tqg yto to IRNA :-::e}.e[ror on6 pto oMq].ou1[o boxA nou 6toXtopi(e- -: :no pLo oMrltrouX(o boxB. Mto r<otvq op65o unoxt- .. *,'tUnou 3 yt' dMo;rtxpo RNA 6Xet tp(o orotle(o - .'-oux[og nou 6].o Bp(oxowoL ovo6txo r1]g oQtrn- : -.: To [6Lo outo orotle(o en(or1g Bp(or<owot Kot ot - -i:.-too unorwqrttrv qg RNA nouprepdoqq il' 3: emopepeiq oMqlenLdpooetq 6toq6pouv orouq , ,: rjnoug eooteptrcbv unoruvrltr.rlv, n opxrl dpoq e(- -- ,,cLvr'1. O TFrIC npoo6dvetot r<o0o6tr<d rlq opern- : r: eire ove[dpqto (unor<tvrp6q tlnou 2 yto IRNA) : -.- :. ouvduoop6 Ue rov TFrrrA (unoxtvqt6g tilnou 1 : 5S RNA). H nopouo(o tou TFrrrC enttp6net orov no- : r, 3',ro tono0Stqoqq TFrrrB vo npoo6e0e[ ornv oqerr]- , : Enerro enLorpore6etot 11 RNA noXutrrepooq lll' : i ouvo{,r[(eL to orobto tnq ow[dpoonq , : - -i tocDreptr<o[g unortvqtdq r0nou 2 nou Xpnotpo- - : .l -wot yLo to yov(6to tou IRNA. H on6mooq ;reto[ti - :,.1 <ot boxB ;rnope[ vo notrfter eneL6q noMd yov[6to ':'. - nepLdxouv 6vo prtr<p6 twp6vto' O TFrrrC npoo6eve- -r (lr oro boxA Kot oto boxB. Auto enitp6net orov -: 3 rro npoo6e0e( ornv oQernp(o. Ie ow6 to orlpe(o --::oi vo npoo6e0e( 11 RNA noLuprepdoq lll' - 5io<popo orouq eoorsptr<o6q unor<wr1t6g t0nou 1 : :o yov(6ro tou 55 RNA) e(voL 6tt o TFrrrA np6net vo -:::6e0eI mo boxA Yto vo enttp6!.reL mov TFrrrC vo 0r eocoreptrol unorLvrlrdq runou 2 rnq pol lll Xpnot- ponotouv roug nopoyovteg ouvoppotroynorlq TFrrrA rot TFrrC oro box,4 rot rc boxC Yto v0 ontoTpore0oouv rov nopoyovro rono06- qorlg TFrrrB, nou npooetrx[etrriv RNA notrupepoorl lll npoo6e0e( wo boxC. O TFrrrA e(vot Svog nopoyowog np6o6eorlg eL6tr<dq Yto rrlv oMq).ou1[o 55 nou npoo- S6verot orov unoKtvrlrrl r<ot oro 55 RNA olq ouvo66g npcltelvq (chaperone) roL yovt6Lor6q puOptmqq H deiper 6rt ;r6trLg npoo6e0e( o TFrrrC, n oKo- ).ou0(o rr.,.lv oupiBdwcov e(vot r<otvr1 pe roug unoxLvqt6q rrjnou 2: o TFrrrB (nou nepL6Xet rrlv otKoupevtr<r1 TBP) npood6veror ornv oqerrlp(o r<ot q RNA nolupepoorl lll ouv66etoL oro odpnl'or<o' Ynortvqt5q t[nou 1 Bp[ot<ov- ror p6vo oio yov(6to Yto to 5S rRNA. bt TFrirA t<ot TFrrrC e(vot nop6yoweq ouvoppoA6yq- oqg (assembly factors), pre provo6txd p6Lo vo Boq0o[v orqv np6o6eol rou nopdyowo rono06tr1or1g TFrrrB orrl oo.lorrl 0eor1. M6LLg npoodeOe( o TFirrB, ot TFrrrA xot TFrrrC lrnopti v' onopoKpuvOo0v ond rov unoKtvrlrrl lcop(g vo enqpeoor-e( q ow(6poor1 6vop[1q. O TFrtB napapivet npoiodedel.tevoq, Kowa o'tqv aEerqpb xat 4 napouo[a rcu enopxe[ 6orc 11 RNA noupepaorl lll v' avayvapbet Kat va npoo6e0e[ oil]v aqfirlp[o. 'Etot, o TFrrrB e(vot o 1'tovodt- r<6q npoy;rottr<6q nopdyowoq 6vop(qg nou onotre(toL on6 tqv nNR noLupepdoq lll. Aurn n oerpo oupBdwov e(qye( nog to ro0o6Lrd orotxe(o rou unoKtvnrq prnopo0v vo nporotr6oouv rnv npdo6eorl qq RNA noLupepooqg oqv oqernp(o, 611l'o611 nto ovo6trd. flop6lo nou Il tKo- v6qro UeroYpoQnq rov ouYKeKptpSvc'-lv yovt6(tov npoo- 6[6etot on6 rov eoortepLr6 unortvltrl, o]}'oy6q mqv neptoXrq opr6orrrg ovo6trd qq oqeqp(oq prnope( v' oMd- (ouv qv onoteleoporLr6rqto qg peroypo<prlg' O TFuC e(vot 6vo peydlo nporeivtx6 otipnlot<o 0r eooreptrol unortvrlr5q r0nou 2 rlq pol lll Xplot- -. :i0uv qv npoo6eorl rou TFrrrC ortg otrtrr1trou1leq rou box' rot -', - :oxB Yto v0 entoTpuTeu00uv Tov nopoyovro ionoOdqorlg TFr' ' -rl npoottrruet qv RNA notrupepooq lll. Aroeqp(o Kour( A Kouri B 2*.4 H RNA notrupepdorl lll Xplorgon0tsiT600 Kooodtro0q '0oo rot AvoDLxoug Ynoxrvltdq 56'l tr
  • 54.
    (>500 kD), ouyr<p(ot;toupey60oug Ue rnv 6to qv RNA noLuprepdoq, nou nepdlet 6(t unoprovd6eg. O TFurA ei vor pr5Loq puoq ev6ro<p5pouooq td(49 npoteivrriv nou nepr6louv 6vo prot[Bo np6o6eoqq voutd.eivxcbv o(dtov yvoor6 olg 6drcru),oq rfeu6opy6pou (zinc finger) (FM- ne Ev6trpa 28.6). O nopdyowog ronoO5tr1or1q TFrrrB onoretre(ror on6 tpetg unoprovddeg. l-lepil,oprBdvet rov [6ro nporeivrr6 nopdyowo TBP nou undplet t6oo orov KewprK6 nopdyowo np6o6eo4g SL1, rov ono(o Xcqoqonororiv or unorlqtdg rrlg pol 1, 5oo Kot orov ow(mor1o preroypo<pm6 nop6yowo TFrrrD, tov ono(o XprlorUonore( rl RNA noLu;repdo4 ll. lleptdlet en(or1g Brf, nou olet((etot Ue ro Ueroyporprr6 nopdyovto TFrrrB o ono(og Xprlouonoefuot on6 qv RNA notrupe- pdo4 ll. H rp(tq unopovddo, yvomri oq 8", e[vot ovo- Lc0oq.r1 ov 6Xer ;repu<dlq onodtorol0e( to 6(xLrovo DNA, yeyov6g nou uno6rlltivet 6n q trsnoupy(o trlg efuot vo evepyonor{oer rn Ueroyporpu<4 quooo}.(6o. O p6loq rnq B" (oog e(vor ouyrp(otprog pe rou nopdyowo o orrl Bor<rqpror<r1 RNA noLuprepdon (FLdne Ev6rryta 19.19). H ovodrni neptofi 6Xet ouptBottx6 p6l,o orouq uno- r<rvqr6g rpkrlg td[r1g qg RNA no],uprepdonq lll. Iro no- pd6erypro rqg Eu<5vog 20.4, undplouv tp(o ovo6trd ororle(o, ro ono(o enronq undpXouv Kot orouq unoKtvrl- 169 yro ro yov[6ro rou snRNA nou pretoypd<powot on6 rrlv RNA notruprepdoq ll. (l-o yov(6to yto pepxd snRNA ;reroyp6gowor on6 n1v RNA noLuprepdorl ll, evcir dMo geroypdrpowor on6 rqv RNA no),uprepdoq lll). To ovoSt- rd ororXe(o Leroupyo0v ;re nop6;toto rp6no orouq uno- r<wqt5g t6oo rqg RNA nol"u;rep6onq ll 6oo ror qg |ll. H Svop[q o' Svov ovo6tr6 unorrulti yto qv RNA no),uprep6oq lll prnope( vo ouprBe( oe pto BpoXe(o Tlepto- Xri nou Bp(oreror opr6otog nptv on6 rrlv orpeqp(o t<ot nepr6Xer p6vo ro orotle(o TATA. H onotetreoporu<5rr1- ro rrlq preroypogrlq ou(dver noLri on6 t4v nopouo(o rtov motle(ov PSE (eyyUg ororXe(o oLL4l,ou{og, proxi- mal sequence element) ror OCT (on6 q 1.6[q octamer = o]fiouep6q) tou evrqxutrl. Or nopdyoweq nou npoo- 66vowor oe owd to orotXe(o oLtr4l.ent6po0v ouvepyo- rrxd. To motle(o PSE nt0ov6v e(vot onopofurlto oe unoxwqr5g nou XcrlotUonotoUwqt on6 rqv RNA noLu- pepdon ll, evri oore( 6teyepnr{ en(6pooq orouq unoKt' v4t6g ng RNA notruprep6o4g lll. To ororle(o TATA npoo6(6et et6x6trlro yto tov tU- no rrlq noLuprepdorlq (ll n lll) nou ovoYvop(etot on6 6vov unorrvrlq snRNA. [lpoo66vetor on6 6vo nopd- yowo nou nepil.oprBdvet tlv TBP, q ono(o finv npoY- potrx6rqro ovoyvtop(et tqv oMrltrouX(o tou DNA. H TBP oMrll,enr6pd ge dMeg npotefveg, etdu<69 Yto rov tUno tou unortvqt4. H Lettoupy(o t4g TBP Kot rov oXetr(6prevov np<oretvc0v e(vot vo rono0etoUv ooqr6 qv RNA noluprepdoq or4v ocperqp(o. Or nop6yoweg Lettoupyo6v rotd tov [6to rp6no Kor yto rouq d6o t[noug unortvrltc0v qg RNA no].u- pepdo4g lll. Ot napayoweq npoo66vowu orov uftoKt- vr1t4 nporcA pnop6,oa va npoo6e0e[ 4 6n 4 RNA no/r.u pepdo1. Ip1;ror((ouv 6vo oripnAoxo npo6vo p- [qq (preinitiation complex) nou roteu0Uvet trlv np6o6eorlrqg RNA noLuprepdorlq. H RNA noLuprepd- oq lll 6ev ovoyv<op((et 1 (6ro qv oMrllouX(o rou uno- rrvr1r1, oLld npoo66vetot oe nopdyoweg ot ono[ot e(vor npoo6edepr5vot L(yo nro ovo6trd on6 r1v qqerrl- p(o. Fro rouq eooJreptxotig unortvqr6g ninou I xot t0- nou ll, or nopdyoweq ouvoppoL6yqoqg e[ootpo]t[' (ouv 6rt o TFuB (nou nepil.oprBdvet qv TBP) e(vot npoo6e6epr5voq l,(yo nto ovodtrd on6 rqv oEeqp(o, nop6Xowog 5rot ttg ntrrlpogop[eg tonoO5tr1or1q. l-to roug ovodtr<otig unoxtvqt6g, o TFrrrB npoo66verot oneuOe(og orrlv neptoXrl nou nepil.ollBdver to xowf TATA. Aur6 oqpo(vet 6rt ove[dpqto on6 q 06o4 rov oMqLouXuirv rou uTroKtvnrn, o(0 nopdyowog(eq) ei- vot npoo6ede;r6voq(or) rowd m4v ogeqp(o Ytq vo xoteu00ver(ouv) rrlv np6oSeoq qg RNA nol,u;rep6- orlg lll. Ie rd0e nepfntooq, 4 Xptoprot(vq npdnet vo 51er rpononorq0e( Kot vo Bp(or<etot oe ovotXrrl 6to- p6prpooq. 2O,5 H Agetnpio yro Tnv RNA l-loAuUepdon ll * H RNA notruuepdon ll yro v' opy(oer q Ueroyporpil onotrel ye- vLroug peroypoqrroug nopoyovreg (ovogo(ovrot TFrrX). o 0r unorrvnr6q rqg RNA notrugepoolq ll ouxvd 6xouv orlv ogerrlpio pto ppoXe(o ouvrrlprlpdvl otrtrntroux(o PyzCAPys. e To rourl TATA eivor rorvo ouorolr6 rov unoxtvlttiv qq R- NA notrugepoolg ll rot onoretre[rot ono 6vo ntro0oto oe A-T orropepeg nou ppiorerot -25 bp ovodtro rqg oqeqpioq. " To DPE elvor rorvo ouoronro rorv unortvrlr6v qg RNA notru- Uepoolq ll nou Dev nept6Xouv rouri TATA. e 'EvoS revrptrog unortvlrrlg yto rrlv RNA notrupepoorl ll nepL' tropBoveL ro lnr xot, ouvrlOoq, elre 6vo rourl TATA elre evo DPE. Mnopei en[o1g vo nept6Xet rot otrtro etroooovo orot- Xe(0. H Boorr<r1 opydvcrloq rnq UeroYpogtr<r1g ouoreuqq yro yov(6to nou ro6tt<onototjv npotelveg onoro].rj- gOqr<e on6 qv ovor<dtrugr1 6tt q r<o0opr1 RNA no).u- prepdor; ll rotq^.Uet Uev rl'l otiv0eorl mRNA oMd de gnope( vo evepyonot4oet rrl UeroYpqrprl Xopiq 6vo np6o0ero erXritopro. H rd0opoq owoU tou erluLl- oporoq o6rlyr;oe orov ro0optop6 ttov yevtro:v peto- ypoqrr<drv nopoy6vttr:v, 11 Baomav peraypaEmav nopoyow@v (basal transcription factors), prtoq opo- 6o9 nptoreivov onopo(rrlrov yto qv Svop(r1 on6 tlv RNA noupepdorl ll oe 6).ouq rouq unoKrvqt6q. H RNA no).uprepdoq ll oe ouvepyoo(o pre ourorig touq nopdyoweg onoretre( tI goorxn pretoypo<ptx( ou- or<eur1 nou e(voL onopokrlq yto rrl pretoyporprl r<dOe unorrv4rrl. OL yevtro( nopdyoweg neptypdgowot rrlg TFrrX, 6nou ro *X, npoodtop((et r<dOe [e1optor6 no- pdyowo. Or unopovddeg rqg RNA no).upepdorlg ll 562 KEOA Al0 20 H MetoYpoQn orouq Europutitteg Boorxeg eworeg
  • 55.
    Kor ol yevrKo(pretoypo<ptxo( nopdyoweg efuot ouwrl- pnudvor geto[0 trrrv europutotcbv. Iqpe(o err(vqoqg Yto rrlv nopouo(oorlqg opydvo- onq rov unortwlrdlv e(vqt vo r<oOop(ooupe og nuprlvo rou unoKtvrlrq qv nro BpqXe(o oMrll,ou{o oqv ono(o prnope( vo evepyoTlorrloet q petoypo<pr1 4 RNA noLu- Uepdon ll. Oeropqtrxd, Svog r<ewptr6g unorwqtnq Uno- pef vo erqpome( oe rdOe r<rirtopo (pro],ov6tt, mrlv flpd[n, 5vog xewptr6g unoxwrltrlq on6 pr6voq rou Ko- rolriyet oe l.(yq 11 r<o06trou geroypotprl oro nAo(oto qg Xpopro{vqg in vivo). E[vot 11 etr6por4 oMrltrou{o nou ennp6net q ouyx6wpoorl rtov yevtxriv peroypoqxdlv nopoy6wrrrv srqv ogerqp(o. Auto( ot nopdyoweq eu- n).6rowot qrrl UnXovtK4 qg np6o6eorlq oro DNA rt ennpSnouv or4v RNA noluprepdoq ll vo evepyonotrloet rn Ueroypoqrl. 'Evog rewptr6g unoxtvqtrlq Lettoupye( pe 1oprr1Lr1 ono6otx6rqto. iortrtr'eg npoteiveg nou ovo- pr6(owor evepyonotnrSg (activators), ox6ptrl prto t6[r1 preroypogu<rilv nopoy6wtov, onoroUwot Ytq ro oroqr6 en(ne6o trenoupy[og (B].6ne Ev6trya 20.9). O' ovStrleve xove(g 6l.eq ot ol'trqtrouX(eq nou eprnL6- Kovror crr4v np6odeon rnq RNA nol,u;repdonq Kor rov yevrrc0v preroypo<prrdlv nopoy6wcov vo 51ouv ouwn- pqOe( orouq neptoo6tepouq r1 oe 6trouq toug unoKt- qr6g, 6ncog tqXriet yto rouq unoKtv4t5q rrlq pol I xot r4g pol lll. Kot' ovotroy(o npoq rouq Bor<tqptoxotig unorrvqt6g, 6tov ouyrp(vowot uTloKl.vqtdq qg RNA nol,uprepdoqq ll, 6rontarc0vetot 6tt ot o1.totroy(eg orrlv neproX4 rowd or4v oEeqp(o neptop((owot oe oxert- xd BpoXe(eg olLrll.ouX[eq nou epntr6rowot m4 tret- roupy(o rou unoKtvqtrl, 6ntog npor[ntet on6 ttg enrmdroetg pretolLd[ecov. H EIK0NA 20.7 defiyet tq 6o- ;r{ ev6q tuntrof rewptt<oti unortvrlrl qg pol ll pe rp(o on6 Tq nto rowd motle(q. l-evtrd, o euKopuort- 169 unorw4rnq yto qv pol ll nopouotd(et peyol'rirep4 6opu<r1 norril.[o on6 tov ow(orotXo Borcrqpror6. Ercr6g on6 ro rp(o rfpro motXe(o, noMd eldooovq orotle(o prnope( en(or1g vo oul.tpetdXouv ornv opydvtoorl tou unoxwrlr{. Ir4v oqeqp(o 11 oMrlloul(o 6ev e(vot ouw4p4pr6- TATM ......N20...... WCAYWYY ......N2+..'.. - - KoutiTATA lnr nuonvoc rou unoxrvnrn .a -r'r'"i +- nou Treprexet T0 KouTt I A I A nuonvoc rou unoxtvnm -loptgro<oulTHTH - evo rour[ TATA -25 bp ovoDtro qg Inr, To xourt TATA eXet rr1 ouvorvertxfi otrtrr1trouXro TATAA. H Inr dXet nuptl-ttdlveq (Y) nou neprpotrtrouv to dtvourtreor[DLo CA oqv otperrlpto. To DPE ppt- oreror ro0odrro qg otperrlprog. H otrtrtttrouX[o delXvet rov rodt- ronoulrtrd rtr6vo. w1. Oo16oo, q npd:tr1 Bdorl tou mRNA ouvn0oq eivaA ror nep$dtrLerot on6 nupqt6(veg. (Aw6 ro10er Kor Yrtr trlv evopr<trlpto ol,L4trouX(o CAT trr:v Borrqptoxtirv unorwqrri:v). H ouyrexptl.t6v4 neptoXr] rotrekot evop rrripro (initiator - lnr) r<ot prnope( vo neptYpqgef orn yevtnj Uopgli Py2CAPyE, 6nou Py owtototle( oe oTrot- odrlnote nuptpu6(vr1. H Inr nept5X*ot ovdpeoo ortg 05- oerg -3 Kot +5. I-lotrLo( unorwrltdg 51ouv ;rto ollqLouX[o nou ovo- ;rd(erot xouri TATA (TATA box). Itoug ovtinepoug eu- ropudlreq ouvrlOo:q Bp(oretot -25 bp ovo6txd qg orpetrlp(oq. E[vot ro povo6tr<6 qvo6tK6 motle[o tou unoruvrlr] nou 61et olenrd oroOepq 06o4 ovogoprxd Ue rnv oqerqp(o. H ouvowertr{ oMq}.ouX(o rou Kewpl'- ro6 orotle(ou e(vot TATAA rot ouv40tog qrol'ouOefuot on6 dtrl.o tp(o (e6yr1 Bdoe<ov A-T (BLdne Evdtrya 19.8). To rour( TATA ouXvd neptBdll.etor on6 oLtrr1l'ouX(eg ntroUoreg oe G-C, nou 0o pnopo0oe vo enqped(ouv tr1 l,enoupy(o rou. Efvot oXe66v novogot6tuno pe ro rour( TATA or4 06oq -10 torv Bor,crlprordlv unortwltriv. Ir4v npoyporu<6rnro, rl provo6u<r1 6togop6 5Xet vo xdvet pe qv ew6nroq tou (-25 6vowt -10, ercr6q on6 rn EUUn, 6nou to row( TATA Bp(or<etot ouvrl0coq oro -90). Awt- roromdoetg preprovopr6vrov vout<} eott6(clv mo rout[ TATA ;rnope( vo 51ouv Oettr4 11 opvqlr4 en(6pooq, ovdtroyo Ue rnv oproloy(o qg ouOewxrlg oMqLouX(og Ue rn ouvorvetxri oM4lou{o t<ot to Bo0;t6 on6xtrto4g qg preroMoyUdvnq oMnloul(og. Tunu<d, ot owtKoro- ordoerg nou etodyouv 6vo (eriyog Bdoeov G-C 51ouv nq Trro ooBopdg entmriroetg. Ot euxoputirteg 6to06touv en(oqg unoxtv4r6q ;1or- pig TATA (TATA-less promoters). Mel.6teg rov oM4- LouXrcilv rtrlv unoKtv4t6v 6qLc0vouv 6tt ro 50% ti neptoo6repo rov unoKtvrltdrv pnope( vo e(vot owoU tou r6nou. 'Otov 6vog unortv4tr1q 6ev nepdXet row( TATA, ouyyd qdpet to motXe[o DPE (downstream pro' moter element, xo0o6tx6 orollefo unoxlv4r{), nou Bp(oreror on6 +28 5coq +32. Ot tuntro( rewptxo( unorwqtdg onotelotiwot efue qn6 5vo rout( TATA ouv tnv neproXri lnr efue on6 trtto neptoXrl lnr ouv to ototle(o DP-E, proLov6tt undplouv ror dMor ouvduoopo( pe etrdooovo orotXefo. 2C..6 H TBP eivor Orroupevrr6q l'lopdyovroq * H TBP e(voL ouororrxo rou nopoyovro rono06rrlonq nou onoue[tot yto qv np6odeorl roOe RNA notrupepoonq oTov unouvrlq rr1q. * 0 nopoyovroq Yto Trlv RNA notrupepool lll elvot o TFllD, nou onoreLe[rot on6 TBP xoL -1 4 TAFs, pe ouvotrxrl go(o -800 kD. c H TBP npoo0Sverot oro rour( TATA oqv etroooovo 0utr0K0 rou DNA. q H TBP oXrlporL(et ;rto oetro yuptr: ono ro DNA rot ro ropnrel raro -800. AGAC +DPE ?S.S H TBP Etvot 0txoupevtrrlq flopoyovroc 553 Boorx6g ewoleq
  • 56.
    Anopokqrrl npoun60eorl yrovo (extvrloet I UeroYpo- qq e(vor vo rpononotqOe( 11 Xpcoprot(vq rot v' ovo6top- yovoOe( orrlv ovoLarrq 6topr6pcpooq. En[or1g, oe 6treg rLg rd{eLg rov euKclpucortrcr:v unoxLvrlrcrlv, to 6noto oxroprepdg voureooull-toticov Bp(or<etot ndvo orov unoxrvrlrq npSner vo peroKrvrloe( 11 v' onotrlor<puvoei (BAdne Ke<pdtroro 28). Ie out6 to orlpre[o, 6vog nopd- yowoq rono0Stqoqq prnope( vo npoodeOei orov uno- xrvr'1rr1. Kd0e rd(q RNA no),upepdonq unopoqOefuot on6 6vov nopdyowo rono06tqoqg o ono[og neprdXet TBP nou oMqenrdpd pre dMo ouorortxd. H ot<poort- X(6o TBP owtorotXe( crro "TATA binding protein", 6q- trodrl "npore(vr1 nou npoo66vet tqv oLLqLouX[o TATA". H ouyxexprll5vq nporeivq opXrxd nrlpe ourd ro 6vopro ener6rl eiye Bpe0e( 6tL npoodev6tov mo r<our[TATA cno yov(ELo rrlq RNA no].uprepdorlg ll. Me- rdner-ro ovor<o).[<p0qre 6rL e[vot en(or1g ouorottrd rorv nopoy6wov-rono05rloqq SL1 Yto rrlv RNA no- ).u;repdorl I (BI5ne Ev6rqta 20.3) t<ot TFrrrB yto rrlv R- NA noAuprepdoq lll (BLdne Evotqra 20.4). Ze outdg rtq Srjo RNA no).uprepdoeg, q TBP 6ev ovoyvtrtp[(eL qv oLr;trouX(o rou KourtoU TATA (er<r6g on6 roug uno- xtvrlr6g rUnou 3 tqg pol lll): ouvenrig, to dvopd qg e(vot ncpontrovqttrd. Entntr5ov, nop6tt noMo( unoKt- vrlrdq qg RNA nolu;repdolq ll dev dXouv r<outLd TA- TA, en(or1q onot-ro0v rqv nopouo(o qq TBP. l-ro rqv RNA noLuprepdorl ll, nopoyowoq rono06rq- oqg e(vot o TFrrD, nou onoreLe(toL ond TBP ouvde6e- ;r5,vr1 oe 6tdgopeg d).Leq unopovd6eg (6tog 14), yvoror6g og TAF (TBP-associated factors, nopoyoweg nou oMrll.enL6porjv Ue rnv TBP). OpLolt6vot TAF e(voL ororxeroperprxo( pre qv TBP, dMot Bp(oxowot oe UL- r<pdrepeg noo6rqteg, eriprlpo nou oqpo(vet 6rt undp- louv nol.Lon).69 nopolLoydg TFrrD. Ot TFrrD nou neprdXouv 6rotpopetrr6g TAF 0o trlnoporioe v' ovoYvo- p((ouv unorrvqtdg pe 6totpopettroUg ouv6uooproUg ouwrlprl prdvov crrotle(ov, 6nog npoovop5pOqre (B1"6- ne Evorqra 20.5). MepLr<o( TAF dlouv toroet6tt<4 6x<ppo- oq. H ouvotrLr<r1 pd(o tou TFrrD tuntxd e(voL -800 kD. Ot TAF rou TFrrD opxLr<d ror6u<onotq01t<ov un6 popgn n.X. TAFIiOO, 6nou ot optO;ro( "00" dqtrorvouv 11 propror<r1 pd- (o rnq uno;iovd6og. l-lp6otpoto, oL TAF tqg RNA no).u- pepdorlg ll petovo;rdornKov oe TAF1, TAF2 t<.o.t<. Ir1 ouyrexpLprSvrl ovoporol.oy(o, TAFl e(vot o peyotr0re- pog TAF, TAF2 o en6pevog oe prdyeOoq r.o.t<. 'Erot, ot otrr6troyot TAF dto<popettru:v eL6o-rv 61ouv (6to 6vopto. H "::.. ' ' 6e(1ver d-rr ot nopdyoweg tonoO5-rq- onq ovoyvop((ouv rouq unoKtvlt6q pe 6totpopertr<6 tp6no rord nep(moor1. Itouq unortvqt6g yto qv RNA notruprepdoq lll, o TFlllB npoo6Svetot 6[ntro orov TFll- lC. Iroug unorrvrl-r6g yto rrlv RNA noLuprepdoq l, o SLI npoodSveror oe ouvduool-td pe rov UBF. O TFIID e(voL onox.,etsrrxd une0Ouvog yto rnv ovoyvtrtptol unoKtvrl- rov yto qv RNA notruprepdoq ll. I' dvov unor<tvr1tr1 ge r<our( TATA, 11 TBP npoodSvetoL et6Lt<d mo xout[ TATA, evc0 oe unortvqtdg ltop(g TATA, ot TAF ovoLopBdvouv ro Spyo rrlg ovoyvtoptorlg dMtov orotXe(rov rov unoKt- vrltcrrv, 6no:g 4 neproXrl lnr rot ro DPE. Ontog t<t ov et- Ynorrvrlr6q qg pol lll C/nruRnorupepooltrt Ynorrvnrdq rnq pol I Or^NAnorusepriqr ' :. Ie otrouq roug unoruvr1t6q, ot RNA notruUsp00€. rono0erouvror ono evov nopoyovro nou nepL6Xet TBP. o6pletoL crro oUpn).oKo Svop(r1q, enL6(olq rrlg TBP er- vor v' oMrl).enr6pooet Ue rrlv RNA no).upepdor1. H TBP 6Xer rrlv oouvrlOtorq L6t6qto vo npoodeve- ror orqv etrdooovo orjloxo tou DNA. (Itr1 ouwptmLxr nlerov6tqro, ot npcoreivsq nou ol.l.qlent6potiv pe rc DNA npood6vowot orr; pei(ovo o0).ot<o). H xpumoML- xr1 6oprl qg TBP unodrlltrtvet 6vo Lemopep5q powdAc yro rqv np6o6eor1 rnq oro DNA. Oncoq rpo(vetot orq'' . ,' . , q TBP neprrurLrrrvet rn [i[o ntreupd tou D- NA, olrlpor((owoq Uto "0610> yiptr; on6 5vo tprlprc rqg etroooovog oriLor<og, nou rdptmetot Yto vo Xopt' oet p6oo oe ourrl rr'1 o6tro. Irqv npd[1, ! eoorsptK- enrqdveto rrlq TBP npoo66vetot mo DNA Kot n UoYC- ).rjrepq e[oreprxrl enrqdveto e(voL dto0SotUrl Yto sno- edq ps dMeg nprotefveg. H 06oq np6o6eoqg tou DNA onoteLe(ror on6 pLo C-tel.tr<q neptoXq, ouwqprlpdv' perolrj rcov er6c,.lv, xot q peroBLrlrl N-teLtr<r1 oupd no" e(vor erreOerpdvrl yr' o).Lr1).enrdpdoetq Ue dl).eg npco- refveg. 'Evo pr6tpo rqq ouw(prlorlq rou prlxovtopot 6vop(qg rrlq Ueroypo<pr1q e(voL drt 11 oM4LouX(o npdc- 564 KE0A Al0 20 H MetoypoQn orouq Euxopu6req YnoKrvnTdq rnq pol ll
  • 57.
    DNA ono rrlvntreupo rrlg orevrlq o0troroq. H TBP onoretrelrot rno duo ouyyevrreg (40% rouroonpeg) ouvqplp6veq neptoX6g, rou errovl(ovrot oe ovotXro xot oroupo pntre. H N-tetrtrfl neptoxn .cLrfier notru rot etxov((erot npootvl. 0t D0o xtrtitvot qq 6Lntrttq :trLroq rou DNA Ue ovotXro rol orotrpo yrpt, H tptoroypotpto e[vot auyevrrn nopoxiipnol rou Stephen K. Burley. Seoqg oro DNA qq TBP e(vot r<otd 80% ouwqpqpr6vq uero[ti (0pqq r<ot ovOptonou. H np6o6eor1 qg TBP prnope[ vo pnv e(voL oui-rBorrl .ls llv nopouo(o orropep<r-rv vout<Leooul1.tor[rrlv. To '.,o u r<Leo ocr: 1-tdtto ouvrl 0 oq oXq por((owot rono Oerov- :og oMr1trou1(eg nLotioteg oe A-T roro t5roto tp6no oore or el,doooveg o0tror<eg vo t<oL-roUv npoq ro toore- cLrd (Bl.6ne Evorqra 70.9), on6te 0o prnoporiot vq flo- cepinod[oouv rrlv np6o6eon rnq TBP. Aut6 ntOovdv epprlveUer yrot( 1 nopouo(o ev6g vouxeooulpor[ou rrov uiloKtvlr( eprno6f(et qv 6vop(q trlg peroypogrlg. H TBP npooddvetot o"rrlv etrdooovo q0Loro xot <opmer ro DNA Kqro -80o, dnog go[verot ornv ':rr:'r " . To t<out( TATA r<o;rmeror npoq 14 pre((ovo o0Ao- <o, dreup[vowoq rr]v e].dooovo orj],ot<o. H nopopt6p- orr.rorl outn neptopl(etot mo 8 bp rou xourtof TATA. Ie r<dOe or<po rqq o),Lr1l.ou1(oq, 11 el"oootr.lv otjLor<o exer ro ouvrl0Lopevo e[pog tttv -5 A, o].].o oTo Kewpo rqq otrtrr1l.ou1iog 51et e0pog >9 A" H nopo;i6pgoroll rqq 6opr1g orqv npoypo-rtr6tqto 6e dtoXotp((et rouq x).rovoug rou DNA, r<oOtrtg 6totqpeirot to (euyoptol.to rcov Bdoeorv. H drroorl rnq Kdpqrnq pnope( vo 6totp6pet ovotroyo pe rrlv oxptBrl oMq).ouX(o tou r<ourLo6 TATA <or olet((erot Ue rnv onoreleoUorLr6tqto rou unoKt- VIT!. Auq q 6opu1 61er opr<etdg LettoupyLx6g npoer-rd- ceLq. H oMoyrl qg opyovoonq rou DNA crro Xurpo oe onoto6r;note ntreupo rou routto0 TATA enttpdneL orouq UeroypopLr<oig nopdyoweg Kot oqv RNA no}'u- gepdorl v' o).Lr1tren16pdoouv orevdtepo on' 6,tt 0o qrov 6uvor6 oro ypoUUtK6 DNA. H xopturl mo Nout( TATA owrorotle( evepyetord oro [erJ].ty;ro tou ev6g rpr-rou nepinou on6 pr(o ol'poel tou DNA KoL owLoroe- p((eror ond pto 0ertr<r1 etr(xotor1. H nopouo(o trlq TBP orrlv eldooovo orjlot<o, oe ,'.: ,,, " H rpuoro),trtrt1 6op1 rlg TBP oe oupntroro pe D- NA ono -40 p6XpL qv orpeqpio be(Xvet pto ropnl rou rourtou TA- TA nou 6reupuver rqv etroooovo ouLoro ere[ onou npoo6dverot 1 TBP H tporoypotpto elvot euyevtrl nopoXtirplol rou Stephen K. Burley ouvduoopr6 ;re dMeq npcotetveg nou npoo6dvowot orn pe[(ovo oU].or<o, 6rlprLoupye[ tLg npounoO6oeLg yto no- Lu oprO peg enorpdg D NA-npu-rreivc0v orrl ouyr<erpt;.rdvq neptoXrl. H np6o6eoq r<oOopqg TBP oro DNA in vitro npoorore0et -1 mpocpq trlg 6tnLrlg 6LLxog oro rour[ TATA, nou runtrd erre(vetot on6 -37 doq -25. H np6o- 6eo'r1 rou ouprntr6rou TFrrD orrlv ow(6poot'1 Svop(qg, oror6oo, rovovrrd npooroteUet rqv neptoxrl ond -45 6o9 -10. M5oo mov TFrrD tDS etrerj0epo np<ote'ivLx6 otipntro- r<o, o nopdyowoq TAFI npooS6vetot mrlv TBP, 6nou r<orotroprBdvet rrlv xo(trr1 entqdveto npdo6eorlq rou DNA, Irrlv npoyportx6tnro, rl 6oprrl qg 06oqq npdo- deorlq, nou Bp[or<erot orrlv N-te].Lr<r1 neptolrl tou TAF1, ;irpre(tor -rqv enLrpdveto rqq etrdooovog oij)'orog rou DNA. Aurq q popLor<r1 onopr(prr1or1 enttpdnet orov TAFl vo eL6yXeL trlv Lt<ovdrqro qg TBP vo npoo65ve- ror oto DNA: yto vo pnopeosL o TFrrD vo npoo6e0ei oro DNA, 11 N-te).tr<1 neprolrl rou TAFl npdnet vo erronrme( on6 tqv entrpdveto np6odeorlq o-ro DNA tnq TBP. E[vor o[toolpre(otro to yeyov6g 6rr no].]'oi TAF protd(ouv pe tor6veg: 9 on6 toug 14 TAF nept6louv prLo neptoXn nruXrlg tmovrr.lv (histone fold), ;roLovdrL mLg nepLoo6repeq neptTlrt.loetg oL TAF 6ev 61ouv ro rorol,otno outrlq tqg nepLoXqq nou e(vot uneU0uvo yro rrlv npdodeorl rou DNA. T6ooeptg TAF 61ouv xo noro ev6oyevrl Lxov6tqto np6o6eoqg DNA: ot TAF4b TAF12, TAFg r<ot TAF6 eivot (pror<ptvo[) o;-r6].oyor to''' 20.6 H TBP E(vor 0xougevx6g llopoyowog 565
  • 58.
    rorovcbv H2A, H2B,H3 r<or H4, owrmo(1<og. (Or ro16- veg gXrlpor[(ouv ro Boou<6 oUpntroro nou npoo66ver DNA mrlv euKopuonKti lprrlpot(vq, BL6ne Ev6tqta 70.3). To etepodr;reprl TAF4b/TAF12 r<or TAFg/TAF6 oXqpor((owor on6 ol.l,qLen(dpooll rtov ow(morlrov unopovd6ov gSoo rcrlv pror(Balv rcov nruXri:v rov roro- v6r.lu).o.yrxo, ouvoppooloitv q Bool prog boprlg nou ;rord(er Ue ro oKrogep5g rov rorovcbv. Mtq rdroro 6op{ nr0ov6v eu06veror yro Un er6rrdg yro qv o},}.r1- LouX(o oLlrlLenLdpdoetg rou TF1lD pe ro DNA. Or rmovx6g muldq rov rorovcbv Xpnorponoro[wqt en(- orlq Kor oe drpepe(g o).LqLenr6pdoerg prero(ri dl,),rrrv nopoy6wcov TAF11. Mepxo( TAFrr Filope( vo Bp(or<owor oe dLlo ori1.r- nloxo ercrdg on6 to TAFrrD. Er6u<6repo, TAFrr "rUnou ror6vrlg undplouv en(or1g ror oe npcoreiVu<d oUprnLor<o TIou rpononororiv rq 6op{ rrlg popor(vqg npw on6 11 peroypogq (B).5ne Ev6tqto 28.9). 2O.7 H Boorri Euoreufi luvoppoAoy€iTot OTOV YnoKIVnTfi * To ovodlro ororXelo xor or nopoyovTeq rrou npoodrlvovror oe ouro ou{ovouv q ouXvorrlro qg 6vop[r1g. + H npoodeol rou TFrrD oro rour( TATA 11 oro lnr elvor ro np6- ro pqpo qq 6vop[qg. * AtrtroL peroypoqrroI nopoyovreg npoodevovrot oro oupntroro pe ro0opro;levq oerpo, enerre(vovrog ro prirog qq npooTo- reupdvlg neproXlg oro DNA. e H RNA notrupepoorl ll opX((er rr; peroyporprl ;rotrrg npoode0ei oro ougn),oro. Iro r<Urropo, unorrvrlr6g prnope( vo Bpe0oUv oe rperq BooLr<oUg rUnoug Xpropror(vr1g. O npoirog rUnog e(vor 6vo ovevepy6 yov(6Lo oe x,trerorr1 Xpco;ror(vr1. O 6eU- repog e(vor 6vo 6uvrltr<d evepy6 yov(6ro oe ovotXrq lprr;por(vq, nou ovogdperot yov(dro oe eypqyoporl (poised gene). Aw6g o rUnog prnope( Uev vo ouvop- pro).oyqoer 14 Boo'u<r1 ouoxeurl, o).).d o6uvote( vo npoXrop4oer crrrl Ueroypogrl rou yovr6(ou Xiop(g 6vo 6eUrepo orlpro yro tqv Svop(4 rrlg Ueroypoqrtq. To yov[6ro Oeppur<oU shock Bp(or<owor oe eyprlyopo4 droL oore vo prnope( vo evepyonoulOoriv op6oog pr6- Lrg ou(q0e( q Oeppror<poo(o. O rp(rog rrlnog e(vor dvo yov[6ro nou evepyonore(ror oe ovorXrrl Xpro;ror(vr1. M51pr np6oqoto 5ro06,ro;re eldlrmeg nL4pogo- p(eg olelr<d Ue rn oupperoxn pn rrodrxonor4tu<ov RNA peroypdrpurv (noncoding RNA, ncRNA) ornv evepyo- no(r1oq rclv yovr6(otv. H r<ordorooq dMo[e r<or orlprepo yvorp((oupre no).).d nopode(yproro oro ono(o q pero- ypoAn ncRNA puOpr((er rn Ueroypoqrl r<owLvrrtv 4 enr- ro).umd;ievcov yovr6(ov nou rro6rronoLoUv nporeiveg. 'Evo np6o<poro nopddeLypro neprypd<per q ou;rpreropl rrov ncRNA nou (errvoriv ovodu<d on6 rov unorrvrpq tou yovr6(ou fbpl tou S. pombe orrl oro6ror<11 ogo(pe- oq 6Uo orcroprepc0v voux,.eooogor(rov on6 rrl ouyKeKpr- p6vq neproXrl rou unoKrwlrrl. H nopoyroyrj ourcirv rrov Lenoupytxrirv ncRNA (nou ovog6powor en(or1g Kor oq rpumxd ooro0rl perdypocpo n CUT - cryptic unstabte transcr)pts) nr0ov6v elvor nro 6todedo;r6vq ouyr<pnu<o pe trg oplu<69 ercrrg4oerg. Ie preydlo opr0p6 evepyc0v unortvrlrciv nopdyowor prerdypogo nou npo6plowor ovodrrd on6 roug unor<rv4r5g (yvoord rog ovo6u<6 pre- rdyporpo rou unoKrvnrri 4 PROMT - promoter up- stream transcripts). To PROMpT peroypdgowor ror npoq lq 66o r<oreu06voerq oe ox6on Ue rov xq0o6rr6 unorrv4rrl xor [orog no((ouv pu0proru<6 pd],o or4 preto- ypoen. Or noMo[ p6],or rotv ncRNA orrl prl0prorl qg peroyporp(g 0o nopouoroorofv orqv Ev6trya 30.5. Anopofu4rrl npoun60eorl yro rn 6ro6txoo(o rqg 5vop(qg e(vor vo Aeroupyrjoouv ge ro0optoprdv4 oerpd or Boorro( pu0ptorxo( nopdyoweg c6ore vo ouvop[o- i,oyrloouv 5vo orign).oro oro ono(o 0q ouv6e0e[ 4 RNA noAupepdorl. H oroLouO[o rov ouprBdwov ouvorl(eror mqv EIK0NA 20.11. M5).rg ouvdeOe( 11 noLuprepdo4, 4 evepy6qtd qg e).5y1eror on6 UeroypqgrKorlq nopd- yoweq nou npoo66vowor orov evrq4ur1. 'Evog unorrvrlrlg yro l.1v RNA no),uprepdorl ll ouyyd onotel.efuor on6 Srio trinoug nepLoXrrlv. O r<ewptx6g unorlqrig nepdXer r4v (6ro rrlv ogerqp(o, nou runrxd routonorekor on6 qv neptoXrl lnr ror ouXyd yenoveUer g' 6vo r<our( TATA n 5vo ororXe[o DpE, ev6 prnope( en[- orlq vo neprdXer rr enrnp5o0ero, Lry6repo rorvd qror- 1e(o. H onoreleoportr6qro ror q et6rx6trlro Ue rnv ono(o ovoyvcop((eror 6vog unoxwqrqg, oo16oo, e!op- rcirwor on6 BpoXe(eq oLL4loul[eg nro ovo6rxd, nou ovoyvcop(owor on6 pto drogoperrrrl oprd6o Ueroypo- qu<riv nopoy6wov, yvcrlorc0v @g evepyofioLqtiq (activa- tors). l-evrxd, or oLLql,ouX(eg-m61or Bp(oxowor -100 bp ovo6u<d qg oqerqp(oq, ov ror preprx6g gop6g ond- Xouv neproo6repo. H np6o6eoq rov evepyonorqrcbv oe ourSg rrg 06oerg ;rnope( vo en4pedoer ro oX41rotropr6 rou oupn).6rou 5vop(r1g oe drdrpopo ord6ro. Or unoxr- vr1r6g e(vot opyovropr6vor pre Bdorl qv oppi rou ouv- 6uooproU ototXe(tov. Ardgopo ororle(o gnope[ vo ouverogS,pouv orq Aeroupy(o rou unoxrvqrj, oMd xo- v6vo 6ev e(vor onopofurlro yro 6l,oug roug unorwrlr6g. To npciro B4trro oqv evepyono(4oq ev6g unoxrvqrrl nou nept6let rout( TATA xot Bp(oreror oe ovorXr{ lpol- ;tot(vq evepyonoofuot pr6Lrg o TF11D npoo6e0e( oro row( TATA. Aut6 prnope( vo evroXferor on6 ovo6u<d ororle(o nou 6pouv pSoco ev6g ouvevepyonornrn. (O TFrrD ovo- yvop(Efl en(onq rrlv otrtrr1LouX(o tnr orrlv oEeqp(o, ro DPE ror (o<og r<or dMo ororXe(o rou uTroKrvnrry). 'Orov oro o0gntroxo npooreOe( rqr o TFrrA, o TFrrD gnope( nM- ov vo npoo6e0e( oe pro neproff nou errefveror or6prl nto ovo6rrd. O TFrrA prnope( vo evepyonoujoer rqv TBp o(powog rrlv r<otooroLri nou nporolekqt on6 rov TAF1. O TFrrB npooddveror xoOodrxd rou rouroU TATA, di- nAo orqv TBP, ener,re(vowog 6"ror rg enoqdg r<ord prrl- Koq rnq prog nl,eupdg rou DNA on6 -10 6<o9 +10. H 566 KE0AAAI0 20 H Meroyporp4 orouq Europurilreg Boorxfg eworeg
  • 59.
    + TFllE & .Evo 0uunl,oKo evopqnq yroTIV RNA notruuepoorl ll ouvoppotroye(rot oroug unorrvqreg ono uto repopxnu6vn oxotrou0(o :rrltrenr6pooeov ue ueTqyporprxoug nopoyovreg. 0 TFLrD onoreeirot ono rqv TBP ouv ro ouv6edepSvo TAF, onoq qo(verot oTo Tlovo ;xluo. l-ro troyouq ontrouorguorlq, oro unotrorno oxnuoro 11 TBP tpo(vetot uovn uq, xopig tov TFrrD. l-lpooopuooptlvo ono: M E Maxon' J' : Goodrich rot R. Ti1an, Genes Dev. I (1 994): 51 5-524' xpuoroMtr( 6ogri nou <po(vetot mqv E|(ONA 20'12 6e(- )yet 6n n TBP q(4got(et enoqdq orqv elr'dooovo oUI'o- io ro0oax6 on6 to row( TATA xt 6plerot oe eno<pr1 pe rr1 pe(ovo q0l.oro qvodKd on6 to xout( TATA, oe Uto neproXrd yvoorn oq BRE. Ito oplo[o, o o;t6troyoq no- pdyowoq tou TFIB npoYUoronoe( etdx6g yo trlv oMrl- iouXio enoq6g Ue rov unortvrlti mrlv neproXti BRE' eur6 to Bripro ntore0etot 6tt enrlped(er xo0oprorxd rrlv ro0r6proorl tr1g noLtr6trlroq rou unortvrlr{, 6nlo6ti npoq ro noti 0o e(vot orpoppr6vrl 11 RNA nol'upep6or1 rot nor6g 0o e(vor o rtrri:vog erqroye(o. O TFrrB nt0ov6v no- pSxeirnv ent<pdveto nou ue q oepd qg ovoyvcop((etot lnO .n, RNA noLuprepdoq, on5te prnope( vo op((et rrlv rmerj0uvorl np6o6eoqg qg notruprepdorlq. H rpuoroMtrci 6oUn tou TFrrB Ue rrlv RNA no}'upe- p6oq 6etXvet 6tt tpetq neptoX6g tou nopdyowo oMq- Lntopo6J pre ro 5v(upro' 'Ontog ono6(6etot oxqpotrr<d ornv EIKONA 20.13, prto N teLu<d xop65lo rpeu6opy0pou or,6 .o, TFrrB 6pxetot o' enoqri pe to 6v(upro row6 orrl 06oq e[66ou tou RNA. Aw6 nt0ov6v nopeprBofuet ornv 6(o6o tou RNA r<ot enqped(er q pretdBoorl on6 r1v oventtupj 6vop(4 mr1 Stoquyri rou unoKrvrlrrl' 'Evog entpr[<uopr6voq,,66*11oS' tou TF1IB eto6pX*ot 96oo oro evepy6 r<6wpo rrlq notrugepdonq. H C-teLtrd ne- proXq oM4Lent6pd pe qv RNA nolugepdoq.xot rov T- F1;D yto vo npooovorotr(oet ro DNA. Ko0op((et en(or1g tlv nope(o rou DNA ere( 6nou 0o 6tr0et o' enoqtl pe toug nopdyoweg TFrrE, TFrrF t<ot TFttH, toug ono(oug nr0ov6v'eubuypoprpr((et pre to otignLoro trov Bootxriv nopoy6wtov. O nopdyowoq TFrrF e(vot etepo-ot<togep6g nou onotel,efuot'on6 6rio t0noug unopov66u:v' H peyol'ti- reprl uTlopovddo (RAP74) 61et prto e[optrirprevrl qn6 to nie'evepy6tqto e)'trdorlg nou 0o pnopoUoe vo eprntr6- Keror orrlv onodt6ro(r1rou DNA r<otd qv 6vop[q' H pt- rp6tepq unopovddo (RAP38) 61et rdnoto oprotroy(o pe ng neptoX6g tou Borct4ptor<o0 nopdyowo o nou 6pXov- roi o''enocpri Ue rnv r<ewprr<r1 nol'uprepdorl xot npoo66- 20.? H Bootrrl luoreurl IuvoppotroYeirot mov Ynonwrn fil
  • 60.
    Auo onorferg rourpuoroyouq ougntr6xou TF;B_ TBP-DNA derXvouv ol o TFIB npooD6veror roro 1u1xog Tnq Ks_ roppevlg ntreupog lou DNA. 0r Duo rtrdrvot rou DNA Ue rpoorvo x0l r[rplvo, i1 TBP pntre ror o TFriB rrjrxrvo rot pop. H tporoyporpio e[vor euyevlrfl nopoXtirprlorl rou Stephen K Burley. vetor o<prXrd orrlv RNA noLuprepdoq ll. O TFrrF nrOovdv g6pveL qv RNA noLupepdorl ll oro ouvopl_roioyo,jprevo orjpnl.or<o rrlq Ueroyporpqg xor nopdler ro pdoo yLo rqv np6odeorl. To orjprn).oKo rrlq TBp r<or ro.rv TAF * : pe(v' oM4lenL6pd ;re rri CTD oupd rrlg RNA nor."*. poong xoL 4 o).).r;tren(6poor1 pe rov TFrrB iorr;g e..: en(ot|g o4powrx4 6tov ro 6rpep6g TFrrF/notruLrep::_ ouv6e0e( pe ro otiprnl.oxo. H npdodeorl rqg no).ugepdorlg enerre(ver rrc :.. oetg nou npoororerjowor xo0o6tr<d rou + 15 orov ().-'. vo expoye(o Kor rou +20 orov r<).rr.lvo ;rr1_enUoysro -_ 5v(upro enexre(ver ro oUpni.oxo oe 6).o ro gqroc :: - enerdfl mo ovo6Lx6 dpto undpXer trtpo enrnpooii:- npomoo[o. Tr y(vetor orouq uroKrvrlrdg 1op(g r<outf TATAr A::. po(r4ror e[vot or [6ror yevu<of UeroypoerKof nopo,, : ,_ teg, geto(U dMo-rv xor o TFrrD. H nepLoXrl lnr nope ,. ro orotle(o rono06rr1oqg, oro ono(o npoo6due-o, . -- FirD [.r5oo qg txov6r4tog ev6g 11 neprooordpcrrv TAF . ovoyviop[(ouv oneuOe(og r4v lnr. !],],or TAF rou -tr len(or1g ovoyvrop((ouv to morle(o DpE xo0o6tr<c :-:ogeqp(og. H TBp l,eLroupye( oe owoUg rouq uno(,,_. r5.g nep(nou dncDg oroug unorrvr;r6g qg RNA no).:*:, pdonq I rot roug eooreprrorig unorrvqrdg rrlg F.,,_ notrupepdorlg ilt. Orov undplet rour( TATA, xo0opi(eL rrlv ewor.:- rrlg o<perrlp(og. H ono,trorcprl rou nporotre( ovur!:.. : orq 05o4 Svop(qg, 1-ro).ovdrL 4 dnoro ouvo).rxq UaLa;* rnq Ueroypo<pnq e(vor olerrxo pur<p(. l-1pdypol, KCr:: _ or unorrvrlr5q Xop(q r<our( TATA 6ev dlouv 1-rovo6L<.: ogeqp(eg; 6ror, 11 dvop[4 npoyproronore[tor peoo ;.puo ouoroLX(o oqerrlpurrv. To xour[ TATA euOuypc_.. (er rqv RNA no).u;repdoq pdoro r;g oMrl).en(dpo;'.: Ue rov TFrrD r<or dLAoug nopdyoweg dtoL rr-lore vo <a, _ vder mr1 oroorrl 0do4. H npdo6eoq r4g TBp oro TA-_ e(vor ro r<[pto Xopor<rqpLorrr<d orqv ovoyvo,rptorl r: _ unorrvqtq, oMd 6rjo trreydi.or TAF (TAF1 r<or TAF2) a orlg dplowor o' enog4 pe ro DNA rowo orrlv oes:-. p(o xr en4ped(ouv qv onore).eoporLrdrqro rr.lQ o,1.. dpoorlg. . . H ouvopl.ro).6y4o4 rou oupnLdrou dvop[rig rlq r" NA no).upepdorlg ll epr<pov((er ;uo ev6rogdpouoo owit:. !n le q petoypo<pq rov npoKopurr;trrrv. H BorcrqpLc*- RNA noAupepoorl ouoroortxd e(vqr dvo ouvercrrx6 our:. 0poro1lo pr evdoyevrl trov6r4ro vo ovoyvop((er Kcl ,. lnpoodSvet ro DNA rou unoKrvnrrl. O nopdyowoq lnou e[vor onopo(r4rog yro rrlv dvop(4 oMd o1r yro :r . enrprlxuvo4, yiverot rpnUo rou ev(U;rou nporoI or:: npoo6e0e( oro DNA, nopdrL opydrepo pnopei vo 5.. ororot. H RNA no).uprepdoq ll npooddvetor orov unci.. vqrrl prdvo ogdrou nporlyou;rdvcrlg npoo6e0c_. [eXoprmo[ UeroypoerKo[ nopdyoweg. Or peroypo<p L,r : nopdyoweg 6to05rouv ovd).oyo pd).o ;re ro BoxrqpLc<: nopdyowo o, 6r-ilo6r1 enrpdnouv otr1 Boo.Lxrl noAuu. pdorl v' ovoyvrr;p(oer eL6rro ro DNA orrq oMrl).ouXLot rou uroKrvrlr4, oMd dlouv ovomri{er ;reyo}.0reprl o,,,r- {opr1o[o. l-lpdyporr, oL nopdyoweg e(vor xup[rr-lq unr_ 0uvot yro rr;v e16rxdrnro rnq ovoyvoprorlq rc- unoxtv4r4. Mdvo optoprdvor ou1lperdlouv o, efloe.. nproreivrlg-DNA (xor pdvo 11 TBp xor prepu<o( TAF npo. potonoloriv eno<pdg eL6Lr<dg yro rrlv oMqtroul(o). Iur,i Ko0oDrrd DNA 0 TFIIB npooDevetor oro DNA rot epleror o,eno_ Ql Ue qv RNA notrugepoorl rovro orrl 0eo1 e{oDol rou RNA r<or ro evepyo revrpo, ro onolo rot npooovorotrL(et novrrl oro DNA. Avnnopopotrtrere Us Tlv Etrovo 20.12, nou beLXvsr rr1 doprl rrlg notrupepoorlg roro q peroyporpq, 568 tGOA/lAt0 20 H Meroypog4 orouq Europutirreg I Merox[v1or1 Ev(rjpou
  • 61.
    ndlq, ot oMrlhenr6pdoetgpreto(ti npoteivdlv e(vot or1- Lrowrdq Yto rn ouvoptrro6yr1oq rou outrrnl6rou' ' tn viiro, ouvopprol'6yr1or1 prnope( vo TlpqYporonot- r10ei or<6po t<ot p6vo orov KewptK6 unorwqrq' oorooo in vivo ourl q ow(6poor1 dev enopre[ Yto rn peroYpo- on. rogtirc onono0wot rot ol'Lr1}'ent6p6oetg U' svepYo- ;"',n;A;, ovoyvtop((ouv ro nio ovo6trd Totxtt:ol evepyoilotnrdq oMqLent6poriv pre q-Bootrt1 ouoKeurl or 5,o,popo orddto rotd tq ouvopgo6ynorl tns (9I5- ne Evotqta 28.5). geto[ti rov UeroYpoqu<tilv nopoy6worv nou e[wtnpr 6e6e;r6vor orouq evtoxur6q rot tou unoKvrlrrl' ---ili"L"u-oioi nopdyoweq nou qtdvouv aro oti1.rtAc oo Jropqns e(vot or TFIE r<ot TFrrH' Apouv oro re)'ewoio *aO-itr1q 6vop[r1q. H np6odeoq tou TFIE nporolei il;;rn' ina rieiioxns nou npoor.oretetot ro0odu6 r<otd or<61.11 1r,o orpo,pf'-r1q 6tnl'riq d'Ltrog' doq to +30' o ri n tfrli o provo6tr6q yevx6q ;retoypoqrr6q nop6- ;;; ut .oLr.ii.Lgq ove(opqteq ev(upru<6q evepy6rrl- irs, trti"gO dl'tov ;rto ATP6orl' eLtraols-1ill no)rtx6trltog Kot Uto evepy6tlto r<wdorlq TIou Unopet vq ,pro,ilpr),,,iroet tr1 CTD rqq RNA nolu;repdorlq ll (orq oep(vn 5 rrlq emonemtdxqg enovdtrqrprlq)' O TFrrH e(vot 6voq oouvqOtorog nopdyowoq nou en(oqq.prnope( vo noiql, *anoio p6tro or4v entpt(xuvorl' ot oMrltrenr6pd- ;;G;", trre to DNA xq0odtKd tqq oqetrlp(og etuot ono- ootmteq vto rn 6tqquYrltrlq RNA noLuprepdorlg on6 tov ;;^,"n-,'i o TFrrH inions etrrntrSretot orrlv ent6t6p0oor1 tou DNA (Blene Ev6rrya 16'4)' ' - -:" gudi ypoprprtr6 erpoye(o, onopofurlteq. npouno0S- ot,iv* in'uetor<ffior1'rr1q nouprepdorlq e(vor q udp6- truot1'tou ATP, 11 nopouo(o TFrrE t<ot .n tu"qY-6-ll,o eiu<6or1g tou TFrH (nou nop6letot o19 rtq ufiouovo- ;;; Fb- rot xeo;. Aw6q oi npounoo5oerq nopor<dp- nrovrot o' 6vo u.,"p,ito'1'5vo exgoye(o' Aut6 ,r,oatlii,ut, 6tt or TFrrE r<qt TFrrH e(vot onopo(tnror Yl'o irlv onoorfuo(r1tou oUn O*t vo gnop6oet vo Eer<wn- oe, n pe.ooluqon rnq RNA no}'uUep-dor1g' H evepy6tq- ro eltrdorlg rnq uTloUovd6oq XPBB tou TFrrH e(vqt une0Ouvr1 Yto rrlv npoyprottr<ri onodtdtofq tou DNA' - ;-RNA'.olupepdorl rowoor6retot oe ;reptr<6 yov[- 6to 6tov op{(er rn UeroYpoQn' (To onot6eotrro 6e o,o,p6pr, noii'oto ttlv oventtuxq.eygpEn qq Borcrr1- p,ooni RNA nol,uprepdonq nou ovotr60r1Ke ornv Evot4- ii i s.'t z, ;toov6tt o prqlovtopr6g e(vot..5to<poperr6q)' Ie noMd yov(5to, n RNA noLuprep6orl ll teppoil(et pe- ; ;6 u*bn ot omoo4' To Bpqx6 RNA npoi6v qno6o- Ut"o, VpnVopo. l-to vo oupBe( entprqxuvor'1 U6oo orn 'ut-oVpoEion provd6o, onome(tot 5vo orSpnl'ot<o Ktvo- !O" *tu r<otreftor R-TEFb' To R-TEFb neptsXet r1v rwd- on COKS, q ono(o e[vot U6troq tqq ou<oy6vetoq rov o*oot.fx nou eL6yXouv rov Kurrctptr<6 x6xtro' O R-TEFb ,ftoorpopuLtOvet nepom6pto tq CTD (orr1 oeptvrl 2 qq emonemtdtrc'1q enovdMlqrlq)' M6Xpt on-gepo 6ev xoto- ,oo,iut o0te yiot( onot're(tqt outi 11 en6poorl oe uept- *o,ii' ,"o*,rr1t6q ror 6Xt oe dI'ouq o0te ncirq puOpti- (etot.--'H',pro,popuL(tloq tqq CTD e(vot onopo(tqq yto t1v onel'eu05pcoon qg RNA nolupepdorlq ll on5 rov unortvrlt( ooi -oug ueroYpoqtKo.Uq nopdyoweg 6tot dlme vo trrnope( vo xdvet rr1 petdBoorl ornv entprlKu- ,Ui"n lropq(, 6norq <po(verot^orrlv EIK0NA 20'14' To np6tuno rptootpopu)'(tlonq rnq CTD rotd t1v entp{ruv- ;n;fu;iorrol-,,*o' e[6yxetot rot Kq'rol0erot on6 notr- igs-nprt",utr5g rwdoeg Kqt Qooqordoeq' ot nepto- ,Zitpt, Boou<oi uerqYpoEtKoi nop6yoweq oneLeule- il";;"ion6 rov unor<wqtri oe our6 to.cnd6to' o Me- diatorol.l.rl}'ent6p6er6tr<dgeno}.upepdoqnourpdpet 2O.8 H'EvopEn AroAou0eiTor on6 K60opon Tou YnoKlvnT6 Kl EntPfiruvon * 0r TFIE ror TFrrH onoLtottvrot yto r1v rfl[r1 iou DNA Kot rn Ue- rorivqorl rrlq notruUePoonq. " rio ro [erwrloer rr ro0opol rou unortvqrrl rot 11 entpflruvorl nprlnet vo rpototpoputrttl0e( 1 CTD + Ie uepLro0q unoxtvlrdq, Yt0 v0 lepuoT(oet q 0ventruxlg evopEn Xpe to(eror eninpoo0errl tpo'rorpoputritloq rrlq CTD' * io o*ilir.pl rov toroviiv rpononorouvrot npooroLpo roro 11 perorivrlorl rrlq RNA nohuUepoonq' * H CTD ouvrov((et rrlv ene[epyoo(o rou RNA ue Tn ucToYpoQr]' =, Av ouUBotiv ptrdpeq oro DNA, 0o entdLop0trl0otiv roro nporf- Ulon To geroYPorP6Pevo Yovi6to' " 'O f i,,U nopdXet ro ouvdeopo yLo evo ouptnhoro ev(0ptt'tlv ent- 6toP0tlonq. An6 q ortYLi( nou 0o olqprottororiv o^tnptotoL vourl'e- oiio,noi at"uoi r<ot ;relpiirlv oneeu0epooq.tqg RNA nouprepooqg on6 tov unoxtvrlrl preootroBouv UeprKo .ri,o[ bnuo-o. Aw6 to ord6to ovo;rd(etot xaaaporl ,o, uno*rur1rr1 (promoter clearance) rot e(vot to rp(otpio preU,qOUeuo'$6po no, r<oOop((et ov 0o preroypo<pe( ;;;i;;(6t oe evplvopo'q 11 5vo.evep16 vov(6to' Aut6 ro Brlpto eL6yXetot on6 evLoXur6q' (Ouprq0elte 6rt to ^p iI,l,l- Bn u"'xLeta( q q pretoy pocpq q To Pool p^t:: uo, q'pre-oiponrl tou r<.etoro0 ouprntr6rou oe ovotxro' Bi.O"t rr,iinru 19.3). ot neptoo6tepot yevtr<o( peto- vpl,p*ot nopdyoweg onottotiwor pi6vo yto r1v np6o- Oton t,fq R'tlA noLuprepdonq fiov. unoKtvqtrl' o)'d geptr<oi 6pouv oe Uetoyevdorepo ord6to' Ot petoypo<ptt<ot nopoyoweq nou nOo96.evow11 1' evrcrlutdq ouv(Oorq eL6yXouv rov unoKtvnrn oxt lt-11? on ino,pll, ol).d pdlov pr6ool ev6q-ouvevepYonorntrl nou ,,poo66vero[ orov unoruvrltq 'Evog on6 rouq nto r<oLvouq ouvevepYonotqt6q e(vot o Mediator' 6vo rt9Ltl preyd).o nolu prep5g npoteivtr6 orj pntroro' ouwnpn Uevo onL .n (rjpq Stlq tov ov0polno, nou evonote( oqgoto on6 noMo0 g UeroYpoQtKo 0q nopoyoweq npoo6edepr6- ,./ouq o' evtcrlwdq. Tdoo to yov(6to oe eYpnYopon ooo . r, i" *tpvl yovidto xpetd(owot r1v oMrlLen(6poorq 20.8 H'EvupEn Arolou0eitot ond Ko0opon tou Ynoxrvnq xt EnrElxwol 5t I Boorx6c ewoteq I
  • 62.
    Gr:G G v,i'Aip $*ro Pol ll PP Y.iersg s RNA pol ll Ysprsgs RNA Pol ll Aeqolevf pol ll rou DNA Ynoxwqqg npooflKuoq r[q pol ll ond ro otiurloxo rpo6vopqnq H Qooqoputrioon qg Ser5 ond ro TFllH Por10o ornv npood)xuon ev(rjpov nou rotrUnrouv ro 5' 6rpo rou [erqypdqou Poly A site Anoqooqoputritoon ond q QtooQordon SerS (n.x. Scpl) Un qoogopu)rroprSvq CTD: <po[vetot 6tt 4 qoogopu- l.(roo4 6tor<6mer ourti qv oM4lenidpoorl, H CTD epntrSretot dpreoo { 6ppeoo orqv ene(epyo- o(q rou mRNA rord 11 orivOeorl rou Kot ogo0 onetreu- Oepo0e( on6 rrlv RNA noLuprepdon. Kd0e 06o11 tproorpopul,(roonq rqq CTD XprlorpeUet og o4pre(o ovo- yvdlpronq r1 oyr<upo66tnonq yto rn ofvdeon dtrl,tov npcoreivc0v Ue rrlv noluprepdoq. To 6v(upro oxrlport- oUoU rnq roL0mpog (tpovogepdorl rou Youovultt<oti, iexivllro rnq pol ll H rptorpopu)r(oor1 on1 Ser2 ond ro P-TEFb evepyonote( rqv enqgrlruvorl, q ouppoErl xor qv noluodevutr(trlq H Q(DoQopu)(oon orn SerT und Utq dyvrooq rwdorl Dev SXet Udxpt mryUnq rdnoro yvomd pdtro oqv drgpootl rtrtv yovt6lov nou rtoduonororiv nporetveq Enrp{xuvol on6 rqv pol ll Iuppotpti guanylyl transferase), nou npoo0Stet to rotdLotno G fio 5' drpo tou veoouweOetg6vou mRNA, npoo6Sve- ror or CTD nou 61er qoogopuLtco0e( arrl oepfuq 5 (r1 npdnrl <proo<popul,(trlon nou KotqLUetot on6 rov TFrrH). Aur6 (oog e(vot oqprowtx5 enet6ri enttp5rlet oro 6v(u- Uo vo rpononor{oer (rt 5tor vo npoqror6q;e0 ro 5' dxpo pr6l,rq ow6 ouwe0e(.'Eneno, q goorpopul,(oorl tqg oep(vr1q 2 on6 to P-TEFb odqye( oe npoo6trr<uor1 orrl CTD puog oprd6og npoteivcbv nou ovoptd(owot S- l-truxo(utrioon; 0toorloputritoorl on6 )dk8/Srb10, ERK %, )DCZCDKl) Anorpoorpoputrioorl onri q qoorpordol Ser2 (n.X. Fcpl ) noqooqoputr(ooq ond m QooQqrdqn SerT; Kon{, notruodevutr[toorl xor repporroprig l-lpdodeol rtov nopoydvttov roluodevutr[oonqmrgqooqoputruog6veg efiovqtrnuerq Ser2 ue nenttdu]tonpo]rtvtro0g 6eoUoUq oe trans 0roudpqoon EIK$NA 20.14 H rpononoirlorl rou enronemrdiou CTD r4q RNA no)upepooqq ll rord q UEToYpoQ1. 0rov q CTD qq RNA notrupepoonq ll ero6pXeror mo o0gntroxo npo6vopEnq 6ev e[vor Qooqoputrroudvn. H Qtr:otpoputr(oon rototrofnov oep(vnq dnptoupYe( O5oetg npdodeoqq t6oo yro ro 6v(upo ene[epyooioq Tou mRNA 6oo ror yro ng uvooeg nou rorotrUouv nspotTdpo tptootpoputr(tlon, dntoq neptYpdqtTot oIo oflUq. AvqTfntoorl on6 Trends Genet., vol.24,P. P. Gardner ror J. Vinther, Mutation of mRNA target sequences..., pp. 262-265. Copyright 2008, ge qv odeLo tou Elsevier [http//www.sciencedirect.com/science/iournal/01 689525]. Avqxurtroon rnq pol ll Enrp{xuvol on6 r4v pol ll 570 KEoAAAI0 20 H Meroypoqn orouq EuKopudTeq Y$prsts RNA Pol ll
  • 63.
    l:= oi ono(eqpre q oetpd roug npoo66vowot orouq -::cyoweq ouppo<pr1g. Auq 11 6tepyoo(o (otog e(vot : :c rponog Ytq ro ouwovtopr6 tqg pieroypoqrlq ge rr1 - -:corpq. Tetrtr<d, rdnoto ouorortrd u1g ouoreuqg r<o- -".: :otruo6evutr(toorlq nou Xprtotponotoriwot xotd tov -::-orrogd rnq UeroYpq<pr1g eniorlg npoo66vowot oe l-) rporogopul,tcoprSvq orrl oep(vq 2. t-leptdpyoq, out6 - -*Soiver 11611 on6 rrlv 6vop[r1, ;rdMov 6tot trlore 1 R- ',: i:oluu0pdoq vo e[vot Srotprq Yto rtq owt6pdoetg . rlepyoolog tou 3' dr<pou pr6].tq [er<tvrloet q oJv0e- , - -t e[o6oq on6 tov nuplvo pt6oto ttrlv nuprlvtrolv n6- : - . enlor1g etrSyXetoL on6 tr; CTD ror nL0ov6v - -..ov((etot Ue rnv ene(epyoo(o rou 3' drpou O)'o --- : uno6qtrcbvouv drt 4 CTD iooq e(vot tivo yevtr6 . (.wpo yLo tq orjvdeoq dMtov 6tepyoottov Ut rn Ue- -:.:orpq. Itrlv nep[mcoon rou crXrlpotLoprof qg ro]'ti- :tr-( Kot rnq ouppoqnq, n CTD I'ettoupye( 5ppeoo, -: rtryowoq q ouvopprotr6Ynon rov nporttvtKov ouFl- -' : (rov nou ovol.optBdvouv ttg owt6pdoetg. Iqv nep[- *il rrlg 6qprtoupy(og tou 3' dr<pou, (oog ouprpret6leL r*.30 ornv ow(6Poor1. -o rplot;to orlpre(o 6tov r<oOop((etot ov (r<ot n6te, ' -,' nep(moon Utoq noLuprepdoqq oe eyprlyoporl) 0o ..:ooore[ 6vo yov(6to e(vot 11 xa)apo4 rou uftoKtvt]rtl, :-'.:5q 11 oneLeuOdpoorl rou ev([;rou on6 rov unoxt- - - Mdltq our6 ouprBe( xot oneLeuOepol0otiv ot nopd- :,.eg dvop[4g, enttp6nerot 11 petdBoorl mq qdoq : *iKUVonq. To pretoyporptx6 otiprntroxo onoteefuot - rcv on6 r1v RNA notrutrrepdoq ll, toug BootroUg no- :-.oweq TFrrE rot TFrrH, nopdYovreq enrprlxuvolg - .g o TFrrS) Yto vo e;-ino6(oouv rl'lv or<otdMr1trq noU- :* <ot d).o to dv(upro Kot rouq nopdyoweg nou e(vot -:cc5e6ep5vo oro CTD. Aut6 ro o0;lnl'oxo np6net trr;- : r 'vo UeroYpdqet 6vo er;.roye(o Xptoprot(vr1g, dtoptSoou '!v vouKleoorogot(ov. OL6r<i.qpo to yov(6to pnope( vo 3ploretot oe ovotXrrl Xptoprorivrl, t6(trlg ov 6ev evot no- .u ireydtro. EvoMorrtxd, oe ovotar4 lpcopror[vq ;rnope[ : bpioretoL pr6vo 11 neptoxrl yJpro ond rov.unortv4r4' ','eprr<d yov(6to, 6nolg to yov(6to tqg Mutr<r1q Auotpo- p(oq (Muscular Dystrophy - DMD),5Xouv g6ye0oq Ue- yoBdoeov ^o, yi' out6 'q preroyporprl. touq 6topre( noiL6s tipeq. To or<roprep( tcov totovrov np6net np6- ;"d uo -rponono,r1oo0v -optop6vo npoo<optvd ono- ouvoplloloyotivtot- rt 6netto vo ['ovoouvopUo- ilrnaJ;;o expove(o (BL5ne Keq6)'oro 10 r<ot 28)' To lO,o io orcro;iepriq e(vot 5to<popettx6' tiXowoq owtKoro- il;;,;;;;"ovtrr1 H3 ue rnv evotr)'o^tt^1 uopan H3.3"' - i,i1-,,grro p' 6vo notrot6tepo prow5lo' 11 npdrtq no- ),ru;;A;n no, eo cpUyer on6 rov urloKtvnrrl evepye(oq ouirurr-nc. K[pto ]'ettoupyto trlq e(vor vo.dtootpotr(oet J-i'iio*Lnpo to yov[6to 0o e(vot oe rotdmoon ovot- ;,i; il;u;.(vqq. MetocpSpet ;ro(( qs ev(uprtr<d orip- nl.oro yto vo -pot-,onoitloet rLq tot6veg Kot v' ouoO,o;rop.pOoet rn Xpopor(vq H rm6vr1 H2B uq(oro- roL OuroUtrn p,ouo-ou l,ntnoutrtv(tr;or1 (monoubiq u iti na- ii""i *ti" evepyd getoypo<p6prev-n. xpouor(vr1' Aut6 e(vot onopo(trlro Yto ro 6e0tepo gnuo' rn preou(oot1 rqq ror6vrlg H3, nou pre q oepd rrlg elvgt anopoiulrn y,o.r1u npoo6Lruoq ttrlv ovodtoptop<pcorriv rng Xpo)Ue itnnq G).ent Evor4req 10.4 t<ot Ev6t4ra 28'11)' Itilitpovo pe ro nto np6ocpoto Uow6I9, xdOe nolur' pepdoq' xprloigonote( 5vo otlprn)r,oxo ovo6tog6p9oonq ;t xdi,;.Gq ;ro(( pe uro tor6vn o1 ou1o66 npotet- "n y,; J oqotp5oet 6vo Etgepdq H2NH2B, oqrivowog 6vo'e(ogepSq (orn 06o'4 rou orroprepo0g), nou e(vot eurol,6tepo v; onoporpuvOe( npootoptvd' Aut6q ot rpo- nonotrloetg e(vor en(orlg onopofutlrcg Yto rnv enqvoou- uoplroi6y,'1orl rou ol(opepotiq tou vou rtreooulpot(ou mo'DNA 1.retd q 6tdLeuo4 t4q RNA noLuprepdorlg (BLdne Evoulta 10.10). 'Onog ovo<p6popre atry Ev6t4ta 20'7, to motle(o otrtrqtrouX(og tou DNA nou onotel'o0v tov rewptr6 unoo,uqtq' e;rqov(ouv orlprowtr<( erepoy6veto, 11 ono(o prnope( v' onotetre('rn Bdo1 Yto rnv et6u<6trpo rou uTIo- r<wrp{. 'Evo on6 out6 to cnotXe(o e(vot yvtoor6 Ue rov opo xoupn[ naioqq (pause button): np6renot Yto ptto nLotioto o"e GC otrtr1trou1(o nou tunu<d Bp(orerot ro- 0o6xd t4g oqeqpiog. Aur6 to orotXe(o 5Xet Bpe0e( o' 6vov onpoo66r<qto preydtro opt0pr6 ovomu(torriv Yovt- 6(cov qg DrosoPhila. H yevrrcl dtepyoo(o rrlg 6vop(qg e(vot nop6proto pe q 6tepyoo(o nou rqtol'ferot on6 q Bolcqprorrl RNA noLuprepdoq. H np6o6eorlqg RNA notru;repdo4q 6npt- oupye( 5vo rLetor6 o6pntroro, nou 6nemo peto'rp5nerot oe ovotXr6 oUgnLoro mo ono(o ot r.tr'cilvot rou DNA 6Xouv 6toXtoptore(. Itrl Borrcrlpror( ow(6poor1, o oxn- prolopr6q rou ovotxro0 ouprnL5xou oLoxl'r1pc0vet rnv onopofu4rrl 6oprri olloyri tou DNA. Mto 6torpopd orrlv europutottrc] ow(6poor1 e(vot 6tt pretd on6 out6 to ord- 6ro ononeftot nepomSprr: onoetr(rcrloq tou erpoye(ou' T6oo mo Bor<tripro 6oo rot orouq euxopuri:teg, undpXet dpeort o6vdeorl preto[ri qg RNA nouprepdorlq oo,.iq evepyonotrlorlq qq ent6r6p0roonq' Aur6 opXtrd 5ywe owtl.r1m6 enet6rl ro geroYpoutrr6vo yov(6to ent- 6rcplrisvowat xard nporfulor1. 'Eror, avaxaA0tp1qxe 6tt or6Xog e(vot pt6vo o xLc0voq erproye(o rou DNA, evri o xL<tvoi prq-er1-roye(o entdtopOrilvetot Ue rnv (6to to1itrl- ro 6n<og rot o r6ptoq 6yrog rou DNA' 'Orov r1 RNA no- LrU"pa'"n ouvow{oeu BI6Bn tou. DNA o-rov r}'ci'lvo il;y"(o, orogot6 enet6( 6a.gnope( vo xpqoqronoqoet rq npoBq;rotu<5g oMrl}'ouXbq oq erpoyeio,yto vo Kq- ili, e i"t, -d (euy6ptopo ttov ou prnr1 prrr;tonrcbv Bdoetov' nuiO tgnvei rnv ei6tr<6qto qg en(6poonq Yto rov-rtr<i- "" L^u"vtf" ftux6v gldgn orov ^I9Y9. prrl er<poye(o 6ev nopr1,,r,oOiq"i rrlv npo6l'oon rnq RNA.notruprepdonq)' jin o,"pvoo(o trlq enr6t6p0tr:or1g epntr6retor o yevtr6g uri"wt,fiOg nopdyo*og TFllH, o ono(og uq(orotot oi ruoLl.oo,o6q uopQ6q nou onorctro6wor on6 uto Kev- ip*n *p,oxn ouv6e8epr5vr1 pe dlleg unoprovd6eq' ' O fff ri-g1et rotvri )retroupy(o t9o9 mnv 6vop[r1 -nslrt-oYpo6ng 6oo rot orqv ent6r6p0toor1 tov Btro- BOv.'Ot iSteq unoprov66eq el'rdorlq fiPB xot XPD) 6n- lrroupyo,lv tqv opltrrl <PuooL(6ocrlltypoqqg rot ono6tfidooouv to pNA oe prq 06orl BLdBrlq' Ynoprov6- ;t; G;6vopo XP t<trldtronoto0wot on6 yov(6to trov ?S,S H 'Evop!r1 Axo)rou0e(rot on6 K(Oopon rou Ynorwqtrl rot Emg
  • 64.
    ofloiov geroMd[erg npoxo].o6vto v6oqpro pelayypot- otxrj fr1polepp[a (Xeroderma pigmentosum), nou Xo- porcrrlpf(eror on6 npo6td0eorl yto ropr(vo. O p6].og rov unoUovddrrrv TFIIH orlv entdt6p0oorl tou DNA ovq.hie'rqr Lemopeprirg ornv Ev6rrya 16.4. H l,enoupy(o qg entdr6p0oroqg nr0ov6v onotte( rponono(r1or1 q ono66gnon rnq croproqpr5vqg RNA noLuprepdorlg. H peyd).r1 unogovd6o qq RNA nol.upe- pdonq 6rooildtor 6tqv ro 6v(upo mopord oe O5oetg BIdtsnS on6 unepuirdrl orcnvoBol,(o. H otiv6eorl pero[[ rrlg ouoxeurlg peroypogrlq/enrdr6pOtoorlq Kot rrlg 6td- oroonq r4q RNA nol,uprepdorlg nopopr5vet o6teuxp(vt- or4. l-h0ov6v q nol,uprepdoq np6ner v' onoporpuvOe[ pr6l,rg oroprorrloer. 2O.9 Or Evroxurdq flepr6xouv Apgi6popo Erorxeio nou YnoBon0o0v rnv'Evop[n e 'EvoS evtoXuqg evepyonole I rov unoxtvlrl nou pplorerot nto xovro rou ror pnopel vo pp[oreror oe onoto6tlnore onoorool elre ovo0rro e[re roOodrro ono rov unortvrlrfi. * Mro AUS (ovodrul evepyonorllrrl otrlntrouxio - upstream ac- tivating sequence) rou (u;ro1.ruKrlTo ouUneprQepeTor oov tvr- oxurng, otrtro treuoupye( povo ovo6rro tou unonvqrl. e 0t evLoxureq oppon(ouv ouprntroxo on6 evepyonou'lTsq n0u otrtrr1trenrbpouv opeoo ll egpeoo pe rov unorrvqrq. Irrq nporlyoUpreveg nopoypdtpoug, o unoKtvqrnq nopouordorrlKe oq onopovroprdv4 neproXrl uneU0u- vrl yro qv np6o6eon rnq RNA nouprepdonq. Om6- oo, or euKopuolrrro( unortvr;tdg 6ev evepyotiv ono- po(rr1ro on6 p6voL roug. Ttg neproo6tepeq eop5q, n evepy6rrlro ev6g unoxrvrlrrl ou[dvet ewun<ootord on6 qv nopouo(o ev6g evtoluq nou Bp(oxetot oe drd<popeg onoordoetg on6 tov rewptr6 unoxtvrlrq. Kdnoror evtoXur6g trettou pyotiv pr6ocr: oll.r1tr ent6pd- oeu:v pey6trou BeLqveroriq, qq rdqnq rrov Serd6tov kb. Aw(Oero, dLi,ot trettoupyotiv pSotr.l oLl.qLent- dpdoeov tr-ur<po0 Belqver<orig r<ot pnope( vo Bp(- oKowor opretd rowd mov rewpu<5 unortvqni. 'Evo on6 ro npdlro rowd ororle(o nou neptypdgrq- rov rowd orov unoKrwlui rpov 4 oMqLouX(o or4 06oq -75, 4 ono(o orlpepo ovoprd(eror r<our( CAAT, on6 tr1 ouvorvetrrrl ol,L4louX(o qg. Iupd Bp(oretot rowd mr1 06oq -80, oll.d ;rnope( vo tretroupy4oet on6 ono- ordoerg nou norxfi.ouv nolti on6 t4v oEerlpfo. Aet- roupye( Kor Trpoq lg 6rio roteuOUvoetg. Ot entmdloetg peroMd(eov mo rowi CAAT onoteLoriv 5v6et[q yto ro yeyov6g 6n ro0op((er onoqootortrd r4v onoteLe- o1lotx6rr1ro rou unoKrvrlr4, evcir 6ev enrlped(et rnv er- 6rr6qrd rou. 'Evo derirepo rorv6 ovodtrd morXeio e(vor ro r<ow( GC orrl 06o11 -90, nou nepdXet qv oMr;- LouX(o GGGCGG. Or evroXurdg ouXyd nepdXouv notr- l,onl.d owtypogo ourof rou ororXe(ou, efue orq pto efue orrlv dMq r<ore60uvor1. To rout( GC efuor en(oqg oXenrd row6 ororle(o xowd orov unor<rw1r1. H dnogr1 6tt o evrolutqg 6rorpdpet on6 tov unoKt- vqtrl owrrorontp((er 6Uo loportrlptmrrd. H 06or; rou evtoXurq oXelrd Ue rov unortvqtrl dev e(vot oro- 0epr1, olL6 prnope[ vq norKil,er no].0. H EIK0NA 20.15 6e(Xver 6rr o evtoXurlg pnope[ vo Bp(orerot qvo6txd, roOodrxd n U6oo o' 5vo yovi6to (runtrd mo rwp6vto). Enrnl.Sov, prnope( vo l,eroupye( ror npog ng 60o ro- reu0Uvoerg (our6 or1;ro(ver 6tr gnope( v' ovompogeQ oe oXSoq pe rov unomvrlrrl. Xerptopo[ tou DNA 6e[- ;3rouv 61 6vog evrolurr]g prnope( vo 6reye(pet oTlotov- 6qnore unoruvqrrl rono0erqOe( yUpco rou, or<6pr1 rot 6er<ddeg lrLrddeg Bdoerg trrorpud npog onotodqnote rore[0uvo4. 'On<og o unor<rvqtr1g, o evtolwrlg (rl o dMog eow6g ElK0tlA 20,15 'Evoq evtoXurrlq Unope( vo evepyonouloer Svov unoxrvrlrrl ono ovodurl n ro0odxrl 05o1 xot 1 otrtrr1trou1(o tou pnope[ v' ovoorpoqel oe oX6o1 pe rov unorrvqrrl. 0 ovodrxdq evrolurrlq evepyonoreI rov unoxrvl14 Ynouvrlqg 0 xo0oduoq evroXurrlg evepyonotei rov unoxtvrlrq 572 KEOMA|0 20 H Meroyporprl orouq Europu6reg ; dworeg
  • 65.
    - - - r,cq onootconllrllq) e(vot 6vo nolupep5q oroLle(o - - - : .oreohot on6 Bpol6o orotle(o DNA Snou npoo- ,, -.:oi dtorpopot t[nor petoypotptrov nopoy6wt':v' - ., c-1utdg l.rnope( vo e(vot ontro( 11 otivOetot, ovdtro- r ..i :ov optOp6 rov orotXe[ov Kot rov t0no tolv peto- - . - .- *, nopoyowov. 3. *etoypogtr<o( nopdyoweg nou npoodtivowot o' , l, --i0g 6Lorp[vovtot oe OetLNoUg ror opvrittr<o0g - r: r, ovreq. Ot 0ertro( 6Leye(pouv tq pretoypotp4 .','EpYonotnr6g - activators), evco ot opvqttro( 11v . :-str.ouv (xorooroheig - repressors)' Ie roOe :.::*rvq ortYpl, dvo r<rirropo 0o nept6Xet 6vo prtypto . .-:.:ccptt<tr.lv nopoy6wr.ov tt<ovtrlv vo npoo6e0o[v o' , '.. t','toxurfi. Av npoo65vowot neptoo6tepor evsp- - -: -;eq on6 xotooroLeig, ro croLXe[o 0o eivot evt- , - -c Ie ow(Oeq nep(mr.':o1, 0o e(vot onooLonn- _. : I. cq dl).oq tp6nog dtdxptoqq ttllv preroypoqtrrltv - :: r . clrov Boo((etor orq Lettoupy(o toug' H npo-rq - :i-,apilo;lBdvet toug npaytrtatxolq uepyonoLqr{'q ",: activators), ot ono(ot Spouv t6oo pr6oo np6ode- r-: ti ouyxerptpr6veg O5oetq -rou DNA 6oo xot 6r1pt- - -: . ivroq enocp6q ;.re tq Bootr<r1 ouoreurl otov -:, .irn, e(re dpeoo (on6 ;r6vot roug) e[te, nro ou- r 'oou-) ouvepYoilotrlttrlv 6ncog o Mediator' ot no- - -.:, isq outrlg qq tolrlg trettoupyo[v e(ioou t<oLo , '. -cve[o DNA n Xpoprot(vqg. Arjo entnp6o0eteq td- :. : . . oOyonotntcilv eXouv ewel'trlq dtotpoperrxd pnxo- - ..l svspYonotnonq. H npoltrl nepLLopBdvet evep- :-:.nrsq nou snrorpoteUouv 6v(upro rponono(qo1g -- r,oopor(v1g t<ot otipntroro enovodtop6pQoong -- : ,.cLogor(vqq. l-1oMo( evepYonotnrSq orqv npoYpo- - . ::-rc Lertoupyotiv t6oo tr-lg npoyprortroi evepYo- - - -^:tq ooo xoL pr6oo npoodLxuo'r1q tpononotqtolv -- - r.c0JUor(vlq. H tpitrl td(r1 nepfioprBovet opxtre- .-, ..oouq UeroYpoQtKorjg nopdyovreq (archi- '.-'-'a transcription factors) , H provodtxrl Lertoupy(o -: - : iivor v' oMd(ouv rn 6oUn rou DNA, tuntrd vo to . :.'-r:ouv. 'EroL pnope( vo ep0ouv r<owd 60o oplrr<d r- - -trKpuop6vot geroyporpLr<of nopdyoweq, Steuxo- - , - .aog rl ouvepYoo(o toug, Itrlv en6pevq ev6rlro' : - ::ilooouue nto npooexttxd nirlg ouvepyd(owot o' - r, .',/loxurrq ot 6to<popetLr6q td(etq rov evepYonot- ---, (ct rcDV Kclroorotr6tov. Ito Kecpdtroto 28, 0o e(e- -:r:-.it rl ueroYpogrrrl pri0ptorl pre neptoodtepeg : . - JLPUTL' I:r (rj;rr1 undpXouv crrotleio ov6).oyo Ue rouq evt- . , - iq nou ovopld(owot ovo6tx6g evepyonorqrl169 ; ,-r lAouXieg (upstream activating sequences - UAS)' '-::.t vo Leroupyrloouv Kot ortq 6Uo roteu0fvoetg -. -.:tpopeg onoordoetg ovo6trd rou unoKtvltrl, 61t - --: (ot 6tov Bp[oxowoL ro0o6tt<o 'EXouv pu0prtortr6 : -' : Jiog rL ere( npoo65vowor ot puoptmtreg npolte'l- :: :cu evepyonoto[v ro ro0o6tt<d yovi6to' -iLpd;.roto ovoo0orootlq oro ono(o q oL)'q)'ouX(o - - - I rLo{urll otpotpe(rot on6 to DNA r<t 5netto evoopo- --,3.ct oe ronoto d).).r1 06o11 tou yovtdttill-totoq 6eG . - - ., orr rguototroyrr<q uetoYpo<prt prnope( vo 6torr1- pqOe( e<p6oov undpxeL evtoXurrlg, ove[oprlrc ] : :- ' or<pLBrl ew6ntorl tou (opr<e( vo Urlv undpXouv irc'';rii oro DNA nou peootr'oBe( B).6ne Evorqta 10'12) At i': yov(6Lo B-ogotp(vqq tonoOetq0e( o' tivo pdpLo DA no, ntpi6xti 6ro, er,oxur1, in vivo I ueroYpoQn rou O'ouEnde( ndvr,-l on6 200 <pop6g, toro Kt ov o evtoxurlq Bp(onerot opr<et6g kb ovobtr<d q r<oOobtrd rnq oQer!- pioq, o" onoto6rlnote npooovotol'Lo;r6 Ax6ptri 6ev 6Xet ovoxol.urp0e[ oe notd on6moorl netiet vo ]'ettoupye( o evLo'1urr1q. 2O.lO Ot Evtoxurdq nelroupYo0v AufidvovToq Tn IuYKdvTpoon rov Evepyonornrdv Kovrd oTov YnoKlvnTn * 0r evtoxursq ouvflOutg trettoupyo0v povo oe cis Stouoptptlorl 0t 0x00n ue Tov unoKtvllTn-oroxo * Ko0e evtoxurrlq unoypetirverot vo ),etroupyei p6vo orov ppe- Oet xovro oTov unoKtvllTll Or evtoXutdg Lettoupyoriv npoo6Svovtog ouv6uo- opro0q 1-reroypocptr<cov nopoy6wt :v, e(re. Oettxriv eire opvrlrir<cirv, nou etreylouv rov unoKtvqrq r<ot xot'en5- Kroon rrlv 5xgpoorl rolv yovt6(ov. O unortvqrrlg e(vot ri 06o11 6nou, srriv ovotXrrl Xptoprm(vq,.npoo65vowot or Boorr<oi peroYpoqtKo( nopoyoweg 6tot oore 11 R- NA no).upepdorl vo Bpe( tov unortvqtq l-lr'rrg dvog evtolutrlg 6teya(per trlv 6vop[q o' 5vov unoxtvrltrl nou'1-rnope( vo Bp(or<etot oe onotodrlnore ondorooq npog onotodfnote ntreuPo tou: H Lettoupy(o tou evtcnlurrl nepil.o;rBdvet otr-I'q)'ent- 6pdoetq pre rr1 Boorxq ouoreurl o--ro orotle(o rou Kev- rpu<o6 unoxtv4tq. Ot evtolut5q e(vot nol'upepq, 6no-tq Kor ot unoKtvrltdg. KdnoLo orotle(o Bp(or<owoL tdoo o' evroXur6g preydl'ou Bel'qver<o6q 6oo t<ot o' evtoXur6g nou.[p(orowot t<owd orouq unoKtvrlrSq' Kdnoto [eXo- p,*d -o,Xt(o nou Bp(oxowot t<owd oroug unortvqt5g 1-rotpd(owot Ue rouq onopoKpuop6voug evtolutdg rrlv rrov6qto vo Letroupyoiv on6 dtdqopeg onoordoeLg Kot npoq onoro6qnote npooovotoLto;-t6 'Etot q 6tdt<pt- o,1 1r"-oi,l evtcrlw6v preydLou t<ot ptr<poti Bel'rlver<orjq 6ev e(vot ooQnq. O ouotooru6q p6).oq tou evtolutq (ootg e(vot 11 o0- Enorl rnq ouyr<6wproorlq rou evepYonotrlrrl rowd otov ,noo,u,'l-tn oe dtopr6ptptrlorl crs. t-loMd netpdproto 66et- [ov drt to en(ne6o qq 6xqpoorlq tolv yovr6(ov (611]'o- O(, q tol:qro rrlq pretoypo<pqq) e(vot ovdtroYo ile rov xo0op6 opr0pr6 to:v 06oeov npdo6eo4q rou evepYo- noulrq. Ooo neptoo6tepot evepYonotnr5g e(vot npoo- 6e6epr5vot oe Uto 05or1 evtolurq, t6oo uqJn^'6repo ro enine6o tqq 6rcPPoorlg. To pLBootoprtr<6 RNA rou evtoxurq rou Xenopus :tt."i,; 0r EvLoxurdq Aeuoupyouv Au[ovovrog rr1 luyr6vrpolol rtrlv Evepyonorrlrtirv Kovro orov YnoxLv-l- 573
  • 66.
    At#EvtoXurr'19 'Evog evtoyurrlg pnope(vo ),etroupyqoetrpepvovrog nporeiveg rovro orov unorrvqq.'Evog evroxurlg rr evog unoxrvlrnc oe Xoptoro xurtrlro DNA dev otrtrr1trentOpouv 6nog oro (f), oIIo pnope( v' otrtrrltrenrdpooouv orov ro 6uo popro elvor ouvdedeg6vo o),u0.- droro 6ntog oro (B). /aevls pnope( vo evepyonor{oer rn Ueroypog4 on6 rov unoKrvnrn rnq RNA no).uprepdonq L Aurrl 11 66- yepon e(vot oletrrd ove[dpqq on6 q 0dor1 r<r e[o- rolou0e( or6pro K[ ov o evroXurqg ogorpeOe( on6 ro Xpop6oro;ro ror tonoOerq0ei go(( Ur rov unoxrvqrq rou o' 6vo rurllr6 nLoopr(dro, evci ow[0ero, no6et 6rov o svroXuqq Kor o uTroKr.v4rrlg Bp(oxowor oe 6ro- gopettrd ntrootrl(6ro. Av 6prog o evroXurrig ronoOe-rr1- 0e( o' 6vo nLoopr(dro ouvde6ep6vo o),uor6ord pr, 6vo de6repo nLoop(6ro nou neptSXer rov unoxrvrlr4, 11 6vop(4 e(vor oXe66v 16oo qnore).eoproru<{ 6oo ror 6rqv o evrolutrlg Kor o unoKrvqtrig Bp(orowor oro (6ro r<ur),u<6 ;r6pro 6n<rlg go(vetor mrlv EIK0NA 20.16 (;roLov6rr, oe our4 r1v nep(nroon o evroxur]g ene- vepye( orov unoKrvrlui rou oe dropr6pqoorl trans). Au- t6 uno6rlLrirver 5rt ro x0pro XopoKrnprorrr6 e(vor 11 ew6nro4 r1g nporeivrlq nou e(vor npoo6e6epr6vrl fiov evroxur4, q ono(o ou(dver rlv nt0ov6tnro rou evroXurri vo 5).0er o' enorp4 Ue Uro npo:reiV4 npoode- 6epdvq orov unortwlr4. Eg6oov nparreiveg npoodedep6veg o' 6vov evroxu- 11 oe on6oroorl oprer6v kb on6 6vov unorrwlrl oMq- ).enrdpo0v dtrleoo pre npcoretveg npoo6e6e;rdveg rowd ornv oqernp(o, 11 opydvrrlon rou DNA np6ner vo e[vor opr<erd eu6Lu<rr1 rilore vo enmpSner orov evroXwq vo Bpe0e( rowd mov unorwqri. Aw6 npounoO5ret 6n to evdtdpeoo DNA oplpror((er pto Ueyqln nOrlfud". T6roeg 0r1Lr5g rilouv nopornpnee( oneuOe(og orqv nep(m<oor1 rtov Bomrlprorciv evrq4urriv. Tt neprop((er qv evepy6r4ro ev6g evrqXurl; Tunxd enevepye( orov nlqouiorepo unoKr.vrprj. Ie opropr6veg neprmc0oerg o evroXutrig Bp(oreror pero[6 60o unoru- vqrcbv oMd evepyonoref p6vo rov 5vov qn6 roug 613o Ue Bdon er6rrSq nprr:te'tvu<5g oM4lenrdpdoerg prero(ri r<ov ouprnl6rov nou e(vot npoode6e;r5vo oro 66o qror- Xe(o. H 6pdoq rou evroxurrl pnope( vo neprop[(etor on6 6vqv provorq - 6vo ororXe(o rou DNA flou nopep- no6(er qv en(6poon rou evroxur{ oe unorwqr6g ndpo on6ro govoq (BI5ne Ev1tqra 10.12). 20.l I H 'Ergpoon Tov fovt6iov lxeri(eror u€ Tnv AnopeouAioon o H onoge0utr(too4 oro 5' drpo rou yovr0(ou eivor onopoir4nl yro q per0yp0qrl. H pe0ul(cooq rou DNA e(vor 6vo on6 oprerd pu0pr- oru<d ouprBdwo nou enqped(ouv 14 6poor4pr6rqro ev6g unoruvrlrn. H preOuL[ooq orov ufloKrv4rq pnopei vq nopegno6[oer q peroypog4 xor q ogo(peorl rov peOuLoprd6cov e(vor onopo(rqrq npoun60eorl yro vo evepyonor40e[ o unoxrvqfig. Aur6 ro rporv6prevo dler ge).eq0e[ ro]d oe unorrwp5g 16oo qg RNA no].upre- pdonq I 6oo xqr qg RNA nol,uprepdoqg il. Irqv npoy- 1-rorrr<6rqro, q pre0uL(crloq e[vor 5vo ovrrorponr6 pu0gronr6 ouprBdv. Evepyonore(ror on6 rpononor- 4oerg rtrrv rorovciv, prero[ri dMcov qnooreru],(oon Kor pe0uL(oor1 (BI6ne Ev1nya 28.1 0). Me0u).(ooq en(o4q ougBo(ver Kor oq enryeverrx6 rporv6;revo. Irqv nporerpSv4 nep(moon, n rponono[- rloq gnope( vo ouprBe( er6xd oro oneptrroro(rodpro rl ro odpro, o6qyriwog oe 6togop6g prero[ri rrov 6rio oMrl- l,opr6pgov or4v en6prew'1 yevrd nou prnope( vo npoKo- L5oouv 6rogoprig mqv 5r<gpoon rov norprrriv ror pqrprxcbv oM4lopr6p<pcov (B),6ne Ev6trya 29.8). Ie our6 ro re<pdloto e[erd(oupre ro pdoo pre ro ono(o 11 preOu).(rooq en4ped(er rn Ueroypqgrl, ro ono[o e(vot ove[dprrlro on6 ro ov or pe0u].oprd6eg npom60q- 574 KEOAAAIO 20 H Meroyporp4 oroug Euxopuoteg Boorxeg evuoreq
  • 67.
    ndqn us Msplnam P8tho . r, n oeotp6Orlrov rog pu0prtortr6 q entyevertr6 ou;t- t-., i ;re0ul(oor1 oe unorwqt6q qq RNA notrupepooqq ,,,u[olur, oe dtvoureoti6to CG (ovoq5powrot rot 6t- ,."-ol!oiio,o "CpG,). H r<otovoprl tolv. pre0uoprd6tov .-l.orl vo e(etome( xpqotponotowoq riv(utrto neptopt- , -.u .o onoio n6mow b6o"tq-*6xoug nou neodxoyy -: 5rvoux.treot(6ro CG" Arjo r[not neptoptopou ouYKpt- .l,.roL ornv , '' To roocrtrt(oprepq nou Xpnot- -.:noLqOqxov oto ne(pogo elvot ev(upto to ono(o Ir.-oru tqv (6ro oLr1ou1(o-mo1o.oro DNA' o)'Ld .,cuv 6to.popertrd onotetrSo;toto ovdtroyo Ue rnv Ko- lf,Tool Pre0u(tooqq tou DNA' --i;;(rt- Hpall n6met trlv o)'rllouxto CCGG (vod- .rroi,l olLnLorXlo p6vo tou ev6q xtrtrlvou tou DNA)' :-Joo ov q 6e0tep1 C e(vot preOutrtcop6vrl' to 5v(upto :, irnoptl n6ov v' ovoyvop(oet q 06oq' Aw(0eto' to ,,lrJ" Mspl n5nter trlv (5ro 05or1-or6xo ove[dpqto , .. -n, xotdmoorl pe0ul'(rooqq orrl ouYKe-Kptp6vq C' :-'renrq, to Mspl lrnopei vo Xpnotponotn0e[ toore v' l, "vtrpi*"tv 6treq ot oMrlou{eq CCGG rot to Hpall ..: vo ro0optote( ov e(vot ge0utrttolt6Ye.S . - ev to ONA e(vot ope0u}'it''lto, to 6rjo 6v(upro 0o 6n- - ...lryqoouu ttg (6teq (oveg neproptotrrot'^Ito geOu- ' --.uo DNA, 6pcoq,'oi -pot'onotrl;reveq.06oetq 6ev -.-::cwoL on6 to Hpall' I-to r<d0e tStoto 06oq' 6vo pe- .'.-..po x).dopro Hpall owtro0tord 60o rldoproto M- :: -r ' 6(vet 6vo noPd6etYPto' -ctr'o yov(6to nopouotd(ouv 6vo np6tuno oto : t.c rl i<otdoroorl geOu)'(oor1q eivot oroOeprl ortg ne- , - ro.tptq O5oetq oM6 notril'et oe 6Meq' Kdnoteq 06- -'-, ;i;;, preoultrrrpr6veg q opreOuL(oteq oe 6trouq toug -.. ;. ALLeq O5oetq 1;rio' petov6trlto) e(vot pre0utto;r6- ': ce Loro0g orouq ono(ouq to yovl6to 6ev errppd(e- -: ovo e(vot opeOu}'(oteq oe toroUq 6nou to yov[6to . ..,-tutpVO. 'EtoL, 6vq evepy6 yov(6Lo prnope( vo nept- : lQe[ og unotrre0u].tco;r6vo' ffi Ztivn onorletmtxd mo HPall E ffi owxoOtmo rtq (tirveq tou ttlslt ffi ES- zOues qnor[etmtxd oto Msol | ] I I = Ueeultogdveq Odostq I L llr ZoVTI omv iDro 0eorl - L l =oue0u(orn06on .l 0r edoetc KoBowot oveqoprrlro ono n1 geOul[oon pi{aMg."ffi t Meouhtog6vo Mspl oxl [eoulrr,lu6vo[ ]lo ccdc -_-cgcq H oue0utr(olu 06on KoPeTot U npilrr r,rre I sa?fffraacffi';'-ffi;ffiGGCC lle H pe0utrttoP6vr1 0doq 0ev rdPerot To 6v(upo neptoprogotl Mspl n6nret oheq rLg otr- ,-ouxieq CCGG, e(re e[vot f-ie0u]'Ltogeveg oi1 deurepn C' e(re - ., ;vii,; Hpall nenret p6vo ro ope0utriolro rerpoueprl CCGG' E|K0NA20'lEToonotetr6ogotoflqn6UnqueMsplrotHpall ;;ffiil;i ue nlenporpdpnorl to:v rtroopdrov neptoptopot oe nr1xu1. I-letpdproto pte to 96pgot<o S-o(orund(vq nop6Xouv efrfr"on on66eiEn Yto ro yeyov6q 6tt 11 onopre0u)r(oor1 fi;;i; oonvno". oe 6rqpooq tov^vovt6(tov' To <pdp- Io^o ,uoruo.tivetor o'o oNR orq 06o11rqg r<ur6fur1q [;i;t";;ffi vo 1-reeumoee(' Aur6 o6qve( orqv e;re6vt- ililt6r"ra;-rr'e6o",u oro DNA pretd qv owrvpoq{ (pMne Etr<6vo 11.10)' Ot tpoworuntreq entmdroetq rnq 5-o(or<wt6(vr1q ne- prLoprBdvouv qv enovtov( oLl'ovtirv *ly ^:TT.::l b,o,poponohonq rov t<urtdp.tr:v' l-to nopd6etyuo' uurKq ^,:tiobo endyowot on6 np6Spopro pr1 purr6 ry11>: to qdppor<o evepyonoe( en(oqq yov(6.to oro otonnpo voou6otouo X, yeyov6q nou ougQove( ;re r1v t66o 6tt f,"dilffi ite,Jxr"ns oxet((etot ;re q 6pocnrlpt6- rnrorou xpouooouoroq' -_ !_' i*rOqind -o ,o e[etd(oupre qv xotdoroorl ueou- )r(toorlg ttr:v p6vt;ttov yovt6(tov, ;rnopo[ge v' owtnopo- |l.L" tir" ils' en,mooe,q rnq etoqY oYrlq peo uhtto;r6vo u [;;fudou DNA ot 'eo r0rropo [evtor6q' T6roto .,r,pouo* deftvouv pto ooqrl ox6oq: to-pre0utrtropdvo VouiOto e(vot ovevepy6, evr'il, ow(Oero' to otrle0u(toto yov(6to evePY6. ...!'- noia eivot 11 Srroorl qg unope0utrttopr6vlg nepto- fiq; Ito 60potopro youi6i" o-o"tpotpffiq rtov epuOpo- et6dlv r<urrdpov Tnq r6tog, q unopreOutr(tooq neptopf. (etot oe 06oerq nou ercrefvowot on6 -500 bp ovo6xd ;J.;il;;o ht o ro 60o evnr.ro o vovi6to 6tog -500 [p ^"e"ri,^a on6 to 6e0rcpo' oSoetq unope0u]'[<oonq ,Iapi" * tt o).6xlr1 pr1 tr1v neptoxq' ou ;rnepiloptBovo- uirJu' oo, tou otoorrl lroroq ovdpeoo crro 1?:i9:: I ::; i,oxn .ns uno ge0ul(trrorlq ou prnhtet. Fl.qv fl eptoxn ili}*,*a euotoorlo(oq orq Dndoq I (Blene Evot4ra io.l'l).'nrtO'to onot6l'eopro orrlp((et qv dnorl'rq 6tt q ,""lrlgriotr1 e(vor xopoKrnptoru<6 proq neptolrlq nou nrpi61r, 5vo pretoypoq6;revo yov(6ro n Yov(5ro' 'Onolq u"-"lilga ollov6q fin xpouqr(vq' cpo(vetot 6rt n [i"rol" ui,eulopo6tov gxet((etot pe tr1v xov6tnro ue- toypogf,q nopd ;re qv (6to rn UeroYpogn' To np6B)rr1po ornv eppqve(o tqq yevtr<dq oX6onq u".oi,i unoueeulioong oi t'"pyono(nonq ttov yovt6itov tiuo, b., oto tpow6pevo etr'tntr5retot p6vo pto petovorn- io tL" rrtgutuo;r5vtov OSoecov' l-lr0ov6v 11 rotdotoorl -is deli-"rlg e(vot r<p(otpr1 oe ouYKeKpu6veg 06oetg 11 oe pto neptoprog6vrl neptol4' En(onq' n el'dtcoon 20.11 H'Extppoorltov [ovtd(tov lxeti(eroL Ue nlv Anope0utrfoon 575
  • 68.
    oro BoeU6 Ueeul(oong(ri or<5Un r<or 4 nLrlprlg oqo(pe- orlrov pe0ul,oprd6cov on6 rdnoro rgnpo DNA) e(vor ;r5- poq Uroq doprrrlg olloyrlg onopoirqtqg yto t4v e[5].r- En qq peroyporp4g. Iuyxerpt;r6vo, r1 onotrre0ul(oor1 mov unorrvqrrl [ocug efuor onopo(tr;q yro vo rov roromrioer 6ro0rior- Uo yro r4v Svop[q rqq Ueroypogrlg. l-to nopd6erypro, mo yov[6ro rnq y-oqorp[v4g 4 nopouo[o rcov ge0ul,o- prd6cov m4v neproXrl yripot on5 qv ogetrlp(o, peto{0 -200 Kqr +90, KoroqrdAler rq preroypoqrl. H o<po(pe- orl rcov rptc0v pe0ulopddtov nou Bp(or<owor ovo6rrd rnq qqrrnp(og, 11 rcov rpuiv ge0u).opd6ov nou BpG oKowor xo0o6rxd r4g, 6ev o(per r4v roromoLrl. H ogo(peor1 6ltrtv rtov pre0uloprdsov, 6prcog, ennp6net fiov unoKrv4rq vo ),enoupyrloer. Iuvenc0g, npoun6- 0eor1 yro rn Ueroypoqq iocog elvor q nopouo(o ptog opeOul,(ror4g neproplg orov ufloKrvrlq (B).5ne Ev6t4- ta 20.12). Qor6oo, undplouv e[orp5oerg oe outq tr1 yevrxrl oX6on. Meptrd yovidro er<rppd(owor or<6pr4 xor 6tov e(vot nolri trreOuLrcopr6vo. H 6norq oX5or1 perolti ;reOuL(oo4g ror 5r<<ppoo4g trov yovr6(rov 6ev e[vor ou<ouprevrrrl, oMd yevx6g rov6vog e(vor 6rt q ;re0uL(cooq onorp6ner qv 5r<<ppooq rcrlv yovr6itov, evri q ono;re0ul.(coor1 e[vor ono- pofuqr4 yro rrlv 5r<gpoor1. 2A.l2 Or Nnoi6eq CpG Eivor Pu0prorrroi Ir6xor * 0r vlor0eg CpG nou pplorovror orouq unoKrvrlTe g rotv rdLoou- ororo exqpo(o;revov yovrd[rrrv e(vor onope0u],urrpdveg. * N1o(6eq CpG pprorovror en[oqg Kor 0T0uq unonvlrdg opr- oprlvu.rv yovLdirov pe roroerdLrrl exqpool. * To ov0p6nrvo yovL6lropo nepreXer-29,000 vqo(6eq CpG. * H Ue0utrlooll luog vqo[dog CpG onorpener qv evepyonoqol rou oXel(opevou unoKrv1Tl. * Koroorotrrl nporotreiror ono nporelveg nou npoodevovror oe pe0utrLopevo (euyri CpG. H nopouo(o vqoidrov CpG (CpG islands) oug 5' ne- proX6g opropr6valv yovr6(ov ouv66eror pe rtg enrmri- oerq rnq peOul,(ooqq mrlv 6r<<ppooq rorv yovL6[ov. T6roreg v4o(6eg touronororiwor on6 r4v nopouoio ou lq pSvq g nu xv6ultog rr1 g 6tvo u r<}.eorr6rxrl g o).).r1- ou1(og, CpG (Cpc : 5'-CG-3'). To 6rvour<Leor(dro CpG ouvowdror oro DNA rotv onov6ui.rrtrrlrv oe oulv6rqro ;r6Lrg -20% ttlc, ovogev6- pevrlg ond qv ovoloyfo rotv (euycov Bdoerov G-C. (Au- td (orrrg ouprBo(ver enerdrl 6rov ro 6rvoux).eor(6ro CpG e(voL ;le0ul.rcopr6vo orr; C, 11 our6protrl onopr(vrrlorl trig prd0uLo-C q getorpdner oe T, 4 ono(o ov enr6top0o0e( eoqoLprdvo erodyet ;uo pretdMo(r1 nou ogorpe( ro 6r- vour}.eor(6Lo). Ie oproprdveg neptoldg otm6oo, 1 n_- rv6rrlto tclv 6rvoui<LeorrS(ov CpG grdver orrlv nc.- B),endprevrl trprl, dqAobrl ouorocnKd OexonLoord(et: ouyxpLttxd pe ro undl,orno yovr6(ropro. To brvourcLec:.. 6Lo CpG oe oureq rLg neproXdg yevrr<d e(voi ogeOu,L*- TO. Aurdg or nlorjoreg oe vrlo[6eg CpG SXouv ;r6ori r:i. prerlx6rrlro oe G-C -60%, nol0 preyotrilrepq ond :: yevtr6 DNA (pdoq ttprl 20%). l-lo(pvouv rq piop<pr1 r*- pdrrov DNA ;u'p<ouq 1-2 kb.Ito ovOprrtnrvo yovr6(o*: undpXouv -45,000 r5-roreg vrio(6eq. Opro;rdveg Bc- oKowor oro enovol.oprBov6pevo o-ror1e(o Alu, nL0o,,:, e[oLt(og rnq uqun].nq neprexlr6rlrdg toug se G-C, - oi.),q).ouX[o rou ovOprinrvou yovrdrcrtprorog entBepc . rrrveL 6rt, ov elorpeooriv or ouyKeKprp5veg vqo(6eq, or:3, pSvouv -29,000 vr;o[6eg. To yovr6(cr"lpo rou nowrKC- nepLdlsr fuydrepeg, -15,000. l-iep(nou 10,000 ond ..: npoB).endpreveg v4o(6eg Kor oro 6Uo e(6r1 qo(veroL c ewon((owor oro n).o(oro otrI.qAoulurtv ouwqprlpdv*. petol0 rrr.lv eL6crtv, yeyovdg nou unodr;Lrrtver 5-rr ou;:: (oog e(voL ot vrlo(6eg ;.re puOtrlrorrr<r1 orlpoo(o. H 6ol- rqg Xpropor(vnq onq ouyr<er<ptpr6veg nepLoXdg epec, - (et oMoydg nou oXer[_(owor pe rr]v dr<gpooq r<,DV yc. - 6(rov 6rov or vqo(6eg CpG e(voL ope0uA(roreg (BAo.. Ev6n1ra 28.11): lopor<r4p((eror ond 1o;rqL6repq oL- .. r<6wparor1 trlq Lordvqg H1 (nrOovdv oqpo(ver 6l 11 6c-- e(vot ).ry6repo oupnoy4g), trreydlo Bo0p6 oxetu).[cocr : tov dMotv torovciv (Xopor<rrlptorLx6 nou ouvrlOcoq qt r[(eroL Ue rrlv 5r<gpool rtrrv yovr6[ov) r<or nopouo(o e :- oov unepeuoioOrltirlv orq DNdo4 r1 oXeddv ono, . Loypr6.vorv on6 or<ropepfl rorovc6v (r<ord ro ovopevou:- vo yro evepyo0g unor<Lvrpdg). H nopouo(o peOuLro.rp,:- vorv 05oeir.lv CpG onor<tre(er on6 to vour<Leoocupror.: qv nopoMoyrl tor-6vqg N2A.Z. Aprerdg eop6q, v1o(6eg n),orioLeg oe CpG (errvo,, pr6}.rg ovo6u<d rou unoxLvrlrrl rL enercre(vowor xo0o6Lr: pSoa orq ;reroypo<p6trrevq neprolq dnou oro6tor: ,,OBrlvOUv". H ::..rr' .l . . : , Orgy6p(vet rqv nUXVdrrlrO r*. 6tvout<l.eol6(rov CpG or prcl <(yevlKr1" neproXrl rou yo,,'.. 6urr;rorog Kor or puo vr;o(6o CpG nou ovoyvc,-lp(mn<i ond qv oMrl).ou1io tou DNA. H vrlo[6o CpG nepLBo; et r1v 5' neptoXrl tou yovt6(ou APRT, ro ono(o extppc. (eroL 16too0ororo. O).o ro dLolerptorLr<d yov(6ro nou er<gpd(ovri. t6Loor-iotoro 6Xouv vrlo(6eg CpG, nou ovrrnpoocL- neUouv nep(nou ro 50% tov vqo(6otv. Ot un6),olnec Bp[or<owoL orouq unoKrvr;r6g yovr6(rrrv ;le roroetdLr<- ptiOptoq: nep(nou 50% outrrv rotv yovL6(ov dlouv vr. o(6e9. Irrg ouyrexprpr6veg nepLnro-loerg, or v4o(deg e.. vor otrle0u).(oreg ove[oprrlto on6 r4v rordorool rri 5xqpoorlg rou yovt6(ou. Iuvencbg, 11 pre0ul(cooq tcu. vrio[6ov CpG 6e oler((eror pe rn Ueroypo<pu<q r<orc. oroon rotv rmoeLdrr<otv yovr6(ov. H nopouo(o opeOt. L(ori,.rv vqo(6ov nLorjotcov oe CpG (oog e(vot ovoyKo.: oMd 6ev e(vor enopr<r1g ouv0r1<q yro Ueroypoqn. Apc 11 nopouo(o opreOuL(ortov vqo(6urv CpG pd).ov unc 6q).olver 6rt 5,vo yov(6ro e(vor 6uvqrrxd evepy6 nopc 6tt ovon6geurro 0o preroypo<pei. l-lo).L5g vrlo(6eg nc- 576 KE0AAAI0 20 H Meroypog1 mouq Euxopudrreg Boorxdg 6woreg
  • 69.
    Y-0Q0tptvn iililiillllili 500 1000 bP APRT iii lliiil liliilllllfillfillfrlilllilililliil l1 i I 5', E[rirvLo E[tivLo -:: .': H runtq [uKVoTr]To Yto 10 DtvouKtreoT[0to CpG rro DNA rov 0rlloolx6v ervot -1l1 00 bp, onoq Qo[veTot Yto T0 ,ovtDto qg y-orpotptvrlg. Ie pto ntrouoto 0e CpG vn0(00, 11 nurvo- .-ro ou[ovet oe > 1 0 6tvourieor(dto/1 00bp. H vrloLdo oro yov[6to :PRT (er<tvo -100 bp ovodtro 0n0 Tov unortvlq Kt tKTttvtTot ..100 bp Us00 0To yovtdto, Ko0e xo0eq ypopprl ovrtnpootdlleutt .,/o DrvouKtrtoT(bto CPG i[vor opre0u].(oreg oto (aro pre0u].ttovowot oe Kurrclpt- <eg oerpdq oe r<oMt6pyeto (r1 oe ronotouq xopx(voug)' Aurd 0o pnopo0oe vo oXer((eror Ue rnv ovtt<ov6-rqto :urci.lv rtov oeLprrtv vo ex<ppdoouv 6leq ttq Letroupy[eq rou XopoKrrlp((ouv rov tor6 on6 tov ono[o npo5plov- :oL VnopXer dvo oo<p6g nopo6etypro Yto rrlv toXupq qdoq peto(6 qg ;re0u).(oor1g tou unor<tvqrrl Kot rrlq 3,Kepoonq to:v yovt6[tov: ot vrlo(6eg CpG pre0u].L6vov- oL oro oSpovonotq;-rdvo Xpopr6oopro X ttlv 0r1troo-rt- <rrrv (B).dne Evoqta 29.5). H pre0ul.(roor1 qg vqo(6oq CpG pnope( vo enqped- Joi rrl Ueroypoprl. Or 60o nL0ovo( prqXovtopro( e(vot: . H peOu).(toon Utoq 05oqg np6odeoqq yto ronoLov nopdyowo prnope( vo epno6(oet rrlv npdodeorl tou' Aw6 oupBo(vet orrlv nepimolol qg np6odeo4g oe ;.tto puO;rtmtr<r1 nepropl 6togopettr<r1 on6 rov unort- vr1rrl (BL6ne Evoqta 29.9). . H Fe0u(roor1 trrnope( vo nporoltloet qv np6o6eoq eL6txtr.lv xotomotr6rov oro DNA. KotootoLrl nporoLe(tot on6 onotovdrlnote on6 :oug 6tio t0noug npoteivolv nou npoo66vowot oe pe- :utrLopdveg oMqLouX(eq CpG. H nprorelvq MeCPI yto ,'o npoo6e0e( oro DNA onoLre( tqv nopouoio opretolv -re0uLoprd6ov, evc0 q MeCP2 Kot pto ott<oySveto oXett- loprevo,tv npirtte[vcbv prnoporjv vo npoode0orjv oe 6vo *dvo pre0ulttrr;r6vo (erjyoq Bdoeov CpG. Aut6 e[r1ye( '.'Lor[ q 6vop[r1 rnq UeroYpo<pr1q npo0no0Stet pto ne- cLolii eLerjOepq on6 ;re0u)r.(coor1. H np6o6eor; nporei- ,'cov onotou6rlnote on6 rouq 60o rinoug eprno6((et r1 reroypo<p1 in vitro on6 6vo nuprlvtr<6 er<1itrtopro. H MeCP2, q ono(o t<otoot5Met dpeoo rrl UeroYpo- cr-1 oltrqLentdptrtwoq pe o[pntroxo orov unortvqrrl, in[or1q oMqtrentdpo Ue ro Koroorotrttx6 otiprn]'oxo Sin 3, nou nept5Xet evepy6rqreg onoqraTul[oonq srowir (BLene Eu<6vo 28.24). Auq q noporqpqoq ouvdeer ilio i ir,ouq rotooto.ttt<cilv tpononot{oeov: trre0u}titrurl tul DNA r<ot onooretutr(toorl rov tqrovtilv. Evdr or unoxtvqr6g nou nept6Xouv vrlo(6eq CpG (-60% CpG nur<v6trpo) 11 6ev SXouv epntrouttop6 oe CpG (-20% CpG nurv6tqto) yevtrd nopouotd(ouv qroxn ouoXdttotl ;reto[0 qq r<ordmooqg qq pe0u- L(toorlg rouq Kot rnq UeroYpqQnq, ot unorwqr5g pttog rpfur1g td(r1g <po(vetot 6tt pu0g((owot on6 tr1 pe0u}'ttrl- on rov CpG. I-lep(no u to 12o/o tcov yovtd(cov tou ovopri- nou nept6Xouv rtq xol,oti;reveg (qoeeve(q) vqo(6eq CpG ("weak CpG islands"), ortq onofeq 11 nurv6trlto tov CpG efvot -30o/o, evdtdpreorl peto[ti rolv 6rio dMtov rd[ecov unortvqrcilv. Autd to yov(dto Xoporcrrlp((owot on6 ro'Xupr1 ow(mpotp4 ouol6ttoq peto(0 qq peOu- L(rrlorlg tcov CpG rou unoKtvnrri r<or t4q np6odeorlq qq RNA no)ruprepdorlq ll. H onouo(o ;re0u}'opd6ov ouv66etot ;.re tr1 5x<ppoorl rcov yovt6(tov (ri touLdXtorov Ue rrl 6uvot6rpo yto 5x- qpoon). Qsr6oo, 11 r<otdoroorlrqq pre0ul[tooqg 6e gno- pe( vo 0eo:pr10e( cr.rq 5vo yevx6 pr5oo yro tov 5l'eyXo rqg 6r<rppoolq rov yovt6(rov. It4v Drosophilla melanogaster (ror dMo e(6r1 Arm6pov), to DNA e[vot etrdXroro peOu- tr rcopr5vo (nop6n 6Xet ovoyvroptoref pto pe0u].otpovo<pe- pdon, 11 Dnmt2, tqg ono(oq 11 oqproo(o 6ev e(vot ooAnq), evc0 oro vqgorrildrl Caenorhabditis e/egans xot orn (6pn 6ev e(vot xo06trou ;re0utrropr6vo' Ot dMeg 6to- qopdq prero[ri ovevepyoti n evepyo0 lproprotffiq qo(- vowor ow(orotleg pe ttg 6to9op6q ot ono(eg nopo- qpo0wot oe e(6q nou eprrpov((ouv preOutr(toor1. Iuve- nd,g, oe owo0g rouq opYovtopotig, 6trot ot p6trot rrlg pe0utr irrlorlq mq onovduLotd 6Xouv owtKqrooro0e( on6 rdnotov dMo Pr41ovtotrr6. Tpetg oMoySg nou ou;rBo(vouv mo evepyd yovbto efuot: . Mto unepeuoioOrFn 06or;(erq) eyroO(orotot t<owd orov unorwrltq. o H Xptouorivq groq neptoXqg, ouprnepil.ogBovop6vng Kor rnq petoyporp6prevrlg neproXrlg, y(vetot nto euo(o0r1t4 mr1 Dn6or1 l. o To DNA rng (6ro9 neptoX(g e(vot unoge0ul'ttrlp6vo' 'OLeg outSq ot oMoy6g e[vot onopofurlteq yto tq preroypo<9r1. 5'*r* e*$ E(rirvto E[titvto 12 2O.13 tlegirntlln An5 rg rpetq euKoputr:ttr6g RNA nol'utrrep6oeg, 11 R- NA nolupepdoq I pretoypdqet rRNA, q RNA no}'upe- p6o4 ll pretoyp6ger 6o;rtxd yovfdto yto mRNA xot nopouotd(et q preydt6teprl no u<t tr'o pto p<p(o npoi6wcov ror 4 RNA noluprepdoq lll peroypdqet ;rtrp6 RNA. To 6v(upro 6Xouv nop6proto 6opt(, pre 56o preydLeg unopo- vd6eg ror noMdq pxp6tepeg unoprov66eg. MeptrSg unopovd6eg e(vot row6g xot oro tp(o 5v(upo' : !-r':-l tll
  • 70.
    Kop(o on6 nqrperq RNA nolu;repdoeg 6ev ovoyvo- p((er oneu0e(og toug unorrvqrSg qg. Mro r<orvr1 opXrl e(vor 6n or Ueroypoeu<o( nopdyoweg 6Xouv rrlv r<ripro eu0[v4 yro rrlv ovoyvri:proq rov ororXeiov Xopo]firlpr- orxtig otr},r1}.ou{og xd0e ouyrexprprdvou unorwqrl ror ge q oerpd roug npoo66vouv tqv RNA noLuprepdoq Kor rrlv ronoOerqoouv oosrd srnv oeernp[o. Ie r<d0e rUno unorrv4rrl, to orcrogep4 rrov rmovc0v np6ner v' orporpe0o6v ri vo geror<wr10oUv. 'Enemo ouvop1-roLoye(- tot ro o0pnloxo Svop(r1g on6 pro oepd owrdpdoeov or4v ono(o [eXtoproro[ nopdyoweg ouvdriowor (n oqn- vouv) ro orlpmLoro. O nopdyowog TBP ononekor yto tqv dvop[q ror on6 ng rperq RNA no].u;repdoeg. Ie rd- 0e nep(mcoor1, nopSXer pto unopovd6o ev5g Ueroypo- gtr<ori nopdyovro nou npoodSvetor rovrd orrlv ogeqp(o. 'Evog unoruvnrnq rnq RNA noLu;repdonq ll onore- i.e(ror on6 n).r1Oog BpoXS<ov morle[cov or4v nepropl ovo6rrd on6 rqv o<pen1p(o. Ie r<d0e orotle(o npoodd- vowor Svog 4 r1 neproo6repor Ueroypogrxo( nopdyov- teg. H Bootrf ouoreuri, nou onoreLe(ror on6 roug nopdyoweg TFII, ouvopprotroye(tor orqv oqerqp(o rr ennp6ner or4v RNA noLuprepdo4 vo npoo6e0e(. To rour( TATA (ov undpXeQ xowd orqv oger4p(o Kqr rl ne- ptoXrl Svop[qg (lnr) oxpgriq ornv oqerqp(o eu00vowor yro rrlv enil.oyrl qg ocoonlg ogerqp(og orouq unoKrvn- 169 yro r4v RNA noluprepdorl ll. H TBP npoo65vetor dpreoo oro row( TATA. Iroug unoruvqrdg Xcopig TATA Bpior<eror rowd orqv oger4p(o ;r6oal t4g np6o6eorlg rnq ornv lnr 11 ro0o6rrd oro DPE. Merd qv np6o6eoq rou TFllD, or 6Mor yevrro( UerqypogrKo( nopdyoweg yro rrlv RNA noLupepdoq ll ouvopl.roLoyoriv q Boou<r1 peroypogrxrl ouoreuq orov unoKrvrlq. 7U.o ororXe(o Tou unoKrvnrrj, nou Bp(oxowor qvo6rxd qn6 ro Kouri TATA, npoo66vouv evepyonor4r6g nou oLL4Lenrdporiv pe q Boorrrl ouoreurl. Or evepyonorqrdq xor or Boou<o[ nopdyoweg onel,euOepri:vowor 6rov q RNA nol,u;repd- or1 [exlqoa qv enrp4r<uvor1. H CTD rqg RNA nolu;repdoqg ll rpoogopulrcivetor rotd trlv ow(6poor1 6vop(49. l-1opri1er 5vo orlpe(o eno- qnq yro nporeiveg nou rpononoro0v ro RNA prerdypo- rpo, ;reto[ri d],].rrlv ro 6v(upro oxnponoUo6 14g 5' xo).Unrpog, nopdyoweg ouppogrlq Kqr ro oriprnl,oxo ene(epyoo(og rou 3' dxpou. Ko0c0g rl RNA noluprepd- or1 trleroruvekor 6ropr6oou rnq Ueroypocpu<rig provd6oq, ro orcroprepri rov torovdlv np6ner vo rpoTronorq0o0v yro vo enrp6qrouv rrl 6t5leuoq. Ot unorrvrlr6g pnope( vo 6reyep0orjv on6 evroXur6g, oMqLouX(eg rxov6g vo 6pouv on6 preydLq on6oroorl Kor fiouq 6Uo npooovoroLtoproUq, efue qn6 q pro efue on6 qv dMrl nleupd ev5g yovrdiou. Or evrolurSg en[- orlg onoreloUwor on6 opd6eg ororle(trlv, oMd e(vot nro ouUnoyr[q. Kdnoro orotXe[o Bp(orowor 16oo oe unoxwrlr6g 6oo ror oe porprvorig unor<rvqt6q. Ot evr- olurdg nr0ov6v Lenou pyo0v ouvopl.roLoyr6wog 5vo nporeivrr6 oUpnl,or<o nou otr).r1l,enr6pd pre npalrefveq npoo6e6epr6veq qrov unorw4r{, ovoyr<d(owoq ro no- pegBoM6pevo DNA vo oX4pror(oer uOq}.d". Or vqo(6eg CpG neptSlouv peydtro opr0pr6 6rvou- 578 KEOA Al0 20 H Meroypotprl oroug Euxopurirreg rLeorrd[rov CpG r<or oulvd neptBd].].ouv rouq uno(.- vqt6g ttrrv rdrooUoroto errppo(6pevo.lv yovr6(ov, n:- p6rr en(oqg Bpior<ovroL Kor orouq unorrvqrdg rL' , pu0;rr(6pevcov yovt6(tr;v, H vrlo'[6o nou nepfiopBo,r:. 6vov unorrv4q np6ner vo e(voL otr-re0utr(orr1 o-rme : unorrvrltqg vo pnope( vo (er<Lvrloer rl Ueroypoo- Mro et6tr<r1 npcorefvr1 npoo66veror oro pe0u].Lopo,: 6tvoux).eor(6ro CpG r<r epno6((er rqv Svop[r1 rlq -.- roypo<p49. BrBAroypogio 20.2 0r Europuorueg RNA l-lotrupepooeg Eivor l-lotrupepq 'Ev(ugo Avooxdnrloq Doi, R. H. and Wang, L.-F. (1986), Multiple prokaryotic RNA poly- merase sigma factors. Microbiol. Rev. 50,227-243. Young, R. A. (1991). RNA polymerase ll. Annu. Rev. Biochem. ffi. 689-71 5. 20.3 H RNA l-lotrupepqq 'EXsr 6vo Arpeprl Ynorrvqrr1 Avooxdnqorl Grummt, l, (2003). Life on a planet of its own: regulation of Rl,lA polymerase I transcription in the nucleolus. Genes Dev. 17. 1 691 -1 702. Mathews, D. A, and 0lson, W. M. (2006). What is new in the nu- cleolus? EMB0 Rep. 7, 870-873. Paule, M. R. and White, R. J. (2000). Survey and summary: transcription by RNA polymeras- es I and lll. Nucleic Acids Bes. 28, 1283-1298. 'Epeuva Bell, S. P., Learned, R. M., Jantzen, H. M., and Tjian, R. (1988). Functional cooperativity between transcription factors UBFl and SL1 mediates human ribosomal RNA synthesis. Science 241. 1192-1197. Kuhn, C.-D,, Geiger, S. R., Baumli, S., Gartmann, M., Gerber, J.. Jennebach, S., Mielke, T., Tschochner, H., Beckmann, R., and Cramer P. (2007). Functional architecture of RNA polymerase l. Cell 131,1260-1273. 20.4 H RNA notrupepuol lll Xpqorponorel Tdoo Ko0o0uouq 0oo rot AvoDrroug Ynorrvrlrdg .l . .... .---. - Geiduschek, E. P. and Tocchini-Valentini, G. P. (1988). Transcr:- tion by RNA polymerase lll. Annu. Rev. Biochem. 57, 873-91r Schramm, L. and Hernandez, N (2002). Recruitment of RNA po.- merase lll to its target promoters. Genes Dev. 16, 2593-2620 Bogenhagen, D. F, Sakonju, S., and Brown, D. D (1980). A co'. trol region in the center of the 5S RNA gene directs specific ir.:- ation of transcription: ll the 3 border ol the region. Cel/ l: Galli G Hofstetter, H., and Birnstiel, M. L (1981). Two conservE: sequence blocks within eukaryotic tRNA genes are major pr:- moter elements . Nature 294. 626-631.
  • 71.
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  • 74.
    Euppogri Kr EneEepyooio TouRNA ffi eLooyoyrt ffi fo 5'Arpo rou Europuarnrou mRNA 06pet Kotrunrpo * Mto 5' rotrunrpo oXrlporr(eror npooOerovrog pro G or4v retrrrrl pool rou peroypotpou Ueoo evdq deopou 5',-5',. e H dtepyoo[o oXlportopou rotrunrpog ouppo(veL roro 11 peroyporpl, ron nou rooq ent1peo(er rrlv enov6vop[r1 rrlg peroypotprlg. * H 5' rotr0nrpo tlv neproooreptov mRNA e(vor lrovope- 0utrulpevl, evtir opropevo Utrpo Un rtodtronolqlro R_ NA e(vor rprpe0utrrurpdvo. + H doilrl rotrunrpog ovoyvop((eror onri nporeivrroug nopoyovreg nou enrlpeo(ouv q oroOepdr4ro, Trl oup_ potp1, rr;v eloVtoyn rorq gerogpool rou mRNA. ffi O, Iluprlvrreg Iuppotreq Iuppoqnq ervot BpoXereg Atrtrq- trou1(eg * 06oerq oupporprlg eivor or otrtrqtrou1reg nou pp(orovrot opeotog Otnlo ono ro dpro e[ov(ou-rvrpov(ou. l'lotp- vouv T0 ovopo roug ono rq 06oq roug ouyxpurro pe ro rvrpovro. 582 KEOAAAI0 21 lupporp1 rr Ene(epyoo(o rou RNA @ Kenneth Eward/Photo Researchers, lnc * H 5' 06q oupporpnq oro 5' (oprorepd) orpo Tou rvTpo- viou nepiloppovrr ul ouvorvrTrrr1 otrtrr1trouX(o GU. e H 3' 0eon ouppoqriq oro 3' (6e(ro) orpo rou rvrpoviou nepiloppover rr1 ouvotvelq otrtr4trouXio AG. * 0 rovovoq GU-AG (nou opyrro ovopo(drov rovovoc GT-AG ovorpoprro pe qv otrtrr1trouX(o rou DNA) opr(eL on ouro ro oro0epo 6rvourtreor(Oto e(vor onopo[qro ong 6uo nptireg ror 6uo reLeuro(eg Oeoerg Tov rvTpo- v[tr-rv oro npodpogo mRNA. e Ynopxouv etroooovo rvrp6vro ouyxprnxo pe ro pei(ovo lvrpovro nou orol,ouOouv rov rovovo GU-AG, * To etroooovo rvrpdvro orotrouOouv rivo yevrro rovovo AU-AC pe pro 6rorpopelx4 opo6o ouvorverxtirv otr),1- trouXuirv oro 6pro e{tov[ou-rvrpoviou. 0L Iuppotr6g Iupporpr.lg Aropo(ovrot ovo Zeuyrl * H ouppoqrl e[oproror ;tovo ono qv ovoyvdrprorl (eu- ytirv ouppotrrirv oupporprl g. e '0treS ot 5'xor ot 3'0eoetg oupporprlg e(vor tooduvopec treuoupyrro. * Enrnprio0ereg ouvrrlp4g6veg otrtrr.1trouXreq onq 5, xoi ffi
  • 75.
    3' 0doetg ouppoqnqdrorplvouv ng treuoupyrrdg 0e- oerg oupporpr1q gero[u oprer(rv otrtrrrlv nr0ovdrv Oeoe- tov oro pre-mRNA. ffi U Iuppoqrl rou pre-mRNA l-lpoXrrlpeiAropeoou Evog Ao- o0u e H ouppoqrl onoue[ tLg 5' ror 3' 06oerg oupporpflg ror pro 06ori dtortrddoolg ovodrro rqg 3' 06orig ouppo- arls r H otrtrlloux(o drortro0toolg elvor ouvrrlpnU6vn or1 (upq oIIo dXr rooo oroug notrur0rropoug europutir- Teq. * Orov ro rvrp6vto rone( orqv 5' 0eoq ouppoqrlg, oXrl- po((erot gro doprl oe oXrlpo Ldoou xot ro 5' oxpo ouv- dderor orq 2' 0eoq prog A mr1 0rlo1 drortrodroorlg g6oo oro lvrprivro. * To rvrp6vto onetreu0epdrveror und poptpr; trooou drov ronei oqv 3' Orior1 oupporpflg u 6neuo ro oprorepo ror ro 6e[r6 e[rivro ouvdeOouv pero[u roug. ffi fo snRNA ELvorAnopolrr1ro yro n1 lupporprl * To n6vre snRNP nou epntr6xovror orrl ouppoqrl eivot ro Ul, Ul, U5, U4 rot U6. * To snRNP po(r pe optog6veg otrtreg nporeiveg oX11ro- r((ouv ro otogor[dro oupporpnq. * Otro ro snRNP errog on6 ro U6 nepr6Xouv Uro ouvTrl- pqp6vrl otrtrqtrouX(o nou npoodever lq npoTeivrq Sm or ono[eg ovoyvrrlpl(ovror ono ovlodrporo nou nopo- YovTor 0e ouroovooo voorlporo. €& leopeuon Tou llpodpogou mRNA orrlv 0d6Iupporprlg * To Ul snRNP nporotret rrlv 6vop(r1 qg oupporprlg pd- trtg npoode0e[orrlv 5' 0eori ouppoqrlg peoo_r prog ovl- 6pooqg (euyoptirporog RNA-RNA. * To oupntroro 66opeuorlq neprdXet ro Ul snRNp npoo- dedegevo oqv 5' 06011 ouppoqrlq Kor Trlv rpoTeivl U2AF, 11 ono[o npoodrlvsTor 0e Uro ouororX(o nupiprdr- vov pero[u rou orlgelou drortrodoiorlg ror qg 3, Oeor1g oupporprlg. & Irouq notruxurropoug europu6reg, or npotreiveg SR no((ouv onopolqro p6tro orrlv tivop[r1 rou oXnUoTr- opo0 rou ougntrrlxou ddopeuolg. " To (euyopogo rov 0eoeov ouppoq1g enuuyyoveror g6oto opropou rov rvrpov(ov 11 rotv e[o.rvrov. ffi n 06oq lXqpolo;lorj rou ltogoldtou Iuppoqnq e To oUUntroro 66ogeuorlg e[etrfooerot oe np60popo oo;ror[dro oupporprlg (oupntroro A) nopouo(o ATp. * H npoodeon rov U5 ror U4lU6 snRNp perorpdner ro np66popo oroporidro oupporprlq oe tirptpo orrrportdro oupporprlg (ougntroro Bl ). e To oupntroro 81 enerro perorp6nerot oro otipn),oro 82, oro onolo ro Ul snRNp onetreu0eptiveTor yro v0 enttpdtl.tet oro U6 snRNA v', otrtrntrentbpooet ge rrlv 5' 0eor1 oupporprlg. * H U4 onoporpuveror on6 ro U6 snRNp enupenovrog oto U6 snRNA vo (euyopooer ge ro U2 snRNA yto vo oX4gorfoer ro roro],ulrrj r6vrpo yro q ouppoqrl. e Kor or dtlo ovrLdpooerq trans-soTrponoulorlq ougpoG vouv 0T0 evepyonorlpevo oroporblo ouppoqnq (ouU- ntroxo C). * H ovr[dpoorl oupporplg e(vor ovnorpenrrl oe otro ro 0nUoro. *q&3 'Evo EvotrLornrd lorpor0ro Iuppotprlg XprlorponoLei ALo- gopenro snRNP yto vo Ene[epyoore( r0 ,,E],oooovo,, lv- rpdvro * Mro evo),trorrur1 odr5g oupporpnq Xplotponorei pro otr- trq opo6o snRNP, q ono(o 6Xer rorvo pe ro rupro orolro- 16ro ouppotprlg ;ulvo ro U5 snRNP. * To rvrprivro orriXor eXouv geyotrtirepeg ouvorvenr6g otrtrqtrouX(eg ong ougpotreg oupporprlq, Xopig v' oro- trouOorlv ntoro roug xov6veg GU-AG n AU-AC. * To Uei(ovo ror ro eLdooovo oropor0ro oupporprlg por- pd(ovror olgovlro0g nporeivrroug nopdyovreg, ge- rolu otrtrrov rtg npolreiveg SR. ffi n Iuppoqn rou pre-mRNA llt0ovov'EXer Kotv6 Mryovr- op6 pe r' AurororoLulro lvrprivro rqq 0po6og ll * To rvrprivro qq 0po6og ll err6pvovrot on6 povo roug onri ro RNA p6oto evog ouppdvrog ourorototruurrlg ouppoqnq. * 0r ouppotr6g ouppog1q Kor o U1x0vrop6g oupporpqg rt0v rvrpovlrov qg opodog ll e[vot nopolroro Ue Tr.l oup- poqrl rtrlv nupqvxcirv rvrpovltrrv. * 'Evo rvrpdvto qg ogodoq Il drntrtirveror oe pro 6eurepo- royrl 6opr1 nou d4proupye( evo rorotrulro r6vrpo ro ono(o 0upr(er q Dogr'1 rou nuplvlrou rvrpovrou U6-U2. ffi U lupporprl e(vor Xpovrrd ror Aeuoupyrro lu(euypevl ge l-lotrtro Bqporo oqv'Erqpooq rolv l-ovldurlv * Iuppoql gnope[ vo npoyporonorlOer xoro q 0ropxero qq ueroYpoQnq n uero q peroyporpl a 0r UnxovroUoi qg peroyporpqg roL qg ouppoprlg ervor quotro ror tretroupyrxo evonollpevor, * H oupporpl ouv6derol pe qv e(oyu.ryr1 rou mRNA xor rov 6treyXo qq moOepoqrog. r H ouppoQrl orov nupilvo pnopeI vo enrlpeooeL q gero- Qpoorl Tou mRNA mo runopontroopo. @ H Evotrtrorlrrl Iuppoqn E(vor Kovdvog,'OXr E(orpeol, oroug llotrururropoug Europudueg * Erdrro e[covto 11 otrtr4trouXieg e[ovulv pnope( v' ono- rtrerorotiv 11 vo oupnepilrlq0oUv oro mRNA npotovro Xprlorponouivrog evotr),orrt16q 0eoerg oupporpdg. e H evo),),ornrr1 oupporprl ouvetoq6pet oq doprrrl r<or treuoupyrrrl norril,[o rtrlv npoiovrov rov yov16(tov. * 0 ro0optoprjg rou rp0trou oq Drosophila nepilogpover pt0 oepo ouppdvrtov evotrtrornrrlg oupporprlg oe yov(- 6to nou roduonorouv 6lo0oXuo npoiovro ;uog o6ou. -ffi H Iupporprl Pu0p(eror ond EvtoXureg ror Anooronlrrlg nou Bploxovror Tooo oro E[drvto'0oo ror oro lvrp6vro e H evotrtrorrun ouppoqn ouXvo oXer(erot pe oo0eve[g 0doerg ouppogr.lg * 0t otrtrntroux[eg nou neprpotrtrouv evo],],oxlro e[tirvro ouXvo e(vor nto ouvrrlpqp6veg e[etrulro on6 ng 0trtrr1- trouXieg nou neprpotrtrouv ro rdroouorolro e[tirvro. e ErdLxeS otrtr4trouX(eg e[ov(ov ror tvrpov(rrlv evroXuouv 11 xoroore),trouv rrlv eniloy( qg 06oqq ouppoqrlq. e H en[0poo1 rcov evroXurtiv Kor Tov onooro:nlrtirv qg ouppoq1g DropeootropeLrot onti er6Lr6g yro rrlv otrtrq- trouXlo npolreiveq nprlodeolg oro RNA, notrtr6g ono ng onoleg pnope[ vo puOp((ovror ovonru[roro ro/r1 vo erqpo(ovror roro-erdrro. e H roxurnro peroypoqrlg enrlpeo(er ogeoo rrlv drpooq qg evo),),orlr1g oupporpfl g. ouvey[(eru or4v enopevq oeAido 583
  • 76.
    dffi" &lxtj 'w 0r Avndpooerg frans-ouppoqrlgXplorponorouv Mrrpo R- NA * 0r ovrLDpooerq ouppoQnq ouvrl0oq npoypcronorouvror ;-rovo in cis perolu ou;rpotrrirv oupporpqg oro [dto ;ropLo RNA. e Trans-oupporpll ouUBoiveL oro rpunovooriruoro Kor T0 orouLrlrro onou puo ppoXero otrtrqlouXio (SL RNA) oupponrerol oro 5' orpo notrtrtirv npodpoprrlv mRNA. * To SL RNA 6Xouv doprl nou poro(et ge q 05o1 npoo- deoqq Sm rov U snRNA. To 3' Arpo rorv mRNA Alptoupyo0vror pe l-leg1 ror l-lo- truoDevutr[oor1 * H otrtrltrouxio AAUAAA e(vor 6vo onuo ndUnq yto rl drlgroupy(o evoq 3' notruodevutrulprlvou oxpou mRNA. * H ovr[dpoon onoue( 6vo nporeivrrrl oupntroro nou ne- preXer evov nopdyovro etOrrdrrlroq, gro evdovourtreo- oq rot notru-A notrupepooq. o 0 nopoyovrog erdrxorr;rog ror q evdovourtreool n6- nTouv To RNA ro0odrro qg AAUAAA. * 0 nopoyovrog eL6trorrlrog rot 11 notru-A notrupepoorl npoo06rouv - 200 rordtrorno A oro 3' drpo. * H oupo no)tu-A etreyXer q oroOeporrlro rou mRNA xr enlpeo(et q peroqpoorl. * H noLuodevutr[roor1 oro xurropon],oo1ro noi(er potro oqv epppuirq ovomu[1 rou Xenopus. H Ene[epyooio rou 3' Arpou rou mRNA Ervor Kpiorlrl yLo rov Tepponop6 11g Meroyporprlq * Yndpxouv drorpopot rponor repponouou rnq usToypu- qflg ono dLotpopenr6g RNA notrupepooeq. * 0 0XIU0TtoU6g rou 3' orpou rou mRNA oqpotodore[ rov reppoloiui r1g peroypotpflg ond rrlv Pol ll. 0 lxrtfrorropdq rou 3' Arpou rou mRNA tov lorovrilv Anouef ro U7 snRNA & H 6KQp0on rcov mRNA rtov rorovdrv e[oprorot on6 rr1 peroypoqrl xor puOp((erot roro rr1 droprero Tou KUTTo- prrou rurtrou. o To mRNA rolv lorovtirv dev eivor notruoOevutrrrol.rtlvo: ro 3' orpo rouq drlptoupyelror gdoro luog ovr[dpoolg n6fig nou e[optorot on6 q 6o91 tou mRNA. a H ovr(dpoorl neQrlg npouno06reL npoodeol rou SLBP oe pro doprl proXou-0r1trroq ror (euyoprrrlro rou U7 snR- NA pe pro yeuovrrl povortrovrl neproXrj. * H ovrldpoon ndQqq rorotrueror ono 6vov nopdyovro rorvo ;le ro oupntroro notruodevutr(trlor1g. H luppotprl rou tRNA llepilol.rpovet Konrl rr Enuvoouvde- o1 oe :exolproreg Avldpooerq * H RNA notrupepoorl lll repuort(er rr1 geroypotpq oqv otrtrntrouxto notru(U)4 nou pp(orerot geoo oe Uro otrtrn- trouXio ntrouoto oe GC. c H ouppoQrl rou IRNA npoyporonotelrot ;le 6to6oXLxe: ovnOpooerg ronqg rot ouv6eoqg, * Mro evDovourtreoorl rdper ro npodpopo IRNA roL c.: 6uo oxpo rou rvrpovfou. * H onetreu06po0rl Tou rvrpov(ou dnuroupyei 6tjo ptol- IRNA pe oouvrlOtoro oxpo nou nept6Xouv 5'-0H roL 2 - 3' ruxtrrro qtoorpop116. * To 5' -0H drpo qootpoputrtoverot on6 pto notruvo,- r),eoldxfl rrvool, q ogddo rou rurtrxo0 QooQOptK:, ovo[yet on6 pro tproorpodreotepoorl onodtdovroq 2'-q.:- oqoptro tetrtro orpo rot 3'-0H opddo, ro drpo i,-' e[ovlov ouv66ovror ono pro RNA trryrooq xor ro 2 - rpulorpopuo otpoLpelror ond pto tprrrorpotootl. H Anovqoq orrlv llopouofo proq iedtntrtop6vlg l-lporra.- v1g IXer((eror Ue rn Iuppoqrl rou IRNA o H lrel e[vot ;uo eotorepxrl nupqvrrrl UeUBpovrx! np';- Tervn Us rrlv N-retrrrr1 neproXl T1q oTov outro rou :: rot q C-retrtr1 neptoxrl Tlq oTov nuprlvo. H C-re),,1<: neproX1 6Xer evepyoqreg rLvoolg n evdovouxtreoorl * H npoodeorl ptoq [edrntrotp6v4g nporeivqg oqv N-."- tru1 neptoXrl evepyonotel rrlv C-retrrrr1 evdovourtreo:- Ur ouToqtrtoQoputr(oor1. * H evepyonotqptlvl evDovourtreooq ropet ro HACl rl- NA (Xbpl oro onovduLroro) onetreu0ep6vovrog e,: rvrpovro ror 0rlgroupyrovrog e(6vto nou ouvdtlo','ri. pero[u roug ono pro IRNA trryrooq. * Movo ro ouppoprlvo HICl mRNA ;inope[ vo USToQC]- orel o' evov peroypotprxo nopoyovro o ono[og evep].- nore( yov[6Lo yLo ouvodoug nporretveg nou poq0o-, oro 6tntropo rov [edtntro;r6vtrlv nptoreivtirv. * H evepyonotluevn lrel endyet on6nrooq orov ro rL.- ropo nepr6Xer nopo notrtreq [eDrntropeveg nporeivec H l'lopoyoyrl rRNA l-lpoUno0ereL lupBovro Kon1g ror f :- prtroppovet prrpo RNA * H RNA notrupepool I repuon(er q peroyporprl o evr, reppolorfl 1 8 pooeorv. s T0 Ueyotro rot ro ptrpo RNA onetreuOep6vovrot pe x:- n1 on6 6vo rorv6 npridpo;ro rRNA. To 55 rRNA pera- ypoqerot IeXtoproro. * H ouodo C/D rov snoRNA elvor onopolrr1Tl yro T'i rpononoqorl qg 2' Oeoqq qq prp6(qg pe npooOq<- lrLog pe0utro;ro6og. a H ouo0o H/ACA rov snoRNA e[vor onopo[rr1Tl yro T" perorponl rrlg ouptd(vr1g oe qeu6ooupt6[v1. * Ie ro0e nep[nroor1, ro snoRNA (euyoptirver pe pto o,- tr1trouX(o RNA nou nepreXer q pooq-oroXo tlore l'c drigroupynoer pro runrrrl 6oprl nou elvor ro un6orpogc yro rqv rponono[qor1. llepilqtpq iasE{ YicfJt! ffii 21.1 Erooyuryri To RNA no[(et rewprr6 p6].o orrlv 6r<gpooq rov yovr- 6(tov. Aplrrd 1oporcr4p(ornKe oq 6vo ev6r6preoo rnq npolreivoo0vOeorlq, qA d 5lftore ovoroL6rp0qxov nol.- Ld dMo RNA pe 6opuxoUg rj Leroupyu<oUg p6Aoug oe 584 lGOAllAl0 21 Iupporprl rr Ene[epyoo(o rou RNA 6rd<popo ord6to qq Srrppoolq rov yovr6(u:v. H ou-- preroXrl rou RNA oe noMdg Aenoupy(eg nou oeopor. orrlv 5r<<ppoorl rov yovr6(iov unomrlp((er rrl yevtKr dno!.rq 6l q (or1 prnope( vo SXer e[e].rX0e[ ond dvc', "K6oUo RNA" orov ono(o ro RNA opXLKd rlrov tc evepy6 ouorotLr6 yto rr1 6roqpqorl Kor 5r<rppooq rcu,, yeverrrcbv nl.qporpopttrrv. Itq ouv5Xero no).).59 onc
  • 77.
    ourdq rrq Leroupy(egunoBorl0ri0qxov ri ovoLrirp0rl- xov qn6 npoteiveg, p' enox6trou0o trlv otifioq rqg euell((og r<or pdMov Kot. rnq onotetreogotx6rrltog. 'OLo rq RNA nou ;retoypdtpowot on6 r' ow(orotXo yov(6ro yro vo y(vouv ti:ptpro r<ot Lemoupytt<d Xpetd(ov- rot nepon6pco ene[epyoo(o. Atorer<otrrp5vo yov(6to undplouv oe 5Leg ng KorrlYop(eg ttrtv euxopurrlttrdlv opyovtopd:v. Awtnpooo:neUouv 5vo noLrJ prxp6 p6pog trov yovrd(tov trov ontroUorepov euKoputotcilv, oLtrd rr1 Ueyoll nlerov6rqto ttov yovtd(cov oro yovt6triproro rov ovtirepctlv euxopuror6v. To yov(6to notril,ouv noAU orip<pcovo Ue rov opt0pr6 xot to ;rrlxog rov twpov(tr:v, 6pr<og 6vo runrr6 yov(6to ttov 04loonrri:v 6Xer 7-8 e[c0- vro nou eme(vowot oe Uto neptoX4 -16 kb. To e[6vto e[vor oletrd ;ru<p6 (-100-200 bp) evtil to twp6vto q4e- rrr<d;reydl,o (>1 kb) (06ne Ev6tr1ra4.7). H ooul.tEcovio pero[U rlg dtoxeroUpdvnq opydvro- onq rou yovt6(ou Kot rnq 6tor<exopr;r6vqg opydvooqg rou ow(qrorXou mRNA npouno06tet enefepyoo(o tou npcotoyevoUg npoi6woq rnq UeroYpoqnq. To Tlporo- yev6g pretdypocpo SXet rqv (6to opydvoor'1 pre to yov(dro xot rolefuor pre-mRNA. H o<po(peor1 to:v twpov(tov on6 ro pre-mRNA og(vet 6vo tuntx5 oyyeAtorp6po - 2.2 kb. H 6repyoo(o ogo(peo4g ttov twpov(ov rolekot ouppo- gri rou RNA (RNA splicing). H ogo(peoq rov twpo- v[cov e(vqr r<[pro pr5pog rrlg nopqyoyrlq RNA oe 6trouq rouq euKopurireq. I-lop6tt to dtoreroppr6vo yov(6to e(- vor olerrrd ondvto orouq neptoo6tepoug l.tovot<6rro- poug/otrryor<6ttopoug europuriteg (6ncoq n (6pn Saccharomyces cerevisiae), 11 ouvoLt- r<q ovotroy(o unoerttpd rq oqproo(o rc,rv rwpov(ov eneL6rl ro neptoo6tepo 6Lorcer<oprp6vo yov(6to xro6Lronototjv oletri<d oqOoveq npotelveg. Iuveno.rg, 1 ouppo<pr1 epn5r<eroL orrlv nopoYoJ- yr1 peyolrjteprlq ovoloy[og otrLxoi m- RNA ond ro ovogev6pevo pe Bdoq qv ovd).uot1 tou yovtdtcrtprotog, (otog 50%. Mro on6 rtg npu:teg ev6e([etq o1e- iLr<o pe q gUorl rnq ooupeov(og og npoq ro p6ye0og preto(ti rolv nuplvt- <r.,-rv yovt6(r.,.lv KClt Tov npoi6wtrlv toug ;roug ovrr.lrepouq euKoputr.lteg 6601- <e on6 rrg rdL6tqteg rou nuprlvtt<oti RNA. To pr6oo prdye06g tou e(voL noL0 geyorirepo on6 to prdyeOog tou mR- NA, eivor no).ti oo-roOdg r<t 6Xet noLti rotr un).or<6tepr1 otrtrqlou1(o. Awtr<o- To RNA rpononote(tot orov ruprlvo pe npoo0ilreg oro 5' rot 3' orpo roL ueoor ouppoqrlq yto Tlv oqo(peo1 Tolv tvTpo- iLov, To ouUBov oupporptlq npouno06ret 3oouol rov oup$otrtlv e[ov[ou-tvrpovlou rot ruvOeol rorv oxptrlv rtov e[ov[tov, To titpLpo nRNA UerotpepeTor Utoo rov nuplvtro.tv no- J0lv oT0 xurropontroopo, onou pteroqpo(eroL. Eqd)vlo1 lwp6vrol Eqovto2lwp6vto2Eq6vtoSlwp6vto3Eqovro4 lwp6vto4 E(civto5 a I ut.oypo,ln Y I Tpononoiqorl tou oxpou l-lotru-A mo 3 oxpo V tAzoo {:uppo,pr1 -,*g!!9-l*[0,:,= I 0r ouu0otreq e(oviou-tvrpov[ou onove Y _-----_ 't- j I To eidrvLo ouvd6ovrot I A h200 nYPHNAI lVletorroPd romp((owoq rlv eupe(o rorovoprl r<ov peye0dlv rou, rotre(ror erepoyevdg nup4vrx<i RNA (hRNA) (hetetu geneous nuclear RNA). H quou<q Uopeti rou hnRNA e(vot 5vo ptBovouxleo- npoteivx6 otopot(6to (hnRNP), 6nou to hnRNA efuot npoode6e;r5vo oe Uro oprddo dgOovtov nporeivrirv. Opt- opr5veg npotetveg prnope( vo 61ouv 6opru<6 p6Lo orq ouoreuoo(o tou hnRNA. Apretdg e(vor yvcoor6 6tt en4- ped(ouv qv ene(epyoo(o rou RNA r] 6reuxotrUvouv rqv e[oy<oyr1 rou qn6 rov nup{vo. H ouppogrl npoypororoe(rot pr6oo orov nuprlvo, po(( pre ng 6trl.eg tpononott'1oetq nou ouptBo(vouv oro veoouwe0erpr6vo RNA. H 6repyoo(o tqg 5rqpoorlg ev6g 6roreroprpdvou yovt6(ou nopouotd(etot orqv EIKO' NA 21.1. To ;retdyporpo onorcrd r<olr3mpo oto 5' dr<po, ro rwp6vro o<potpo0wot t<ot ro 3' dt<po nol'uov6evu- hr.6vetor. 'Enero, to RNA petorp6petot ;t6oto ttov nupq- vxriv n6pov oro rurtop6ntroopto, 6nou e(vot 6tq06ot- pro yro gerdgpooq. Avo<poprd pre rtg 6tdcpopeg owtdpdoetg ene[epyo- o(og nou oupBo(vouv orov nuprlvo, 0o O5tropre vo yvcopG (oupre n6te or<prBriq npoYUoronote(tot q ouppotp4 oXenxd pe ng dltreg tpononot4oetg tou RNA. Apoye, o'uppoqri ouprBo(vet oe pto ouyxer<ptpt6vq 05or1 pr6oo orov nupqvo r<ot ouv66etot pe dMo ou;rBdwo - fl.X. 'rrl pretoypoEq r<o/q q preto<popd on6 tov nup{vo oro KUr- rop6ntroqro; En(oqq, rt ow(rcruno orrlv 5rrppoorl tcov prq dtoreroprprdvcov yovt6(ov 61et 4 6lstr!4 ouppotptig; Avorpopu<d pe rnv (6to qv ow6pooq rnq ouppo- E[r0vto 1r- Y- I -l- lMerooooon V" :a:_--:: KYTTAPONAA'[,1A Kotr0ffipo oro 5' oKpo Azoo 585
  • 78.
    tptig, 6vo on6ro x[pro eporrlporo e(vor ndlg eMyleror q et6rr6rrlrd rqg. Tr 6toogol.i(er 5rr ro drpo rd0e rv- tpov(ou ovoyvop[(owot ovd (eUy4 Sror cilore on6 ro RNA v' orporpe[ror q ocr:orr1 o].].q].ou1(o; To twp6vro r6Bowor on6 5vo np6Spogo ;r6pro pre r<o0opropr6vr1 oerpd; H rop[provoq rou RNA XpnorUonore(ror yro rq p60proq qg 6r<cppooqg ro:v yovr6(ov prSoco 6rdr<prorlg peto(0 rrov 6to05or;rolv npodp6prov ri oLLoyrlg rou nporUnou oupporpfg; Emdg on6 rr1 ouppogrlrou RNA yro rnv ogo(peor1 rov twpov(o:v, ene(epyoo(o yro vo rrtprprdoouv Xper- d(owot en(or1g noMd pr4 ro6xonorqru<d RNA nou no(- (ouv p6l,oug oe noxll,eg muX6g qg 6r<gpoorlq rov yo- vr6[ov. 21.2 To 5' Arpo Tou EuKopuoTtKo0 mRNA Odper KoA0nrpo * Mro 5' xotrunrpo oXrlporl(eror npoo0drovrog pro G oqv retrr- rr1 pool rou geroyporpou ;.uloo evog deogou 5'-5'. e H 0repyoo(o oxlpologou rotr0nrpog ou;rpo(ver roro q pero- yporprt, rort nou (otog enqpeo(er qv enovdvop[n Tnq UeTo- Yp0arlq * H 5' rotrunrpo rtov neptooorepcov mRNA e(vor povopeOutruo- gev1, ev6 oprol.ulvo pxpo 1ui rodrronorlrxo RNA e(vor rpr- pe0utrulgevo. * H 60[rl rotrunrpog ovoyvrrlpl(erot ono npotreivrrouq nopd- yovTeq nou enqpeo(ouv q oro0epo-rr1ro, rq ouppoqrl, qv e[oyrrryrl rorq perorppool rou mRNA. H peroypogrl (er<rvd pr' 5vo rpr<proorpoptx6 voux).eoo(- 6ro (ouvr1Oog noup(vr1, A n G). To npcoro vouxl.eor(- 6to 6Lorqpe( tqv 5' rpretrtoeoprr<r1 opiddo rou Kor 6q prou pye[ to ouvrl0topr6vo <proo<po6reoreprr6 6eopr6 on6 rqv 3' 05o4 rou ornv 5' 05or1 rou en6gevou vou- Bp[orerot oe dtreq rtq r<otrrlnrpeg I Bp(oreror oqv ro)Unrpo 1 Bploxeror oqv xotrUnrpo 2 rheol6(ou. H opXrrq oALqLoul(o rou preroypdgou gnope( v' ovonopooroOe( og: spppYopNpNpNp . . . MoLoro0ro, 5rov to r6prpro mRNA rorepyoore( rn vitro $e 6v(upro nou ro ono6opro0v oe (eXoprord vou- xLeor(6ro, ro 5' dxpo 5ev ono6(6er ro ovopev6pevo rpr- rpcoorpop116 vourleoo(6ro. Avr[0ero, neprSler 60o vouxLeor[6ro nou ouv66owor 1.r' 6vov 5'-5'rpreoogopr- 15 6eopr6 n en(onq 6ro05touv puo pre0u).oprd6o. H refu- x4 Bdoq efuor ndwo plo youov(vq nou npoor[Oeror crro ou0ewtx6 g6pto RNA ;rerd rq geroyporp4. H npooOrlrq qg 5'rcLxr1g G rorol0eror qn6 5vo nupqvx6 5v(u;ro, qv youovuLuo-rpovogepdon (GT). Ito 04l,oonrd, r1 GT 61er 6Uo ev(upur<5q evepy6rqreg. Aenoupye( 16oo rog tprEcoorpordoq o<porpcirwog 60o groogoprr6g ogd6eg on6 to GTP 6oo Kor oq youovrr l,u).o-rpovorpepdorl ouwrlrowoq q youov(vr1 pe ro ou- 0ewrx6 5'rpr<prrlogoprr5 dxpo tou RNA. Ir4 (rip4, our6g or 60o evepy6rqreg ov4Louv oe 6rorpoperrd 6v- (upo. To v6o xotdlorno G npoor(Oeror oro 6xpo rou RNA ge ow(orpogo npooovoroLrop6 on' 6Lq t' 6IAq vour<Leor(61o. SGppp + 5 pppApNpNp. . .-+ GpppS-SApNpNp. . . + pp + p Aurn n 6opr{ r<o}.efuor xotrrlmpo (cap). E(vor ro uno orpoUo yr' opxerd ouprBdwo ;re0uL(oo4q. H ElK0tlA 212 6efiyer r4v n^,npn 6oUri puog xoA0mpog og6rou npoorE- Oorjv 6l,eg or nr0ovSg peOuLopddeg. To nro oqprowx6 ouprBdv efuor q npooOrlxrl puog provo6rrrlg pre0u).otrr66og orn e6on 7 rqg reLx4g youov(vr1g, q ono(o roro.l,feror on6 rrlv 7-peOuLorpovogepdorl rnq youovfuqs (MT). flop6rr q 6ro6roo(o rqg r<dLurlqg ;rnopef vo flptry- poronot40e( in vitro pre ro0opd 5v(upo, rovovrxd n owi- 6poon npoyporono€kor rqrd rn Ueroypoerl. Ap6orog prerd qv 6vop(q rnq Ueroypoens, n Pol ll oroprotd -30 vouxl.eoil6Lo roOodtrd rqg ogeqp(og, nept;r6vowoq vq enrorporeu0orjv ro 5v(upro r<dLur[4q nou 0o npoo06oouv rnv r<otrrinrpo mo 5' drpo tou ovo6u6pevou RNA. Xrop(g our4v rrlv npe oroo(o, ro ovo6u6pevo RNA e(vor eudltoro d en(0eoq on6 5'-3' e[rovour].edoeg; 5vo rSroro rfoL(dropo prnope( vo rdva ro oriprnl,oro Pol ll vo n6oer on5 ro DNA ergoye(o. Iuven<iq, 11 dtodtxoo(o qg rdi.ur.f4g efvor orlgowxrj rilore 4 Pol ll vo nepdoer oe nopoyoyu<r1 enrprlruv- orl yro vo peroypdtler ro un6lorno yov(6ro. An6 oufi qv dnorfrrl, 11 el5tu[q rou prlxovr- oproU norio4q yro rnv npoo0r]r<q r4g 5' xoA0- nrpoq owrnpoooneUer 6vo orlpre(o eL6yXou yro rrlv enov6vop(r1 rnq Feroypogqg on6 rqv oplu<ri 05oq noUorlg. ElKOllA 21.2 H xotr0nrpo (0noqpdootr,, ro 5' drpo rou mRNA ror eivor pe0uhuop6vl oe opxer6g 06oetg. 586 KE0AAAI0 21 Iuppoqrl r<t Ene[epyooio rou RNA Boorxeg eworeg
  • 79.
    I' 6vov ntrq0uopr6europutottxc0v mRNA, r<d0e pr6pto nepr6Xer ;r6vo puo ;reOutroprd6o orqv retrtni youov(vr1, nou yevxd ovoE6petot trtg provopreOutrttopr6vq r<oL0mpo. Aw[Oero, optopt5vo pupd prrl rodxonoqnrd RNA, n.X. exefuo nou epdSrowot oq ouppog4 rou RNA oro oo- pcn(6ro ouppoqqg (BI6ne Ev6rrya 27.6) preOuLttirvowot nepordp<o t<ot nepdlouv tperg preOutroprddeg orrlv rcLrq youov(vr1. Aurri q 6oprri r<otrekot tptpeOul,trog6vq rol[- mpo. To 6v(upo yt' out5g ttg entnp6oOeteg preroEop6q peOutrop66tov Bp(or<owot oro Kurrop6ntroopto. Aur6 (ooq drooqoA(er 6rt p6vo optopr6vo e(et6u<eug6vo RNA rpononoro0wot nepor6pco ortg xo}Umpeq rouq. Mro on6 lg r<Upteg l.eroupy(eg rou oXrlgottol.to0 xol.Umpog e[vot 4 npootoo(o tou mRNA on6 ono66- pqoq. Iqv npoy;rorx6qto, q ev(upxti ogo(peor1 qg xol,0mpog e(vot 6vog on6 roug rfptoug pr4XovtqroUg yro q p0Optorl rnq ovorfxtroorlg tou mRNA oro euKo- puortrd r<[rropo (F],5ne Ev6tqta 21.11).Itov nupfvo, ro erepodrpep6g np6o6eonq ornv rotrUmpo CBP20/80 ovoyvrop((er qv xolUmpo rot npoo66vetqt oe ourqv. Aur6 6reyeQet r4v o<po(peorl rou npd:rou twpov(ou rot, prSoco dpreorlq otrl.ql,en(6poonq Ue rn Unxovn e[oytoy{q rou mRNA (otiprnLoro TREX), 6teuxolUvet qv 5[o6o rou mRNA on6 tov nup4vo. 'Orov to mRNA gtdoet oro xurrop5ntroopo, pto 6toEopertrq oprd6o npulteivdtv (elF4F) npoo66vowor offlv rotrtintpo nporotrri:woq rqv 6vop(4 rqq perd<ppooqg. 2'1.3 Or llupnvrrdq IuUBoAdq lupporphq Eivor Bpoxeieq AAAnAouxieq a 06oetq oupporprtq e[vot ot otrtrrltrouxteq nou pp(orovrot optl- oog drntro ono ra opLo e[otv[ou-tvrpov[ou. 11o(pvouv ro ovol.lo roug on6 q 05011 rouq ouyrptrtro pe ro tvrpovto. * H 5' 06on oupporprtq oro 5' (optorepo) orpo rou tvrpovlou neprtroppovet rr1 ouvotvettq otrtr1trouXio GU. * H 3' 06oI ouppoQnq oro 3' (de[t6) drpo rou rvrpov[ou nepr- troppover rr1 ouvotvertrtl otrtrltroux(o AG. u 0 xovovoq GU-AG (nou opxtro ovopo(orov rovovog GT-AG ovorpoprro pe rrlv otrtrr1trouXio rou DNA) op((et ort ouro to oro- 0epo Drvouxtreor[Dto elvot onopo[qro ortg duo nptitreg rot 6uo retreuro(eg 06oetq tu-tv tvrpov[tlv oro np6dpopo mRNA. * Ynopxouv etroooovo tvrpovto ouyrptnro pe ro ge(ovo tvrpd- vto nou orotrouOouv rov rovovo GU-AG. * To etrdooovo lvrp6vto orotrou0ouv 6vo yevtro rovovo AU-AC pe gro 6rorpoperrrrl opo6o ouvotvertrtlv otrtr1trouXttilv oto opto eItov[ou-rvtpov[ou. l-ro vo eortdoouUe oro proproxd ouprBdwo nou e;intr5- KOVTor orn ouppoQl ttr;v nuplvtr<c0v twpov(olv, np6- ner vo e[erdooupre q gUorl ttov 06oeav ouppaEt]q, 6q).o6rl rov 6tjo op(ov e(cov(ou-twpov(ou nou nept- LoprBdvouv lg 06oerg OpoUorlg xt enov6vtoonq. Ot ouprBoldg peto[ti e[ov(ov xot twpovf<ov pnope( vo r<o0opLoroUv ouyrp(vowog qv oMlLouX(o voux.treo- rL6(rov rou mRNA Kot rou dogtr<otl yovt6(ou. Aev undpla ercretopt6vq o;rol'oy[o rl oupnA4potr rrr6rlro ;reto[rj tcov 6Uo drpov evSq twpov(ou. Ot oupBol.5q, tr:01600, 6Xouv ro}'d ouvrrlpqprSveq, clv Ktrl oXenr6 prxp5g, ouvowerr6g oMql,oulieg. E(vot egu<r6 vo npoodrop(ooupe 6vo et6tr6 dr<po oe xd0e rwp6vto Ue Bdon q ouwrip4crq tov ouprBoLcirv e(ovfou-twpovG ou. 'OLeg or oupBol,6g e[cov(ou-twpov(ou efuot nopotr- ).oy5g r4g ouvotverxqg oMrlLouXfoq nou dfuetot oro ndvco pr6pog rnq EIK0NAI21.3. To rjryog r<dOe ypd;r;rotog unod4trdrvet rrl ou1y6r1- ro proq ouyr<erptpt6vns B6onS oe xdOe ouvotveurcl 05- orl. YqrrlLi ouwripqorl nopoqpe(tot pr6vo pt6oo mo rwpSvro orrq unonO6preveg ou;rBol6g. Aur5 op(er rqv otr,r1trou1(o ev6g yevtrori twpov[ou rrlg: GU...,.. AG To rwp6vro nou op((etot rqt' out6v rov tp6no [ert- vd pe ro 6tvourtreot(dto GU rot teletrilvet pe ro 6wou- rLeor(dro AG. 'Etot, ouXvd l,6yerot 6tt ot ouprBoldg orol.ou0o(v tov xov6vo GU-AG (GU'AG rule). Iqprerdrme 6tt ot 6uo O5oetg 5Xouv dtogopertr5g oMrltrouX(eg on6te op((ouv rov npooovototrtotrl6 rcov drp<ov rou rwpov(ou. Ovotrrd(owot on6 t' optmepd npoq ro 6eltd xotd prrirog tou twpov(ou rog 5' 06o11 ouppogriq (peprr6g Eop6q rotre[tot oproreprl 06on n 06o11 d5qq) ror 3' 06oq ouppogrlg (en(or1g rotre(tot 5e[rd 06oq ri 06on 6611119). Erlpretox6q preto]']'d(etg orrq ouvqtvertr6q oMqouX(eq nopeprno6((ouv rrl oup- pog{ 16oo in vivo ooo rot in vitro. To neptoo6tepo twp6vto ot<olou0oriv tov rov6vo GU-AG. Oar6oo, 5vo ;lrp6 xLdopro twpov(tov 6ro05tet puo 6togopetu<n opddo ouvowertrcilv oMrltrouXtrilv mo 6pro e[roviou-twpov(ou, 6ncog <po(vetot oro r<dtto pr5- poq rnq Erx6vog 21.3. Autd to twp6vto oplrxd nept- ypdrpqr<ov crlg etrdooovo twp6vto nou orotrouOotiv tov rqv6vo AU-AC L6yco rcov ouwqpqprdvov 6tvoux.treott- 6(rov AU-AC Kot oro 6Uo drpo xd0e twpov(ou, 6ncog qo(verot oro p6oov rrlg Etx6vog 21.3. Tq pe((ovo t<ot eldooovo rwp6vto neptypdgowot rotrUtepo og rwp6- vro rfnou U2 r<or U12 ;re Bdoq rtg 6torpn6g pqXov6g oupporp4g nou XpnotUonoto[wqt yro rrlv ene[epyoo(o roug (Btr5ne Ev6tryra 21.9).'Ero; evri: optopr5vo twp6- vto qo(vetor v' qxolou0oUv tov rov6vo GU-AG, orqv npoyprorx6rqro ene(epyd(owor 6nog to twp6vto tti- nou U 12 (pMne r<fuo pr6pog r4g Etx6vog 21 .3). 21.4 Or lupBoAdq luppogriq AroBd(ovror Avd Ze0yn e H ouppoQn e[oprorot povo ono rrlv ovoyvdrptol (euY6v ouu- Botrtitv ouppotplq. c'0trsS or 5'ror oL 3'06oetg ouppoQnq elvot tooduvopleg tret- roupyrro. a Enrnpoo0ereg ouvqplp6veq otrtrltrouxieq ortq 5' rot 3' 0d- oeLq oupporpnq 6rorp[vouv rtg tretroupytt<6q 0eoetq 0uppoQ]lc pero[rj oprerdrv otrtrtov ntOovtirv 06oeov oro pre-mRNA. 21.4 0r Iuppotreg Iupporpnq Atopd(owoL Avo ZeuYTl 587 Boorxeq eworeg Boorx6g eworeg
  • 80.
    Aproreprl (5') 0eon Irlpotoouppoq1g yto ge((ovo rvrprivto (rUnou US n GU-AG) Ilpe(o 6LoxtrdDooqg o -74--54 3 2 1 1 2 3 4 5 Ae[td (3') 06o1 IrlUqTo ouppoQrlq yto e)dooovo rvrprivro (ilinou U1 2 n AU-AC) Iqgeio 6tortrd0rrlo4g o I C. Aeird (3') 06011 IrlUoTo ouppoQnq yro eJdooovo twprivro (runou U1 2) nou flep0dLovror on6 GU rot AG mo dxpo Irlpe(o drortrddroorlg o Ae{ro (3') 06ort To orpo rov nuprlvtKov tvTpovll]v Koeopl(ovTor on6 rov K0v0v0 GU-AG. To etroooovo rvTpovro Koeopi(ovTor U60o 6r0Qops- TrKd)v 0uv0rveTrK6v otrtrntrouxuilv oqv 5 Oeoq ouppotplg, rq Oeoq dLoxtrodrrrolg xot qv 3' 0do1 oupporpqq. 'Evo runx6 mRNA Or1).omrroU 6ro06rer noMd rwp6- vro. To Boorr6 np6Blqpo rnq ouppogrjg rou pre-mR- NA onoppder on6 rrlv onl.6rnro rov 06oecov ouppogrlg Kor nopouor6(eror mrlv EIK0NA 21.4. Me de6op6vq rqv nLrlOcirpq oMnlouxrcbv rou rwpov[ou nou rorprd(ouv pe rn ouvorvelrr; oll,ql,ouX[o rrov yvtiorcov 06oerov ouppognq tr 6roo<pol.[(er 6n 0' ovo- yvrop((owor xor 0o ouppdnrowor ro ocomd (e[yr; 06oeo:v; IlpSner vo ouv6e0oriv rordMrll.o (eUVn GU- AG nou on6louv noL6 (opro;r6vo rwp6vro e(vqr > 100 kb). A0o trjnor prrlXovrol.rcilv 0o ;rnopotioe vo e(vor une(Ouvor yro ro (euydpropro rcov xotdMrllov 5' xor 3'06oetrlv: o H oriv6eorl rorv O5oeov or' dr<po ev6g ouyxer<prpr6- vou rwpov(ou 0o pnopofoe vo e[vor 1.uo eyyev7g tdt6rqta rou RNA. Aur6 0o npoun60ere to ro(pto- opo er6trriv otrtrr1trouXr6v 4 6oprirv, rdtr nou 61er nopotqp4Oe( oe opropr6vo yov(6ro ew6purlv, oMd ''t.::i:::-. .-..----x 1 i; :.i:r!r;ir*ili; ,r,:;ili;rl :;ti.tii:t,r:;ty l] tt]i-ti!l+tir,rlr ait.i;t':r.t 0t ou1tBotr6g oupporplq ovoyvtr.tpf(ovrot p6vo 6rov 6Xouv rov xorotrtr1tro ouvduoolro (euyopo-rporog, 588 KEOAAAI0 21 luppoQn xr Ene[epyoolo rou RNA Aptorepq (5') eeon Aprorepn (5') 06on
  • 81.
    6e go(vetot votoX0et Yto ro neptoo6tepo euKopuoJ- uxd yovbto. o'OLeS ot 5' Kot ot 3' 06oerg 0o prnopo0oe pev vo e(- vot Lemoupytxd too6tivopreq otrI'd 11 ouppo<pf oro- )touOe( xav6veg nou 6toogol'((ouv 6tt ptto 5' 06o11 n6wo ouv66erot Ue rnv en6;revr1 3' 06oq oro RNA. Orjte ot 06oetg ouppogqq oUte ot neptBdMouo'eg neprox6q 51ouv rdnoto ouprnl,4ptrlpotx6trpo toq flpoq rqv oMqLouX(o, yeyov6q nou onoxtre(et to 1.tow6tr'o nou unooq p(ouv ou prntr 4 p<oprorx6 (euydpcopo prero(U tov dxprov rov twpov(tov. l-letp6proro nou Xprlottrronotorlv np66popro RNA 6e(1youv 6n onotodrinote 5' 06oq oup- poqns prnopei Oeoprltxd vo ouv6e0e( pre onoto6qnore 3' 06o11 ouppogrlg. l-to nopd6erypro, 6tov to npcilro e{6vro rqg nprirrtrtrlg preroypoqtrulg govddog UeroYpo- grlg rou SV40 ouv66etot pe ro rpiro e[cirvto rqq B oqot- pivr;g rou nowu<oU, to uBpt6rr<6 twp6vto ogopekot yto vo 64puoupyr10e( puo t5treto otiv6eorl ptero[6 rou SV40 e(ov(ou Kot rou e(ov(ou rqq B orpotp(vr1g. flpdyprott, ourn n owotrLo[tpr6t4to e(vot n gdon Ylo rrlv telytr<r1 nayi6euoq g e[cov[cov no u neptypdqn Ke nponYo u p5vtog orqv Eu<6vq 5.8. Tdtoto nepdptoto ovodetxvUouv 60o yevxd onpe(o: o OL 06oeq ouppaEriq e(vat yevxiq: Aev 6Xouv etdrx6- rnro yro (eXroptord np66pogo RNA, ofte to [eXor- prord np66pogo RNA g5pouv et6rrdg ntrqpoqop(eg (n.X. 6ewepotoVri 6oUri) onopofurlteg Yto rn oup- poen. Om6oo, oe optop5veg neptntrioetg 6Xet ono6etX0e( 6tt et6tr<5g nprore[veg nou npoo6Svowot mo RNA (n.1. hnRNP A1) npo6youv to (euydptol.to mrg Odoetq ouppog{q prSoro np6odeonq oe Yetrovt- x6g nr0ov5q O5oetg ouPPorP{q. o H ouoxeui yrc rn ouppaEn 6ev e(vu torc-etdmfi: 'Evo RNA ouvrlOog ouppdmetot oo:md oe onot- o6rinote r0rropo, ove[dptqto on6 to ov ouvrlOtrrq ouw[0erqt pr5oo oro ouyrer<ptptdvo r<0rropo. (Itqv Ev6t4ra 21.12 0o e(erdooupe rtg e[orp5oetq ortq ono(eg nopoulpotiwot toro-et6trd pot(Bo evoMq- rcttruig ouppogrlg). Mol.ov6tt ot 5' t<ot 3' 06oerg ouppoqrlg go(vowot 6poeg on1 ouoreurl ouppoq(q, notot rov6veg 6too<po- Ai(ouv 6l 11 ovoyvrilpton rov Odoetov ouppo<prlg nepto- p((eror 5rot dlme vo ouppdmowot pt6vo ot 5' rot 3' fuoerg tou [6tou twpov(oui To twp6vto orpopotiwot pe xdnoto et6u<r1 oetpd on6 5vo ouyrexptp6vo RNA; H ouppo<prj e(vot lpovtrd ou(euyp6vq Ue rn Usro- ypqqn ft.1. notrl.d ou;rBdwo ouppo<prig oLoxtrrlpdrvov- ror nporoU q RNA notruprepdorl rptdoet oro t5Log tou yovt6iou). Iuvendlg, efvor e6)loyo vo uno0Sooupe 6tt r1 trEf,cypogn nop6ler ;lo o6pri oepd ouppoqnq oe ro- teri0uvorl 5'-3' (6rllo6n, rqpefuot xdnoto noepd npore- por6qtog"). Aeftepov, 1-tto Leroupyxq 05oq oupporp{q orry1vd neptBdlletot on6 pto oerpd orotle(cov oM4lou- foq nou evtopjouv ri r<otoorS].].ouv rrl 05or1 (B].6ne Ev6r4ta 21.13). 'Etot, oMrltrou{eg 16oo rov e[cov[trrv 6oo xot ttov twpov(tov treroupyo0v en(oqg og pu0ptort- xd orotle(o yto rnv enttroyri tov 06oetov ouppog(g' l-to v' ovoyvtrlptore( onoteleoporLrd ono rc -- ,.1' .- opr6 oupporpnq, Uto ).ettoupyLxq 05or1 ouppoQrc ::- ntr vo 6to06ter q oomrl oMrll'ouX(o, peto[u c)-'*. er6Lxdg ouvotvertrdg oMrlLoulieq rot ototlelo flou t!c- 6civouv q ouppoprl to ono(o r<uptoplotiv 6vowL o)'- Lrov ororxe(ulv nou rlv t<otom6Mouv. Iutrtroytr<d, ouroi or unxovtopoi 6rooqol((ouv 6tt to oriproro ouppocpqg 6roBo(owot ovd (e0yr1 Ue pto o{ertKd ypop;rtr<r1 oetpd. 2l "5 H Xuppqqr'r Tou pre-mRNA llpoxopei Arqudoou Evdg Adoou a H ouppqQn onotre( rtq 5' rot 3' 06oetq ouppqQrlq rot gto 06- or1 0Lortro6oor1g ovodtrd rnq 3' 0eonq oupporpflg. * H otrtrntroux(o otododtoolg e(vot ouvqprlg6vq ott'1 (uUn oIIo 6Xr 16oo otoug notrux0uopoug euxoputlteg. . 0T0v ro wrp6vro xone( oqv 5'06o11 ouppoQnq, oplpor((etot Uro 0opn oe oXflpo trdoou xoL to 5' orpo ouvD6etot oq 2' 0e- on Uroq A mn 06on 6toxhd6oonq p6oo oto tvtptivto. . To tvrp6vto one)eu0ep6vetot un6 poprprl trdoou 6tov ronef aqv 3' 06orl oupporpnq rt 6netro to optotepri roL to de[t6 e[tirvro ouvde0ouv peto[U roug. O prrlXovropr6q qq ouppo<prlg 6Xer loporctqprore( rn vitro Xpqor;ronotcilwoq ouorqpoto el'eU0epo xurrd- pov oro ono(o twp6vto pnope( v' ogotpeOoriv on6 np66po;ro RNA. I-lupr1vu<d exluA(opoto ouppdmouv roOopd np66popro RNA. Aur6 6e(lyet 6tt 4 ouppogfl 6e ouvS6etot uno1ceortKd ;re tr1 6repyoo(o rrlq Uero- ypoqns. Iuppogq prnope[ vo ouprBe( oe RNA nou 6ev e(vor o0re roLugprdvo o[te no].uodevu].ttr:p5vo, nop6- r. oro rUropo owd to ouprBdwo t<ovovtr6 npoYpo- ronorotivtot pe ouvrovtopt6vo tp6no Kot n onotetreotrtottr6rrlto rnq ouppoqrlg enqped(etot qn6 rn Ueroypoqri xr dMo ou;rBdwo ene[epyoo(oq (B16- ne Ev6tryo 21.11). Errlv o66 r4g EIK0NAI21.5 nopouotd(owot tq ord- 6ro r4g ouppogrlg in vitro. Oo e[etdooupe rrlv ow(6poor1 on6 rq orontd rcov feloprmdrv RNA nou pnope( v' ovoyvtoptororiv. Qor6- oo, in vivo, RNA nou nept6Xouv e[drvto 6ev uqiorowot og eLeriOepo p6pto oMd ouyt<pototiwot on6 rn ou- oxeurj oupporpqq. H EIK0NA 21.6 defiyet 6tt to npcino B4go qg ow(- dpqonq ouppogrlg e(vot 1.tto nupqv6Eil.q en(Oeo4 tou 2'-OH mqv 5' 06o11 ouppoqrig. To optmep6 e[cirvro no[pver rn Uopqn ev6q ypogpxotl prop(ou. To 6e[t6 pt6- pto rwpoviou-e[ov[ou oXqgot(et prto 6ogt1 oe oplpo tr6oou (lariat), 6nou to 5' dxpo nou dq;rtoupyefuot qro drpo tou twpov(ou tout6Xpovo trans-emeponote(rot drors vo ouv6e0e( pr5oco ev6q deopro0 2'-5' ge gto Bdoq ;16oo oro rwp6vto. H Bdorl-m6Xog e(vot ;rto A nou Bpi- oKeror gdoo oe puo oMqlouX(o yvooni toq 06o4 6lo' xtr66toot1g (branch site). 21.5 H Iupporprt tou pre-mRNA l-lpoXopeiAropdoou Evdq A6out 589 Boorxeg 6vvocg
  • 82.
    5'r3' 5'0dq 3'060n GU UACUMCAG Pyr6 N Py66 PysT Pu75 A Pye5 ffiffii[I-0H :li -0 'lP :* i ,rj-ir'il E{rirvro 2 :f!dii0l-O:' r' "fi E{(rvro 2 ElK0llA 21.6 H nupqvxrl oupporprl npoypotonou[rur p6otrl 6uo ovrtdpdoeov trans-eoreponoqonq otq ono[eg pro oUd6o 0H enr- r[Oerot o' 6vo rproorpodreorepxo 6eopd. ).erQr1 ouvqpnonq ornv oMrlLouX(o rnq e6o'nq 6toxL6- dcoonq enrpdner rn Xpnorpono(qor1 nopepgepcbv oAtrq- LouXrdrv (xoloriwot r<purpdg 06oerg, cryptic sites) ov 4 ouOewx4 6tor<ld6ooq onoLor<pe[ r1 ;reroLl.o10ei. H ey- ywrpo ornv 3' 05or1 ouppo<pr1g rpo(vetor 5r e(vfl snpov- rrrl, ener6{ n Kpugri 05or1 Bp(or<eror ndwore rowd orqv ouOewu<r1 06o4. Mto rpugrj 06o11 1p4oq-ronoefuor g6vo 6rov 11 05oq droxld6tooqq 6Xer odpovonorqOe(. 'Orov pno Kpugn oMqlou{o 6ror<}.ddroorlg Xpnorponorekor xcn' ow6v rov rp6no, 11 ouppogrj ouve((eror xovovrrd ror ro e[ri:vto ono6[6ouv ro (6ro npo't6wo pe rov dypro ri- no. Zuventiq, o p6Log qq 06or7q SLaxAddaor1q e[vat v' avayvupi(a qv nA1o6areprl 3' 06o4 ouppaE4q aq otG Xo y@ o0vdeoq pe rnv 5' 06o17 ouppacprTq. Aur5 pnopei vo eppqveuOe( on6 ro yeyov6g 6n npoyproronoefuor puo oMqleni6poorl ;rero[ri rolv nporcivu<riv oupn],6rov nou npoo65vowor oe out6q ng 6Uo 06oerg. de5'@ 3, L==.lL ' E.- ., UACUAAC AG I2'5', AnoDtortrd6oon tvrpov[ou 5',Gr-i 'u_*UACUMC AG 3' . : :r.:. H oupporprl npoyporonoLelrot oe 6uo oroDLo. l-lpdr- ro roperoL ro 5'-e[drvro rr enerro ouvdeeror pe ro 3'-e[ovto. Ito 6efrepo Brlpo, to e].e[0epo 3'-OH rou e[rov(ou nou oneLeu0eputOrlxe on6 qv npotq ow(6poor1 enn(- Oetot oro 6eopr6 orrlv 3' 06o11 ouppo<prlg. nopoqp(- ote drr o opL0;r6q rov qotoqodreoreprrolv 6eoprriv ouwqpefuot. Aplrxd unqpXov 6rjo 6eoproi 5'-3' ong 05- oerq ouppog4q e[cov(ou-rwpov(ou. O 6vog 61er owrr<o- rosro0e( on6 ro 6eop5 5'-3' pero[0 totv e[o:v(o:v, evot o dl),og on6 to 6eopr6 2'-S'nou olrlprot((er to Ldoo. 'Eneno, ro Ldoo "ono6[oKlo6((eror,,, ono6(6owog 6vo ypoprpu<6 rwp6vro ro ono(o ypnyopo ono6ope(ror. Anopofurlteg otr-I.r1).ou1(eg yro rn ouppogq e(voL or Bpole(eg oMlloul(eq olq 5' r<or 3' O5oerg ouppo<pr1g Kor orn 05oq 6tor<)d6roor1q. Ko0rrtg ro peyol.rirepo U6- poq rnq oMrlLouX[og ev6g rwpov(ou pnopei v' qno].or- <pe( Xop(g ow(rruno orn ouppoerl, ouvdyeror 6tr 6ev onorefuor rdnoro er6u<4 6ropr6pqoon ;rdoo oro rwp6- vLo (q to e[otvto). H 06oq 6rori.d6oor1q no((eL o4prowrr6 p6).o mqv ovoyvtrrpto4 qq 3' 05or1q ouppocprlg. Ir4 (Upq, 11 05or1 6ror<Ld6oroqg e(vor e[otperu<d ouwrlprlpr6vrl r<or 6to05- rer rl ouvorverrr<r1 o).).r1).ou1(o UACUAAC. Aw(Oeto, oroug nolurUrropoug eur<opurrtreg 6ev e[vqr r<o],d ouv- qprlprdvr;, oMd 6e(Xvet ;uo npot(pu1oq yro noup(veg r1 nuptprt6(veg oe 6torpoperLr<6,9 Odoetg r<or 6rorrlpei ro vourd.eot(6to-or6Xo A (B).6ne Eu<dvo 21.5). H Odq 6ror).d6oor1g Bp(oxeror 18 6cog 40 vour<).eot[- 6ro ovodu<d qg 3' 06oqg ouppocp4q. Irn ([Un, peroMd- [etg r1 ono].oupdg orrl 06o11 6rox].d6oonq nopepno6((ouv q ouppo<pr1. Ztoug no).ur<Urropoug eur<opuroreg, q 6L- 590 KEOAAAI0 21 Iupporprl rt Ene[epyoo(o rou RNA 2 1.6 To snRNA Eivor Anopqirnro ylo Tn Iuppoqi a To n6vre snRNP nou epntrdrovror orq ouppoqrl e(vor ro U1 U1, U5, U4 xor U6. a To snRNP Uo(r Ue oproUeveg otrtreg npotrelveg oXrlpon(ouv ro oopor[dro oupporp1g. * '0tr0 ro snRNP errriq onri ro U6 nepr6Xouv 1uo ouvqpqpevl 0trtrntroux(o nou npoodever Trq rpoTeiveq Sm or onoleg ovo- yvropl(ovror ond ovnotirporo nou nopoyovror oe ouroovooo voorlporo. 3', {e ls' Boorr6g eworcg I
  • 83.
    Or 5' ror3' 06oerg ouppo<priq r<ot q oM4l,ouX(o 6ro- x,l,66oor1g ovoyv<op((owqr on6 ouqrorrrd rnq ou- oxeurlg oupporp4g nou ouvoOpo[(ovrot oXrlpror(- (owog rivo peydLo o0pnloxo. Aw6 ro orjpnloxo g6pva rowd rrg 5' r<or 3' O5oerg ouppogqg nporori oupBe( onoto6{nore ow(6poor1. To oUpn}.oro ouvqp- poloye[ror 6ro6o1u<d ndvo oro pre-mRNA. Apr<erd ev6rdgeoo prnope[ vo rouroTror.4Ootiv prSoo qg x].o- oporono(4 o4 g ou prnl5xrov 6rogopetrxo 11 preydOou g. Iuppog1 ouprBo(ver ;r6vo 6rov ouvoptr_roLoyqOo0v 6)to ro ouorolrd. H ouoreurl oupporpqg 6ro06ter 16oo npcoreiVeg 6oo xor RNA (exc6g rou pre-mRNA). To RNA no(pvouv r1 popei pru<p<itv 1.top(<ov nou ug(orowor rog prBovoux.treo- npcoreivu<d oo;rot[6to. T6oo o nup4vog 6oo xor ro KUr- rop6nl,oo;ro rov euKopuonr<cilv xurrdpov nepr6louv noMd 6ror<prd prxpd RNA. To p5ye06g rouq xu;ro(ve- ror on6 100 6ro9 300 Bdoerg oroug ovrirepouq euKo- puri:reg, evri m4 (rip4 ercre(vowor oe -1000 Bdoerg. Arogdpouv noLrj og npoq rrlv og0ov(o toug, on6 10s irog 106 ovd r<Urropo Srog ouyxewprioerg 16oo 1oprr1- ).eq rriore vo pn Unope( v' ovrXyeu0oriv d;reoo. To RNA nou neprop((owot orov nup4vo r<ol.oUwor prxp6 nupqvrxd RNA (small nuclear RNAs, snRNA), evri exe(vo nou Bp(or<owor oro xu-rrop6nAoopo plxp6 xurropontroopolx6 RNA (smalt cytoplasmic RNAs, scRNA). Kovovu<d ug(orowor cog prBovouxleonporei- vrx6 oor;.rori6to (snRNP r<or scRNp). yndpXer en(o4g pro xorqyop(o pru<priv RNA nou Bp(orowor orov nuprl- doxo r<or rqloUwor snoRNA (plxpri RNA rou nupqvi. oxou, small nucleolar RNA), ro ono(o eprnldKowor orqv enelepyoo(o rou prBooropu<ori RNA (B).6ne Ev6t4- ru21.20). To snRNP nou etrmfuirowqr orrl ouppogri, pro(( ;re noA.69 dMeg npolreiveg, q4qprot((ouv dvo ;reydAo oro- pmrdror<6 orjgnLor<o yv<rror6 <og oopori6lo ouppog{q (spliceosome). 'E1er onopovco0e( ond oum4proro rn vrlro ou p porpn q coq p rBovo u r<Leonproreivu<6 oco 1_ror(6 ro 50-605. To otol.rot(6ro oupporprjg nr0ov6v oXqtrror((etor mo6rord xoOdrg ouv66owor rq snRNp. E(vor peydLo orol.rorf6ro, pre preyo].Urepn Ud(o on6 ro ptBoorr:gduo. H EIK0NA 21.7 ouvotp(er ro ouorctrrKd rou ocol.rorrd[- ou oupporprlg. To ndwe snRNA onoreloUv ndvo on6 ro 6vq r6ropro qg pd(og rou ocopon6(ou ouppogrlg. Mo(i pe rtg 41 nporefveq Ue rrq ono(eg ouv66owor, ov- nnpooone0ouv oXe66v ro 1.uo6 qg pd(og rou. l-tep[- nou 70 dtreg npcoreiveq nou Bp(or<owor oro oopor(dro ouppogrlg ovogSpowor <og nop6yoweg ouppogfq (splicing factors). l-'lepd.oprBdvouv nporetveg onopof- rryreq yro rr1 ouvopl.rol,6ynon rou ocotrrorrd(ou ouppo- gnq, rnv np6odeori rou oro RNA un6orpogcl Kor rnv xcnooreu4 ev6g r<5wpou pre Bdoq ro RNA yt, owr6pd- oerq trans-eoreponoi4o4g (transesterification). Ercr6g on6 owSg ng npcoteiVeg, nep(nou S0 enrnp6oOereg nprrlreiveg nou ouv65owo. oro orrrprot(dro ouppogrlg Oecopefuor 6rr evepyoriv oe dlLo ord6ro rrlg dr<rppoo.qg rov yovr6(rov, yeyov6q nou unodrltrc6ver 6n q ouppogrj (oog ouv6detor pe dMo Bri;roro or4v dr<gpooq rov yo- vr6(cov (BMne Ev6rrya 21.11). EIKONA 21.7 To otoporidro oupporprlg eivoL -12 MDa. ll6we s- nRNP onoretrouv oXedov ro pro6 Tlq Ud(oS rou. 0r undi,orneg nptrlrelVeg nepiloppdvouv yvtrlmoUg nopdyovreg oupporp(g, ro- Otig en(o4g Kor rrpoTelveq nou epni,6rovrot oe dUo mddro qg 6rgpoorl g rov yovr6[rrrv. To oropor(6to ouppog4g 6rlgroupye(ror ndvo mo or6,poro np66popo RNA ror nepvd prdoo puog evdrdtrre- orlq rordoroonq ornv ono(o nept6Xer ro ypogptr6 ;16- pro rou 5' e(ov[ou ror to 6e(t5 ].doo rwpdvro-e[rivro. MSoo oro orjprnLor<o Bp(oreror pr6vo 5vo ptxp5 pdpoq rou ouppopgdvou npoi6vrog, 5v6er[r1 6rt ouvrl0rug oneAeu0epcbveror opr6ooq prerd t4v ronrl r;g 3, 06o119 r<or 14 oUvdeorl rov e(cov[<ov. To orrlpro{6ro snRNP eprnfuixowor oqv rorooreu4 rrlg 6ogng rou orrlproldiou ouppogrlq. 'Onog Kcu ro pr- Booco;rdrro, to oropor(6ro ouppo<pr]g e[optdror qn6 oI- trrlAenrdpdoerg R NA- R NA r<or o).l,r1Asnr6 pdoerg npo- reivqg - npolretv4g. Opro;r6veg owr6pdoerg nou nepr- Lo;rBdvouv ro snRNP npounoOdrouv dpeoo (euydpogo pero([ rou RNA rouq Kor. rou RNA nou ouppdmetor. lMeg owtd pdoerg npo uno0Srouv ovoyvrilproq prero[ri ttov RNP { pero(rj rov npcoreivdlv toug r<ot dMrov ou- ororxriv tou orrlpold(ou oupporprjg. H orlpoo(o rov snRNA prnope( vo eLeylOe( dpreoo orn (UUn p5oo qg np6x).r1oqq preroMd[eo:v oro yov! 6ro roug 11 oe owr6p6oerg rn vftro ouppogrig ;16oo qq oroXeuprSvrlg ono66;rqoqg [eXroprorci:v snRNA oro nu- pqvx6 erlUtrropro. H odpovono(4or1 tov nriwe snRNA, [eyoprord r1 oe ouv6uoqr6, onorprinet q ouppocprl. OLo ro snRNA nou epnl,6rowor orrl ouppo<pr1 ovo- yvrop((owor oe ouwrlpnUdvq popgrl oe 6l,oug toug euxopucirreg, ou gnepil,opBovoprdvov r<ov rputcirv. Irrl (6Un r' ow(orotlo RNA ouyyd e[vor preyo],rirepo, oMd or ouwnprlpdveg neptoXSg nepil,oprBdvouv XopoKrrlpr- mxd nop6proro Ue ro snRNA tov nolur<Urropov euKo- purorciv. Tq snRNP nou epnl,6rowor qrrl ouppog4 e(vor ro U1, U2, U5, U4 ror U6. Ovoprd(owor orjpgcovo Ue rq s- nRNA nou nept6louv. Kd0e snRNp nepr6let dvo provo- 6116 snRNA ror oprer6g (<20) nporeiveg. To U4 ror U6 snRNP ouvqOcog Bp(or<owor go(( oq dvo ocopro{dro 6r-snRNP (U4IUO). O xorv6g 6opur<6g nuprlvog xd0e s- Ailteq 30 rtporeiveq, l7% mc uotoc 5 snRNA, aao/ *^ ., ,o trg,, 1td(0! 70 nqoqvowec ouppoqns,sai. ;jXf#ffi*'nlq [o(oq iar" irlC ul(.s 5E1
  • 84.
    nRNP ofloretrefuot on6;uo o;rd6o orcr6 nporeivdlv, or ono(eg ovoyvco p((owor ro ouroclwrorirproto qwr-S m (anti-Sm). Iuwqprlprdveq oMqLou{eg orrg nporteiVeg onoteloriv ro or6Xo owrilv roJV owroopdrrov, Or un6- Lorneg nporelVeg e(vot llovo6rxdg yro rd0e snRNP. Or npo:teiveg Sm npoodSvowor orrl ouwrlp4;r6vr1 oMq- LouX(o PuAUg-oGpu 11 ono(o undpXer oe 6Lq to snRNA ercr6g rou U6. To U6 snRNP neprSXer pro opd6o npo- reivdlv nop6pouov pre ng Sm (Lsm). Opropdveg nporetveg rov snRNP nr0ov6v epntr5r<ov- ror dpeoo orn ouppqqn. iMeq (oo4 SXouv 6oprrorjg p5louq 11 e[vot onopo(r4reg yro rrl ouvopprol6yrlon n yr' oMqlentdpdoerg pero[0 rov ocol-ron6(ov snRNP. l-1epfuou ro tivo rp(ro rov npoteiv6v nou epnfuixowor orn ouppoq4 efuot ouoronxd rcov snRNP. 'OLo r<or ne- ptoo6repeg evde[(erg uno64],d:vouv 6u to RNA 6Xer dpeoo p6Lo or4v r<otdtruor1: ouvend:q, or neproo6repor nopdyoweq ouppoEr1q Unope( vo 51ouv dopur<oUg p6- ).ouq n vo oupr;rer51ouv fin ouvopprol.6yrlorl rou oo1.to- n6(ou ouppogfq. 2 L7 Adop€uon Tou fip66popou mRNA oTnv 006 Iupporpfiq " To U1 snRNP nporotre( qv evop[1 11q oupporprlg pdtrLq npoo0e0er oqv 5' 06oq oupporpqq U6orrl luog ovr(6poo1g (euyoptirporog RNA-RNA. u* To ouUnl,oro 66opeuoqg nepL6yer ro U1 snRNP npooDe6epS- vo orrlv 5' Oeoq ouppoQnq rol rrlv nporelvl U2AF, 1 ono[o npooddveror 0e Uro ouoTorxlo nuprprdrvdrv pero[u rou orlpe! ou 6ror),o0cooqq Kor rnq 3' 0eo1g oupporprlq. e Iroug notrururopouq europutlreg, or nporeiveg SR nor(ouv onopo(qro p6lo oqv evopfr1 rou oXr11-rolopott rou oupntrri- rou deopreuorlg. * To (euyoptolro rorv 0doeov ouppoqlq enuuyXdveror pulotrt oproliou rov rvrpovlov r1 ralv e[trlvirrtv. H ovoyvrrrpron rcov ouvorvelrcbv orlpdro.lv ouppogrlg nepil.oprBdver 16oo RNA 6oo rot nporelveq. Opropr6- vo snRNA 61ouv oMrl).ouXieg nou e(vor ouprn).r1popro- nr6g trrrv ouvqtverrKcbv oL).q).ouXrov 11 prero[rj roug r<or to (euydpc,.rpo ;rerofri tou snRNA Kor rou pre-mR- NA r1 peto[ri rov snRNA no[(er orlprowrx6 p6].o or4 oupporprl. To U'1 snRNP tou ovOptonou neprdXer trg r<ewptxdg nporeiveg Sm, rperg er6u<69 yro ro U1 npurrelveg (U1- 70k, U1A r<or U1C) xor U1 snRNA. H deuteporoyrlq 6o- prrq rou U1 snRNA qo(veror orrlv . l-1ept61et opxetSg neproX6g. H 06oq np6odeoqg Sm e(vor ono- pofuqrq yro rnv otrLr;l.eni6poorl ps rrq r<orv6g npcoret- veg snRNP. Or neptoXdg nou ovoyvop[(owor on6 ttg [eXoprm5g 6o;rdg p(oXou-0r1Lrdg nop6Xouv 06oerg np6o6eoqg yto nporeiveg nou e(vor provodrx5g yro ro Ul snRNP. H np6odeorl rou U1 snRNP orqv 5' 05or1 592 KE0AAAI0 21 Iupporpn xr Ene[epyooto rou RNA ouppoqqg e(vot ro npri.lro prtpo o-cn oupporprl. Tc -- RNA oMq).enL6pd pe qv 5' 0dor1 oupporprlg pe (e". :- popo pero[0 tou pov6r.trovou 5' dr<pou rou Kor 4-6 3:- oeov rrlq 5' 06oqg oupporplq. MeroMd(etg orqv 5' 06o4 ouppo<pqe r<oL oro U' -- nRNA gnopei vo lplorpronoLrl0otiv c,rryre vo eley):: dpeoo ov e(voL onopo(rr1ro ro (euydprop-ro rouc - onore).dopioro ev6g rdrotou nerpdprotog nopouoro(:, - ror ornv H oMrl).ou1[o oyprou t0nou :-: Odor1g ouppogqg mo pre-mRNA rou 125 odevo.:- (euyoptrrver oe ndvte ond rrg 6(L O5oerq pr ro ;' snRNA. 'Evo pretdL).oyUo rou 12S RNA nou 6e oupc:- ftrtror dler 6uo oMoySq orqv oMrl].ouX(o: to r<ord.: - no GG orLg 0eoeLg 5 r<ot 6 rou twpov(ou 5.Xouv oMcl: oe AU. Av eLooX0e[ puo prerd].].oEn mo U1 snRNA * ono(o onoro0Lmd ro (euydpropro mq 05oq 5, q oupc:. qq pnope( vo ouprBe( xovovLrd. ALleg nrprnroJCs.: mrg ono(eg ow(morleg preroLi.dleLg drlptoupyou, : oro U1 snRNA rrrore vo 6ronLoro.l0e( ov e(vot rKovrq .l rotooreil.ouv q prerdMo[r1 orrl 06o11 ouppocprlg, c,.:- detrvrjouv ro yevrrd rov6vo: q ou;rnLqpcr;1lorrrorr:: piero(U tou U'1 snRNA Kor rnq 5' 06o119 oupporprlg eL',: onopofuqtrl yro rrl ouppogrl, oMd q onoreLeoporL<c- rlro rrlq o'upporprlq 6ev r<o0op((eroL onox.l.eLorLxd cr : rov opr0;rd trov (euyrirv Bdoeotv nou oXq;ro-r[(owor, H ow(6pooq (euyoprrtl.ioroq rou U1 snRNA ;-re r-. 5'06o11 ouppogrlg oro0eponore(toL ond nporeiveq. - nporelvq nou npoo6S.veror orrl 05or1 6Lor<Lddcoor : (branchpoint binding protein, BBP, yvroorq r<oL og SF' oM4).enL6pd pe rrlv oM4).ouX(o rou orlpe(ou 6rontrc- 6oro119, evo q U2AF (5vo e-repodrprep6g nou onore,s.- ror ond rqv U2AF65 r<or U2AF35 oe noLux0rtoc: europurotrr<d rUrropo q rqv Mud2 orn (6Un Saccha- romyces cerevisiae) npoo6SvetoL orq oumorX(o nupt! - 6rvcrrv prerolU rqq otr).r1Lou1(og rou oqpe(ou 6ror<Lc- 6ooqq Kor rou mo0eporj 6rvour<LeotL6(ou AG oro :8- Log xdOe rwpov(ou. An6 pr6vo-rou, r<ov6vo on6 ourd i: ouprBdwo np6o6eoqg 6ev e(voL opr<erd ro1up6, r,lmc- oo rl ouvepyoo(o roug o6r1ye( oro oxnUorrop6 evoc oleru<d oro0epo6 oupnl.6rou nou ro).e[tor ot-ipn].ox: 6do;reuoqg (commitment complex). Iro rrjmopo rotv 0qLomLrov, ro orjprnl.or<o -:,- opeuorlq e(vor en(or1g yvoor6 otg oripntroxo E (,,E,, y:c ,,e?rly>, npt,ttpro). O ol4porLo;r6g rou 6ev ononel ATP (ow(Oeto on' 6Lo ro dtl.rtpro Brlporo xord q ouvoppic- ).6yqor1 rou ooUor.6[ou ouppo<pr'19, B],6ne Evotqrc 21 .8). Aorcoo, or ouvorverrr<5q o).).r1LouX(eg orLg 0e- oerq ouppognq oro yov(6Lo rrov 0lAoortrurv e(vor 1c- Lopd ouwqpqprdveq xot, ouvencog, yro ro oXnUoloUC rou ougnL6rou E onoLrorjwoL enrnp6oOetor nporetvt- r<o( nopdyoweg. Or nopoyoweq nou no((ouv rewpu<6 p6lo 16oo oe ourqv 6oo Kor oe dMeg 6repyoo(eg ouvopl-ro).dyqo1c rou oro;rotr6(ou ouppogqg efvor ot nporeiveg SR, prrc ou<oydveto nopoy6wolv oupporpqg nou nepL6louv dvc rq 6Uo goriBo ovoyvrllpLorlg RNA oro N-reLrr<6 drpo xo. pto neptopl "unoYpogri" nLoUoLo oe noMonLSq enc- voLrlqUetg rou 6rnenrr6(ou Arg/Ser (r<o).e(ror neproxr RS) oro C-re).Lr<d dr<po. Aurd ro ;ror(Bo ovoyvrirproqc Boorxfq 6womg
  • 85.
    . To U1snRNA 6Xet pLo (euYoptlp6vrl 5opri nou di'lptoupyer oprer6g neptoXeq. To 5' orpo iopo;t6vel povortrovo xot (euyoptivet pe rqv 5' Otlot1 u ppoQIlq. RNA e(vot uneU0uvo yto etdtrrl yto qv otrr1- LouX(o np6o6eoq mo RNA. En(or1q, n flepto- Xrl RS ;rnope( vo npoo6e0e( t6oo oe RNA 6oo xot oe dMoug nop6yoweg oupporprlq prSoo otrrlLenr6pdoeov preto(ti npotetv6v, nop6lowoq 6tot np6o0etrl ur6Mo" yro 6td- rpopo p5p4 tou oupnl'6rou E. 'Onog nopouotd(erot orrlv EIK0NA 21.10, ot npu:tefveq SR npoo66vowot oro ouororx6 70 kD rou U1 snRNP (r1 Ul np<otetvq 70 kD neprsXet en(or1g prto neptoXrl RS, ol d 6e Oeo- pefuor rog runu<r1 npcotetvq SR) yro vo evtg4ti- oouv rl vo oroOeponot{oouv to (euydpogd 'rnq Ue rrlv 5' 05or1 oupporpqq. Ot npcotetveg SR npooS6vowot enbqg ornv U2AF nou efuot 6eopreupr5vq ornv 3' 06o11 ouppotpr]g (pto ne- proXq RS undpXet entonq t6oo qrnv U2AF65 6oo rot orlv U2AF35). Autd ro 6(rruo npto- rcivrrdlv oMrllentdpdoecov 0eolporiwot rp(ot- Uq yro ro oxrlpqrtopr6 rou oupntr6rou E. Ot npoteiVeq SR onopovdrvowot pro(( pe to o6pr- nloro Pol Il t<t 6Xouv rqv rov6rrpo xrvrltu<tig 65o1-reuoqg tou RNA ornv o66 ouppotprlg. Iu- vencig, nt0ov6v l.eroupyofv cog evoprcrrlptot nopdyoweg rnq ouppoqriq oto rfrropo ttov notrur<Urropolv euroPuotd:v. nepLoxn B ^A^ HH JY IbrfiD $B-H$' ffiH HF.B s&E npoooeontrsf. t&ftKg #"# "g sfry ocu ccu'oii uuo cuGAccc " rFffi ,,^^*' % r rnc c un u u" oooo"oo oot^ EETiS:l AC ^dAdcouccA u u cA u ApppGme 5' c E s'E[rr]vto GU A A GU A lvrPovto 3' cG I UA I UG I UA I , H ZeuYdptouu tvtPov(ou GC GC nu'oru Tunu<5q npcoretveq SR 6ev rco6tronotofwot sro Yo- vt6(orpto tou S. cerevisiae o0te e(vqt ovoyr<o(eg Yto rn ouppogrl oe ow6v rov opYovtopr6 6nou ro origoto ouppo<pr1q e(vot oXelrd opretdBtrrlto' Aw(Oeto, e(vot on6Luto onopo(tqteq Yto rn ouppoqrl oe 6).oug toug notruxfrropoug europucireg 6nou to or]proto ouppo- griq onoxL[vouv notr0. H e(5].t[4 tov npu:teivriv SR oroug notruxrirropoug europutiteg nt0ov6v ouvetorp5- per oe ouppoErl uUnInS onotel'eopotr6tqrog Kot nt- m6tqtog oe Xol.opd ouwrlpqpSveg 06oerq ouppotp4g' H ovoyvrirptorl tretroupytr<drv 06oeov ouppoEriq r<otd rq dtdpr<eto rou oxrlporl.opo6 tou oupntr6rou E qxo- )rou0ei 66o nope(eg, 5nog <po(verot orrlv EIKONA 21'11' Erov S. cerevisiae,6nou oXe66v 6Lq to yov(6to nou ne- pdXouv twp6vto 6tox6mowot on6 6vo provo6tr6 pxp6 rwp6vto (prlroug -100-300 vouxtreond(rov), or 5' rot 3' O5oetg ouppqgnq ovoyvop((owot tout5lpovo on6 to Ut snnNp, BBP rot Mud2, 6n<oq npoovorp5pO4r<e' Auti q 6repyoo(o ovoq6petot oq oprop6g Iwpovitov (lntron definition), 6ncog <gofvetot oro optorep6 pdpoq qg Et- x6vog 21.11. (l-lopoqprlore 6tt o prqXovropr6q optogoti rwpov[tov toXUet yto prtrpd rwp6vto oe nol'ur6rrqpo eu- ropuo:ru<d r<rirropo rt 6tot q etx6vo lpqorpronote( r4v opol.oy(o rov nopoy6wtov ouppoqdg tcov 04).oortr<drv nou egntrSrowot srrl 6tePYoo(o). Aw(0eto, mq notruK0rropo euKopucottx6 yovt6ttil- AAoo"oa' nePoXn n poro ro twp6vto e(vot preydtro n 61ouv nol'0 getoBl'r1- 16 p6ye0oq. En(or1g, nepr5Xouv noM6g ol'I'r1}'ou1(eg nou pot6(ouv Ue npoYportxdg Odoetq oupporprlq' Aur6 duoXepo(vet ri rot r<oOtmd ov6qtrtq q ou(euy;r6vr1 ovoyvtirptorl rov 5' t<ot 3' 06oeov ouppogriq. H )'Uorl owori tou npoBldpotog e(vor 11 dtepyoo(o rou opl- opof e(oviov, 11 ono(o o(tonote( to ;uxp6 p5yeOoq tcov eiov(ov oro rcittopo tcov ovrilteprov europutrltcirv (pui- rog - 1 00-300 voux).eot(6to). 'Ontog cpo(vetor oT o 6e(t6 p6pog qq Erx6vog 2'l ' 1 1, xotd tov oprog6 tov efcov(ov, to etepo6tptep6g U2AF npoo6Svetot orrlv 3' 05oq oupporpdg xotto U1 snRNP (euyopcirvet pe rnv 5' 06oq oupporprlq ro0o6tr6 qg oi'- l,rllou{og rou e(tov(ou. Ir1 ouyxerptp6vrl 6tepyoo(o ouverog6pouv nptotetveg SR nou npoo66vowot oe et- 6u<5g oMqlouX(eg tov e[tov(tov preto[rj rcov 3' Kctt rov xo0o6xrilv 5' 05oerov oupporpriq. M5ool ev6g dyvtrlorou prrlXovropoti, ro oUpmtroro nou oXrlprotf(owot r<otd grl- xog tou e[ov(ou peton(mouv oo oUpntror<o nou ouv65- ouv rrlv 3' 06o11 ouppoqriq otrlv ovo6trri 5' 06o4 ouppogrlq Kot rrlv roOodtr<r1 5' 06o4 ouppoqrlq ottg en6lreveg ro0o6tr<5q 3' 06oerg ouppoqrlg 6top6oou rolv twpov(tov. Aw6 eyro0tord tqv uenttpemtr{" 6rqp6p- qoorl nou enup5net vo ouptBotiv to peroyevSorepo Bn- Uoro ouvopUoL6yrloqg rou oto gon6(ou ou ppoqf g' H ovooro)rri ourlg qg ;retdmoorlg e(vot 5vo p6oo yro rn pU0prto4 qg enrloyriq ouYKeKptgdvtr:v e(tov(tov rord tr1 pu0g(6prevq oupporprl (B).6ne Ev6t4ru 21'13)' 3'1"? A6oueusn rou [lp66poUou mRNA otlv 066 ttnpoqrts 5S lleproxn C
  • 86.
    Ul snRNA dyprouflinou rqt 'l 25 pre-mRNA h. : ... i,. ' .:.J5[ ,: :, flH g3' flfl fgqr JIH Y ,:R,F;@FrurubeFkut-5 5' Ep$vro G U G A G G lwp6vro 3'Hq $l 06on oupporpftq 125 qdevolori U 1 snRNA dyptou rUnou xor gerdUoygrivo 1 25 pre-mRNA i1r '. "',rCSl 3', 5' E{<0vro G U G AA iJ lwp6vro 3' irl g: gli :riii*?:t *ir;1 f rrflf *{ir: -t MeroMoypdvo U1 snRNA (0r Uer0ll0yU6v0 125 RNA ;'':.i:::. 3', iiiiti,lflg:: iiit i"il 5rRliil ' : .,.C.A.U,,Ui-r..AU 5' 5' E(tirvro G U G A A u lwp6vLo 3 tlK0NA 21.9 MeroMd(etq nou KoTopyodv rn treroupy(o rnq 5' 06o4q oupporprlq KoTqor6trtrowor U6oo ovTrppoflrorrKd)v UeTotrIo- [eov mo U1 snRNA ftou orroKoeroroUv ro (euydptolro Tov Pdoeov. E[tivto reuu Te).u<d, o pu'11ovrop6q opropoJ e[ov(ov nou 6roLieoolc- Befuor on6lq npore(veq SR nopSler en(onq 5vo prlXc- vrou6 nou enrrp6ner (euydprr;pro Kor oUvdeorl U6oo r.( ouppoenq pi6vo peto[rl yerrovrrov 5' ror 3' 05oea, ouppoQriq. 21.8 H O66q Ixnpono;"ro0 rou Etopon6iou Iuppogfiq * T0 ouuntroKo dSoueuonq e€rtr(ooeTor 0e np00p0u0 o0ll.10:- Dro ouppoqqg (oupntroro A) nopouo(o ATP * H npoodeon rov U5 ror U4lU6 snRNP perorpSrer To npur- 6popo otolrorrdro ouppoQrlq oe dlprUo otopor(6ro 0uppo0-i (o0Untroxo B1). e To ouUn),oro B1 6neuo perorpeneror oro oupntroxo 82, c.: onolo ro Ul snRNP one],eu0ep6veT0r yro v0 enrTpriqrer 0.: U6 snRNA v' otrtrltrenrdpdoer Ue qv 5' Oeol ouppoqrtq * H U4 onouorpuveror ono ro U6 snRNP enupenovrog oro Uc snRNA vo (euyoptioer pe ro U2 snRNA yto v0 oxlpolosr:: rorotrunro revrpo yro tr1 oupporprl. * Kor or 611o ovldpooerq lrans-eoTeponoinqq ouptpolvouv o.: evepyonorlp6vo orol-ror[0ro ouppoqnq (oupntroro C). * H 0vT[6p00rl oupp0Qnq tlvor ovTroTprnrrl oe otro ro BrlUorc M6Lrq oplpotrme( ro oriprntror<o E, r' dl,o snRNP ror or nopdyoweq nou eprntr5r<owor fir1 ouppoe- oL).qen16poriv pre ro oripnlor<o pe ro0oproprdv'- oetpd. H llil,ri:l;i ;:= l,: 6e(Xvet To oUoToTtKd ttlv ou1-- ni.6rc,rv nou ovoyvop((owot xo0rrrg npoXrope( q c,,- r(dpooq. Iro npcoro Brlpro, to ono(o e[optdtor on6 ro ATF Eltirvro -ry -Au 06on luorory(q Pyg' 6uddd0onq Odoq ouPPotPtlg sPyssAGFm lvrpdvto 5', 0dor1 ouppoqrlgH Ul ror ot nporetveg SR xoOop((ouv qv 5' 06o1 ouppoqrlq '- ':il, SR :'. H U2Af npoo66veror oqv oupri Py roL qv 3' 0dor1 oupporpr'1q, gto etbtrrl npoTsivn ovoyvrop((er m Oeon drortroDoonc SR" 0t nptoreiveg SR ouvd6ouv ro oUpntroro mrg 5' rot 3' 06serq ouppo9ng 6me vq oxnuqrrme( ro oUpntroro d6opeuorlq (E) BBP ,U2AF65i35 GU ::UACUMC --Py =AG I SR *"U24ffi,/35:- -, ------t {@_UACUMCI EIK0NA 21"10 To oupntroKo d6opeuoqg (E) oXrlpori(eror pe q drodoXxrl npooerlKn rou U1 snRNP orrlv 5' 06o1 oupporpqg, qg U2AF m1 ouororX(o notrunuprprdrvrirv/3' 0604 ouppoQnq ror qg nporelVlq ouv6vrrlolg SF1/BBP. 594 KEOA Al0 21 Iupporprl rr Ene[epyoolo rou RNA Boorxeg eworeq
  • 87.
    I o*r^rn,uouor,, V 0tou6oourou tvrpov(ou 0propdg rvrpov[tov Elrivto lvrpovro E(rivro wGUwUACUAACT PyxAGrw 5' 06o1 Odoq DrortrdDoorlg 3' 06011 0propdq elov(trrv E[tivro lvrpdvro E[rirvro xAG wq gg w utrQLJMC x Pya AGwsslE GU = 5' 0do4 06oq Dtortrd6ooqq 3'0dor1 5' 06o11 I or,,nrrr,,oooor,, V 6ro[doou rou e[ovfou UACUAACE Merdnroo4 ond d1,flentbpdoerg 6ro96oou tou e[orv(ou oe otrtrltrentdpdoerg 0rog6oou rou tvrpov[ou YnopXouv 6uo odot yto qv opXtrrl ovoyvtirprol Ttrv 5' Kor 3' 06oeov ouppoQnq, e(Ts Us opropd rvrpov[rov e(Te Ue oproUo ,:-,.iJV. -: .12 snRNP ouv66etot pe ro U1 s- - : ' P ndvco oro np66popo mRNA 95- :* rpdodeonq ornv oMq).ou1(o rou r-..erou 6ror).d6coo4q. Aur6 en(onq -i:ri.opiBdveL (euydpopro UeroqU rnq :'-'.-ou1(og rou U2 snRNA Kor rrlq : .'.1),ouX(og rou onpe[ou 6rox].d6tr:- -'.: To or-iprnloxo E petotpdneror - ..3v oro np66po;ro ocopor[6ro oup- : ri-q nou e(voL yvcoor6 oq oUp[tro- .3 A xor our6 ro Bqpo onorre( - I:c,n uor1 tou ATP. -c orlpn).oro Bl oXqpor((eror p6- : t',,o rprpep6g oUprnLoro on6 ro -: <eL U4lU6 snRNP npoo6ege( oro - --:^oKo A. To 81 Oeope(tor drr e! : :'"rpor[6Lo ouppo(pnq enerdrl ne- : I :.ai To ouotottrd nou onotrofwor : .1v ovr(6poon rnq ouppqerlq. -,:J oneleuoepooe( ro U1, ro oUp- - :(c ourd perotpS,neror mo o0pr- - :<c E2. H o<po(peoq rou U1 e(vot :-::o[rqr1 otote ro und].orno ou- . H ovT[dpoon ouppoglg npo- -,. ir Storpuo orodro roro T0 oilolo o . --: :1.rog rou otrllrofldlou 0uppoQnq ne- - :-i:','er qv otrtrqtreni6p00rl Tov ouoToTr- , l- ovoyvop((ouv Trg ouvolverrreg otr- - . ,.. IUpntroro E IU[ntroro A IUpiltoxo 81 Itlpntroro B2 Idpntroxo C1 1 U2 IXqporLoprlg tou ougntr6xou deoueuonq oro ono(o n U1 eivor (euyopouevn ge qv 5 0€orl ouppoQng tlpoo0nKn U2 yro v0 (euyoptioer pe rn 060r1 6rox),odooqg nopouofo ATP ' ^46t!n@ *AU" + Iuvdeoq tov U4,6/U5 rpL-snRNP Anetreu0dproorl rov U1 ror U4 IXrlpouopdg rou rorotrurrxorj Kdvrpou oro onolo ro U6 (euyopriver Ue rnv 5' 0don oupporprlg, ro U6 (euyopriveL en[o1q ge ro U2, ro U2 nopogdver (euyopopevo pe q 0dor1* ^F@ - ^u-- 6rortro6oror1q. ro U5 o),trr1trenr6po ror ge ro 6Uo e(6vro U6oo rnq Onroq rou & , # To npdrro gnUo rnq trans-eoreponoqorlq ' ffi; xlBe-0,,'r5'06o11ouppogr1s,o1r1pori(eror U2 Gqw "" To trooo UACUMC:PycEAGr?Em To 6eurepo BnUo tr1S trans-eoTeponorn0nq Krlperot 11 3' 0eor1 oupporprlg, ouvddovror ro eIdrvro m.'5AGu IUgntroxo C2 ?:.fi H Odog IXr;poloUOU TOU IorgorLd[ou Iuoorc.t 595
  • 88.
    srcntx6, 16(ro9 roU6 snRNA, vo Bpe0oUv 5hLo orrlv 5' 05or1ouppoqrlg. H rorol.urxrl ow6poo4 nupodotekot on6 r4v one- Aeu06pooq rou U4 nou en(oqg ouprBofuer rord q prerd- Booq on6 ro oUpnloro B'l oro B2.To U4 snRNA (orog "eyr<Ao9((er,,to U6 snRNA 6cog 6tou ttotoore( onopokq- ro. H EIK0NA 21.13 6e(yyer rg oLl,oy6g nou ougBo(vouv mrg oLtrrllenr6pdoerq (euyopdrporog Bdoeov gero(U rov snRNA xord 11 ouppogrl. Iro U6/U4 snRNP, puo ouveXriq 6rcrooq 26 Bdoeov rou U6 (euyopdrver pre 6uo [eXopror6g neptoXdq rou U4. M6].rg onoproxpuv0e( ro U4, 11 neproXrl rnq UO nou onetreu0ep<ilveror prnope( nL6- ov vo uro0erqoer dMq 6opq. To nprito pr6pog rqg (eu- yopcirver ;re ro U2, evtil ro 6er3repo oXqprot((er puo evSotrroprorrl qoupr5ro. H otrl,qLenbpooq prero(ri trov U4 r<or U6 efuot ol.rorBo[o ootiprBoq pe rqv oMqlen(6po- o1 pero[ri trov U2 Kqt U6, on6te 4 one].eu0dpoor1 rrlg U4 eMyXet rqv xov6r4ro rou ooUorr6(ou ouppo<ptjg vo npoXcoprloer ornv evepyonorngdvn rordorooq. l-ro L6youq oo<pfvetog, q err6vo 6e(1vet ro RNA un6orptol.ro o' errerop6vrl poperl. Iqv npoyponr6q- ro, 6prcog, r1 5' 06oq ouppog(q Bp(orerot xowd orqv oM4lou{o U6 oprdoog 6(n},o orqv 5' ntreupd rnq oI- LrlLouX(og nou e(vor npoo6e6epr6vq oro U2. Auuj q o),- LqLouX(o rou U6 snRNA (euyoprirver pre oAlr1trou1(eg 5'(opLmepfl) 06on ouppoQnq EIK0NA 21.'13 To (euydpogo U6-U4 eivor ooripporo pe ro (euyo- proUo U6-U2. Orov ro U6 ouvdeOei mo ourl.roridro ouppoQnq, (eu- yoprlver pe ro U4. H onetreu06proon rou U4 enrp6net ULo dttroyrl orn druu6pQoon rou U6. 'Evo u6poq rnq otrtrrltroux(oq nou onetreu- 0eptir0nre oxnUor(eL pro rpoupr6ro (yrpr) evdl ro dtrtro (poQ (euyo- pdlver pe r1v U2. Mro yerrovrrrl neproxn rou U2 eivor 11611 (euyopop6vrl ge q 0eoq dtodddooqg, q6pvowog ro U6 ovnrpu- or0 oq 6rortro6oor1. l-lopoqprlme 6r ro RNA unomptrllto 6ev 6Xer ro ouvl0top6vo npooovorotrropr6 ror qofueror oe rore00uvorl 3'-5'. To Ul (euyoptiver pe qv 5' 06srl ouppoqnq 5' 0dorl ouppqqrlq 3'AU'GAU..5' 5L-U A C U AC ------------AG--- 3', 06011 0Lortro0roor1g n To U6 (euyopdrver ge qv 5' 6dor1 oupporprlg U6 3'GAGACA.5' 5,AGGUAUGU 3, 5' Odon ouppoQnq E[tirvro 1 at +1 Eqdvro 2 at +1 H oupporprl Xplorponorel pro oerpo ovrr6pooa-, (e uyoptirporog pero[0 rtov snRNA ror rorv 06oetov 0uppoQrlc rou twpov(ou nou Bp(or<owor r<o0o6rxd qg ouwrlp-- p6vqg GU mqv 5'0rior1 ouppo<pr1g (prerol.LdleLg nc- evroXriouv 6vq rSroro (euydpulpro Betrtr<r-lvouv rnv or:c- re).eopotrrdrnro rnq ouppocp4g). Iuveno-rg, xord rqv nope(o rqq ouppogrlq, npoy;-c' ronoLorjwor op rer6g owrS pooerg (euyopr.,.r ;roroq [rer c- (U turv snRNA Kor rou RNA unoorpcirprorog. O. owrbpdoeLg ourdg ouvotlr[(owot orrlv C. snRNP dXouv oMrlLoul(eg nou (euyoprrrvouv tdoo -a ro un6mpull-ro 6oo xot prero[ti toug. En(oqg, 51ouv ;-rc- vdxi.toveg neprol6g oe Oqlldg nou Bp(or<owot t<owd o: oL).q).ouX(eq tou unoorproporoq Kor no((ouv onUowrK. p6).o, 6norq ouvdyeroL on6 ro yeyov6q 6rr preroMd(e.c ortg Oqlteg roropyorlv q ouppogq. To (euydptopro preto[rj rou U2 Kor rou otlpe[ou 6Lc- r<),d6oror1g xor prero[ri rov U2 r<ot U6 6rlprroupye( pric 6oprr1 nou por6(er pre ro evepy6 r6wpo rov ouro-oup- pont6pevorv rwpov(tov 11g opd6og ll (B).5ne ELrovc 596 KE0AAAI0 21 IuppoQrl rr Enelepyooro rou RNA J 5', Korotrurtxd
  • 89.
    2 1 15oillv Evorrlra 21 '9). Auro eye(pet rlv nt0ov6trlto :c rorotrurrr6 oustottx6 vq e(vot Utq 6ogn RNA nou :opoyetot on6 qv oMrlleni6poorl U2-U6 To U6 (eu- ,iopover Us rnv 5' 05oq ouppotpqq Kot Pto 0r1l'rd mo U5 snRNA Bpioxetot or<ptBtog 6(nLo on6 ttg npolteg 3ooerg Kot rov 60o e(irlv(tov. Motrov6tt ot 6ro0Sotpeq avbe([etg unodetrvJouv 6tt o prllovtopr6g r<ordtruorlg reoo oro otrlpor(6to ouppoqrlq Boo((etot mo RNA, q :uveto<popd tov npotte[vtiv 6e pnope( v' onox]'etore(' ',lio uno{.rq<pto npote(vr1 e(vot t1 Prp8, ;rto prey6trq npo- :eLvr1 trptoprqroq nou 6rlpuoupye( dpeoeg enorp6g rot -a rLq 6rio O5oetg ouppqqrlq, 5' t<ot 3', pr6oo oro ooJpo- :5ro oupporpqg. Kot ot 6[o owtdpdoerg trans-eoreponoiqorlq npoy- - oronoto Uwoi oro svepYoTlo tn Udvo otr: prot(6to ou ppo- :11g (o0prnLoro C) p6LLq olorLqpo0e( pto oetpd :ro'6toro(eov tou RNA. O oXrlpiottop6g tou Ldoou -q 06oq dtortrddororlg roOop((er q lplotprono(qoq :'^c 3 06onq ouppotp4g, enetdrl ri 3' ouvotvettr<4 oMq- ':.X[o nou Bp[oxetot nto xowo oqv 3' n]'eupd tqq :.:nd6too4q y(vetoL o or6Xog Yto rrl 6eUrepq trans- . -apono(qoq, To orlprowtr<6 oupndpoo;.to oro ono(o xotoLqyouge :.. r. ort to snRNA rqq ouoKeuqq ouppaqnq aM4Lent- - :: -tv t6oo peralu rcuq 6oo Kot pe ro RNA unootpupa ,.: -t (euyapds;tarcq Baoeotv xat auttq ot aMrllent6pa- -. : €nLTpenouv aMayiq or4 5opr1 ot onobq pnope[ va -..auv Kowa rLq oilddeg nou awtdpolv 4 ax6pa xat va :' -.o u pytloouv xata)'urLxa xiwPa. -op6rt to ocopot[6to ouppo<pr1g prd].]'ov efvot f-ito -..:q Ulxovn RNA, 6noq to ptBooto;rdtto, yto vo Let- -: -:v(oet lpetd(erot noMo0g npoteivtr<otlg nopdyov- -:: Extetop6veg ovoL0oetg pretoLl'o[eov nou -ttruorollotrloqrov orn E[un ovoyvtrlptoov t600 to :',- ooo Kot ro npoJTeivtxd ouoroflr<d (to ono(o ovo- -.rc(oi Ue ro oKpovriprto PrP: pre-mRNA processing, ,-rieoyoo(o pre-mRNA). Apr<etd on6 to npoi6wo ou- - - , :Lov yovt6(tlv SXouv prot(Bo nou ro rowonototiv trlg : - :', eveLo RNA eltrooov e(opttiprevcov ono to ATP, ot :-:.eq e(vot rp(otpreq Yto Uto oetpd ovo6tord[eotv mo - -*:;i6Lo oupPotPrlq. + Prp5 e(vot rp(otpr1 Yto rnv np6o6eor1 tou U2 m1 :: ;- Sror<.l.d6coor1g xotd q pretdBooq on6 to o0;rnLo- . - f mo A. H Brr2 6teurol0vet trlv onel'euO5ptr-loq - - , J 1 roL U4 xotd tn pretdBoorl on6 to orlprntroro 81 - : 32, H Prp2 e(vot uneJOuvn Yto tqv evepyono(1or1 -:, :.1;.tott6(ou ouppoqrlg r<otd tr1 trletotponq rou , -- .i.o<ou 82 oe o[prn}'or<o C rot 11 Prp22 Forl0d . -., :neleuOdpooq tou r.,-lpt;'rou mRNA on6 to oopo- - : : ouppotpqq. Entnl,6ov, opropr6veg RNA etr<doeg -: -:-v pd).o orrlv ovoxtit<trtooq tov snRNP Ytq rov : -: -i,/o r<Urtro ouvoppro).6yrlonq rou otr-11-'tott6(ou ---*-vll, : -:o to euprl;roto e[qyoriv yrot( xpetd(erot udp6- - r- :ou ATP oe 6td<popo orddto rrlg ovt(6pooqg - -:::c.cqq, polov6tt ot (6teg ot ovtt6pdoetq trans- , - .:cno(qonq 6ev onottoUv ATP. I-1op6tt oro ooUo- - - : ;..rppogr1q ouprBo(vouv 6to6oXtrdq ovodtotd(etg rou RNA, e(vot o[tooqtrle(oto 6rt q 6to6troclo ctr ''' Tot noq e(vot owtorpentrl prerd rqv nprr-ltt1 roL 5a-::- pr1 ow(6poorl trans-effiepono(r1or1q. 21.9'Evo EvqA orrtro Ecouori6ro lupporpfiq Xpnotponolei AlogopeTlKd snRNP Ylo vo EneEepyoorei To <tEAdooovorl lvrp6vto o Mto evotr)ronxn o66q ouppuQnq Xplotponore[ Uto dt]rn oud- dq snRNP, 1 ono(o 6Xet xotv6 pe to rupto otoporidto oupp0- qrlg privo ro U5 snRNP. . io tvtp6vto ot6Xot 6Xouv ueYolUtepeq ouvotvetxdq olJrrl- trouXieg oilq ouUBoI6q ouppqqnq, Xolpig v' orolou0ouv nt- md roug rovdveg GU-AG n AU-AC. . T0 uei(ovo rot to eldooovo otrlpor6to ouppoqrlq uopd(owot orlpovrxoug nptrlreivtrofg nopdyovreg, gero[6 dtrtrtov ttg nPtrlreiveq SR. To twp6vto GU-AG enu<potoUv (>98olo rov ougBol'ti:v ouppoq(g oro ov0pdrnwo yovr6(topto)' E(orp6oerg e[- ,oi o, 1r4 t<ovovtxSg ouprBoL6g ouppog(q AU-AC rt dMeg nopohLoy6g. l-1ol,ot6tepo, ro twp6vto out(g -qg "dooorog r<otrlyop(og ovoq6powov oq twp6vto AU-AC oe owtdtomotr'n npoq to "ge((ovo" tvrp6vtq nou qrolouOo0v tov t<ov6vo GU-AG rot6 q ouppo- <p(. I(;repo nou 6tol,eurdv0qre n pnxov{ ene(epyo- oloq tOoo t<ov pret(5vtov 6oo Kot rov etrooo6vtov rwpov(ov e(vot ntr6ov oo<p6g 6r ourrl q opotroy(o yto tr1v etr6ooovo rorqyop[o twpov[tov 6ev e(vot on6truto orptBrlg. Me o6nV6 trlv epnetp[o nou onorcrf04re on6 tqv notr01povr16peuvo pe owxe(pevo to pre[(ov otopot(6to ouppo<priq, n Unxovn ene[epyoo(oq tov e]'ooo5vcov w- tpov(tov 6ioLeur<dv0r1re yp(yopo. AnoreLefuot on6 U11 r<ot U12 (q4eil(owot pe to U'l rot U2, owtorofioq), 5vo r<orv6 U5 nou propd(etol pe ro pre((ov oopotfdto oup- pqenq Kqt ro U4atac Kot U6atac snRNA' H ow(6poor1 buppoEnq ouotocnxd e(vot nop6ptoto ge rqv ow(6po- on ytq to pre((ovo rwp6vto Kqt to snRNA 61ouv ovdtro- youg p6trouq: to U11 (euyopdrvet Ue rtq 5' 06oetg oupporprlg, to U12 Ue rrlv oMrlLouX(o rrlg 05oqg 6to- drddooqq rowd orrlv 3' 06on oupporpqq Kot ro U4atac Kor U6rt"c nop5louv ovo)ioyeg Leroupy(eq rotd tr1 ou- vop;rol6yr1or1 Kot rrlv evepyono[r1or1 tou otrll'ton6(ou ouppogrlq. 'Onoq ono6e(X0nre, q enil'oyr1 tou rtinou rou oo- pron6(ou ouppo<prlg en4pe6(etot en(or1g on6 ng oMrl- ioul(eq dLl'tov rprlprdrov rou tvrpov(ou, on6re un6pXouv optop6vo twp6vto GU-AG nou ouppdmov- ror on6 to otopor(6to ouppoqriq rfnou U12. Mto tolu- pr1 ouvotvettrri oLLqLoul(o oto optorep6 drpo r<oOop((er rov U12-efoptdrprevo tUno tvtpov(ou: 5' ,i:,.j 'Ev0 Evotrtroxlr6 loporioro luppoqnq Xplotgonote(Atorpopertxo snRNP Yto vo Ene[epyoore(ro "E],com"l ' :: ' I 5Ei
  • 90.
    GnUAUCCUUU,..PyAGc 3'. ItrlvnpoYportK6rnro, ro ne- proo6repo U12-e[optrirgevo twp6vto 6Xouv oro 6rpo rouq rnv oMqLouX(o GU...AG. Entonq, 6Xouv pto el,ot- pertrd ouvrrlprlpr6vq 06oq 6tortrddtootlq (UCCU- UPuAPy), 11 ono(o (euyopcilvet Ue ro U12 (BL6ne Etx6vo 21.3). Aurq 11 6togopd mtg oLl4LouX[eg qg 05or1g 6rox).66oo49 e(vot 4 rripto Srdxptoq preto[ti Uet(5vcov xor eLooo6vrov rwpov[rov. l-t' our6 to tr6yo, to pe(ovo rwp6vro r<otroUwot U2-eaptdpeva, wti) ta eAdooova U 1 2-eaprdpeva, aw( yn tw p6vLa AU -AC. Or 6uo r6nor twpov(tov ouvundpxouv oe notrfi,o yo' vrdtcill.roto Kor rrq neptoo6tepeq gop6q Bp(or<owot oro (6ro yovi6to, To U12-e(optcirpevo twp6vto ouv4Oog ne- prBdMowot on6 U2-e[oprdrpevo twp6vto. H <pu].oyo- v(o ourrilv rrrlv rwpov(cov uno64Ldrvet 6tt (ooq xdnote rupropXotioov ro AU-AC U12-e(opt<irprevo twp6vto, oL- trd oe peroyev6orepo orddro qq eE6LtEnq enxpdrqoov ro rwp6vro pre drpo GU-AG nou e(optriwot on6 ro U2. H rorvrl e[6r[q ttov ouorqpdrov unoypoppr(etot on6 ro yeyov6g 6tr XpqoqonotoUv ovdloyeq op66eq (eu- yopropr5vov Bdoeov peto[0 trov snRNA Kot rou pre-m- RNA unompcbprotog. EntnLSov, 6tror ot onopo(rrltot nopdyoweg ouppoqrlq (n.X. npoteiveq SR) nou petre- rjOqrov p5Xpr rcilpo e(vot ovoyro[ot yto t4v ene[epyo- o(o r5oo rov rvrpov(rov t6nou U2 ooo Kot rov rwpov[olvilnou U12. Mro o[roorlpre(toq dtogopd peto[ti ttov U2 xot U12 ninov rwpov(tov e(vor 6tr to U1 rot U2 go[verot v' ovo- yvrop((ouv ove[dpqru ug 5' rot 3' 06oerg ouppoqrlg oro pre(ovo rwp6vro xffid ro crXrlprottopr6 tov oupntr6- r<ov E ror A, evdr ro U11 rot U12 npcilto oXqpror((ouv 6vo oriprnLor<o ru 6nero 6pXowot 1-to(i o' enorpq pe ng 5' ror 3' 06oerg oupporpfg yto rrlv 6vop(r1 tqq ene(ep- yoo'(og rrov eloooSvcov twpov[tov. Aw6 6tootpotr[(er 6rt or 06oerg ouppo<prjg oro eldooovo twp5vto ovoyvcop[- (owor rout6lpovo on6 to prqlovtopr6 optopo6 twpo- v(ov. En(oqq, 6rot onogefyetot I "otiYxuol, ttov pr4Xovrilv ouppoqrig rord r1 gerdBoorl on6 rov optopt6 efov(rov orov opropr6 rwpov(cov yto rrlv ene[epyoo(o rov prer(6vtov rot etrooo6vtov twpov(ov nou undplouv oro (6ro yovbro. 2 l .l O H luppocpri Tou pre-mRNA llr0ov6v 'Exer Korv6 Mnxovrop6 U€ T' AuToKoToAuTtKd lvrp6vro Tnq OU66oq ll * To rvrpovro uq 0uo6oq ll err6;tvovrot on6 p6vo roug ond ro RNA peoo evdg ougpovrog ourororotrurtrlq ouppoqnq. * 0r ouu0otr6q oupporprlq Kot o pnx0vtou6g ouppoqqg rov tv- rpov[rov rnq opodoq ll eivot nopo;toto Ue Trl ouppotptl Tov ru- pqvrr6v rvrpovlrov. e 'Evo tvrpovto rqg ogoDoq ll Stntrdrverot oe gto deureporoytl 6oprl nou dlUroupyei 6vo rorotrurtxo r6vrpo ro onolo Oupi(et q 6oUrt rou nupqvtrou tvrpov[ou U6-U2. To rwp6vto rcov yovt6[cov nou rtodtronotofv nporct- veg (orr1v npoyl.tottr6tnro, 6Lcov ttov yovt6[tov erc6q rov Truprlvrrrirv yovtS[tov TIou rodtronoto0v IRNA) droxp(vowor oe rpetq yevrxSg r<otrlyop(eg. To twp6vto rov nupnvrKdrv pre-mRNA ovoyvop((owot g6vo on6 rnv nopouo(o ttov dtvourLsott6(trlv GU...AG mo 5' ror 3' dr<po Kot rnq 06onq 6roxld6<ooqg/ouororXioq nup4.u6rvrirv rowd mo 3' dxpo. Aev 61ouv row61+ porrrlprorxd 6ewepotoyoUq 6oUiq. Aw(Oeto, tq u- rp6vro qg oprdSog I xot qg oprddog ll nou Bp(orowcu oe opyov[dro xor Borcrrjpro (rwp6vto otrrddog I undg louv en(or1g mov nupf vo ptovorUrropo:v/otrtYorfrre pov euKoputotc0v) to[tvoproUwot oigqrovo Ue rnv eororeptrrl opydvtoo4 toug. Ko05vq 6tnl,c0vetot oE 1.uo tunrxq 6eurepotoyrj 6oprr1. To rwp6vto oprd6og I rot oprd6og ll 6Xouv rrlv ofie oqpre(coq trov6rlto vo err5gvowot on6 p6vo roug on6 ro RNA. Aut6 xoLefuot ouroouppog{ (autosplie ing rl self splicing). To twp6vto optd6og I efuot nto xor- vd on6 ro rwp5vto oprd6og ll. Yndplet ptrpri oX6orl peto[( rcov 6tio rd[ecov, oMd oe r<dOe nep(moorl ro RNA npoyproronore[ to (6to qv ow(6pooq ouppoqrlq in vitro, Xrrlp(q vo Xpetd(etor ev(upru<69 evepy6rqteg nou nop6lowor on6 npcorefveg. 0o16oo, e(vot oXe66v B6- Boro 6rt ot npoteiVeg e(vot onopofuqteq in vrVo roOtirg Borl0oriv oro 6(nLopro (B).6ne Keq0roto 23). H EIK0NA 21.15 defivet 6tt to twp6vto rot tov tpttil rd(ecov emdgvowot pr6oo 6uo StoboXtt<triv trans-eore- ponorrloecov (6ntoq ebopre npo4youpr6voq Yto ro nuprF vrrd rwp6vro ornv Etr6vo 21.6). Inlv nprirrl owL 6poon, n 5' ouUpolri e[ovfou-twpov[ou 66Xerot en(Oe- oq on5 puo eLerl0eprl oprddo udpo(uL(ou (nop6Xelot on6 puo eocoreptq 06oq 2'-OH oro nupnvtKd twp6vto xor ro rwp6vro oprd6og ll, f on6 6vo eLeri0epo vourle- ot(6ro youov(vr1g oro twp6vto opd6og l). Itrl 6ewep4 ow(6pooq, ro eLe0Oepo 3'-OH oro drpo rou one),eu 0epcopr6vou e[ov(ou enm(Oetot or4v 3' oupBolrl rwpov{- ou-e(ov(ou. Yndplouv o1.tot6tr1teg preto[ri tov twpov(ov rqg oprd6oq ll rot rqq ouppogtig rou pre-mRNA. To pme Xov6pror<d twp6vto oprddog ll err6gvowot p6oo rou (6rou 1-nlXovtopro6 pe ro nupqvtrd pre-mRNA dtoptSoou ev6g trdoou nou ouyKpore(tot 1.t' Svov 6eop6 2'-5'. 'Orov 6vo pregovtopr6vo RNA opd6oq ll enod(erot ln vtf- ro ltop(q entnp6o0eto ouoronrd, prnope[ vo npoYpcnG notrloer qv ow(6pooq ouppogrlg. Aw6 oqpro[vet 6rt ot 6uo owrdpdoetq trans-eorepono(qoqg nou go(vowor orrlv Ex6vo 21.15 npoypqtonototiwot on6 rrlv 6to rqv oM4l,oufo RNA tou twpov(ou opr66og ll. O optOgoq rrov Etootpo6teoreptrciv deqrtirv ouwqpefuot ornv owi- dpoorl rr 6tot 6ev onome(tot e[roteptni nopoXq ev6p yerog. Aur6 0o prnopo[oe vo e(vot oqpovttr6 XopoKrnprortr6 orqv e[#rt[q trlq ouppo<pr1g. 'Evo rwp6vto ogd6og ll onor,cd 6ewepotoyq 6o94 nou nepdler opr<et5g neprol6q ot ono(eg oXq;rot(ov- ror pr5oro (euyoptop6vov ;t(oXtov xot l.tov6xtrtovtov 0q- l,u6v. H neproXq 5 6toXrr:p(etot on6 56o Bdoetg on6 rrlv neprol4 6, 11 ono(o nept6Xet 6vo rotdtrotno A nou 6olp((et qv oprddo 2'-OH yto tqv npdnrl trans-eorepo- 598 KEOAAAI0 21 Iuppotpn rt Ene[epyooio rou RNA Boorxcg eworcg
  • 91.
    rov o opLep6qrov 6uvrlnKov unoorpoprdtcov), or npo- re'iveg 6nor(ov 6Lo r<ot Trro orlpowrK6 p6Lo, 21.11 H luppogli eivor XpovrKd Ko! Aetroupyrrd Eu(euypdyn p€ tloAAd Bfiporo ornv'Ergpoon Tov fovr6iov o IuppoQrl gnope[ vo npoyporonoul0ef rord rr1 dtdprero u1g petoypoqrlg d pero q peroypoqrl. . 0t unxovrouo(qg peroyporpfig ror qq oupporpnq e[vor guoxd xor treuoupyrd evonorrlp6vor. o H ouppoQtl ouvd6eror ge qv e[oytrlyrl tou mRNA Kor rov dtreyxo rnq moOepdqrog. r H ouppoQn orov nup4vo pnope( vo enqpedoer q perdqpoorl rou mRNA oro xurrop6n),oopo. An6 xop6 ovoyvrop[orqre 6tt q ouppo<prj rou pre-mR- NA ouprBo[ver row6lpovo Ue rn lreroypo<pri, nop6rr or 6uo owt6p6oerg prnope( vcr Trpoypqronorrl0o0v [elcopr- oro, in vitro ru 5Xouv peLerrlOe( og (eXo:ptorSg 6repyo- o(eg crrqv 5r<gpooq r<ov yovr6(cov. Kripteg nepopcnu<69 ev6ef(erg nou unoornp(ouv our6 to yeyov6g npoq].- Oqv on6 rrq flopcnnp{oetg 6tr notrtrd oupBdwo ouppq- gqg oLoxLqprivovror nprv rrlv o).orl,rlptr:on rnq peroypogrlq. l-evrrd, ro rwp6vro nou Bp(orowfl xowd oro 5' dxpo rou yovr6(ou oEorporiwor rord q Srdp- Kero rrlq peroypoqrlg, evd: to rwp6vro nou Bp(oxowor xowd oto 16l,o9 rou yovrS(ou ene[epyd(owot xord rr'; drdprero rrlg preroypoqrlg 11 r<or prerd q pretoypogrj. Ercr6g on6 q Xpovxri o6(eu[r1 prero(ri peroypotp4g xot ouppogrjg, nt0ov6v undplouv rt dll.or L6yor yro rq tremoupyu<q oUvdeorl outcbv rov Boou<tilv Stepyooui:v. 'Owoq, ot prlXovSg yro rnv 5'- rdAuQrl, qv oqo(peoq rwpovirov, or<6pro Kor rnv noluo6evuL(toorl oro 3' drpo (BL6ns Ev6trya 21.16) ep<pov((ouv rpuotr5q oMrl).ent- dpdoerg Ue rrlv r<evrpu<r1 pnxovn r4g peroyporprlq. 'Evog r<ow6g prqXovtotrr6g efuor vq Xprlorponororiv 14 pe- yd).q C-reLu<q neptoplg qg peyotrrjrepqg unopovd6og rqg Pol Il (yvoorrl og CTD) rog 06o4 yro ro <p6prco1-ro 6rdgopulv nopoy6wrov ene(epyoo[og tou RNA, nop6rr orrg neptoo6repeg neptmdloerg 6ev 6Xet ro0optore( ov r; np6odeorl e(vor d;ieoq 11 dropeool,oBekor on6 opr- opr6voug rorvo6g npcoreivrxoiq i RNA nopdyoweg (BL6ne Ev6trya 20.8). Mro r6roro rpuou<rl evono(qor1 droo<potr(er rrlv ono- tel,eoprotxrl ovoyvcirproq ovo6u6pevov o4 prdttov oup poeiq yro ro (euydptopo yetrovrrciv l.enoupyrxri:v 06oeov ouppo<p{q xord 14 ;reroypo<ptj, 6torqp6wog 6ro1 1.uo o6prl oepd ouppog4g oe rore60uvoq 5'npog 3'. H ovoyvcOprorl rov ovo6u5pevov oqprdrcov ouppo- tp4g on6 roug nopdyoweg ene[epyoo[og rou RNA ror ro 5v(upro nou ouv66owqr pe ro oriprntroro enrpqruv- orlg Pol ll enup6nouv en(or1g oe ourorig roug nopdyov- req v' owoyovroroUv onoreLeoporxd dMeg prq er6xdg nponelVeg np6odeo4g oro RNA, 6nog or npoter'veg h- nRNP, nou Bp(or<owor nowof orov nuprlvo yro rrl ou- oreuqo(o rou RNA. Av 4 ouppoEri tou RNA encogel,e[ror on6 q pero- ypoqn, ytot( vo prrl ou;rBower ror to ow(orpogo; l-1pdy- 1.rorr, ou(ov5;reveg ev6e([erg unomrlp((ouv 6rr rdrr r6roro 6wtog ouprBoiver. 'Oncog nopouord(eror qrnv EI- K0NA 21.17, ro 6v(upro qg 5' rdlur,l.rrlq rpo(veror 6l Boq- OoUv vo [enepome( n opXrxri UeroypoerKq nofor1 : .',,:. ... I Iu(eu[1 rrlq peroypotprlq [0 r-. ovridpoorl 5' Kotrurflq. H UeroypoQn p5oo q-q P: ll opXrro oroporo rovro oqv oqerqplo Tlq Usrc- ypoqrlq. H youovutro-rpovoqepoorl (GT) rot q - pe0utrorpovorpepool (MT) enrorporeuovror o : o0pntroro rqq Pol ll yro vo rorotruoouv q 61proi"-- yio luog xotrunrpog oro 5' oxpo rou veooXlpori.:- pevou peroypotpou onou npood6verot 6vo etDii: oupntroro. Aur6g or ovrr6pooerq enupenouv o:-. Pol ll vo nepooer oe nopoyr,lyrrl enrprlruvol, 600 KE0MA|O 21 lupporp1 rr Ene[epyoo(o rou RNA Boorxeg 6wong
  • 92.
    |:rppo,p,l I H nporeivnnPoobSvetoL { oto ouuntroro oupPoQnq .*+F*".,:, ---- i- I H nPulteivrl noPogdvet on1 t ouUgoIn e[ov[ou-e[ov[ou .....,"..,,' ; To oripnhoxo (EJC) * ryvryl*qv:lll , RNA ot oilofeq epnL6rowot ornv eEqYoY( tou mRNA l-lporpov6q, outo( or ;rllovtopro( ;rnopeivo eveplofv or vepyortrd npodyowoq qv e[oytrlyn tou mRNA nou npodpleto, on6 rr1 peroypoqrl rot tr1 oUylpovn Fetn petoVplqri depyoolo rrlq ouppoqriq' Aw6 forr:q [erv6 vop(q r<otd rn UeroYpoqrl. To oign)'oro rp6odeoqq orqv rol0mpo CBP20/80 go(vetot 6r npoo6Svetot dge- oo orn pnxovn e[oytr:yr1g mRNA (ori;rntroro TREX) r<otd tp6no'nou'e[optoror on6 rq ouppoqq cilsre v' orporpS- oer to npdlro twp6vto xowd sto 5' dr<po yto vo 6euro- l.rjvet rqv e(oyoyrl rou mRNA. 'Evog Bootr6q nopd- yowoq t o, 6,opeooloBefuot orqv e(oycoyrl tou mRNA e[vot o REE (orrf (6Pn rotrefuot Aly, Yralp), o ono(og ono- reLe( pSpog tou EJC t<ot oMrllenrdpd dge.oo Ue rnv nprtdtun TAP (orq (6Un, Mex67p) nou peto<p6petto mR- NA 6nog rpofuetot orqv EIK0NA 21.19' To otiprntroro EJC 61et 5vo np6o0eto p6Lo oq ouvo- 669 rou mRNA ercr6g tou nup(vo, YeYov6q ;re preydtro o*(,.rrr,o orrl oro0ep6tnro rou mRNA oro rurrop6- ntroqto. Aut6 ouprBo(vet enet6q 5vo EJC nou SXet ouy- r<por4oer optoptSvo ov6prol,o mRNA. prnope( vo enrorporcUoet 6Moug nopdyoweq ot ono[ot npodyouv rrlv o<po(peo4 rnq npoororeutrrlg r<oLrimpoq on6 ro 5' 6xpo rou mRNA pe q 6pdoq ev(0prov onoxdLu$r1g' 'Onog qo(vetot ornv EIK0NA 21.20,to EJC rovovwd ono- 1ro^p,irttot on6 to gerorloUgevo ptBooto;r6tto rotd u1 btdpr<eto tou npcitou r0r<trou gerd<ppooqg .oto rurrop6- ntroopto. Qor6oo, ov o' 5vo ene(epyoopr6vo mRNA et- orn ou$oln e(ov[ou-eqov[ou I I o EJC npoo66ver npoteiveg nou epntr6rovroL ornv eqoYurYn' evrontol, onoddpnon rou RNA : To EJC (oupntroro ouUpotrnq e(ovitlv) evonorl- :,-r r0VT0 orn ouu0otrn ouppoQrlq olq ouveneto rnq ovriopoolq ,: litlq, . : , .l orov unor<wr1rr1. Ot nopdyoweg ouppo<pr1q 9o(- . :. ort no((ouv rdnoto p6l'o oio vo 6teurol'Uvouv -- . inL;.rrlruvon rnq ptetoyporprlq Kot o oxrlUottopr6g -, 3 oxpou tou mRNA e[vot ooqrog ro0optmtr<69 yto - -. :epUorLop6 trlg pretoypo<pqq (BL6ne Ev6trlta -- -5r Iuvenc,:q, 11 petoypoq4 r<ot 11 ene(epyoo(o tou :',: e[vot e(orpettxd ouwovtopdveg oro r<[rropo rov - :'...,r <urropolv euxoputotrov. - ene[epyoo(o tou RNA ouv66etot ]'ettoupytxd 6Xt --,3 ue rlovo6trd preroypoqtr<d ouprBdwo, oMd eni ---: Jt ro0o6tr6 Bqpoto, 6n<oq q e[oycoyrl tou mRNA . : : eleyXoq qq oro0ep6qtog. An6 rotp6 yvorp((ou- -. .., .o RNA nou 6Xet unocne( otelq ene[epyoo(o t<t .1:<oou0e( vo neptdXet oprop5vo twp6vto 5e ;rnope( : "!o10ei onoteLeopottxd, yeyov6q nou (otog ocpeil'e- -: :ro cpotv6pevo KoroKpftnongtou otollott6lou oup- :::qg 96oo mov nupqvo. H eu66oor1 rqq e[oytoyqq -, - "nRNA on6 q oupporprl ovodetrv0etot pe tqv 6ve- r'- trrov nuprlvo RNA Xop(q twp6vto nou npo6pXetot --: oDNA n pre-mRNA nou 0' ono6oloet tow6orlpio : "q xotd q ouppotprl' To RNA nou 6xet nepdoet pr5o<l _-: 5Lo6troo(og qg ouppo<pqg e(dyetot nto onotee- - -:ttrco on6 to RNA nou npo6pXetot on6 to cDNA, ye- :.cq rou unodrlltovet 6rt q dtepyoo(o trlg ouppoqqq ::-0o r1v e[oYtoYq rou mRNA. Onrr.lg <powetot orrlv :.:".1r:ri ':-r ':, ndvo'l orrl oupBoLq . : lviou-eEoviou evonor(Oerot 6vo et6tr6 o0prnLoro, yvol- r : oq oripnhoxo oupBoA{q e[tovitov (exon iunction :omplex, EJC). Aw6 to otiptnoro qo[verot 6tt entorpo- -.-.i dp,.oo opto;rdveg nporelVeg nou npoo66vowot fio fl I KyTTApon^ArMA I @ l#r#lX.**, Mto npotreivrl REF npoo66verol o' dvov nopoyov- ro ouppoQrlq rot nopol.tdvet Uo(i Ue ro ouppoltp6vo npoiov RNA H REF npoo06veroL o' tivov nopoyovro eEoytoVrtq nou npoo6eve- T0r 0T0v nuplvtro nopo. H rporsivn REF (Aly) e(vor p6pog rou EJC 0 nopdyowoq Uerogopdq TAP/Mex npood6vetor cqv REF tou nupqvtro! ndPou ?,t :t H lupporpn e[vot Xpovu<o rot Aerroupytro lu(euypdvrl ue l-lotrtro Brlporo orqv'Errppoon roi' [:' i *' 601 E -&ilvto lvrPdvto EEOvto,
  • 93.
    Anetreu06poor1 prpootogor[ou ond ronpoopo xrodrrovro repgolo;rorj EJC ; Entmpdreuorl ev(upou ogo(peor1 + Trlq Kotrt1flTpoq Ano66pr1or1 rou mRNA EI(0NA 21.20 To o[UntroKo EJC ouvd6erq ouppogn Ue Tnv NMD. To EJC enrmpoTeuer ettionq rporefveq Upr ov nopoge(ver oro s[o- ydpevo mRNA. Merd qv eqoyoyn ono rov rupnvo, ro EJC np6ner v' onogorpuvOef U6oo Tou UeToKrvougevou prpootogor(ou orov flpdlro yupo qg perorppoqq. Av 6v0 EJC nopogeiveL mo mRNA trdyo rqg nopouo(og evdg npdopou rtodrxovfou repponoporj, ro onofo onetreuOep6ver ro prpoorrlpdlo, ro EJC 0o enrorporeUoer enmpdoOereg npoTetveq, rt.X. rr1v Upf, or onofeg pe q oepo toug 0o enrorporet1oouv ro 6v(upo ogoipeoqg qg xotrunrpoq (DCp). Aurd endyeL qv onopdrpuvoq qq xotrunrpog mo 5' drpo xor n1v onoddprpoq rou mRNA oe roreu0uvorl 5'-3' mo runop6doogo. oo10e( 6vo np6opo rrodx6vro repporroproU e[or(og oq- perorqg perdMo[4q 11 evoMorcnruiq ouppogrlg (BI6ne Ev6tryta 21.12), to prBoo'<o;rdrro 0' onoonoqre( on6ro mRNA erpoye(o nporof <prdoer oro xovovx6 rto6rr6vro repporropro0, ro ono[o ouvrl0og Bp(oreror oro ts^.eu- rofo e(c0vro. H o6uvopr(o rou prBoo<o1.ror[ou v' onoB6.tret ro ofpnloxo EJC nou Bp(oxeror gerd ro np5opo xo6r- x6vto repporroproU enrp6ner vo enrorporeu0orjv 6v(upro nou orpoporiv rqv xoLUmpo, endyowog roXe(o ono66- pnon rou mRNA. Aurn n 6repyoo(o rolefuor g0op6 rou mRNA 6ropeootropo0pev4 on6 oouv0enx6q pl6Beg (nonsense-mediated mRNA decay, NMD). Awrnpooro- ne0er dvo prqlovrop6 enmrlprlorlg rou mRNA o ono(og onorpdna q perdrppooq roloBtirv npu:reivdrv on6 mR- NA nou rp6per rivo np6trrpo rodx6vro tepprorroprori. H NMD 0o nopouoroore( tremoprepc0g atlv Evotrpa 22.g. 21.12 H EvoAAomtKr{ Euppoeri Elvor Kov6voq, Oxr E[oipeon, oTouq lloAur0rropouq EuKopu6Teq * Erdrro e[tirvro 11 otrtrr1trouX(eg e[trlv(ov gnope[ v' onoxtrs . orouv r1 vo ougnepilrltp0ouv oro mRNA npoiovro Xprlorpot .: - tilvrog evotrtrorlreq 0eoerg ouppoQ1q * H evoLl,orlrl ouppoqn ouverorp6pet oq dopxrl rqr ),euo::- yrrrl norrr),[o rov npoiovrov rov yovr6(rov. s 0 xoOoprogdg rou tprltrou oq Drosophila nepd,oppdvaL ;.: oepo oulrBovrov evotrtrorrtxrlg oupporpqg oe yovldro nou r:- 6uonorouv dLodoXrro npoiovro proq odo0. Orov 6vo 6Lorer<oprprSvo yov(6to ;retoypoge( o' dvo F- NA nou ono6(deL 5vo provo6u<6 tUno ouppoll;rdvou r- RNA, 6ev undpler oodgero orrlv ovoyvtiproq rc, e[,orv(ov Kol rov rwpov(rov. To RNA rov neproodrepa. yovr6(ov, toor6oo, or<olou0oUv np6runo evoA,lorrr. x{g oupporp{q (alternative spticing), q ono(o oup_rBc.. ver 6rov 6vo ;rovo6u<6 yov(6ro ono6(deL neptoodrepe: on6 pLo otri.ql.ou1(eg mRNA. Me ng tey3rLr5g rnq cD: x).rovono(qoqg peydqg xi.(proxog Kor rnq oMq).oLJX- oq, Syrve oogdg 6tr ndvo on6 ro g0% trov yovr6il-, nou erqpd(owor oro O4looru<d ouppdmowor evci.- Aornr<d. Iuvenog, 11 evoMornrrl oupporp4 6ev eii,c. on).c69 ro onotS,l.eo;ro LoOri:v nou npoyporoflora - Unxovn ouppo<pr1q oMd pr6pog rou npoypd;r[-loroQ gr- epoolq rov yovr6(ov nou o64ye( oe noMon).d yo,;. 6rord npoi6wo ond rov (6ro yeveru<6 r6no. YndpXouv drd<popot rUnor evoMor<nxr1g ouppogr : peto(0 d).).rr;v 11 ouyr<pdrr1o4 rwpovirrtv (intron rete^- tion), r; enil,oyq evotr,orcrtrrrlv 5' 06oeotv ouppogrlc * 6wo(q r1 nopdLerrpq e(olv(rov (exon inclusion or sk c- ping) ror 4 oporBo(o onor<).er6gew1 enil.oyr1 totv evoMc. t<ttt<tr.lv e[olv(ov t::]i;...,:;,i :r r ',, To (6to npoToyeve i gerdyporpo ;rnope( vo unoore( nepLoo6tepoug ond dvc, rUnoug evoMorcrrr<r1g ouppoqrlg. T' o;rotBo(o onox,,e.. 6pevo e(trrvro r<ovovrrd pu0pr((owoL ge roro-er6rx6 rpc- no. Meprrdg eop5q, oe ourrlv rqv nolunLor6rq:: npoor(Oetor to refur6 np6runo 5r<ppoorlg TroU KorcUgc.- vetoL on6 rn Xpnon 6rogopetrrcov orlpre(rov 5vop(qg r: geroypogr;g 11 qv nopoyrrtyq evotr),orcru<ov 3', dr<po.lv H evoMomLrq ouppo<pr1 prnope( vo enrlpedoer r", 6r<rppoorl rov yovr6(otv pre 6Uo rou).d1rorov rp6noug, C Svog e(voL q 6r1;rroupy(o 6optrrlg norxL).(og oro npoic,- ro rov yovr6(o.lv oupnepil.oprBdvowog r1 nopotre(no.- rog optop6veg rodrr<onorqrrr<d,g o).).r1Lou1(eq - 6lprroupyrrtwog evoLLomrxd nLo(oro ovdyvororlg v,: 5vo rprrlpo rou yovr6(ou. Aur6 ouXvd rpononore( q ),e.- roupyrxq rdr6qto trlv r<rr.ldrronoroUpevov nporeivo, FLo nopd6etypo, ro yov(6ro CaMKlld nepr6,1er rp(o eE.- vro nou ouppdnrovtot evq^.^.oKtLxd, 6nrog <po(ver: oTllv rr ':i.l .i ,l- ,.,'. TO yOv(6tO er<rppd(erOt oe 6).Oug o1:. cap Ptpooopdlo EJC I cap 602 KEOA AI0 21 Iupporpn rr Ene{epyooio rou RNA llprirtog y0pog gerdqpoolg Boolxeq evvoreg EJC I
  • 94.
    KorqKpoTnorl rvTpov(ov Evotrtrorrudg 3'06oeLg ouppoQnq 'Evroqr/ropdtrerq,1 e[tovltov :lK0NA 21,21 Arorpoperlro(rUror rvotrtroKTrKr1g ouppotpqg. 1:.' roug xurtoprro[g r0noug r<ot toro[g rrrlv Oq].oon- . *, Av nopo).er<p0otiv Kor ro rp(o evoMorrrr<d e[utvro, -: rRNA xro6xonore[ Uro Kurropon].ool-torrrrl rrvdorl -:, eooeopulr<r.lveL nol d nporeivtrd unoorpo.lproro. 3 :,,, oupnepd.oprBdveror ro eqovro 14, q r<rvdorl prero- :.:.ror orov nuprlvo eneL6q ro e(utvro 14 neptdlet 6vo l-*o nupnvrKoU ewonroproU. Aw6 ennp5ner orqv rrvd- - .: pu0p((el rrl peroypotprl orov nuprlvo. Otov ou1-r- -.:.,ogBovowor Kor to eforvro 15 r<or 16, r<drt nou - - ,r0cog ouprBo(ver oroug veupoveg, q r<rvdon Koreu- i, , i:or orrlv Kurroprrq pepBpdvq dnou enqped(er er6r' .:-c Sro0Loug r6wrov. Ie d).i.eg neptnriooerg, ro npoi6wo evoMoxrrr<r1g :-:cog(q 6,1ouv ow(Oereg ).etroupy[eg. Aur6 toX[er :-:.omtrd yro 6).o ro yov(6ro nou epnlSrowor orn :-:-Lot1 rqg on6mooqg. Kdoe yov(6ro er<<ppd(et tou- ' l,..croV 60o roopropg6q, pto nou npodyer rrlv on6- *ol.l Kor. pto dMq nou npoorore6er ro rdrropo ond ''-::roJorl. Oeropefuor 6l 11 ovoLoy(o rrrtv toopop<pritv -, . rv rov puopumo.rv qg on6mioorlg xooopi(er ov 0o . -.3r:rroeL to r<Urropo. -'J evoMorcrrxrl ouppogn prnope( en(oqg vo enrlped- ::. Srdcpopeg rdL6-rrlreg rou mRNA ouprnepil.opBdvov- -:: q nopoLe(nowog optopdvo puOprmrrd orotle[o -:- RNA ro ono(o prnope( vo pretoBd).).ouv no).U ro , ::vo q;-u(orqg rou mRNA. Ie noM6g neptmoroerg, xri- prog oron6g qg evoMor,rx{g ouppogrlq (ocog e(vor vo e[ovoyrdoer 6vo ouyrerprp6vo noooor6 rov nporoye- vriv preroyp6gov vo gdpouv 6vo np6opo xro6u<6vro 11 rcodrr6vro tepgortoprori Sror rilore ourd ro gerdypogo v' ono6oprlOoriv yprlyopo. Aw6 nr0qv6v owrnpooo- neUer pro evotrl,or,rrrrl orporqyu<r1 preroypogrrqq p6e- Utonq yro rov 6).eyXo qg orpOov(og erdrrciv mRNA oro rUrropo. O ouyxexprl.uivog prqlovrog6g Xpqorpronoe(- rqt yto vo enneulOe( oporoororu<r1 5r<gpoorl yro nol- ----*-":iffirg:-T' 6A ryryffi + rrdxsuon orn ueuopdvn dB*M@,r 6c wffi Evotrtrorlrrl ouppotpn rou yovrdlou CalVlKll: 0Lo- qopertro evotrl,orlro e[tlvto roreu0uvouv Trlv rLvool oe 6rorpo- penro rurroprro Dropep(oporo. ----+- IT6xeuorl orov nupqvo + Evr6nro4 oro xunopdn),oogro ?1.J4 H Evotrtrorrtrl Iuppoqn E(vot Kovovog,'OXr E(o(peo1 oroug [lotruxuncOC-a !-,::,--.: 603 17- Agorpo(o onoxtreuigevo e[6vro Evotrtrorlrdg 5' 06oerg ouppoqrlg Iuv6uoolrrl eniloyn eqoviov Evotrtrorlxoq unouvnrnq/ouppoQn Evotrtroxrrxrl noluoDevutr(oon/oupporprl
  • 95.
    Lot3g pu0pror6q qqouppoqrlq oe erdtxorig Kurropr- rotig dnoug rj tororig. Ie pto t6toto priO;uoq, 6vog er- 6tr6g 0eru<6q pu0prmqq rnq ouppoqrig ;rnope( vo enrlped(er rr1 6u<r1 rou evoMorrxrl oupporp{, o6qyc0v- roq o' 6wo[q ev6g e[trlv(ou nou nepr6Xet 6vo np6opo ro6x6vro rcpprorroprori. Aur6 droXere6er 6vo x).dopo rou mRNA rou npoq ono66gr1or1, per6vowog Sror 14 ouyx6wpoorl rcov nporeivc0v. Iuvenrig, 6tov n ouy- rSwpcoorl ev6g r6rotou Oenro0 puO;uord rrlq ouppo- rp4g ou(operdrveror;16oo oro rfrropo, q ouyr<6wptrloq rou ow(ororlou mRNA 0o orpo<pe( npoq rrlv ow(Oer1 roteU0uvoq. Motrov6l SXouv Xoporrrlprme( noAAd ouprBdwo evoMor<ru<r1g ouppocpr]g rr dXouv xo0oprore( or BtoLo- yrxo( p6l,or rov npoi6wcov evo.trl,ornr<r1q ouppo<p4q, ro nro Kqrovonr6 nopdderypro nopopr6ver 11 od6g roOopt- ol.rori tou <prJLou orn D. melanogaster, nou neprlopiBd- ver oMqLenrdpdoerg peto[ti yovrd(cov 6nou ouprBdwo evoMorcrxrlg ouppocprlg 6rorp[vouv r' opoevxd on6 rq 0ql,ur<d. H o669 no(pver rn Uopgn nou nqpouord(eror m4v EIKONA 21.23, 6nou q ovooy(o rcov X Xproprooolprd- rov oq npoq ro ouroocopxd Xprr:l.roodrpoto roOop(er qv 6r<<ppooq rou yovr6(ou sex lethal(sx/) r<or or oMo- ydg orqv 6r<gpoorl nepvoriv 6ro6oXrd g6oco rrov dM<ov yovr6(tov mo yov(6ro doublesex (dsx), to retreuro(o orqv o66. H o669 [er<rvd ;re gu].o-er6xr] oupporp{ rou sx/. To e[cbvto 3 rou yovr6(ou sx/ nepr6ler 6vo rco6rx6vro rep- prottoprof nou onorp6ner rrl o(v0eor1 l,erroupyrxrlq Avotroy(o X:A: 2:2 Ot- I * Irafira-Z Apvlrrdg pug;ttonlq rov yovrdltov n1q opoevtxflg 6torpoponoqoqg Apvlrr6g pu0Urornq rov yovrd[tov qg 01truxr1g 6rorpoponoirloqq / n( I Sxl off I Tra (xotroBo) ftKSl;P.f1.?3 0ro0opro;rogrouqUtrouorrlD.melanogasternepiloppdverprooddorrlvono(odrotpopeurdouppdvro0uppoQrc :- -:- ronorouvror oro 0qtruro. Kotrupporo oe onoloDqnore oroDro orrlg o6ou odrlyouv oe ovonru(r1 opoevuou. Errov((eror | 0upp0.- -- -, pre-mRNA nou puOp((eror ono qv nptorelvl Sxl, 11 onolo onortre(er Trl Xplol qg evotrtrontrlg 3' 0eo1q ouppqqlq, Korl 0uppc:- -- - : sx pre-mRNA nou puOpl(eror ono ng nporelveg Tra ror Tra2 oe ouv6uooiro pe dtrtreg npotrelveg SR, or onoreg enlped(ouv 0elL.: , - ro[1 rou evo],trorlrou e[ovrou. 604 KEOAAAI0 21 luppoqrl rt Ene(epyoo[o rou RNA rj ) Y ? Dsx npioteiVqg. Aut6 ro e(trrvto ouprnepil.oprBdvercL -: - RNA nou nopdyerot or' opoevrKd, evc,r nopo.).i.-:-- oro 0r1trur<d. 'Eror, p6vo ro Oqlur<d nopdyouv .*, -: -- refur1 Sx/. H nporetvq neprdler d<pOovo Boou<d c* . : l. : rot on6 ouq qv dnoqtrrl protd(er pre dMeg r:c* 1,:: nou npoo65vowot oro RNA. H nopouo(o qg npore[vqg Sxl oMo(er m ;-:: :: - rou yovr6(ou transformer (tra). N Etr6vo 21 2:3 1. , . 6tr our6 neprl.oprBdvet ouppo<pr1 prog oroOecr: : i= onq o' evoMorcnr<69 3' O6oetg (orlpreuoore o:L :---: : rp6nog e<poppr6(eror orn oupporprl t6oo ro- s, : r- Korrou tra).To (6ro np6runo oupporpqg roXJeL:::- - t' opoevrxd 6oo roL yro ro Or1).uxd r<or ono6i53. ..: : NA pr' 5vo np6orpo r<ro6u<dvro tepporrol.rori, H n::: - - : rqg npo.rre'lvr1g Sxl ovoord].,trer rrl Xpnorponorrl- -: ovodu<qg 3'06oqg oupporp4g pr6oor rqq noo:Ii- : rnq orn ouororl(o nuprl-udrvtov or1 05oq 6Lor).:l * - : tqg. Av nopo).eL<p0e[ouq q 05oq, 0o XpqoLl-ron: -:: - en6prevq 3' 0do1. Aur6 nopdyer 6vo mRNA e.1.,: : ro 0qLurd nou xro6rxonoe( ;lto npu:re'lvq. Iuvencog, 11 Sxl puOp((er r1v (6ro rrl oUQCCi- -. yto vo 6toorpoL[oer rqv 5xrppooq rnq oro 0r1],u < r . : - - tra nopdyet pto npo:te[vr1 pr6vo mo 01].uxc 3--: - Sxl, 11 nportelVq Tra elvot 6vog puOprorqg rrQ ]-::: enq. To tra2 6,Xer nop6proto Lenoupy(o fio 8.'.-. : - ).d er<<ppd(eror en(or1g Kor or'opoevrxd) , OL .:- . .: Tra rot Tra2 e(vor nopdyoweg ouppogqq SF -:- .-. vepyotiv dpreoo mo prerdyporpo-mdloug O. -'= .- Tra2 ouvepyd(owor (mo O1Lur<o) enrlpeolc,::: -
  • 96.
    r-opoeq rou dsx.Ito yov(6to dsx, to Olluxd ouppd- -3.rv rrlv 5'05or1 tou twpov(ou 3 pe qv 3' 05or1 ou- -:- rou rwpov(ou. 'Etot, 11 pretdtppoorl teppror((etot : o rel.oq tou e(orv(ou 4. T' opoevtxd ouppdmouv qv : -:eoq rou rwpov(ou 3 r<oteuOe(ov m4v 3' Odoq tou tv- --:,,'iou 4, nopotre(nowog dtot to e[iovto 4 on6 to mR- ',: Kr 0nrrp6novrog vq ouvexrote( 11 pietogpootl I :leoou rou eforv[ou 6. To onot6].eopo rlq evoMo- - .(ng ouppocp4g e(vor 6tt 6totpopettr6g npolre[veg Dsx - : <iOe cp0lo nopdyowor.: ro npoidv ttov opoevtrtr.lv - : : e uno6 [(er rr1 0r1].u xrl <pu].etu<r1 dtotpopono(4 or1, evot -: :oor6v rc'rv 0rlLuro-lv xotoordMet qv dr<rppoorl eL6t- . * . yro ro opoevtx6 yovt6[ulv. 2 t.l 3 H lupporprl Pu0pl(eror on6 EvtoxuTdq ror AnooronnTCq nou BpioKovlot T60o oTo E[6vro Ooo ror oTo lwp6vro . - svotrtrorrtrut oupporpn ouxvo oxer((etoL pe ooOevefg 0eoetg : - cporprlq , l. otrtrntrouxteg nou neptpotrtrouv evotrtrorrlro e[(tvto ouXvo ..,,0r nro ouvrqprlpeveg e[etrtrrtro ono rtg otrtr1trouX(eg nou .e cLpotrtrouv ro rdroouotottro e[titvto. . l,SLxdq otrtrntroux(eq e[trlv[tov rot tvrpov[ov evtoX0ouv r1 ro- -:retr),ouv t1v enLtroyq qg Oeolg ouppotpnq. . ' en[Spoorl rov evroXur6v Kor Tov oTlootonrlrdtv rrlq ouppo- :'q 6LoUeootrope(rot ono et6tr6g yto qv otrtrr1trouX[o nporel- ,:q npoodeonq oro RNA, notrtrdq on6 rtg ono[eg pnopel vo :,3p[(ovior ovonru[roro ro/q vo exqpo(ovrot toro-et0xo. ' - rcxui1ro geroypoq1g eqpeo(eL dpeoo rrlv 6rpoo1 rnq . , ctr,ortrrlq ouppotpflq. - i,,oMorrrxq ouppo<prl yevtr<rilg olet((etot pe ooOe- ,.1 0eoerg ouppogrlg, rdtt nou orlpoivet 6rt to or1- / Uoro ouppoqrig nou ewon[(owot Kot oro 66o 6xpo rrrlv twpov[tov onor.l.(vouv on6 ro ouvowerxd orl;re ro ouppo<prlg. Aw6 entrp6net q pU0gtorl ttrlv oo0e- vc0v orlprdrov ouppog4g on6 6td<popoug trans- Epomrxorig nop6yoweq nou yevtrd e(vot yvooro[ og pu0lttot6g eva&axtur1q ouppaEfig. Aw(Oetq on' 6,tt 0o un60ere xove[q, oo16oo, ot ooOeve(g O5oetq oup- pognq yevxd e(vst nto ouwrlprlprdveq oro yovr6tripo- ro rtrrv OqLoontrciv oe oX6oq pe rtg t6toouorottxdg 06oerq ouppoqrlg. Auq q nopotiprloq owtt(Oetqt rrrlv owil.r1r[r1 6tr 11 evoMot<ctr( ouppoqrl 0o pno- poUoe vo nporUmer on6 Ld0q r;g Unxovnq ouppo- gr]g xor unootrlp((er 6tt noLLd ouptBdvto evoi.- Lorcnr<qg oupporprjg nt0qv6v e(vot e(efurcrtx6 ouwrl- prlpr6vo ciore vo dtoqpqOe( 11 pU0ptoq qg 6rrppo- onq roJV yovr6(trlv mo en(ne6o rqg ene(epyooiog tou RNA. H priOptoq qg evoi.l,orcru<r1g ouppotprlq e(vor nep(- nl,or4 6repyoo(o 11 ono(o nepil.opBdvet preydLo opr0;r6 frans-6poonrcirv puOpumcbv qg ouppogqg nou npoo- 66vowot oro RNA. 'Onrog rpo(vetot firlv EIKONA 21'24, ouro( or pu0puor6g nt0ov6v ovoyvtop((ouv orotXe(o R- NA r5oo oro e[cilvro 6oo rot oto twp6vto rowd or4v evollorcnrf 06o11 ouppoqrlg r<ot qoxo6v Oettrti rot opvqrurl en(6pooq orrlv entl,oyrltrlg evoLLorctrrlq 06- onq ouppognq. Ot pu0gtor6g nou npoodSvowot oe e[r6vro yro vo evtoXtioouv qv enil.oyrl e(vqt Oettro( pu0pumSg rnq ouppoerlq r<ot t' ow(orotlo cis-6poort- rd motXe(o ovoq6powor rog e(ovtro( evtoXutdg qg ouppogqg (exonic splicing enhancers, ESE). Ot npcrt- reiveg SR ovqrouv orouq nto r<oLd lopor,itrlptoprdvouq puOptordg nou npoo66vowot oe ESE. Aw(0eto, dMeg npcorefveg, 6nog or hnRNP A t<ot B, npoo6Svowot oe oMqlouX(eq e[cov(tov yto vo Kffioore^ouv qv enAoyti rcov 06oeolv ouppogrjq. T' ow(motXo cis-dpomtxd morXe(o e(vor yvoud rrlg e[ovtxo( onootonqrSg qg ouppo<priq (exonic splicing silencers, ESS). flopopo(- og, notrLdg np<oreiVeg nou npoodSvowot mo RNA enqped(ouv r4v enr).oyq ttov O5oe<ov ouppoEfq pSoto Anoouonnrnq qq ouppqQrig rov twpov(tov (tss) EvLoyurrlq Anootonlrrlg EvtoXur1g rnq oupp0Qrlq Tns ouppoQnq rnc ouppoqrlq rov e[ovirrrv rov e[ulvttrtv lov tvTpovtov (ESE) (ESS) (lSE) 0L otrtrntrouxLeq rov e[ovlrov xor rotv rvrpov[ov prnopel vo enlpeooouv rrlv eniloyrl qq 06olq ouppoQnq tretroupytirvroq - . .,.qureg 1 onoouonqr6g r1q ouppoQrlq. levrro, or npolrelveg SR npood6vovrot o' evtoXur6g rt1q ouppoQnq rov e(ov[tlv Kot ot npu]- ', ..: rnRNP (n.X. or otroyevereg A xot B rorv nptureivdrv nou npooDevovTor 0T0 RNA) 0e onootonlrdg e[trlvttrtv. Atrtreg nporeiveg nou - - - levovror oro RNA (RBP) treuoupyouv oq puOproreq rnq ouppoQqg peool qg npoobeolq Touq 0' evtoxuT6g 1 onootonlr6q rr1-q ouo- ,--: Toiv rvtpovltov. / ?1.'13 H lupporp1 Pu0p(etor onrl EvroXur6g ror Anooronrltdg nou Bp(orovtot Tooo oro E[6vto'0oo xot mo lvrptivto 605 eworeg
  • 97.
    rov rwpovrKc0v oLLrlLouXrc6v.T' ow(ororXo 0errrd ror opvqrrrd cis-6pomrxd ororle[o ro].orlwor twpovtxo[ evtoyutdq rl anooon4r6g qq ouppaEryg @tronic sptic- ing enhancers, lSElintronic splicing sl/encers, /SS), ov- rroro(Xcoq. Ir4v nolunLor6qro outrj dplowor vo npoore- 0otiv or enr6pdoerg noMriv puOpuoroiv rqq ouppogrlg. To nro yvcoord nopo6etyproro e(vor or pu0pum6g qg ouppoqrlg rov orKoyevercriv Nova ror Fox, nou npoo- 66vowor mo RNA rr evrolriouv 11 r<oroordlAouv rnv enAoy4 rrlg 05o49 ouppocprlg, ovdLoyo pre q 06oq np6o6eorlg oe o16oq pe ro evol.Lorirrr6 e(civro. I-ro nopdderypro, 6nog go(veror orrlv E|KONA 21.25,1np6o- 6eor1 rov Nova xor Fox oe oMql,oul(eg rwpov(<ov nou Bp(or<owot ovo6rrd rou evoMorcru<ori e[ov(ou yevrrd odqyei oe roromolq rou elcov(ou, evdr 4 np6o6eori rouq or oMq).ou1(eg rwpov[ov nou Bp(or<owor Kq0o- 6u<d rou evotrl,orcnroU e[rov(ou ouXyd evroXriel rrlv enr- l,oyri rou e[ov(ou. T6oo q Nova 6oo ror 11 Fox ergpd(owor 6rogoperrrd oe 6rogoperrroUg rmoUg, t6(tog orov eyr<5goLo. Enopdvrog, 4 roro-er6u<rl pU0pr- on rnq evoMorrtrqg ouppogrig enmuyldvetor pr6oo qg rmo-erdrrqg drgpoorlq rov trans puOptorcbv rqg ouppog4g. Aev e(vor nLripcrlg rorovo4r6 nrig 5vo erdrrd oupr- Bdv evoMor,mu<qg ouppogqg pu0pr((eror pr6oo 6rdEo- p<ov Oetrxcilv xor opwlru<riv puOpumtiv qg oupporprig. Oeop4rrxd, or pu0ptar6g Lemoupyoriv evrolrlowog q rqtomdLLowoq rnv ovoyvcilproq ardrrrirv oq prdrcov ouppogrig on6 oproprdvo Boorrd ouororu<d rnq UnXo- vqg ouppognq. Or nro Korqvorldg neprmcboerg e(vor or nporeiveg SR xor or nporeiVeg hnRNA A r<or B yto ro 0enx6 xor opvrltrxd p5tro toug or4v ev(o1uon ri xo- toorolli u1g ovoyvrilprorlq rov O5,oeov ouppogqg, ov- norofX<og. H np6obeo4 rov nporeivriv SR oroug ESE npodyer 4 aroOeponore( qv np6odeoq rou U1 mqv 5' 05oq oupporpqg r<ot qg U2AF m4v 3' 06oq ouppo- rprjq. Iuvenrirg, 4 ouvop;roL6yrloq rou oopoud(ou ouppog{q y(vetot nro onore},eoprolxri nopouo(o rcov npoteivcbv SR. Aurdg o p6Log-r<ov npcoreivdrv SR ei- vor orlpowrK6g 16oo yio rnv r6rooumorrxrl 6oo xor yro rrlv evoMorcu<f ouppogri, xo0rmriwog nq npo- reiveg SR ovoyro(oug nopdyovreq ouppogrlg xcr puOprmdg evoMorcrxrlg ouppogrlg. Aw(Oero, or npo- reiveg hnRNP A/B go(veror 6n npoo66vowor o..ro RNA Kor. owoyov(owor qv np6odeorl rov nporeivri:v SR ror dLl.rov Boou<cilv oumotrrc0v rou oorpond(ou oup- pogriq xord -rqv ovoyvc0ptorl ).etrou pyrrcbv orlpdtcov ouppogqg. Or npotetveg SR 61ouv trlv rxov6trlro vo 6e- 4reriouv 6vo pre-mRNA mqv o66 ouppogqg, evri or nporretveq hnRNP owoyrov((owor our4 14 6repyoo[o. Ae6oprdvou 6rr or nporciveg hnRNP e(vor e(orpetrrd dgOoveg orov nup4vo, ncbg ot npotetveg SR owoycovi (owor onoreleopotu<d ng hnRNP orov nupqvo yro vo 6reuroLtivouv rn ouppogrj; I-'lporpovcbg, our6 enmuyXd- veror pr6,o<o ouyxcouoUoti r.1g preroypoqfq Ue rrl oup porp{ ;16oo orov nuprlvo rou rurrdpou (BMne Ev|trlro 21.7). Zuvenrilq, q dtepyoo(o qg ;retoypogrlg nr0qv6v eqped(et r1v evotrl,or,cu<4 ouppocprl, rdrr nou 6wcog rqXUer. H evqMoKru(4 ouppoqri go(veror 6l enqped(e- ror on6 et6roUg unor<rvqr6q nou Xprlorponororiwor yto vo npoo0rloouv rnv dr<rppoorl rrov yovr6(rov 6nog en[- onq Kor on6 trlv roXUr4ro rnq peroypqgrlg ro16 rr1 rpdoq enrprqr<uvonq. Arogopenro( unorrvqrdg prnope[ vo npooe],rUouv 6rogopertrdq opd6eq preroyporpu<civ nopoy6wrov, or ono(or pe n1 oerpd roug gnope( vo enqped(ouv rrlv eTrr- p{r<uvoq rnq Ueroypog4g. EnoprSvog, o (6ro9 Unxovr- opr6g prnope( vo euOriverqr yro rrlv en(6poon rqq enil.oyrig unorrvrpriv Kor rnq roXriqrog enrpuixuvolg rrlq Ueroypoqnq ornv evoMorcrxrl oupporp4. Or 6ro06- orgeg ev6e([erg unoderrv[ouv 6vo xrv4ru<6 prow6]ro oU;rqcr;vo pe ro ono[o n XoUnIf toXUqro ent;rrlxuvoqg rnq peroypoqnq eo noperXe oe Uro oo0evrl 06o11 oup poqrlq ovo6u6pev4 on6 ro oriprnlor<o r4g Pol ll nou enrprlr6veror opxer6 1p5vo yro vo (euyoprioer Ue rrlv ovo6rx4 06oq ouppogqg nporo6 etrrgovrore( pro r<o0o. U' .,r:r' j'.. '- - 0r orroydvereg npotreivdrv npoodl- qq oT0 RNA Nova ror Fox npooyouv 1 roroorer- trouv qv eniloyrl rrlg Oeor1q ouppotprlq ple rpo : ue[oprdrUevo ono ro ouprppo(opevo,,. H npdode:- qg Nova o' e(6vro ror neprpotrtrovro ovodrro .,- rp6vro ovooretrtret rqv evro[q rou evotrtrorrLx:- e[tov[ou, ev6 1 npoodeol qg oe neprpotrtrouc:: xo0odtr6g Lvrpovtreg otrtrqtrouXreq eXel ro ovrlOe : onoretreopo, H ddopeuol rrlg Fox orqv ovodtxn ,. rpovLrrl otrtr1trouX(o ovoorrltrtrer 11v evro(1 r: - evotr),orrrroI e(orviou evto 1 npoodeorl qq o:- , ro0odrrq tvrpovrxrl otrtr1trouX(o 6Xer ro ovr[0E.: onorel,eopo. 606 KEOA Al0 21 Iupporpq rr Ene[epyoo(o rou RNA ry
  • 98.
    21.14 Ot Avrr6Pooetq Xpnotponoro0vMrrPd frons-ouPPoq6q RNA 6rq owoytrlvtortrrl 06o11 oupporprlq' Aw6 to pow6Lo unoVpouUi(tt pto tremoupyxrl en(mooq qq o0(eu(qq ue.qtO rnq LrerqYpqqriq r<ot rnq ouppqqfq tou RNA mov nup{vo, ore-mRNA ;retoypd<ptov tou 6tou yovt6(ou'-p6)rlov e(- ;; "q;p"ild *aut, enet6( ov {tov. ouxurl to e(druo ruii vo*otou 0o prnopo6oe vo oupntrrlpdrvouv to 6vo ;dII; owi v' ovtit<ouv oe pto provo6x( op66o oup- ntrrlpopronx6rrlrog. bp,opduot letptopro( unortvo0v trans-ouppqqn' >.o nopdday;ro nou nopouotd(erol c*3. r<dtt': pr6pog ;n; ;ft;;s ft.zo, ouunr.npgqoJx:s oMqloux(es et- or11Oqr<ov oro twpovtq aUb nun' To (euydptouo 99- o"rr' UetoqU rov o u pnr1 pul prot rrdrv o)r'r1l'o uXtdrv anu,ouovti'6vo p6pro oX4go-oq H' Aut6 to pt6pto il;pti'vo ouppoQe( e(te in cls, on6're ouppdmowot !g,6;. nou ,fuo, o1-totonotr6 ouv6e6eprSvo pr' 6vo tv- ijouo, eke in trans, on6te oupp6ntowot e(cbvto rov oG,^pio-,Ou prop(tov RNA' Kot ot 60o owtdpdoetg ou1't- Bo(vouv in vitro. '- - M,o 6trr1 nep(moon ornv ono(o pnope( v9 npoygo- ronotrlOe( trans-ouppoqn in vitro ouptBolvet orov nope- x;;;,;i; unomprir;roto RNA on6 t9.ono(o to 6vo f,;;;6x", u. 5' 06on ouppotpriq rot ro 6Lo gto 3' 05o'n "rbpaqiiq pro(( ;re rotdLt'r1Leq r<oOodrrdg oMql'ou{eq i",-"*its'p"ope(vo e(vot efue 11 en6prevrl 5'.06on oup- poqtis eke 6vog evrq4ut(q qq ouppoq(S) :-1'T1I; 1roiu.6qto, out6 prtpeftot rn ouppoqri.p6oto optouou I(*fru'(Bi5ne 6e[id na'eupd qq Erx6voq 21'11) t<ot oitxrt, oit, in vitro, ot 06oetq ouppoqqq cno 6e[td xot t' "p,*"pa bev e(vot onopokrlto vo Bp(ot<owor cno (6to p6pto RNA. --'nu-at' onotetr6optoto 6eftvouv 6tt 6ev undplet ptq- lovtotu<6 r<cbLupro nou v' onoyopeUet rnv trans-ouppo- lp,i inrons, onortre[ouv povr6[o ouppoErig nou npounoO6rouv rnv npoodewtr{ Uttgl.tygJ ev6g o<r;- liIioi"t ,roooqriq rot6 ;llroq tou RNA' 'Evq oogoil- 5. ouppo,pnq np6ner vo pnope( v' ovoyvop(et rtg 5' r<or 3' b6oetg oupporprlq 6torpopettxdrv RNA 6tov Bp(- oKowot row6, l-1op6tt rl trans-ouppoqq e(vot.qndvto oroug notr'u- norroi*s europuciteg, ewqt o Ku!t9.s prrlxovtop6q yto .nu ""tetiVoo(o ttov np66po;rov RNA oe 6pq'ro mRNA oe optolt6vouq opYovtouo6g 6nog ro rpunovooril;toto *o, -L uhUo.cilbq or<ou)'.r1xtq' :to tpunovootirl-toro' 6l'o i" v*to. ,^,pp&go*o, rog notruototpovxd ;retdypotpo nop6proto Ue rov porcrnp(u:v' Oo16oo' to RNA nou Ue- toyp6gqxe 6e gnope(,o pt-otppoct'ei ltop(g npooOrirrl rrili -ol,inLo uxlqq-bonvo6 37 vou rtr'eort6(ov nou <p6pe- tot 1.t6oto irans-ouppotpnq ytq rrl Uerqrponri ev69 noLy- oi*pou,^o0 RNA oe telcoptord Uovootorpovr6 RNA' H ol"r[rouX(o-o6nV6q 6ev xo6trorrorefuot ovo6xd rolv qeXroptorOv getoypotprxdlv pov66tov' Aw(Oeto' geto- Vp&,pt-o, o' 5vo ove(dprqto RNA, nou qdpet entnp6- t'g"itq otrtrr1lou{eq oro 3' drpo r.ou' on.o Uto enqvo- in;"h pou66o'no, ewon((etot rdnou.otrLot cno yo- ,,ioud n EIK0NA 21.27 deftvet 6tt our6^ro RNA E6pet tn, .ilULorX(o-o6qy6 rot oe-ro0o6rn 06on pto otr't1- LouX(o s' egirhq ouppoQris' Ot oMrl)rou{eq nou rodt- r.onoto0v rq mnNh'q6pouv pto 3' 06o11 ouppo<P{q oujorq nptv on6 rrlv ol'Lr1lou1(o nou undplet mo rilpqro mRNA. u 0r ovl[pooeLq oupporpnq ouvqOtog npoyporonototivrot Uovo in cls peto[u ouppohdrv ouppoQnq oro i6Lo ;r6pto RNA * irrr-orppoQn ouuBo(vet oro tpunovootituoTo Kot.T0 oroutrfl- rto onou Uro Bpoxeto otrtrr1trouXio (SL RNA) oupponretot oro 5' orPo notrtrdrv npodPoptlv mRNA' u To SL RNA exouv 6oun nou poto(eL ;re rn 06on npooDeorlq Sm ttrlv U snRNA. Tdoo on6 prqlovtottr<r1 6oo xot on6 e!,etrcrtxr1 or<ont6' r-orppo,pn'6eopr10qre og ev6o;roprorrl ow(6pooq' "o, luo,o*trd ouv(orotoi oe e}'eyx6pevq onoLotg( rr,.rv oLLqLouXttov tcov twpov(<ov oto en(ne6o tou R- ',tn n ,O yeveitxrl oxontd, q ouppo<pr1 ouprBo(vet pr6vo n "i.. nriO orlpo(vet 6tt prnope(vo ouppoQo0v pr6vo cL oMqtrouX(eg nou Bp(orowot oro (6to pr6pto RNA' To ndv<o trrdpoq rnq , r'' r'' ''': : : [s(1vs1 rnv Kovovt- <l rotdmoorl. To twp6vto pnopef v' o<potpe0oUv on6 <cOe pr6pro RNA, enttp6nowoq oro e(<r'rvtd rou vo oup- .o,poir, oMd 6e ouprBo(vet 6nptopnxrl oupporpq e[tr;- ..lcv preto(ti 6totpopettrov prop[tov RNA' Mo]'ov6tt ,urpiiouut 6tt ouprBo(vet trans-ouppo<pr1 preto(0 rov Kovovu6, ouppoqrl ougpoivet gdvo in cis EEtitvto'1 lvrpdvto E[titvto 2 E[titvto 1 t-*= Iuppoqn Unope( vo npoYgoronotrl0ef ln trans 0v {0 tvrpovto neptdxouv ou gntrqpolpolr6q olJtrltrouXteq --::-'r-.jji::::: .i.' H oupporpl ouvrlOoq npoyporonote(rot povo in cis -::o[u e(ov(tr:v nou pp(orovrot oro i5to pdpto RNA 0mooo' trans- - -:ootpfi'pnopei vo ouppet orov 61proupyq0o0v etbtreq xor0- - - e;dg nou unoonlpi(ouv ro (euyoptogo pooeov pero(u tvrpov(utv' 21.14 0t Avttdpdoetg trans-ouppoqnq Xpnstponoto0v Mxpd RNA flf'
  • 99.
    Aro6oXu6q enovotrnqrerq qg povoDogodqyoU ffi I Y O0nVdq 100 Bdoorg 37 Booetov m:*wGUw Aptorep6 rvtpdvto? :eltoptm6g petoypoqrrdq govo0eq ffi I Y AtrLntrouxio mRNA ;g]mx{Qryse Ae(td rwprSvro? I 4t *w odrlYrls G Mtipto oXrlpotoq Y -.4. AG:g I 06r1yoq Atrir1trouX(o mRNA 37 pooeov . To SL RNA n0p6xer evo e[6vro flou ouvdeeTor Us T0 npdTo sEdvr0 evoq mRNA p6orrr lrans-oupporprlg. H ovropoorl nepiloU0ovtr rtq iorsq ovTrdp0oerq Us Tnv nuplvrKl crs-ouppotprl, otrtro nopoyer 6vo RNA 0nUoToq Y ovr[ yro trooo. SL 5' odoq oupporlfg Trans-ouppog( yo qv nopoXrl ev6g odnyo[ yro q gerdqpoorl x 5' 06o11 oupporprlg pA pA pA t_ I Trans-ouppo91 ou(euypevn pe notruobevutrioon yro rnv ene[epyooio J Tou no^uororpovrKou ueT0YpoQou IIII n IWreW IXKTo SL RNA rp00e6Tst evov o6rlyo yro v0 drsuKotruver i1 peroqpoon. Mo(i Ue nq ovTrdpooerq ndqnq Kor noluoDsvui.*- onq, 11 ilpooelKl Tou SL RNA Xprlotponots[Tot er[orlq 0Trl UeToTponrl rotruoroTpovrKd)v UeToypoQov or UovooroTpovtreg povodeg. 608 KEOAAAIO 21 Iuppoqrl Kr Ene[epyoo(o rou RNA Orov o odny6q Kor ro mRNA ouv66owor p6oo *.r: ow(6pooqg frans-ouppoenq, n 3' neproXq rou 06r'. : - RNA r<or n 5' neproxn rou mRNA oqv npoyporrKc:--: onoteloUv ro 5' Kor. 3' puod ev6g rwpov(ou. Orov :--- Bei ouppogq, oXqprot[(eror Svog 2'-5' 6eop6g p5oo. :-: ouvrl0topr6vqg ow(6pooqg preto[ti rcov GU rou 5 r,,:::- v(ou Kor rlq oI nlouX(oq dtox).d6olor1g xowd mc - ; tou 3' rwpov[ou. To 6[o rgrlporo-rou rwpov(ou a.,: o1-rotono).rrd ouv6e6epevo, oMd nopdyouv 5vo ;,c: : oXrlpotog Y ow( yro Ldoo. To RNA nou ouvrroe6per ro 5' e[ovro yro lre-j- ouppo<pr1 ro).ekoL SL RNA (spliced leader RNA). -: SL RNA, prp<oug 100 vour<l,eort6(ov, drntrovowc. :. pro r<orvr1 6eweporoyrl 6oprq nou dXer tpetg pr(o.;c,:- 0qLt6g Kot pto pov6r<trovr1 neproX4 nou prord(er r-: - 06oq np6o6eoqg Sm. Iuvencbg, ue(orovrot cr,c :, nRNP nou onoreLo0v p6q rrlg rorqyop(oq S- . nRNP. Kord rqv ow(6poor1 frans-ouppogqq -rc ! - RNA y(vetor Udpoq rou npoi6wog ouppo<pr1q c -: nopouoLd(eror oro ndvor prSpog rrlq ' . Cno.lg r<t dl,).o snRNP nou egnLdrowor orl oupoc-- (ercr6g rou U6), ro SL RNA <p6peL pLo rprge0u).rrr;i:,- r<o).[nrpo 11 ono(o ovoyvop((erot on6 6vov noc:, ).oyprdvo nopdyovro elF4E ome vo dreuKoluv8:,. - perd<ppooq. Iro C. elegans, nep(nou 70o/orav yovt6[r,-lv rne::-.
  • 100.
    yd(owot trr5ool touprlXovtoprori trans-ouppqqqq' Autd ro yov(6to rorotdooowot oe 60o roqyop(eg' H nptirq nepil,oprBdvet yov(6to nou ono6i6ouv Uovootorpovtrd pet6ypogo to ono(o ene[epyd(owot t6oo trre crs- 6oo ^o, pJ trars-ouppoQn. Ie ow6g rg neptmciroetg, evtir q crs-ouppogrl Xpnotponoekot yto rrlv oqo(peor1 eoore- prrtilv twpov(tov, evcit rl trans-ouppo<p{ nopSXet gto otr- irlLoulio-oO r1yo 22 vo u xLeottd(ov no u n podpletot on6 to St irun ciore vo 6euxotruv0e( 11 pretdrppoorl' H dlLn r<oqyop(o nepil'oprB6vet yovf6to nou ono6(6ouv no]'u- ormpovrrd getdypocpo. Ie ourdg ttg neptmtiroetg, 11 trans-ouppoqrl Xp4otpronotefuot Yto rn petotpon{ tcov noLuotmpovrrrilv getoypdqcov oe povootmpovtrd rt en(or1g nop6ler rrlv oMll'oufo-odqy6 SL yto q pretd- qpqon toug, 6nrog go(vetot oro rdtro p5pog qq Etr6- vog 21.28. Ito C. elegans undpXouv 66o trjnot SL RNA' To SL1 RNA (ro npcino nou ovorqtrf<p0r1r<e) Xp4otponoe(- 'ror ;t6vo yto rl'lv oqo(peoq tov 5' dxptov rov pre-mR- NA nou pretoypd<powot on6 Uovool'orpovtrd yov(6to' flrig Bp(oret to SL RNA rrlv 3' 06o4 ouppo<Prtq Yto rrlv 5vop(r1 rrlq trans-ouppoq{S, rot ndrg onoqe6yet 11 trans-ouppqqrl rov owoytovtop6 11 rrlv nope;rBolq pe qv cis-ouppoqn; H txov6rqto yto m6leuoq ptog tret- toupyu<rlg 3' 05or1q ouppo<p4g nopdXerot on6 rtg npto- tetveg tou SL snRNP. Fto nopddetypct, ro ro0op6 SL snRNP on6 to Ascaris, Uto flopootrtrq vqprot6l4tw0o, nepr5Xet 6uo er6tr5g nproretveq, pto on6 rrq ono(eq (SL - 30 kD) oMql,ent6pd 6peoo ps rrlv npcoreivrl np6ode- oqq orq 06o11 6roxi.d6ooq qg 3' 06oqg ouppoqqg' To SL1 RNA trans-oupp6metot p6vo orqv nprilq 5' ope- rdgpoorq neptoXq, oMd 6ev nopeprBo(ver oe ro0o6trd oulrB6wo cis-ouppoqriq. Aut6 ouprBo[vet enetdrl pr6vo 4 5' otrrerdrppqorn neptoX4 (xotre(ror ourp6vtolutron, B).5ne Err<6vo 21.28) nepfxet pto l,etroupyrxrl 3' 06o11 ouppo<prig, oMd dev SXet rqv ovo6u<r1 5' 05or1 ouppo- qiq ytq vo (euyoprioet Ue rnv xo0o6u<r1 3' 05oq oup- poaiq. To SL2 RNA ttg neptoo6tepeq qopdq Xploq-ronoe(- ror yro qv ene(epyoo(o noluotmpovtxdlv pretoypd<ptlv nou 6toXrop((owot p5oro ;rtog dtoorqprottx4g otrtrqLou- X(og 100 vouxtreottd(cov peto[ri 6Uo yettovtrcilv yovt- 6(tov. I' 6vo prtr<p6 r<l,dopro yovt6(olv 6nou to 6Uo yemovrrd yov(6to e(vot ouvdedeprdvo lcopfg vo nopep- BdMowot 6tomqponr<59 oMrl)rouX(eq, Yto ro dtoXcopt- opr6 roug Xprlotponotefuot to SL1 RNA. Kotd t4v ene[epyoo(o ttov notruotcrrpovtrd:v peto- ypdqorv on' onotod{note on6 outdtq SL snRNA, 4 ov- tidpooq trans-ouppogqq e(vor orevd ou(euypr5vrl pe ng owrdpdoerg n6$nq rot noluo6evul'(ooqq mo t6trog xdOe yovtdtor<rjg povddoq. Mto t6toto o[(eu[q <po(vetot 6rt euvoe(rqt on5 dpreoeg npoteivxdg oMrl}'ent6pdoetq pero[ri rou SL2 snRNP Kot rou nop6yowo 6t6yeporlq qg n6r.pqg CstF nou npood6verot orrlv nLorjoto oe U otrtrr1Lou1(o roOodtrd tou orlpotoq AAUAAA (BI6ne Ev6rrpa 21.15). Awo( ot pqXovroproi enttp5nouv oe oXerr(5pevo yov(6to vo pu0p((owot ouYxpovtopdvo oro en(nedo rnq UeroYpqqrlg (enet6r1 pretoypdEowot o:g notruotmpovtrd perdypoEo) xot vo puOgf(owot [e- lcoprord petd 11 ;retoypoqq (ener6q oL (eXopL::z: ':- vr6tor<6g provo6eq 6toltlp[(owot oq ouventto ric '-:t- (epyoo(oq tou RNA). 2l.l 5 To 3' Arpo To)v mRNA Anproupyo0wot pe n6{,n Kol noAuo6svulioon e H otrtrntroux(o AAUAAA e[vot 6vo ortUo neQnq yto rr1 6lptoup- yio ev6q 3'no),uodevutrto;ttlvou orpou mRNA. * H ovr[Dpoon onotre( 6vo nptlretvtro oOpn],oro nou neptexet evov nopoyovro et6trorrlrog, pto evdovourtreoorl rot noLu-A notrupepooq. * 0 nopoYovrog etdtroqrog rot 1 evdovourtreoorl n6nrouv ro RNA ro0odLxo qq AAUAAA. " 0 nopoyovrog etbtxoqtog rot 1 nou-A notrupepoorl npo- o06rouv - 200 rorotrorno A oro 3' oxPo. * H oupo notru-A etrSyXet rr1 oroOeporqro rou mRNA Kt srrlpe- o(er q perdqpool. + H notruodevutrurlorl oro rurrop6ntroo;to noi(et potro oTnv 0U- Bpurrrl ovonru[r1 rou XenoPus. To 3' dr<po tov mRNA nou UeroYpdgowot on6 qv Pol ll nopdyowot pe n6Qq nou ot<oou0e(rot on6 no- Luo6evutr(ooq. H npoo0rp<r1 notru-A oe nuprlvLr6 D- NA onotpdneror on6 to ovd).oyo 3'-6eo[uo6evoo(vq, 11 ono(o e(vot en(oqg yvoorq o.lg rop6uoen(vr1 @ordy' cepin). l-lop6rt 11 ropduoen(vq 6e mopotd rrl pero- ypoAn rou nuprlvtroU RNA, 11 npooO4r<n rnq nopeuno6[(et rqv suQdvton rou mHNA oro KUTrop6- nlool-to. Aur6 6e(Xvet 6tt 11 noluo6evutr(tr-loq e(vor onopoitqtrl yto rrlv otp(provorl tou mRNA on6 nupqvt- x6 RNA. H oupd no).u-A npooroteUet to mRNA on6 ono66prrloq pr6oto 3'-5' e[rrlvour]'eooov. Irrl ([pn, n oupd notru-A en[or1g prdMov eprnLdr<etot orrlv e[oyco- yr1 rou riprprou mRNA on6 rov nuprlvo Kot orn oro0e- p6qro qg r<oLUmpog. H nopoyotyrl tou 3' dr<pou nopouotd(etot or1v .t:':::.::t. :.: .'1r,. N RNA notruprepdOrl ;retOypdqet pretd rq 06o11 nou owtororXe( mo 3' drpo r<ot oMq).ou1(eg tou RNA ovoyvrr:p((owor otg m6Xot evdovourtreotrurLr<r1g ndqrnq 11 ono(o npoqye(tot qq noLuo6evu).iooqg. H RNA notrupepdorl ouveX((et q preroypo<pr1 prerd qv n6- rl.rr1, oMd to 5' drpo nou nopdyerot on6 qv ndqq 6ev npoorotetietot, yeyov6g nou orlporodote( tov repport- opr6 qg peroypo<pr1q (B).5ne Evorqra 21 .16). Iro neptoo6tepo pre-mRNA, 4 05oq ndqrlg/noluo- devutr(coorlg neptBdMetot on6 6Uo cls-6poortr<d orlpo- to: 6vo ovodtr6 prot[Bo AAUAAA, ro ono[o ouvrlOoq on6Xer 11-30 vour<l.eor(61o, xt tivo t<o0odtx6 motXe(o nlorjoto oe U 11 oe GU. To AAUAAA onotte(toL 16oo yto qv n6Qr1 6oo t<ot yto rnv noluo6evul'(toor1, 6nolg cpo- vepc0vet to yeyov6g 6tt onotrottpq r1 pero).LoEn rou eEoUepo0q AAUAAA ovom6Met rlv nopoYoYl rou no- l,uodevul.urlllevou 3' dt<pou. l; 1:{ To 3'Axpo rotv mRNA Arlgtoupyouvrot Us ileqrl xor [lo],uoDevuhltrlorl 609 Buorx6g ewoeg
  • 101.
    Ev6ovouxtredorl AAUAAA opxereq evepy6rrlreg. 0tCPSF ror CstF onoretro0vror ono opre- reg unopovoDeg. To unotrorno ouorolro e(voL lLovopepl, H ouvo- trrrq po(o eLvor >900 kD. T0 mRNA or00ep0n0re[r0r U6oo rnq notrugodevul(oolg tlojvouK^eoorl * _.--"9 ,4il*orulcit flouodevutr (oor1 I S'Kotrunrpo AAUAAA An . : '.:: ', rr ' H otrtrr1trouXio AAUAAA efvor onopoiqrn yro Trlv n6gq rqq 5'rotrtinrpog tlore vo 6lproupyn0e( 6vo 3'orpo yro no),uodevuL(tooq, H ovdntu[4 ev6g ouorqproroq yro in vitro no].uo- 6evuL[coor1 dvor[e ro 6p6pro yro rlv ovd].uor1 rov ov- 116pdoeov. H 6qprroupy(o rot or Letroupy(eg tou oupn).6xou nou ovoloprBdvet qv 3' ene[epyoo(o no- pouotd(ovtor orlv :.' . ,:i ," . H 6qprroupy(o rrlg ooorqg 3' reLrrrlq 6oUnq e[oprdror on6 tov eL6txo nopdyovto yro rnv n6,Qq xu uyv no).uaevulfuo4 (cleavage and polyadenylation specific factor, CPSFl o ono(og neprdler no}.LonL5g unoprovd6eg. Mto ono rrq unoUovd6eg npoo66veror dpreoo oro proriBo AAUAAA Kor orov ndpoyovra 66yepor7q rnq nEqnq (cleavage stimulatory factor, CstF), o ono[og e[vor eni- orlg 5vo oripnLoxo pe noL).ontrd ouororrxd. 'Evo ono outd ro oumorrrd npoo6Sveror dpeoo oe pro ro0o- du<r1 o).qlouX(o nlorioro oe GU. Or CPSF r<or CstF evtoljouv o 5vog tov d.tro xord qv ovoyvtr;pLoq ror', o4ptdttrrv no).uo6evu).(oroqq. To er6rxd 5v(upro nou e!- nl6t<ovtoL e(vor 1-rro evdovour<Ledon (n unogovd-c tov 73 kD rou CPSF) nou n6nrer ro RNA Kor Uro no- Au(A) notrupep6o4 (PAP) nou ouv0SreL qv oupd nc- Lu-A. H nou-A noLupepdorl (poly-A polymerase) exg -- er6rrrl rorolutu<r1 evepy6rrlto. Otov ouvdud(erol "i .: un6lorno ouorortrd, q ouv0elrq ow(6pooq y(ver:. "- 6tr4 yto RNA nou neprdlet qv oMq).oux[o MU;-:-- - ow(6poor1 noLuo6evul.iuroqg e[e].(ooeror 6Lc-::: - 60o mo6[ov. I-lproro npoor(Oetor 1-uo oXerrro 3c:,i: oMrl).ouX(o o)lyo-A (-10 r<ordl.orno) oro 3' oi<oc :-- 4 ow(6poor1 e[oprdroL n].4ptoq on6 tqv oMr,.:,, : AAUAAA r<or 11 noLu(A) noLupepdorl rrlv 6LeEc,-. ,-: rrlv xo0o6qyrlon rou nopdyowo erdrxdrlrog I:: I"-- tepo md6ro, 11 nupqvrrq nporrelvr1 PABP I .: ., : binding protein) npoo6Sver rrlv oupo otrryo-A 3:.:::- nowoq tqv endmooq qg Sorq ro n).qpeg !rK3i - -200 xoto).o(n<rrv. H (6ro q notru-A noupepor -::- o06tet xotd).orno A mrlv 3' 05oq. O eyyevrc . --:: dpdoqg r1q e(vor vo 6lfororor rd0e cpopo nou n::,-i: ter 5vo voux..eot(6to. 0o16oo, nopouo[o rov CpS= . : PABP ll, Lenoupye( oStdLeLmo, enemefvowoq *.: ::- X<r-)pto,q oLuo(6o nou-A. Merd qv ow(6poor ,-'-:- 6evu).(coor1g, q PABP ll npoo65vetor fiorxao-.r: . : mrlv npo5rcrooq notru-A. Awd ;-re dyvooro lrit.r, --: neprop((er tq 6pdor1 rrlg noLupepdoqq no)..-: :: : -200 npoo0rlreg rorolo(nolv A. Kotd tqv e[oytrryq ttr-lv tr-lpL1-uov mRNA or:c ::. -- pnvo, n rurroponLooprortxrl nporelvq PABP .:-::: veror otrlv oupd nol,u-A. H PABP I d1r uc.: -:,- more[er ro mRNA on6 rr1 6poor1 tcov 3' r<oL 5 e!- , : - xLeooorv, oL}d en[or1q npoo6dveror orov ncc]. : ,- l 6vop(49 qg perdgpooqq elF4G, 6LeuxoAu,,'c.:r: - ;rerdgpoorl tou mRNA. 'Eror oro rurropon).::-: -- mRNA oplpror((er prro xLetmrl Or1LLo mqv c:3 : : - nporeivrx6 otipnoxo nept5ler tdoo ro 5' doc ,.:. : : dr<po rou mRNA (B).5ne Erx6vo 24.22 orqlt E,: --: 24.9). Zuvenrirg, 11 no).uo6evu).(oor1 enqped(e. :::: - oro0ep6r4ro 6oo ror qv 5vop[r1qg prerdgpc:*: :-: rurtop6nlool-ro. Kord qv epBpuirrl ovdntu(r1 rou Xenoprs - -:' ).uo6evuL[tooq 6re(dyetor oro Kurrop6ntr oc - : : i : o<poL((ovroq ro Unrprr<6 6l.eyXo rlq r:. - -- : epBpuoySveoqg. OpLopr5vo ono0qxeupdvo pqrc., : - : NA pnope( efre vo notruo6evu],t<l0oUv ond q', -:'- - noLutr-repdorl oro Kurrop6n),oo1.ro oore vo 6re ,"::i - Enrprlxuvorl rrrir> $sP$o ..- ., -net|trl ,,-.fi; , & 610 KEOAAAI0 21 Iuppoqrl rr Ene[epyoofo rou RNA To o0pntroro 6r[sr0T0r [er0 rnv npoo0{xr1' 200 xoro]o[nov A
  • 102.
    Uerdgpoon e(re v'onoo6evulloeoUv, reppqr(owoq rn petdgpooq. 'Evo er6tr5 cis-6poonr6 qrorxe(o nLorioto oe AU (CPE) orrlv 3' oupd roreu0[ver rqv erdxr] yro 11 peuorxq top(provo4 notruo6evutr(rooq oro rur.op6nlo- opo, evepyonor<ilwog 6rot tq prerdrppooq oprog6vov er- drxdrv p4tpu<rriv mRNA. l-to q pU0puorl qg ono66- Unonq rou mRNA, undplouv roudlrorov drio rUnor cis-6pooru<rirv otrrlLouXrc0v orrlv 3' oupd nou nupo6o- ro6v rrlv onoo6evul(<oorl rou mRNA: 11 EDEN (embry- onic deadenylation element, orotle(o eprBpuir<tiq ono- o6evu),(oor1g), gro otri.r1Lou1(o 1 7 vour<.treorr6(rov, ror ro ororXe(o ARE, otri.qLouX(eg ntrotioteg oe AU, nou ou- vnOoq nepdXouv 6ro6o1u<69 enovolrlQerg qg AUUUA. Mto RNAdoq erdrr4 yro r4v notru-A oupd (poly(A)-spe- cific RNAase, PARN) epnMrerm orrlv ono66prrlon rou mRNA oro rurrop6nlool.ro. Aorpofuirg, 4 onoodevuL[ro- on rou mRNA Bpioreror ndwo oe owoycovrop6 pre 14 oroOeponofiorl rou mRNA: ouMoyu<d xo0op((ouv ro 1p6vo rlpr(ror1g [eloprorrirv mRNA oro xfrropo (BI5ne KegdLoro 22). 21.16 H EneEepyooio Tou 3' Arpou rou mRNA Eivor Kpioryn yro Tov TepUoTtoU6 Tnq MeToypoQfiq e Ynopxouv drotpopor rponor reppolopou qg peroyporprlq ono 6rorpopelxeg RNA notrupepooeq. * 0 oxlUolouog rou 3' orpou rou mRNA o1;roro6ore[ rov rep- polopd nlq Ueroyporprlg ond qv Pol Il. OL nlqpogopieg nou 6ro0Sroupe oXertrd Ue rnv ov- r(6poor1 teppotroporj yro nq europurrrrrr5g RNA no- uprepdoeg e(voL ).ry6repo Lenropepe(g ouyrprrrr<d lre rrq yvoloerg ;rog oXerrr<d Ue rnv dvop[r1. To 3' or<po rcov RNA nopdyowor Ue 6Uo rp6nouq. Opr- op5veg RNA nouprepdoerg teppor((ouv rrl UEro- ypoen oe Uro r<o0opro;r6vr1 oLtrqAouX(o tou DNA irepporrorqg, terminator), 6nrog <po(veror orrlv ., :i. H RNA no).upepdoq lll <po(veror 6l XpqoL- uonoteI ourrl tqv orporqyrxq 6ro06rovrog 1.rro 5ror<prrr1 o).).r1).ou1(o oAryo-dT nou onUoro6ore( rrlv cneLeu0dprr-ron rnq Pol lll rrtore vo repporrcrre( 11 pe- :oypo<pr1. l-to rlv RNA no).upepdoq l, ro provo6rr6 npoi6v qq leroypo<prlq e(vor 6,vo preydLo npdbpopro p6pro nou ne- cr51er ttg oLi.r1).ou1(eg rou rUprou rRNA. O repUorr- ;pdq npoyporonore(ror oe 6uo 6ror<prr6g 06oeLg (T1 <or T2) ro0o6u<d tou roptprou 3' dr<pou. Awo( ot reppo- :rordg ovoyvrop((owor on6 pro et6rr<q npcorefvr1 npdo- 5eoqq oro DNA (Reb1p orrl (Upq r1 TTF1 orov :owu<6). O repgotrol.r6q qq Pol I oler((eror en(oqq pr' ivo ou;rBdv n6Qnq nou dropeooLoBe(tor on6 rqv ev6o- W EIKONA 21.31 H UeroypoQn p6otrl rtrlv Pol I ror Pol ll Xpqotponor- e( erdx6q otrtrqhouXieq (repyolm6g) yro Tov repp0Trop6 qg pe- royporpflg. vouxi.edoq Rnt1p, 11 ono[o n6mer ro RNA nou opX((er vo 6qproupye(ror nep(nou 15-50 Bdoerq roOobrd rou 3'drpou rou ene[epyoop5vou 28S rRNA (BI5ne Ev6t4- ro 21.20). An6 ouri rnv dnoqrrl, o repporropr6g qg Pol I oXefl(etor l.r4Xovronr<d Ue rov repporropr6 qq Pol ll ener6rl ror or 6uo 6repyoo[eg (oog nepil.oprBdvouv 5vo ouprBdv ndqrqg RNA. Aw(Oero Ue rov tep;ronopr6 rov Pol I ror Pol ll, q RNA noLuprepdon ll ouvq0rog 6ev npoyproronote[ 6ro- rpn5 reptrlolopr6, ol)d ouvel((et vo pretoypd<per yro nepfuou 1.5 kb n6po on6 rq 06o11 nou owrororXe( oro 3' dxpo. To outrrBdv n6qnq ot4 05oq noluodevutr(oroqg nop6Xer ro 5vouo1.ro yro rov repprouopr6 on6 qv RNA noLuprepdo4 ll, 6nrrlg go[veror orrlv EtK0ilA 21.32, Auo prow6),o dXouv nporoOef yro rov repporropr6 rqg Pol ll.I0pgtrrvo prero oMoorepx6ltowi)to, n ndqn rou RNA orq 06o11 notruodevuL(ooqg (oog nupodore( optopdveg oMoy6g drop6prpoorlg 16oo oro oUpmloro Pol Il 6oo Kor ornv ronxq 6opu1 rrlg Xpcopror(vqg. Aur6 prnope( vo endyetot on6 owoMoy6g nopoy6wtrtv Kqrd t4v ow(6 poorl nolu o6evutr(oor1 q, n porol,c0wog nori orl qg Pol ll rr 6nerq oneLeuO6ptoon Uq qn6 ro DNA er<- goye[o. 'Evo evoMorcrx6 gow6tro, yvoor6 aq powilo top- ntAr1 ftorpedo model), npore(ver 6u ;uo er6u<{ e[<ovou- rledorl npoo66veror oto 5' drpo rou RNA nou ouvel((er vo geroypdrperor perd rqv n6q4. Ano6opre( to RNA nol,ri yprlyopo, tolUrepo on' 6rr ouw(0eror, on6te npogro(ver ulv RNA noLuprepdoq. 'Enemo, o.Lq- awarcffiffirymw P til Pol lll Poll affi* J xffiyffimffiw B.{ f ?i't* H Ene{epyooio rou 3'Arpou rou mRNA E(vorKproL;r1yto rov Tepgolopo rnc l'.le .:,:::-: 611 Boorreg fvuoreg
  • 103.
    Pol ll gerdro orlpo IIotru-A H rorq er06ret ro onpomdreuro 5'6rpo Anoddpqo4 qn6 eqovouKle6oq Erdyer Inv ofl oorooeponofi or1 n1g Pol ll Aneleu0dpooq qg Pol ll qnd ro eruoyeb EIKONA 21.32 0 oXqpolop6g Tou 3' oKpou rtov Pol ll peroypd- Qov 6reuKotrriver Tov TepUoTroUd Tnq UeTqypognq. Lenr6pd U' enrKouprK6g npoteiveq npoo6edepdveq orqv ropBo[ure].xr1 neptolq rqg no].upepdoqg. H o].- Lrllen(dpooq nupo6ore[ tqv one],eu06proo4 rqg RNA nol,upepdor;g on6 ro DNA, nporoLcirwoq repprorolr6 rnq Ueroypogrjg. Aw6 ro prowStro e(qye( ytor( or 06- oerq repporropro( qg RNA nolu;repdonq ll 6ev e(vor rold roOoprotrr5veg, oMd prnope( vo epgovroroUv oe drdgopeq 06o'erg pr6oo oe puo peydLq neproXll r<o0o6r- rd qq 06o49 nou owrororle( oro 3' dr<po tou RNA. H rUpro nepopronrrl dv6er[4 yro ro pow6].o ropnrtr4 efuor o p6troq r4g nuprlvrxrig 5'-3' e[ovoux].edoqg Ratl orq ([Un n trlg Xrn2 oro Oqtroonrd. H ono].ot<pq rou yovr- 6(ou ouyyd nporole( prerdBooq rnq Ueroypoenq oro en6pevo yovi6ro. Ie oproprdvo netpogoru<d ouorrjpoto, oqr6oo, peroMdlerg rou orlporog AAUAAA nou el,o- oOev((ouv qv n6q4 m4v xovovrrq 05oq noLuo6evu- Liroo4g 6ev nupo6ororiv ovoyroonrd oneLeu0Sptoo4 rnq Pol ll ofte ror perdBoorl oro en6prevo yov[6ro. Xu- venrig, ouui q 6v6er[4, po(i pe oproprdveq tonu<5q otr- Loy6g orn 6opn rnq Xpoprot(vr;9, euvoeI to oMoorepx6 pow6l.o. 'E1er nL6ov r<oroore( oog5g 6r ro 6vo prow6tro 6ev onor.).e(et ovoyroortrd ro dMo. Kor to 611o (otrrg owr- roromp((ouv oprop6veg xp(otpreg muX6g nou oXe{(ov- ror Ue rov rep;ronop6 rnq Ueroypocprlg p5oco qq Pol ll. Me onorov6rlnore 11 Kor pe rouq 6Uo ;rqXovtopo0g, e(vor ooq6q 6n o repporropr6g rqg preroypoqrjg pr5oor rrlg Pol ll e[vor orevd ou(euypr6vog Ue ro oxrlportop6 rou 3' drpou yro ro neproo6repo mRNA rov europur.rrttrri:v ruudprov. 21.17 O Ixnporop6q rou 3'Arpou rou mRNA Ttltv loTovd)v Anorrei To U7 snRNA " H 6rtppoorl rov mRNA rorv rorov6v e[oproror ono rr1 pero- ypotpl ror pu0p((eror roro q dtopxeto rou rurroprxou xu- xtrou. u To mRNA rov rorovrirv dev elvor notruodevutrropevo: ro 3 orpo rouq drlproupyelrot peoor lrrog ovridpoolg ndQrlg nou e[oproror ono q doprl rou mRNA. * H ovrOpoorl n5g1q nporino06reL npoodeorl tou SLBP oe gLo 6oU! Uroxou-0ntrrog ror (euyoprogo rou U7 snRNA Ue Uro y0r- rovrrrl povcirtro-rvrl fl sp roxn * H ovr[dpool neQqg rorotruerot ono 6vov nopoyovro Kotvo l.1t ro o0pntroro notruodevul[r,lor1g. H Broydveorl rov rmovurv etr5y1e-rot r<up(cog pdoo priOpuo4q qg o<pOov(og rrr.rv ow[orotlrov mRNA xoro rq drdpr<eto rou KurroprroU rUr<Lou. Irq prerdBooq G1/S, n ouyr<dwpcrroq rov mRNA trov rorovrrrv oulo- ver >30 gop6q L6yro ou(qpdvr;q pretoypogrlg. Auq q drepyoo(o puOpr((etor on6 ro o[gn].or<o Kux].[vqg El3dk2. H oU[r;o4 rov mRNA tov torovci.lv ot<o].ou- Oe(tor on6 toXe(o ono66prqorl rouq fio rSLog rqg gdor1g S. To mRNA rrov rorovc6v 6ev e[voL notruo6evu].uo;.-rdvo (ercr6g rou S. cerevisrae). Iuvenc6g, ro 3' dr<po roug 6q- puoupyotlwot pe 6to<popertx6 tp6no on6 q ouwovr- oprdvrl ovr(6pooq n6qtrqq/notruo6evu).(orotlg. O oXqprolopr6g rov 3' dr<prr-lv e(oprdror on6 pro e[orperL- xd ouwrlprlprdvq 6opq pr[o1ou-0qArdg nou ewon((eror 14-50 Bdoerg r<o0o6rr<d rou xrridtxov(ou tepprorroptou ror on6 6vo r<o0o6u<6 ororXe(o rorovcbv (histone down- stream element, HDE) -15 vour.treot(6to roOodrxd qg 6oUnq pr(cryou-0ql,rdg. H ndqurl npoyporonorekor preto- (U p(o1ou-0q).rdg r<oL HDE, o<prlvowog ndwe Bdoerg r<o0odLr<d rqq dopulg pr(o1ou-0r1Lrdg. MetoMd(erq nou nopepno6((ouv ro oxrlUorrop6 rou 6(x).covou pr(oXou ovomdMouv tr1 6qpuoupy(o rou drpou rou RNA. Aeu- teporoye(g 6o;rdq nou onoro0Lmo[v q 6[r].tovq 6opq (proi.ov6rr 61r onoporrrltc,:q rrlv opXLxrl o).Lq).ouX(o) onoxo0tmoUv Kor rrl 6qproupy(o rou 3' dr<pou, Auro uno6r1orver 6l o crlrlprortopdg qg 6eureporoyo0q 6o- prrlg e(vor nro orlpowrK6g on6 tqv oxprBrl oMll"ouX(o. H 6qproupy[o rou 3' dxpou rtov rmovtrrv <po(veroL orlv , . Auo nopdyoweg ononorjwor yto rov roooptopd rqg ow(6poor1q ndqUqg: 4 np<oreivq nou npoo6Svetor oro ;i(oXo-OqLrd (stem-loop binding pro- tein, SLBP) ovoyvop((er q 6opu1 1-riolou-0r1).tdq KoL ro 5' dr<po tou U7 snRNA (euyopover Ue Uro noOoro oe noup(veg oMqtroul(o prSoo oro HDE. To U7 snRNA e[- vor 6vo puxp6 snRNP nou onoretreftor on6 ro U7 snRNA (63 vour<Leor(6ro) r<or pro oprddo oprerorv npoteivorv or ono(eg olerf(owot pe lg snRNP nou etrrnL6xowor orrl -e__j-_reG Xrn2 I (A)n 612 KEOAAAI0 21 Iupporprl rr Ene[epyooio rou RNA Boorxeg 6worcg
  • 104.
    A G SLBP UU ,'U jjc ffi,ffi ffic mRNA Lmovtirv Iugntrer(vr1 oupporprl toumRNA (BLene Ev6t4ta 27.6). Movo6tr6g yro ro U7 snRNP e(vor d6o npcoretveg nop6proeg pe rtg Sm, or LSM10 ror LSM11, or ono(eg owroOtororiv ttg Sm D1 rot D2 qrn ouppoerl tov snRNP. Av ovoorqlei ro (euydpropro prero[U U7 snRNA xot HDE, q 3' ene[ep- yoo(o tcov mRNA r<ov totovtilv 0o 6toropol0e[. An5 qv d.tr.q nLeupd, owroro0prorx6g pretoMd(etg oro U7 s- nRNA nou onoro0roroUv q ouprnqpoptonr6tpo ono- xo0roro0v en(or1q Kot rnv 3' ene[epyoo(o. Aur6 unodqtrdrver 6n ro U7 snRNA Leroupye( pr6oo (euyo- pdrgorog Bdoeo:v pe to mRNA ttov tqrovdlv. l-16r[q yro rq 5r1p[oupy(o ev6q 3' drpou ouprBofuet oe moOeprl on6oroorl on6 11 06o11 nou ovoyvop[(erot on6 ro U7 snRNA, yeyov6g nou unodrltrcbvet 6tt to U7 snRNA etrrnMretoL orov xo0optopr6 qq 05ong ndq.trlq. O nopdyowoq nou euO0verot yto rrlv n5Qr1 e(vot q npone[vr1 CPSF73 (cleavage and polyadenylaflon speci- ficity factor 73). Iuven6q, ou'r6 to prdLog qg oroySvet- oq rov peroMo-B-Aor<ropoo<ilv no((et Bootx6 p6Lo orq 64puoupy(o rou 3' dt<pou t6oo yto to nol,uodevulttopr6- vq RNA 6oo r<or yro rq Un notruo6evuluo;.tdvo RNA ttov rqrovdlv. 'EXouv ovoyvoprore( opr<et6g nporetveg nou no[(ouv oqprowtr6 p5],o orrl 6qpuoupy(o tou 3' drpou rrov rorovdlv, pero[6 dMarv 4 CPSF100 rot q lugn].er(- vq (symplekin), oMd or erdu<5q 6pdoerg toug 6ev 61ouv xo0optore(. Aut6g or enrnp6oOereq npolteiveg nt0qv6v nop6louv trenoupy(eg rxptc0ptotog nou oro0eponototiv ro oripntroro ene[epyoo(og rou 3' dxpou. E(vot ev6ro<p5pov 6tr 11 dtotopoXrl rou (euyopdrtrro- roq rou U7 pre trg oLlrlLoul(eg-m6Xoug oro yov(6to rc,:v torov6v 6nog en(or1g ror q dropeool,oBotipevrl on6 siRNA e(dLetqrl dlLrov ouorotx6v nou egnl6t<owot orq 6qploupy(o rou 3' dr<pou rov torovdlv o6qyoUv oe CA-Py7-N2-u -AAAGAGCUGU-3, ttttttllll uuucuccAcn %e5' U7 snRNA nopoB(oor1 tou nSpotog rnq peroypqg{g rot oe TIo- l,uodevuL(cooq Xprlotpronotdrwog 5vo orlpro nol,u-A t<o- 0odrxd rou DHE. Iuvenc(lg, rot' owtorot{o pre to p6}'o qg n6gqg/notruo6evutr(oonq rou mRNA orov repport- opr6 qg preroypogrlq p6oco rqq Pol ll sro neptoo6tepo yov[6ro nou ro6rxonorofv yov(6to, 11 n6Q4 tou RNA nou peootroBe[tot qn6 ro U7 t<otd q 64;rtoupy(o tou 3' drpou Eofveror 6rr e(vot r<pfotprr; yro rov teppottopt6 rnq Ueroypoqnq oro yov(6to ttov torovrilv. neun U7 snRNP : H [rlproupyro rou 3' orpou rou H3 mRNA rotv torovtirv e[opToTor ono puo ouvrr1p1p6v1 qoupr6ro roL ;tto o),trr1LouX(o nou .e-yoptover ue ro U7 snRNA. 2 I .l I H luppogri Tou IRNA lleprAopBdvet Konh rr Enovoo0v6eon ot lextoprordq Avrr6pdoerq a H RNA notrugepoorl lll repuon(er rt,l UeroypoQn orqv 0trtrr1- trouXio notru(U)4 nou Bp(oKsTot ptloo oe Uto otrtrltrouxto ntro0- oro oe GC. * H ouppoQn rou IRNA npoyporonote(rot pe dto6oxtr6g ovrt- 6pooetg ronrlg ror orlvDeolg. e Mro evdovourtreooq roper ro npodpopo IRNA rot oto duo orpo rou rvrpoviou. * H onetreu05poorl Tou lvrpovlou dqptoupyei 000 utootRNA ue oouvrl0toro orpo nou nepr6Xouv 5'-0H rot 2'-3' rurtrtrd qo- oqoprro. * To 5'-0H oxpo rptootpoputrttlverot on6 pto notruvouxtreondtrl ruvoorl, 11 opo6o rou rurtrtro0 Qtrtotpoptrou ovo[yet ono pto rprootpodteorepoorl onodldovrog 2'-tpoorpoptro retrxo orpo rot 3'-0H oUo6o, ro orpo rolv e[ov[ov ouv66ovrot on6 ;tto RNA trtyroorl ror ro 2'-tptrrotpopuo otpopelrot ono gto Qo0Q0T0or1, :: 'i8 H Iupporpn rou tRNA l-lepilopBover Konfl rt Enovoorjvdeol oe :eXoprore_Q Avl6pooet-c 613 Lsm10 Lsm11 Boorreg 6woreg
  • 105.
    Or neptoo6repeg owrdpdoergouppocp(g e(oprcbwor on6 BpoXe(eq ouvorverrK6g oMr.1).ou1(eg xor 6re[dyov- ror p6oto ovrr6pdoecov frans-eorepono[r1oqg mrg ono(eg 4 OpoUorl efvor ouwovropdvq pre q 64ploup- y(o 6eoprciv. H ouppogri rcov yovrS(ov IRNA enmuyld- veror p6oro ev6g drogoperrroU prqXovropro[ nou Boo[(eror oe [eXopror6q owtdpdoerg xonrlg rqr o[v- 6eo4g. [1epftou 59 on6 ra 272 nupqvrrd yov(6ro IRNA orq (UUn S. cerevisiae e(vor 6roreroprprdvo. Ko06vo riler 6vo ptovo6rx6 rwp6vro nou ewon((eror g6Lrg 6vo voux.l.eo- rbro prerd rrlv 3' nleupd rou owxul6u<ov(ou. To rwp6- vto r<ugo(vowor oe ;r5ye0og on6 14 6rrlg 60 Bdoerq. Ooo Bp(orowor oe oXerr(6prevo yov(6to IRNA SXouv nop6trloro oM4l,ouX(o, evdl ro rwp6vto oro yov(6ro tR- NA nou owrTrpoooneriouv drogopertrd oprrvo[6o 6e oler(owor. Aev undpyu ouvaweuxrj aAA4louyta nou 0a pnopoloe v' avayv@porc( an6 ru 6v(upa ouppa- g{9. Aur6 en(oqg roXUer oro 6rorerop;r6vo nuprlvrrd yov6ro IRNA rcov rpur6v, r<ov opuprB(cov ror rrov 0r1Lo- orrcbv. 'OLo to rwp6vro nepil.oprBdvouv pro oLl,rlLouX(o oupnL4pcol.rorxrj pe ro owrr<odm5vro rou IRNA, Aur6 6qproupye( pro evoMorcnxrl dropr6pgoorl yro to Bpo- 1(ovo rou owrxrrl6rrov[ou offlv ono[o ro owu<rrldrr6vro e(vor (euyopropr6vo, q44poil(owog 6ror puo npo6r,roorl rou ouvrlOoug BpoX[ovo. 'Evo nopd6erypo nopouord(e- ror orrlv EIK0NA 21.34. M5vo o Bpo(ovog rou owro6r- rov(ou enrlped(eror - ro un6lorno pr6pro 6roqpe( u1 ouvqOropdvq 6oprr1 rou. H orprBr1q otrLrlLouX(o r<or ro pr6ye0oq rou rwpovi ou 6ev 6Xouv o41.roo(o. Or neproo6tepeg preroMd[erg oro rwp6vro 6ev nopepno6[(ouv q ouppogrl. H oup- paEfi rcu IRNA elaptdtat xup[aq an6 u1v avoyvthpto4 ploq xo:fiq eutepotayolq 6op4q oro IRNA, aw[ yn pn xolr1 aMr1Aouy[a rcu twpov[ou. YndpXouv onpqwr- r<6q neptoX6g oe drdtpopo pr6p4 rou pop(ou, prero[0 d].- ).ov oLLrll,ouX(eg peto[O rou BpoX(ovo-66rcrr1 Kor rou ZeuydpoUo To twprlvro (euyoptirver ge q 0r[trd tou owtrordrrov(ou To rvrpovlo oT0 tRNAPrre r1q (uUlq (euyoprr-rver pe ro ovlrorDrrovro yro v' otrtro[er q dogrl rou ppoX[ovo TOU ovTrKo]- drrovlou. To (euyoptoUo pero(f pLog onortreropevlg poolg oro ploXo ror qq 0qtrrog rou rvrpov(ou rou npodpopou poprou rotrtg onoue [ror yro q ouppotpil. BpoX[ovo D, oMrll,ouX(eg oro Bpol(ovo T/C ror rd(cog oro BpoX(ovo rou owrKo6u<ov(ou. Aur6 Oug((er ng 6o- pm6g npo0no0Soerg nou r[Oewor oro IRNA yro rn pe- rdgpoorl (BMne KegdLoro 24). MoLorqUro, ro rwp6vro 6ev s(vor eweL6g doXero. To (euydpropro prero[ri puog Bdorlg orq 0r1].td rou rv- tpov(ou Kor Uroq o(euydpoqq Bdonq oro p(oXo e(vor onopofuqro yro rrl oupporpq. Mero).1,d[erg oe dtr].eg 06oerg nou enrlped(ouv out6 ro (euydprrtpto (n.X. 6r;pu- o u pyriwog evoMorcrxd np6runo (euyopcil poroq) enrl - ped(ouv 14 ouppogrl. Or rov6veg nou 6r6nouv rr1 6toOeorpr6qro rov npo6p6prov tRNA yro ouppo<pr1 ptord(ouv pe rouq ow(orotloug xov6veg yro rrlv ovo- yvciprorl on6 rrg opuvoor<ulo-tRNA-ouv0erdoeg (BLdne Ev6trya 25.9). I' dvo Oepproeuo(oOqro perd).troypro (0prqg nou onoruyldver v' ogotpSoer ro rwp6vto, droreroppdvo np6Spopo prdpro ouooopeUowor mov nuprlvo. To np66popo pr6pro lprlorponororiwor <og unoorp6proro yto rivo oUmqpro el,eU0epo rurrdpov nou er<Xu),((e- rqr on6 r0rropo dyptou rUnou. H oupporprl tou np6- dpoprou 1.rop(ou prnope( vo noporrlprl0e( L6yo rrlg enor6).ou0r1g pe[ooqg rou prey60oug. Aur6 onnKo- norefuot pSoo qg ol},oyr1g mr1 06oq qg (rrivrlg r<ord qv qLerrpoe6pnon oe nr1rr4, 6nog <porveror orrlv EIN0NA 21.35. H eLdrrcoorl rou pey60oug epprrlverieror on6 rqv eprgdvroq ;uog (cilvrlg nou owrnpoooneUer to rwp6vro. To eLe(0epo rurrdpov erlULrotrro prnope( vo xLo- ol.rotono rr1 0e[ el,5yXowo g rqv rxov6rrlro ou p poE4 g rou IRNA. H in vitro ow(6pooq onone( ATP. O lopo- rcrqprop6g rotv owr6pdoeov flou npoypororroroUwor pe { 1op(g ATP deiXver 6r ro 6uo leyuptord otd6n qq owbpaoqq xataiowat on6 lnEopetxa 6v(upa. Erd)ool ['erXUtrrogo llp6dpogo tRNA NG G -,ffi,# Ufl; :d,6nl ffiffi ./ ffi'8rry, TP ffiIvrprivto Oprgo tRNA CG GA AU GCm AV CG m AU cA UG wAvflK(l)6rK6vro tl ffi noo*,, ffi ry1 turpdvro ffi -r n*o [:'] cH"YdhW+ %, H),exrpoqopqol oe nrlrrrl ' H ouppoqrg rou IRNA Tnq (uUnS in vitro noparo- ),ouOe(ror p6oo qg ovotruolg rou npodpopou RNA xor Tov npo- iovtrov pe qtrerrpoqoploq oe n1rrr1. 614 KEOAAAI0 21 lupporprl rr Ene(epyoo(o rou RNA
  • 106.
    o To npdnognUo 6ev onore( ATP. l-lepiloprBdver n6- rlq <pootpo6ooreproU 6eogoti p6oto gtog dtunrlg ow(dpooqq vouxtredoqq. Kototr0etot on6 pto ev6o- vouxl.edoq. o To 6eUtepo FnUo onore( ATP xot nepfioprB6vet ro oXrlprotropr6 6eopro0. Eivot 1-tto owfdpooq o6v6eoqg xor 11 une60uvq ev(u;lxri evepy6qto neptYpd<petqt coq RNA lryrdo4 (RNA ligase). H ouppoqrl rou pre-tRNA Yto rrlv orpo[peor1 twpo- vfov e(vot onopo(tr1tq oe 6].oug rouq opYovtoproUg, otrtrd 6to<popettro[ opyovtopro( Xprlor;ronotoUv 6torpo- perrrotig prrlXovropro0g Yto rrlv en(teu[4 qq. Ito Bo- rcr{pro, to twp6vto ttov pre-tRNA ouro-ouppdntowot oq ouroKqrotrutrxd twp6vto opr66og I tl ogdSoq ll' Iro Aplo(o rot to Eurdpuo, n ouppo<pr1 tou pre{RNA nepil.o;rBdvet q 6pdoq tptc0v ev(0prcov: (1) gtoq ev6o- vouxtredorlg nou ovoYvopf(er rot nSmet ro np6Spopto Kor oro 66o drpo tou twpov(ou, ;rto fuyxdon nou ouv- 65er to e(drvto tou IRNA t<ot t1 2' 9t:ogotpovoqepd- on nou ogotpe( ro 2'-9rr:o9optr6 mo ouppog6vo tRNA. H ev6ovourledo'q rnq EUUnq e[vot 1-tto srepore- rpopepnq nproteivrl nou qnotel'e(tot on6 6uo xoto- l,uttr6g unoprovddeg, rtq Sen34 rot Sen2, rot 5uo doprrSg unoprovd6eg, trg Sen54 rot Sen15. Ot 5pd- oerq rrlq nopouotd(owot ornv Ell(0NA 21.36. Ot oXett- (6preveq unoprovddeq Sen34 rot Sen2 n5mouv ttg 3' ^oi s' e6oetq oupporprig, owtmo(1roq. H unoprovddo Sen54 (oroq roOop((er ttg 06oetg ronriq "petpri:wog" t4v on6moorl on6 6vo oqpre[o m1 6opr{ rou IRNA' AutS ro orlpre(o Bp[or<etot mov oyrdrvo qq (cbptpng) 6oUnS opi;rotog L. O p6Log qg unogovddog Sen15 6ev efuoiyvt:m6q, ol'Ld ro yov(6t6 r1g e(vot onopo(- rnro yto rn EUUn. To (e(yog Bdoeov nou oplprot[(e- ror ;rero[ti rqg nptirtqs B6onq orq 0r1].td tou owu<trrdtrov(ou xot qq Bdoqq ilou nponYe(tot qg 3' tlt(0NA 21.3? H evdovouxtredon rnq ouppoQnq rov IRNA rolv Apxq(ov r6pet xdOe rtr6vo o' 6vo tpouoxtol-to evdg ;roripou tpotl- orol po-6)rwo-tpouoro p0. 06oqg ouppoqriq e(vot ovoyxo[o yto ulv ron( qq 3' 05oqq ouppoqdq. Mto ev6roq5pouoo 5vdet[r1 Yto rrlv e[6].t[r1 rnq oup- pqgns tou IRNA npoqL0e on6 rg ev6ovouxirsdoeq rov Aplouov. flp6xemot yto opro6q.repri ri ogotxpopepi, oro ono(o rd0e unopovd6o 6Xet 5vo evepy6 r6wpo (nop6tt oro rerpopep6g treroupyotiv pr6vo d0o r<5wpo) nou 16- Ber puo on6 rtg O5oetq ouppoqriq. H unoprovd6o 61et oL- i4l,oux(eg nou oxet((ovrot Us ttq otrI'rlLouX(eq tov evepycirv rSwptrlv orq unoprovd6eq Sen34 xot Sen2 tou ev(riUou rnq EUUq. Om6oo, ro 6v(u;ro tcov oplo(tov ovo- yvrrlp(ouv ro unoorp6ptotd roug pe 6totpopunr<6 tp6no' Awlvq gerpo0vrqv on6oroc4 on6 ouyrexprydveg oMrl- LouX(eq, ovoyvop((ouv 6vo 6optr6 XqpoKrnptorx6 yvro' o16 coq e[6yr<o: go-6ltr<o-e(6yr<o go (bu I ge-hel ix-bu I ge)' H H(ONA 21.37 6e(yyu 6t q r<onq ouprBo(vet oro 60o e[oy- rrigoro. Euvenc0g, 11 ouppocprl rcu IRNA npoqyekot rou 6roXroproproU ttov Aplo(rov Kot rov europutottitv' Av npo- 5ruQe pr6o<o rqg erooyroyrlg tou twpov(ou oto IRNA, ou- t6 np6na vo {tov 6vo no)rU opXo(o ouprBdv. H ouvol,txrl ow(6pooq ouppo<pr1g tou IRNA ouvorf(- (eror orrlv Ell(0NA 21.38. To npoi6wo qg n6tl'r4q e(vot 5uo ypo1rg,n6 rwp6vto ror 6uo ptod g6pto IRNA' Autd ro ev6rdpeoo 6Xouv provodtt<d drpo. Kd0e 5' drpo ro- roLrlyet oe Uto udpo[uLopd6o, ev6 r<d0e 3' drpo oe puo 2',3'-rurtrxri rprrlo<popxrl ogd6o. ' To 6uo prtod IRNA (euyoprirvouv yto to olqpottopt6 ptog 6o;rig nop6protog Ue rou IRNA. 'Orov npoore0e[ ATP, npoyl.rotonotefuot 11 de6tepq ow[6poo4, 11 ono(o r<orol0etoi on6 6vq povodtx6 6v(upro pre noMontr6g ev- (upux5q evepy6qteg. 1. Evepy6tlta xuxLtxfiq EuoEodeorcpdo4q' Kot to 66o oouvriOtoro 6t<po nou nopdyowot on6 tlv evEovourtredorl np6net vo rpononotq0oOv nptv qv ow(6pooq ofv6eoqg. H r<uxl'txrl goo<poptxrl o;rd6o np6ro ovofuet onoS(dowog 6vo 2'-rpoorpo- pu<6 tetrtr6 dxPo. 2. Evepyotqta x:do7q. To npoi6v 6Xet pto 2'-qor- 5',3', FS pS fllS gI'{ hH Phi! h*ffi F$ryh/ H,{ l/- EqoYKouo mm-"-f ;+4-- E[6Yxtouo pd gt /&Ffif,* PS. lr/XopoKog 5' Kontl ll5', Senl 5 - O : Avrtrodtxdvto-tvrpovto (Al) (euyoq pooetlv ..,,r' . ' ::r 0r 3' rot 5' nerfetq oro pretRNA rou S, cerevisiae roroluovrot ono dtorpopertrSq unopovodeg qg ev6ovourtreoorlg' N4ro otrtrn unogovoDo ioolq ro0opi(et qv evrontol rotv Oeoeotv nrlfiq Uerpdrviog rrlv onooroorl ono qv 6ptUtt OoUn. En(onq, ol- povrtr6 e(vot ro (e0Yoq pooeotv Al. 3' Konrl Sen34 ?1."i.* H Iupporprt routRNA flepiloppover Konrlrt Enovoouvdeorloe:eyot0Lc :l - 615
  • 107.
    3', 3', i,l i,n ry"pHry'#kry l?lqt,,, z',-3', P H rproogodteotepdorl ovo(yet ro grrrorpoprxd Oorrrjtrro AtruoLDq A),uo(60 tRNA Boon IRNA Boort oft-r'-r',+ 2'-3' xuxtrrxd groogoprr6 3',-0H, 2'rproogoprxri oqoprrrl oprd6o Kor Uro 3'-OH oprd6o. H 5'-OH oprddo nou nopdyerot on6 tr1v evSovour<).edor1 npSna vo eooeopulroOe( yro v' ono6c0oer S'-rptrl- o<popu<6. Aut6 nopdyet pto 05or1 orqv ono(o q 3'- OH Bp(oreror 6(nlo oro 5'-<proogoprx6. 3. Evepy6tryo ttyxdoryq. H oproronollrrl orepot6qro tr1 g noluvo u ri.eol6rruj g oLu o(6o9 qno ro0(ororor ;r6oco qg evepy6qrog l,tyxdoqg. To pr6pro nou 51er ouppo<pe[ efvot ouveX5g, pr' Svov 5'-3' groo<po- prr6 6eopr6 orrl 06o11rnq ouppoqris, q)ild eil(onq 6Xer gro 2'-<p<oogopxrl oprddo nou enrorlpo(ver ro ou;rBdv ndvo oro ouppoprpr6vo IRNA. Iro reLeu- to(o B{pro, outtj q enrnp6o0eq o;rd6o orporpeitor on6 pro g<r:orpordoq, q ono(o ;rero<pdper ro 2'-Ero- ogoprx6 oro NDP g4rlgor((owog ADP 1,2-xuxLu<rl qrooEopxrl prp6(n. H o669 oupporprlg rou IRNA nou neprypdgrlKe no- pondvo e(vor el,ogprilg 6rogopertrrl on6 qv owforor- Xq o66 rrov onov6u),orriv. l-1poto[ dpdoouv or RNA l,ryxdoeg, puo r<urLdoq nopdyer 5vo r<ux.l.rr6 2,3 rux.l.t- 16 dr<po ond ro opXx5 3'-groogoovoeoreprx6 drpo pr5- oo ev6g 3'-odevuLr<o1ldvou ev6topr6oou. H RNA Lryxdo4 en(oqg 6rogripet on6 ro 5v(utrro rnq (6pnq enetdrl pno- pe( vo ouv66ou 6vo 2',3'-xuxLu<6 EroogoSeor6po ror gto 5'-OH yro vo olqtrror[oer 5vo ouprBoru<6 3',5'-rpro- ogo6eoreprr5 6eopr6, oMd ourdg ot owrdpdoerg 6ev o<ptlvouv nLeovd(ov 2'-<p<oogopu<6. Ktrrovog K),ovoq IRNA IRNA 21.19 H Andvrnon oTnv Ilopouoio proq !e6rnAtrlpdvnq flpolrei'vnq Ix€Ti(srot U€ Tn Euppogfi Tou IRNA * H lrel e[vor pro eooreprrfl nupqvLrrl pepppovLq nporeivl pe qv N-retrrxq neproXl Tlq oTov outro rou ER ror q C-retrrx1 neproxn qq 0T0v nuprlvo. H C-retrrq neproX1 5Xer evepyoq- reg rtvoorlg rr ev6ovourtreoorlg. * H npdodeol ptog [edrntrop6vqg nporeivlg oqv N-retrtri1 neproxrl evepyonorel qv C-retrtxq evdovourl,eool Us 0uT0- tproorpop utr[too1. * H evepyonorlpevl evdovourtreooq rripet ro H,4C7 mRNA (Xbpl oto onovdutroro) onetreuOep6vovrog rlvo rvrpdvro ror dqpLoupytirvrog e[6vro nou ouvdeovror perol[ roug on6 pLo tRNA trryrooq. * M6vo ro ouppoptlvo H/C/ mRNA pnope[ vo perotppoore[ o 6vov peroypotprxo nopoyovro o onoiog evepyonote[ yov[6ro yro ouvo6o0g nporelveg nou po10o0v oro 6[n],topo rtllv [edr- ntro.t ptivrriv np toreiv6v. e H evepyonotlpdvq lrel enoyel ononrool rirov ro rurropo nepr6Xer nopo notrtr6q [edrntropeveg nporretveg. 'Evo oouvrlOroro otiorrlpro ouppoqrlg nou olet((etor Ue rrl ouppogrl rou IRNA e(vor 11 o66q ondwqoqg oe H ouppotptl rou tRNA onotre( (eXtoptoreq evepyorrireg vouxtreoolg u e[ovourtreoorlg. To 6pro e[otvfou-Lvrpoviou xopor- rot ono rr1 vourtreoorl nopoyovtog 2'-3'rurtrrxrj rporotpoprrd ror 5'-0H orpo.To rurtrlro qoorpoprrrl ovoryeL onodrbovrog 3'0H xor 2 q,;- oqoptr6g opo6eq To 5 -0H e[vot tptrtotpopu],urt1uivo. Mero qv one],eu0epoorl TOU rvrpovlou, ro pLoo popro IRNA dLntrovovror oe pLo dol* nopol-roro Us Tou tRNA nou r6po 5Xet ;rLo 3'-0H, 5-P 0po0o1 nou orppoyi(eror ono pro trryroo1. 0.16' KE0AAAI0 21 lupporp4 rr Ene[epyoo(o rou RNA H xrvdoq rpoorpoputrloiv€r To retrrK6 5'-0H 6xpo Boorxeg eworeg
  • 108.
    (e6rn).oprdveg nporetveg (unfoldedprotein re- sponse, UPR) nou e(vor ouw4pnpdvn on6 q (tipq 5r,-lg ro OqLoortr<d. 'Onog ouvorp((etot orqv EIK0NA 21.39, 11 ouooril peu oq [e6rn],otrrdvr.ov npulreivcilv qrov qu^.6 rou evdonl.oopolxo0 6rrr[ou (EA) nupoSore( qv o66 UPR. Aur6 odrlye( oe ou(4prdvq ;reroypoqr; yovr6(crlv yro ouvo6orig nporeiVeg nou Bo4Ootiv oro dft).opro rolv nporetvri:v oro EA. Euvenrig, 6vo orjpo npSner vo perodoOe( on6 rov ouL6 rou EA orov nu- pnvo. O oro04rrlpoq Trou evepyonore( trlv o66 e[vor r1 nporetVq lre1, 4 ono(o ewon[(eror mo EA Ko/n rnv eororeprr4 nupqvu<r1 pepBpdvq. H N-reLu<rl neproX4 rqg lrel Bp(or<etor orov ou6 rou EA 6nou ovrXyeUer r1v nopouo(o (e6rnlcr:pr6vov npolreivriv, prdMov g6oro qg np6o6eonq rnq o' erre0erp6vo pror(Bo. To C-te).u<5 9ro6 r4g lrel ewon(erot e(re oro xurrop6nl.oopo e(te orov nuprlvo (L6yco qq ouveloUg pepBpdvqg rou EA Kor rou nuprlvo) xot 6ro06rer 16oo evepy6rrlro rrvdorlg Ser/Thr 6oo ror erdu<r1 evepy6ulro ev6ovour<tredo4g. H n p6o'6eor1 (e6rn).to prSvov npcoreiv6v n po roLe( ou oori- peuon Uovopeprirv lrel orrl gepBpdvq rou EA, odqydrv- roq qrrlv evepyono(qo4 qg C-relu<r1g neproXlq orqv H neproXrl orov quld rou EA npoo66vet qv [edrnAogdvl nporelvrl H neproX4 mov nupqvo / xunoprintroopo ropet ro H,4C7 mRNA +rc dtrl.q nLeupd qg pepBpdvrlg pSoro owogargopuXrrr- onq. H evepyonor4prdvrl C-relrrrj ev6ovour<Ledon 6xe1 npoq ro nop6v, pr6vo 5vo (oqpowu<6) un6mporpo: np6xeLror yro ro mRNA nou rro6rxonore( rov etdtr6 yro rnv UPR nopdyowo Ueroypo<pfq Hacl orn ([Un (Xbpl mo onovdulrord). Yn6 xovovrr6g ouvOqr<eg, 6rov 4 o669 UPR 6ev e(vot evepyonorrlpdv4, ro mRNA rou HAC| nepr6Xer 5vo rwp6vro 252 voux.treorr6(rov (To Xbpl nept6ler 6vo rwp6vro 26 vourd.eortS(rov). Irn (0prf , ro rwp6vro tou HAC| nopeprno6((er rn Ue- tdgpooq tou mRNA oe l,etroupyu<4 nporeivq, evdl oro 0r1Loonx6 ro rwp6vro rou Xbpl ennp6ner Uev rrl petd<ppooq oLl,d 11 npotetvrl ypnyopo ono6ope(ror on6 ro npcoreooopdrro. :rrlv ene[epyoo(o ouro[ rou rwpov(ou epnL6rowor oouvrlOrmo oumolrd ouppo- qnq. H evepyororrlp6wl ev6ovourl.edoq lrel 6po dpre- oo oro mRNA tou HACI (mRNA rou XbpT oro onov6uLrord) rou<6Ber lg 6uo ou;rBol.rig oupporprlg, ogrlvowog 5vo 2',3'-r<ur<Lu<6 googoprr6 mo 3' dxpo rou 5'e(olviou Kor pto 5'-OH mo 5' dr<po rou 3' e(o:v(- ou. 'Enetto, or 6uo oupBoLdg ouvdSowor on6 r1v tR- NA Atyr<dor1 nou 6po orrlv o66 ouppogrig rou IRNA. 'Eror, ol,6x).r1pq r; o669 ene[epyoo(og rou mRNA rou HAC1 (Xbp1) prord(er Ue rnv o66 rou pretRNA. Om6oo, undplouv oqprowx6g 6roqop6g prero[ri rtov 6uo o6drv. H lrel xor 11 ev6ovouxedoq IRNA 6ev SXouv nop6;roro oprol,oy(o ri orimooq oe unopovd- 6eg. H evepy6rqto ev6ovoux..edonq rnq lrel pu0pri- (etot oe peydLo Bo0p6 oro EA ru 61et pr5vo 5vo un6mpopro (pre-mRNA tou HACI). Aw(0ero, q ev6o- vour),edoq IRNA 5Xet no).),d unomptirporo, 6Lo pre xorv6 6(nLopo rou IRNA, pre ;lrprl npor(p4o4 yr' otr- l,4LouX(eg nou neprBdll,ouv rrg ouprBol6g ouppo- ens. Xprlorponortirvrog 1.rro tritoro o66 nop6proro pre rou IRNA yro rnv ogo(peor1 tou rwpov[ou on6'ro m- RNA rou HAC1 (Xbpl), ro riprpro mRNA prerogpd(e- ror Kor. nopdyer 6vov roXup5 preroypoprr6 nopdyowo pe por(Bo Boorrof-<peppoudp Leur[vr1g (bZip) nou npoo65veror o' 5vo r<orv6 prot(Bo (UPRE) orov unoKr- vr1r1 noLlcilv r<o0o6rrd:v yovr6(rrlv. To yovrbrord npo- i6vro npootore(ouv ro x0nopo ou(dvovtog rqv 5r<gpoo4 nporeivrilv nou Borl0otiv mo 6(nLtopo rov nptoreivrrSv. Av ro oUorrlpo UPR roror.truore( on6 [e6rn].opr6veg npoteiveg, n evepyoilornp6v4 neproXrl rrvdorlg r;g lrel npoo6Svetor oro pr5pro-npoooppromrl TRAF2 oro KUr- top6n).oopro yro vo evepyonorrloer rqv o66 qg on6- nroorlq Kor vo Oovorcboer to r0rtopo. 'Erot, to r0rropo Xprlo$onoe( pto oouvqOroq qrpqrrlyxq ene- (epyoo[og rou IRNA yro v' onowrloer oe (e6rn]lr:pr6veg nprrrreiVeg. 0o16oo, 6ev undpXer epqovnq oX6or; ;rero- E0 qq evSovoux.,edor1g lrel Kqr rnq ev6ovour<Ledoqg rnq ouppogr1q rou IRNA, on6re 6ev e(vor npogovSg n6g 0o pnoporioe vo SXet e[e].tX0e( ow6 ro efet6rreu- pSvo orior4pro. | rulr.ooooon I nnooounon I tnr,rn i,v*oon MeroQpoon i i .] HAC! '! j9 . '.;6P I H onovqol oTrlv nopouo(o prog [edrntropevlq rporrelvlg npoyporonorelror Us illv evepyono[101 erbrxqg oup- roqflg rou HACl mRNA yro Tr'1v nopoytrty1 evoq UeToypoQrKou ropoyovro nou ovoyvop[(er rqv UPRE, UPRE ;i.i* HAnovqorloqvllopouo[oprog:e0rntropevqgl-lporeivrlglXen(erorpeqlupoc:-,,-::',- 617
  • 109.
    21.2O H IlopoyrrrvfirRNA [lpoUno0drer IugBdvro Koniq ror fleprAopBdver Mrrpd RNA * H RNA notrupepool I reppori(er rn Ueroyporprl o' evov reppo- norl 18 pooeolv. * To Ueydtro rot ro prrpo RNA onetreu0eptirvovrol ge ronrl on6 evo rotv6 npo6po;ro rRNA. To 55 rRNA peroypoqerot [eXrrrpr- oro. * H oUodo C/D rtov snoRNA e[vor onopolqrq yro qv rpononol- loq Tnq 2' Oeor1g qq pgo(qg pe npoo0rirl prog ;re0utropo- doq. * H ouo0o H/ACA rov snoRNA elvor onopo(rr1T! yro q UeTo- rpoq rnq ouptdivrlg oe qreudooupr6[vn. * Ie roOe neplnroor1, ro snoRNA (euyoprlver pe gro otrtrqtrou- Xro RNA nou nepr6Xer q pooq-oroXo riore vo dqproupyrloer 1.uo tunrxfl Dop1 nou elvot ro unoorpo;ro yro rrlv rponono(- ton. To r<rlpro rRNA ouw(Oewor oq U6poq ev6g povo6txorl npo-lroyevoUg preroypdgou nou unoBdMeror o' ene- (epyoo(o yro v' onodriloer ro roptpro npoidwo. To np66popro pr6pro neprdxer nq oM4).ouXieg rov 18S, 5.8S, and 28S rRNA. (H ovoproro).oy(o tov drogoperr- r<cbv prBoorr;prrcbv RNA Boo((eroL oe npot;reg prei.dreg ro0((r1or1g nou 6re[4X0r]Kov oe 6roBo0pr[oerq ooKxo- p6(qg ro 1970.) Iroug no).urrirropoug europucbreg, to np66popro pr6pro noipver ro 6voprd rou on6 rnv ro- XUqro xoO((qoqg rou oq 455 RNA.Iroug govorfr- ropou g/otrtyovr<Unopo u g eu xopu c6reg e(vor puxp6re- po (orn (0prl, 35S). To cbprpo rRNA onel.euOeptr:vowor on6 ro np66po- po ;16pro pr6oo evdg ouv6uool-roti ouprBdwrrrv r<onr1g rot owt6pdoeov {,ro}.t6(ouoroq nou ogoLpo0v t6oo ro e[olreptrd 6oo xor ro eooreprKd preroypo<pdpevo dro- orqprotu<d rpqproto (external/internal transcribed spac- 5,ETS 18S ers, ETS ror lTS, owroroiXtog). H Ell(0t{A 21.40 6efi3rer rr1 yevrxrl o66 orrl (0;rq. YndpXouv 6rogop6g orq oerp6 rrrrv ouprBdwrrlv, oMd nop6proreg owrdpdoerg eprnM- Kowor oe 6Loug rouq euKopu6teg. To neproo6repo 5' drpo nopdyowor dpeoo ;16oo ev6g oupBdwog ronrig. To neproo6repo 3' drpo nopdyowor prSoro ron{g nou oroLouOe(ror on5 puo 3'-5' ow(6poon qrofu6(oporoq. Aurdg or 6repyoo(eg r<o0op((owor on6 noMd crs-6po- orrrd prot(Bo mo ETS ror ITS ror dre[dyowor Ue rrl 6pdon > 150 nopoy6wcov ene[epyoo(og. l-lotrLdg prBovouxLedoeg epmL5xowqr ornv ene(ep yoo(o rou rRNA, pero[U d],],rov opropr6vo oumorxd rou e(roool.rqr(ou, ro ono(o e(vqr pro ouvd0poropo op rerc0v efrovouxtreoocbv nou enloqg outrrprerSler mrlv ono66;rqoq rou mRNA (FL6ne Ev6-r4ta 22.5). MeroMd- [erg oe (eXoprord 6v(upro ouvr1O<og 6ev nopepnodi(ow qv ene[epyoo(o, yeyov6g nou uno6r1],civer 6l or evep y6r4t6g roug e(vor nl,eovd(ouoeg ror 6n yro rnv nopo- yoyf rov drprprov prop(rrlv prnope( vo Xcrlorponorrl0o0v 6rogoperxoI ouv6uool.ro[ ndryeov. Yndplouv ndwo nollonl,d owrypogo rnq Uero- ypocpxrlg provddog yro ro rRNA. To owiyporpo opyo- vrivowor rog ruXo(eg enovoA4Qerg (BA5ne Ev6qta 7.8). To yov(6ro nou ro6rr<onoro0v rRNA preroypdrpowot on6 tqv RNA noLuprepdoq I orov nupqv(or<o. Aw[Oero, ro 55 RNA peroypdrperor on6 [eloprord yov(6ro pre rrl 6pdoq qg RNA no).uprepdorlg lll. l-evrd, ro 55 yovi6ro e(vot ouyrewptrlpr6vo, oMd 6roXrop((owor on6 ro yovi- 6ro yto ro r0pto rRNA. Yndpler puo drorpopd orqv opydvroon rou np66po 1.rou 1.rop(ou oro Borcrripro. H oMrlLoul(o nou owrorot- 1e( oro 5.8 S rRNA oXqtr-ror[(er ro 5' dxpo rou preydou (23S) RNA, dpo 6e ouprBo(ver ene(epyoo(o ;rero[ri ou- rdrv rtrlv oLLq).ouXrcirv. H EIKONA 21.41 de(per 6n ro np66popo prdpro nepdXer en(or1g ro 55 rRNA r<r 6vo 4 60o IRNA. Zto E. coli, ro 7 onep6vrq rrn e(vor drdonog ro oe 6l.o ro yovr6[tr;1-ro. T6ooeprg yevetxo( r6not rrn nepr6louv 6vo yov(6ro IRNA peto(ri rrr:v oMrll,ouXrdrv 165 ror 23S rRNA r<or or dMor yevenxo( r6nor rrn ne- E OC lTSl r'eu lTS2 28S 3, ETS Evdovouxtredorl i t Evdovourtredorl I EvdovouxLedoetg t * EErovourtredon E[rovourtredoetq ++ Ev5ovoux.edor1 |re 185 E[tovoudte6oerg ++ 585 : ' To topt;to europuortro rRNA nopoyovrot ono ro nporoyev6g peroyporpo peoto oupBovrov rondg ror rl.ro),rdrogorog. 618 KE0AAAI0 21 Iuppotprl rr Ene[epyooio rou RNA Boorxeg eworeg taa
  • 110.
    yov[dro yro rRNAroL IRNA. To orpLBeq plrog rov pe- royporprrrv e[oproror ono roug unorrvlr6g (P) xot rep;loloreq (t) nou XprtorUonorouvTor. Ko0e npor6v RNA np6ner v' onetreuOepoOer ono ro geruyporpo 1r' evrop6g oe roOe n),eupo. pr6louv 6uo yov[6ro IRNA oe ouqv qv neptoXrl. Mept- r<69 qop6g, peto[ri trlg o].].qLouX(oq 55 rot rou 3' drpou ;rnope( vo undplouv enrnp6o0eto yov(6ro IRNA. 'Eror, ot owr6pdoetg ene(epyoo(oq nou onorroriwor yro rrlv oneLeu05poorl rov npoi6wrov e(oprrirwor on6 ro nepeX6prevo rou ouyrerpql6vou yevenroU r6nou r- rn. T5oo oroug nporopurrireg 6oo ror oroug europuri- reg, rord qv ene[epyoo(o tou rRNA nopdyoweq ene- [epyoo[og ror prBoorol.ur<69 nporeiVeg (iocog xor dMeg nproretveg) npoo66vowor oro np6dpo;ro ;r6pro, on6re un6orpopo yro rnv ene[epyoofo 6ev e(vor ro e],e0Oepo RNA oLLd 5vo prBovouxleonproreivrr6 o(;rnl.oro. 'Onog q ene[epyoo(o rou pre-mRNA, q ene[epyoo(o rou rRNA npoyporonoekor oe ouylpovtog6 Ue rn Ue- roypo<pr1. 'Eror, or nopdyoweg ene[epyoo(oq 6ron].6- Kowor pe ptBoorol.ux6g nporeiVeg oX41"rot((owoq ro prBooroprdrro, ow[ npdlro vo ene[epyd(owor r<r 6nemo vo ouvoplloLoyoriwor oro6roxd ndvo o' ene(epyoopS- vq rRNA. H ene(epyoo[o ror rpononofio4 rou rRNA npouno- 06rer pro ror4yop(o ptr<priv RNA nou rqloUwor snoR- NA (small nucleolar RNA, pmpd RNA rou nupqv[orou). Iro yovr6(opro rou S. cerevisiae Kor rov onovduLorcbv undplouv exorowd6eg snoRNA. Opropr6vo rrodrxonor- o0wor on6 [eXoprord yov(6ro, dMo erEpd(owor on6 nol,uormp6vto r<or nolLd npodpXowor on6 twp6vro trov yovrS(tov-[evrmciv. Aurd to snoRNA ug[mowor no).6nLorq ene[epyoo(o ror orp(povo4. Opro;r5vo s- noRNA onomo6wot yro rnv xon{ rou np66poprou prop(- ou oe rRNA, 'Evo nopd6erygo e(vor ro U3 snoRNA, ro ono(o e(vor onopofurpo oro npri:ro ouprBdv ronrlg. To oUpntror<o nou nepdlet U3 owrororle( oroug urellKoUq r6pnoug, mo 5' dt<po ttov veooXqprott(6prevov rRNA peroyp6tprov, ro ono[o efuor opord oro rltrempovu<5 pu- rpoor6nro. Ae yvop((oupre n p6l,o no(er ro snoRNA orqv n6q4. Oo prnoporloe vo e(vor onopokrpo yro ro (euydpopo pe erdxSg oLL4Lou{eq rRNA tirore vo q1n- gorrme( gro 6eweporoy{q 6oprf nou ovoyvop((eror on6 puo ev6ovour).edo4. Auo ogddeg snoRNA onono6wot yro rrq rpononor- rloetg nou npoygoronoroUwor oe Bdoerg rou rRNA. To p6Lq rdOe o;rd6og ovoyvrop[(owqr on6 no].ti pur<p6q ouw4pqpr6veg oMqLoul(eq Kor rotvd loporcrrlpronrd 6eureporoyo0s 6o[nq. H opd6o snoRNA C/D onotrefuqr yro rrlv npooOrp<r1 tRNA tRNA Pl P2 165 rRNA 23S rRNA 23S rRNA ilt2 165 rRNA 23S rRNA I3OS RNA llpotrivro **Y' 5S rRNA Y tRNA proq preOuLoprd6og orrl 2' 06o111119 prB6(49. Ie ouwq- pqp6veg O5oerg rov rRNA r<ov onov6ul<otrilv un6plouv >100 2'-O-pre0u),opddeg. Aur{ q oprddo snoRNA no(p- ver t' 6voprd t4g on6 6rio prxpd ouwqpr;pr6vo pror(Bo otrL4LouXrc0v nou rotro[wor rou'nd C xor D. Kd0e s- noRNA nepdler pto oLi.r'1Lou/o rowd oro rour( D nou e[vot oupn].r1po1.rotrq Ue pro neprolq tou 18S 11 rou 28S rRNA nou e(vqt pre0ui,uopr6v4. H onri:trero ev6g ouy- rerpql6vou snoRNA nopeprnod(er q preOuL(roon ornv neproXri tou rRNA Ue rrlv ono(q e(vor ouprntr4pco1.rorr6. H EIK0NA 21.42 6e(yyer 6n ro snoRNA (euyoprirver pre ro rRNA yro vo 6qgroupyrloer q 6(xLovrl neproXrl nou ovoyvrop((eror og un5orpopo yro rq peOuL(rrloq. H Ue- 0ul(ooq ouprBofuu prSoo orqv neptoXrl ouprnLqprrlpron- r<6111o9 oe Uro 06oq nou on61er oxprBcirg ndwe Bdoerq 5'rou rowto0 D. l-lt0ov6v, xdOe oupBdv pre0ul,(too4g ro0op(eror ond 5vo 6torpopenx6 snoRNA. Ie owrlrrlv rponono(qo4 epnl,Srowor toutrdltmov 40 snoRNA. Kd0e C+D rout( snoRNA ouv66eru Ue rperq nproret- veg Noplp (oro onovduLrord, gq.rnprMopwn - fibrillar- in), Nop56p r<or Nop58p. Or peOuldoeg dev SXouv XopoKrrlproref nArlpog, nopdrr q rUpro snoRNP npo- re(v4 Noplp/grpnprMop(vq e(vor doprxd nop6proro pre ng pe0utrorpovogepdoeg. Mro dLLq opd6o snoRNA epntr5reror srrlv rpono- RNA-m6Xog 3, RNA-ortlXog EIKONi{ 2'!.42 'Evo snoRNA (euyoprirver Ue Uro neptoxn rRNA nou npdretrot vo peOutrrtll0e[, BoxD' ctD snoRNA box D 21.20 H llopoyroyrl rRNA tlpoUno06rer luppdvro Komjq ror llepiloppdver Mrxpd RNA 619
  • 111.
    00H P/ EIK0NA 21,43 Houprdtr4 gerorp6neror oe tpeudoouprd[vq owr- ro0rortrlvrog rov N1-oorXopurr6 deopo p' 6vov C5-oorXopLnr6 0eopd roL neprorpdgowoq rn Boq oe oX6oq pe ro ooxXopo. no(r1o4 Bdoeov prerorp6nowoq rnv oupr6(vq oe r,Ueu6o- oupr6(vq. Yn6plouv -50 xotdlorno oro rRNA rnq (6UnS r<ot -100 oro rRNA rov onovduLrotcbv nou rpononor- oUwor ;r5oo rpeu6oouprdw(aroqg (pseudouridination). It4 ouyrerprprdvq ow[6poo4 {EIKONA Z1.4tl, o N1 6e- opr6g prero[ri tou oupr6u],rro0 o(5og Kor rnq prB6EnS ondet, 11 Bdo4 neprorpdrpetor r<or o C5 enovoouvd6erot oro odrlopo. O oXrlponopr6g qUeu6ooupr6(vr1g oro rRNA onome( qv oprdSo rrov H/ACA snoRNA. l'lo(pvouv t, 6voprd toug on6 rrlv nqpouo(o truog tprnAsrog ACA oe on6- oroorl rprcbv vourLeor6(rrlv qn6 to 3' drpo Kol. proq Ue- prrdrg ouvt4prlpr6v4g olLr1trou1(og (r<our( H) nou Bp(oreror prero[ri 6uo 6o;rcbv goupr<6rog pr(o1ou-0q- l,rdg. Ko06vo on6 owd to snoRNA 61er pro oM4Lou(o oupn).4pto1.rolrq pre ro rRNA prd,oo mo trrioXo rdOe goupr6rog. H EIKONA 21.44 6e(yyer q 6opri nou 0o no- poy6rov trrdoro rou (euyopcbproroq pe ro rRNA. Mdoo oe rdOe neproXrl (euyopd:prorog, undpXouv duo o(eu- ydpcoreg Bdoetg, on6 lg ono(eg q pro e(vor oupr6(vr1 nou perorprlneror oe rfeudoouprd(vq. snoRNA 5' dr l-1 : .. . To H/ACASnoRNA eXouv duo Bpoxereq ouvTnpl- g6veq otrtr1trouy[eg ror duo 6opeq qouprerog, ro0epro on6 rrg onoleg eXer reploxeq oro pfoXo nou e[vor oupntrqptopolxeg pe ro rRNA. H Qeu0ooupLdivq oXrlpol(erot UeToTpenovToq pLo o(eu- yopotrrl ouprDLvrl luioo orrl oupnLqpotporLrl neptoXrl rou rRNA. 620 KEQAAAI0 21 Iupporprl xt Ene(epyooro rou RNA To H/ACA snoRNA ouv66owor pe r6ooeprg er6u<dg npcoreiveg rou nup4v(orou: Cbf5p (oro onov6uLrr:rd, duorepfuq - dyskerin), Nhp2p, Nop10p r<or Garlp. E(vor orlpowu<6 6l 11 Cbf5p/6uorep(vr1 e[vor 6opurd nop6- Uoro Ue yvroordg ouv0doeg rfeu6ooupr6(vr1g ru 6ror nr- 0ov6v nopSler r4v ev(u;rrrr1 evepy6rrlro orrlv roOodr;yoriprevq on5 snoRNA ow6pooq qUeu6ooupr6r- v(too'r1q. I-loMd snoRNA Xpnorponororiwor en[or1g yro vo xoOo6rlyrloouv rponoTrot{oerg Bdoeov oro IFINA Kor sro snRNA nou epnl6rowor orrl oupporp4 rou pre-m- RNA. Aurdg or rpononot4oerg e[vor r<p(orpreg yro ng Aer- roupy(eg rouq olq owfmorleg owr6pdoerg. 0o16oo, noMd snoRNA 6ev 51ouv e;rgove(g or61oug. Awd ro snoRNA r<olorivror opEava RNA (orphan RNA). H Unop(r1 roug uno64Lcbver 6rL notrtr5g BroLoyrx6g 6rep- yooleg (oog 1p4or;ronoroUv prrllovroproUg r<o0o6qyo6- Uevouq on6 ro RNA yro vo rpononorr;oouv Leroupyrxd dtri.o er<<ppo(6prevo RNA pe nto nou<[],oug tp6noug on' 6,tr yvop(ou;re oripepo. 21.21 IlepiAn{n H ouppogq enmuyXdver qv ogo(peoq rtov rwpov(rrrv xor rq oUvdeon rov e[covirrtv orrlv c0pq-rq oM4].ou1io rou RNA. Yndplouv roui.dlrorov r6ooeprg r6nor ow[- 6pooqg, nou 6ror<p(vowor on6 rg onomqoerg roug rn vitro xot ro ev6rdpeoo nou nopdyouv. To ouorrlgcrro nepil.ogBdvouv ro euKopuortKd nuprlvrxd rwp6vto, ro rwp6vro o;rd6og I r<or II Kor ro rwp6vro rou IRNA. Kd0e ow(6poor1 ouv(ororot oe oMoy4 opydvoorlg, pr6oo o' 5vo (eXo:pror6 trl6pro RNA, dpo e(vor dvo cis- 6pooru<6 ougBdv. H ouppog4 rou pre-mRNA qKo).ou0e( npol;rcirpre- veg oMd 6Xr unoXpeonr5g o6orjg. Anopo(rrlteg e(vor p6vo noLU pu<p6g ouvorvettx6g oLl,rlLouX(eg, evc6 or un6Loneg oMr;l,ouX(eg rcov rwpov[cov e(vor odrdqopeg. Oqr6oo, oLl4Aoul(eg 16oo rrov e[otv(rov 6oo ror r<ov rwpov(ov prnope( vo enqpedoouv 0etxd 4 opvrlnr<d rqv enil.oyrl rnq yerrovrKrlg 05orlg oupporp4g. O).eg or 5, O5oerg ouppogrlq pdMov e(vor roodtjvopeg, 6ncog ror 6Leg or 3' O5oerg ouppocpqg. Or onopo[rrlreg oM4],ou- X(eg 6(vowor qn5 rov xov6vo GU-AG, o ono(og nepr- ypdger ro drpo rou twpov(ou. Anopofu4rrl e(vor enioqq n e6on droxlddrrrorlg (UACUAAC orn (UUn, ri pro Lty6- repo xoAd ouwrlpqprdvq ouvorverrrrl oMrll.ouX(o oro rwp6vro rrov 0qLooru<rrlv). H owidpoorl pe rrlv 5, 06o11 ouppogrlg nepil,opBdver ro oxnpqrropr6 ev6g Adoou nou evdlver ro GU dxpo tou rwpov(ou ;r6ocrl 2'-5' 6e- opo0 trre rrlv A orrl 06o11 6 qg 05or1g 6rorLddolorlg. 'Eneno, ro 3'-OH drpo rou e{ov(ou enn(Oerot orrlv 3' 06oq ouppo<pr1g, 6ror cilore vo ouv6e0ofv ro e(c6vro ror ro rwp6vro v' oneleu0epo0e( un6 propE4 Ldoou. O oXrlgotropr6g Ldoou e(vor unerj0uvog yro rqv enr).oy{ t4g 3' 06oqg oupporprlg. Kor ot 6uo owr6pdoerg e(vor trans-emeponor{oerg ortg ono[eg ouw4porjwot gro- oqodreoreprxo( beotrrof. Apr<erd or6bro qg ow(dpoorlg ononoriv 11 u6p6luoq rou ATP, nr0ov6v yro vo npo. 0 P s /!-'-' I e(rr ")-o.i i( )r" w00H P/ 0 ']* /'*, nspmpogn 180 oC -)> Iuv0dorl qeu0ooupt6tuqg P I o -'- I I RNA-orrlXog
  • 112.
    oenoouv olloydg dtopr6prpotoqgoro RNA rot/n oe nproreivtrd ouorortrd. Evotrtrorrtrd np6tuno ouppo- griq nporotrotiwot on6 npcoteivtrorig nopdyoweq nou efue dteye(pouv q Xprlorl puog v6og 05or1q efue TIopeU- no6[(ouv rn Xpnon qg npoenil.eyp6v4g 06or1q. H ouppogq tou pre-mRNA npouno0Stet ro oxrlUo- roprS evdq oorptou6(ou ouppoqqg - 6vo peydlo oopo- t(6ro nou ouvopptoloye( ng ouvoweru<69 otrtr1ouX(eg oe dropr6p<pcooq r<or6Mql,q Ytq vo ou;rBe( ow[6pooq. To oropoilEto ouppoqrlq 6rlprtoupye(tot on6 tr; 6tepyo- o[o opropori twpov(cov, 4 ono(o nept],oprBdvet ovoyvtil- pron rnq 5' 06o119 ouppogriq, qg 05or1q 6toxLd6toorlq Kor rrlq 3' 06o119 ouppoqrlg. Aut6 toltiet yto 1-ttrp6 w- rp6vro, n.X.rnq (riUnq. Av, toor6oo, to twp6vto efuot preydl,o, 6nrog to twp6vto tov onovduLrotdlv, n ovo- yvdpron tov O5oerov oupporprig orotrouOe( q 6epyo- o(o rou optoproU efov[ov, nou nepil.ol.tBdvet oMq]'e- nrdpdoerg 6ro;r5oou tou e(o:v(ou ovdpeoo orqv 3' 05oq ouppogrlg Kot rrlv roOodx4 5' 06o11 ouppoqrig. Aw6 6nemo peron(met oe ou(euypdveg owtdpdoetg 6topr5- oou rou rwpov(ou yto ro petoyevdmepo mddto rqg ouvopprotr6ynorlq rou otr:1.ton6(ou ouppo<pt'1q. E(te pe rov oprop6 rwpov(<ov e(te pe tov optop6 e[ov[rov, q op- Xrr4 depyoo[o ovoyvd:ptoqq rqg 06oqg oupporprlg 6e- opre[er to pre-mRNA un6orpo;to orqv o66 ouppotp4g. To orj;rntroxo pre-mRNA nept6Xet U1 snRNP Kot opt- qr5voug rp(orproug npcoteivu<otig nopdyoweg ouppo- <pr]q, pero[ti ttov ono(ov o U2AF-rot o nopdyowog nou npooddveror orrl 06oq 6toxl.d6cooqg. Ito nolut<frropo euxopurr:nrd x0rropo, o oxrlportou5g tou outrtntr6rou 65opeusnq onotte( r1 ouprtrreroXri rov npoteivdlv SR. To oo;rot[dto oupporpqg nept5Xet to U1, U2, U4lU6 r<ot U5 snRNP, 6nolg enioqg rot ;reydtro optOpr5 nopo- y6wrrlv ouppoqrjg. To U1, U2 t<ot U5 snRNP nepdXouv 6vo provodtr6 snRNA rot opretdq nptotefveg. To U4lU6 snRNP nept6Xet 6[o snRNA t<ot opretdg nprrlteiveg. Oprou5veq npotefveg e(vot t<owdg yto 5tro to orrrpo{- 6ro snRNP.To U1 snRNA (euyopdrvet pe rnv 5' 06o11 ouppogrlg, to U2 snRNA Us rnv oMrllouX(o 6ror<Ld6o- onq Kor q U5 snRNP ouyrpote(rq 5' Kot 3' 06oetg oup- poqns ;16oo prtog oLLrlLoul[og Oq]'tdq pSoo oro orrlpor[dro ouppoqrjg. M6]lg to U4 qne],eu0ep6oet to U6, ro U6 snRNA (euyoptilvet pe rnv 5' 06o4 ouppocprlg xor to U2, nou nopogSvet (euyopcop6vo, ge rqv oMrl- l,ouX(o 6toxtrd6trroqg. Aut6 nt0ov6v drlprtoupye( ro Ko- rol,utrr6 rdwpo yto rn oupporprl. Mto evollorcrtr<r1 oprd6o snRNP nop6Xet ovdLoyeq Lettoupy[eg Yto rrl ouppoEq t4g U 1 2-e(opttirprevrlg unor<oqyop(oq twpo- v(tov. O xotol.uttr6g nuprlvog Oup((et tov owlorotlo nuprlvo rov ouroKoroluttrriv twpov(ov opddog ll. Iu- venri:q, to otol.tot(dto ouppo<p4g prnope[ vo e[vot Uto Yt- ydwro Unxovn RNA (6ncog to ptBoocoprdtto) orrlv ono(o to Booxd ororle(o RNA Bp(orowot oro rSwpo rnq ov- il6pooqg. H ouppotp4 ouvrlOog e(vot evSoptoptorq. 0o16oo, oro rpunovootilporo Kot ro vrlporr664 orouLqrto oup- Bo(ver trans-ouppoqn (dtoproprorrl ouppotp4). l-lept}'o;t- Bdver ;uo ow(6pooq prero[6 ev6g prtr<poti SL RNA rot rou pre-mRNA. Ito oxoul.rlrto undplouv 6uo tfnot SL RNA: o 6voq Xprlot;ronoteftqt Yto'rn oupporpr; mo 5' drpo ev6q mRNA xt o d).l.og Ytq'rrl ouppogn oe Pto eoorepwi 05or1 cirore vo dtoonoore[ ro notruotorpovt- 16 np66popo RNA. H erooyoyri"rou SL RNA o' ene(ep- yoopSvo mRNA nop6Xet onopo[tqro o41-toto Yto rr'l prerd<ppoo'q. H xov6tlto tepprottoprori r1g RNA nolupepdoqg ll e(vor mevd ouvde6ep6vn Ue ro olrlpottopr6 tou 3' 6rpou tou mRNA. H oM4Loul(o AAUAAA, nou ewon[- (erot 11-30 Bdoetq ovo6md rnq eSonq ronrlg, nopdXet ro orlpro yto xon4 on6 pto ev6ovout<Ledon Kil nol'uo- 6evu(trloq on6 r4v notru-A nol,u;rep6oq. Aw6 evtoXrie- tor ptSoto tou oupnL6Kou nou e(vot deotr;eupr5vo mo nLoUoto oe G/U orotXelo to ono(o Bp(oxerot rq0o6Kd rrlg 06oqg ronrlg. H ;retoypogrl tepproil(etot 6tov 1.tto e[covoux].edoq nou npoo66vetqt oro 5' dr<po tou RNA to ono(o opXi(et vo dr1;rtoupyefuot on6 trlv r<onri npo- <ptdoer qv RNA nol,uprepdorl. Otro to ;ret6ypotpo Pol ll e(vot notruo6evul,tcrlpdvo, er<r6g on6 ro mRNA tcov torovdlv, nou orjte nepr6louv rwp6vro o6re onomoUv oup6 nol.u-A. O oXrlpottopr6q rou 3' 6r<pou rou mRNA ttov torovr.riv e[optdtot on6 ;to 6opq pr(oXou-Or1Ltdg t<ot on6 to (euydpotrro ev6g xo0o6rxori o.rotXe(ou pre to U7 snRNA drme vo npoerot- proore( puo ron{. H 6opri pr(oXou-0q}.tdg nt0ov6v npo- sroteUer to 6rpo, 6nog oro Botctipto. H ouppo<pri rou IRNA neprloprBdvet [elroptmSg ov- ndpdoetg evdovouxl,edonq Kot llyrdoqg. H ev6ovou- rledorl ovoyvtrrpi(et 11 6eweporoYn (n tpttotoyrl) 6oprl rou np66poprou pop(ou rot ndntet rot ro 60o drpo rou rwpovfou. To 6uo prtod IRNA nou oneleuOe- pri:vowor pr6oo qg on6tretog tou twpov(ou ouv65ov- tor on5 qv IRNA Ltyrdoq nopouo[o ATP. Autl 11 o669 u:pf;rovorlg tou IRNA oftonoe(tot on6 qv o66 ondw4- onq oe (edrn).ctprSveg npote(veg oro EA. H ene[epyooio tou rRNA npoypotonotefuot orov nupqv[oro, 6nou to U3 snRNA nupodore( pto oetpd dpdoetov ev6ovoux.treoori:v rqt e(ovoux.Leood:v nou 16- Bouv r<ot qrqfud[(ouv entnp6oOereq o].trr1]'ou1(eg mo np66popro rRNA, nopdyowoq felcoptm6 p$ootogort- r<d RNA. Er<orow66eg 6o9 ltltddeg ptrl r<codxonoqtxd RNA exgpd(owot oro europuconrd xrirropo. Itov nu- pqv(oro, 6uo oprddeg t6totcov pr1 ro6tronotrlttxtilv R- NA, nou ovoprd(owot snoRNA, e(vot une0Ouveq Yto ro (euydpopo ;re rRNA oe O5oetg TIou rpononoto0wot. To snoRNA t4g oprddoq C/D ovoYvtop((ouv 05oerg-m6- Xoug ytq preOuL(ooq, evc0 ro snoRNA qg optddog H/A- CA r<o0op[(ouv ttg 05oetg 6nou 4 oupt6(vr1 perorpSnerot oe tPeu6oou Ptdtvn. BrBAroyporpio :i i rir-:t,ti ttrrl I Lewin, B. (1975). Units of transcription and translation: sequence components of hnRNA and mRNA. Cell 4,77-93. BrB),.:.::: : 621
  • 113.
    21.2 To 5'Arpo rou Euxopuorlrori mRNA 06pet Kohtjnrpo quires intact U4lU6 small nuclear ribonucleoprotein. Cell 46, Avooxdnqorl Bannerjee, A. K. (1980). 5 terminal cap structure in eukaryotic mR- NAs. Microbiol. Rev. 44,175-205. 'Epeuvo Mandal, S. S. et al. (2004). Functional interactions of RNA-capping enzyme with factors that positively and negatively regulated pro- moter escape by RNA polymerase ll. Proc. Natl. Acad. Sci. USA 101 ,7572-7577 . Mcoracken, S. et al. (1997). S-capping enzymes are targeted to pre-mRNA by binding to the phosphorylated carboxy-terminal domain of RNA polymerase ll. Genes Dev. 11,3306-3318. 21.3 0t l-luprlvudq luUBotr6q luppoq1q e[vot BpoXe[eq Aitr1)ouXieg Avaox6nrlorl Padgett, R. A. (1986). Splicing of messenger RNA precursors. /n- nu. Rev. Biochem. 55, 1 1'19-1150. Sharp, P. A. (1987), Splicing of mRNA precursors. Science 235, 766-771. Sharp, P, A. and Burge, C. B. (1997). Classification of introns: U2- type or U1 2-type. Cell 91,875-879, 'Epeuvo Graveley, B. R, (2005). Mutually exclusive splicing of the insect D- scam pre-mRNA directed by competing intronic RNA secondary structures. Cell 1 23, 65-7 3. Krainer, A. R. et al. (1984). Normal and mutant human B-globin pre-mRNAs are accurately and efficiently spliced rn vitro. Cell 36, 993-'1005. 21.5 H luppotpn Tou pre-mRNA llpoxoper Arop6oou Evrig Aooou Avooxdnqcny Sharp, P, A. (1994). Split genes and RNA splicing. Ce77,805- 81 5, 'Epeuvo Reed, R, and Maniatis, T. (1985).lntron sequences involved in lari- at formation during pre-mRNA splicing. Cel/ 41 , 95-105. Ruskin, B. et al. (1984). Excision of an intact intron as a novel lari- at stru ctu re during premRNA splicing in vitro . Cell 38, 31 7-331 . 21.6 To snRNA Elvot Anopo[qTo yro Tn luppoqrl .r1r,.r' 1 ; ii i.i ii i ],l il Guthrie, C. (1991). Messenger RNA splicing in yeast: clues to why the spliceosome is a ribonucleoprotein. Science 253, 157-163. Guthrie, C. and Patterson, B. (1988). 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  • 117.
    Ero0ep6rnTo Kor Xropo0drnon ToumRNA ffiHS ErooVoyn W To Ayyetrrorpdpo RNA E(vorAoro0rl Mdpro * H ooroOeto rou mRNA oqeiLeror oq 6poo1 prpovou- rtreoo6v. a 0r prBovour),eooeq droQ6pouv oq rpoq rrlv npor[pr1orl yro ro un6orptop0 Kor Tov rp6no eni0eorlg. * To mRNA nopouoro(ouv eupe(o drorupovorl ro:v Xpo- vtrrv 11p(0r1q. * H dLorpopun oro0epdrr[o rtov lroplrov mRNA enr;peo- (eL rr1 ouyx6vrptoo4 roug, ouven6q rot ro qoogo rov nporeivdrv nou nopdyovrot oro rUnopo. ffiffi To Europuonro mRNA YndpXouv un6 Moptprl mRNP ono q l-evvr1or1 6o9 ror ro 0ovor6 roug s To mRNA otrtrqtrentdpo g' 6vo peropotrtr6pevo ntrq0u- opd nporreivdv rord rrlv orpilrovor'1 Tou 0T0v nuprlvo Kor Tnv nopoUovr1 rou oro rurropontroo;ro. * Meptr6q nporetveq rov nupqvxtitv mRNP 6Xouv ptitro rot oro rurropontroopo. <e Ynopxouv nopo notrtr6q npulreiveg nou npoodSvouv RNA. 0r neptooorepeg dev SXouv Xoporrqprorel ordgo. * Atotpopelro mRNA oXeri(ovror pe [eXoprordq, otrtro otrtrr1troenxotrunr6peveg, oU0deq pu0Urolx6v nporei- vtilv, DqproupyrirvToq T0 RNA peyroutrdvro. 626 KEOAAAI0 22 lro0ep6rnro rot Xropo06rnon rou mRNA o Equinox Graphics/Photo Researchers H Anoddprlorl rclv flporopurolrdrv mRNA l-lepilop$oveL l-lotrtro'Ev(upo e H onodounq rov porqprordv mRNA opxr(er ue oQoL- peol ev6g rupoQooqoprKou ond ro 5'drpo. * To Uovotprrrorpoputrulgevo mRNA ono6o;to[vrot xoro rr UeroQpool o' 6vo rurtro Duo prlporu:v nou nepiloppo- ver evdovourtreotrurrr6g ewopdg, ttg onofeg orotrou- 0e( n5(l4 rolv rpqporov nou npoxunrouv oe roreuOuv- ot 3'-+5'. e H 3'notruodevutritool gnopel vo dreurotruver Trlv 0n0- 6opr1o1 rtrooporov mRNA nou nepr6Xouv deureporoy'; doun. * To rOplo ev(uuo Uq onoddunolg Leuoupyo0v oq ouu- ntroro nou ovo go(eror orr.rpor(6to ono6o gqorl g. To [lepLooorepo Europuorrro mRNA Anodopouvror Mrloto Auo Movonou6v E{oprtirpevtov ono flo},uo6evu- trioo1 * 0r rpononorrloerq Kor oro 6uo orpo rou mRNA ro npo- ororeuouv ono q 6poor1 rov e[ovour],eootirv. * To 0uo rupro lrovonorLo onoouvOeorlg rou mRNA opXr- (ouv pe onoodevutrltrlo1 11 onoio rorotrueror ono no- tru(A) vourtreooeq. * Mero rqv onoodevutr[oor1 pnopei v' orotrou0rloet e[re oqorpeol qq rotrunrpog ror 5'-+3' n6(rr1 ono ev6o-
  • 118.
    vouKtrooon, siTe rdqJnord eqovouKtreoorl oe K0TeU- 0uvo1 3'-+5'. o To 6v(uuo orpo(peonq rqg rotrunrpoq ovroyov[(etot pe ro oupntroxo 6vop[qg rnq UerdQpoonq yro npdodeon oqv 5'rotrunrpo. * To e[ulotoudlo, ro ono(o rorqluer qv 3'-+5'n6Un rou mRNA, e(voL 6vo peydtro, e[etrrrlrd ouwqplp6vo o[p- ntroro. e H ono66utlotl ouppofvet o' euOtorpuo rutopontrooport- ro oopor[6ro nou ovopo(ovror orrrpor6ro ene[epyoo(og. @ nffo Movonolo Ano6riprlorlg lroXe0ouv Iuyrerprpdvo Mopro mRNA e T6ooepo enrnpdo0ero povondlo ono66pr1or1g enue- trouv q pu0;u(opevq onodoglorl erdLrtlv mRNA. o Atpo[peorl qq rotrUnrpoq ove[dprnrrl on6 qv onoode- vutrltoor1 e[etr[ooeror rlrov unopXer pro lroxpro notru(A) oupd. * H qno66unol rov un notruodevutrur:1.uivtr:v mRNA ro:v rorovdrv nupodorelror ge qv npoo0rlKrl Uroq notru(U) oupoq oro 3'orpo. c H ono66uno1 oprop6vrov mRNA pnoper v' opX[oet ge n6fi ond evdovourtreool erdrq yro qv otrtrr1trouXio 11 q doPr1. * 'EvoS oyvtrlorog opt0gdg uop(tr:v mRNA oroXeuovrot yt' onoddgnon n Heroqpoolrn roroorotrrl ond microRNA. ffi O Erdrrdg Xpovoq HU(urnq Ko0e mRNA Etr6yXeroL ono AtrtrqtrouX(eg q Aopeg M6oo oro mRNA * Elduo crs-ororXe[o evrig mBNA en4ped(ouv rnv roxri- Tnro Trlq onoddp4o4g rou. * frorxe[o onooro0eponoi4o4g gnope( vo enuoXtlvouv rr;v onoouvOeorl rou mRNA, ev6 ororXe(o oro0eponof- lonq vo qv neptop[oouv. * To n),orjoto oe AU ororXe(o e(vor rorvd motXelo ono- oro0epono(4or1g oro 0qloolro oro ono(o npoodevov_ ror notx[],eg npo:reiveg. * Optopeveq npoTe(veq nou npoodevovTor or ororXe(o onooro0eponoirloqg otrtrr.1trentdporjv ge ouoronrd qg Uqxovng onoouv0eoqg roL potrtrov ro enrorporeUouv yr'ono6opr;0r1. e lrorxe[o olo0eponoqolg undpXouv oe oprop6vo notr( oroOepo mRNA. * H roxut1ro ono66p1or1g rou mRNA pnopet v, otrtro[er oe onovrqol npog norr[tr0 oriporo. ffi fo Neoouvre0erpevo Mrlpro RNA EtreyXovroryro Etroruo- poro MSoto Evog llup4vuorj Iuorlgorog Enurlpqorlg * To etrorrotpolrti nupqvrrd RNA ovoyvropr(ovTor Kor K0_ roorpdqovrot ond 6vo ouorrlpo enunplonq e To nuprlvtr6 elooouorto tretroupyeI rooo <o:: .*, ene[epyoo(o rpuorotroytrdv RNA unoorpoigtoTurv,:: ror xoro TIV KoTooTpoqq rov etrorrtrlltoiu6v RNA. s Iu (uul, ro oUpnl,oxo TRAMP entorporeuet ro e[o- orol-rono oe ovtopotro popto RNA xot 6teuro),uver qv 3'-+5' e[trlvourtreotrunrf 6poot1 tou. a Iro unootpriruoro yr' onoDdurlon 96oo rou oupnLrkou TRAMP-e[ooropotlou nepilopBovovrot ptrl ouppoppevo 1 ovrirpotro ouppoppevo npo-mRNA xot tro0og reppo- ropevo perdypoqo qq RNA notrupepoonq ll Xtopiq no- Iu(A) oupo. e To neprooorepo geroyporpo rrlg RNA notruuepoonq ll [oog e[vor rpuqo ooroOq peroyporpo nou roroorp5- govror yprlyopo orov nuprlvo. ffi O'EtreyXoq llororrpog qg Meroqpooqq rou mRNA l-lpoy- goronore(ror ono Kurropontrooportrd luor4poro Enurl- pnortq * 0 unxuvtoudg onoouv0eoqg NMD oroxeuet mRNA pe npoorpo rtodrrovro reppouogou. * H oroxeuorl Tov unooTpouororv rou NMD onouei 6vo 6torqpqp6vo ouvotro nporeivtirv UPF ror SMG. " Ie notrtrofq opyovrolroug, urtrerd0, yro rr;v ovoyvdrprorl npooptrtv rodrrov[trrv reppologou e(vor 11 oouvrl0rorrl doUl n ro oouvrlOroro pqrog qq 3' UTR. Iro 0r1trool- ro, o 1r4Xovto1r6g ovoyvtirprorlg entoqg o[totroye( ror qv nopouo[o roOodutirv oupntrdrtov oupBotrqg e[o- v(rov. * 0 Unxovroltdg onoouv0eo4g NSD oroXerier mRNA Xu.r- p(g evrdg nloto(ou ro:drrdvro repgouogorj ror poot(e- T0r 0' rvo ouvr4p4grivo orjvotro nporeiV6v SKl. " 0 priXovropog onooUv0eonq NGD oroyeuet mRNA pe ordorpo prpooropriuo or4v xrodrxonor4rtrrl nepto_ xn. ffi UeprxO Europuolro mRNA Evronr(ovTot 0r Erdudg l.1e- proXdg rou Kurropou + 0 evrontoprjq rov mRNA e(unrlperer drorpopenr6g let_ roupy(eg oe ;lepovtrt;rdvo rUrropo ror ovonruooripevo eUgpuo. * 'Exouv roroyporpe( Tperq Unxovropo( yro rov evronropo rtov mRNA. * 0 evronropog onorre[ crs-orotXero oro mRNA_oroXo roL lrans-nopoyovreg. * 0 ruproq p4Xovropog evepyou perorpopoq nepilogpo_ ver qv roreuOuvripev4 x[v404 rtov mRNp rord p(rog odtiv rou rurropooxetrerou. @ YrffiS; IleprtrnUn 22.1 Erooyayh ]9 nrun 61er rewpu<6 p6),o mr.1v 6r<gpoorl rorv yovr_ 6icov. ApXu<d XoporcrqpiorrlKe oq 6vo ev6rdpeoo npo_ i6v rrlg npotelvooriv0eo4g. Irjpepo,'rrlor6oo, yvcop[(ou;re no].].d enrnp6o0ero RNA nou no((ouv dogrxorjg q Letroupytroriq pdLouq oe dLtro ord6ro qg Srgpooqg rcov yovr6(tov. H ou1-rprero14 rou RNA oe noM6g i.etroupy(eg nou ogoporiv offlv 6r<gpoo4 rcr.rv yovrdfurv unomnpf(et rq yevtxn dno{q dtr 4 6L4 dtepyoo(o nr0ov6v e[e].rX04re o, 6vqv "r6opo RNA,, orov onoio to RNA oplu<d rjrov ro evepy6 ouororu<d yro rn drorfpqo4 Kqr rrlv 6r<gpooq rov yeverrxc0v nAqpogopuirv. Meroyevdfiepo, or nporetveg unoBofl_ Oqoov ri ovdAoBov onox,,ermtxd noMdg on6 our6g rrg Lenoupy(eg, pre ouvdnero qv orl[4or1 r1g eue]l_ [(oq r<or rqg onote].eopotrx6r4rog. To'regd].oto our6 !io:',:,* 627
  • 119.
    eond(er oro oyyefuoq6poRNA (messenger RNA, m- RNA). Or Lefioupy(eg r<ov d].l,rr:v xurroprrri:v RNA onoretroUv owxe(trlevo dMcov reqotro(rov: snRNA xor snoRNA oro KegdLoro 21, IRNA rqr rRNA oro Kerpdloto 24, miRNA xot siRNA oro Keqdl.oro 30. To unooUvotro lov RNA nou 51ouv droqprloet qv opX6- yovn evepy6rlro r<ordLuorlg e(vor owtre(pevo rou Keqol,o(ou 23. To oyyel,tog6po RNA no[(er rov r<Upro p6].o mrlv Srrppooq rov yovrd(rov nou ro6rronoro(v nptoreiveg. Kd0e ;r6pro mRNA g6pet ro yeverrr6 r66xq yro rrl orivOeoq ev6g er6xo0 notrunemr6(ou rord q prerdrppo- on. En(onq, 6vo mRNA g6per enmp6o0ereg n).qpoqo- p(eg, 6ncog n6oo ouprd 0o prerorppd(etor, n6oo 1p6vo 0o enrBuiver rot oe nor6 orlpe(o rou rurdpou 0o pero- rppd(erot. Awdg or nLqporpop[eq prerog6powqr un6 Uo pen cls-oro r1e(tov r<or oXer(5 prevov n pcoreiVcirv. l-l ol.- Mg ewon((owor oe rpqporo rqg oll,r1l.ou1(oq rou mR- NA ro ono(o dev epnl,Sr<owor oneu0e(oq orrnv rro6u<o- no(4or1 npolrefvrlg. H EIKONA 22.1 6eiyyer oprotrrSvo qn6 rq tunxd 6opr- xd loporcrqptonrd rov mRNA orouq ilpoKopucbreg ror rouq euKopurilreg. To drpo ttov Borcrrlprordrv mRNA 6ev rpononoro6wor pretd q pretoyporp{, on6re op1(- (ouv pre ro 5'tprgroorpopx6 voux.leorbro nou Xprlotpo- nore(ror orrlv 6vop[q rnq Ueroypogrlg r<or re],euivouv pe ro refur6 voux).eor(6ro nou npoo0Srer q RNA notru- Uepdon nprv rov reppotropr6. To 3' drpo noMciv mR- NA rou E. coli opl;ror[(et pro 6oprf goupr<6rog nou epnl,Sreror orov evdoyevrl (rho-ove[6prrlro) repport- o96 qq peroypoqqg (BI5ne KeEdLoro 19). To euro- purortxd mRNA onorroUv rol.0nrpo rot oupd noluo6evulrxoU ev6oro preroypdgowor (B).5ne Keqd- Loro 21). Or neproo6rspeq Un xtrldrxonorqnr<5q, pu0pu- mrr6g nlr.qporpop(eq rp5powor mtg 5' ror 3' oper6- epooreq neprolSg (untranslated regions, UTR) rou mRNA, oproprSveg 6prrlq ewon((owor Kor or4v ro6xo- noqnr<f neproXri. Evri 6Lo ro pr6pto mRNA e(vor ypopr- prrrdg otrL4Lou1(eg vour<Leot16(rov, pnopeI vo oXqponoro0v 6eweporoye(g xor rpmoroye(q 6op6g ge ev6opropror<6 (euydpopro B6oeov. Aut6g or 6opr6g pno- pe( vo eivor onl,5g, 6ntog or 6op6g piolou-Oqtrldg (stem-loop) nou Tropouotd(owot orrrlv Eu<6vo 22.1, i nro oriv0ercg, pre 6roxi,o6roprdveg (branched) 5oU6S { (A) 5'UTR s'UTR 3'UTR Xoporrrlprolro Tolv npoKopuoTu6v ror rov euKopuoTrrorv mRNA. (A) 'Evo runtro porqptor6 mRNA. Aurd e(vot 6vo povc- olorpovtro mRNA, oporq ro porqptoro mRNA pnoper en(o1g vo e(vot ror notruororpovtro. l-lotrtro porrrlptoro mRNA retretdrvouv Ue p,c 6opfl p(oXou-0qfuoq. (B) '0tr0 ro europuo-rnro mRNA opx((ouv pe puo rotrunrpo (m?G) roL oXe66v 6tr0 retrerorvouv pe gto notru(A) 0up0 t notru(A) oupo entrotrunreror ond nptorelveg [poly(A)-binding proteins, PABP]. To europuolro mRNA pnopeivo tlXouv 1-uo fl xot rept00.- repeg neproX6g Deureporoyoug dogflg, runLro otrg 5'ror 3'UTR. (f) Iro 0r1troonro, ro mRNA nou rtodrronolouv lg rupLeg toroveq ov, qq notru(A) oupoq eXouv evo p(oXo-01trro. (B) (r) M(oXog-011trro KroDrxonoulrtrl neptolrl (0RF) 628 KE0AAAI0 22 lro0epoqro ror Xopo06rnoq rou mRNA 3' UTR
  • 120.
    (euydpopo vourtreotr6(ov preto(0onoporpuotrrdvov neprolti:v rou pop(ou. H 6repe0vrloq tov prqlovto;rtirv Ue rouq ono[oug onoKpurnoyporpoUwot ot puOptoru<dg nLqpo<pop(eg rou mRNA ror ror6ntv Xprlotponorofwot on6 ro pqlovrop6 nou e(vqr une60uvog yto rnv onod6- Unon, rn ;retdgpoorl Kqt rov ewontopt6 tou mRNA e(- vor onUowr.K6 ne6(o qg oriylpovqg poptorfg Bto],o- Y(oq. 22.2 To AyyeArog6po RNA Eivor Aoro0ri M6pro r H oord0ero rou mRNA orpeitreror oq Opdon p$ovouxtreoodrv. o 0r pt9ovouxtreooeq drotpdpouv oq rpoq rrlv npoqrloq yto ro undmpopo rot rov rpdno en[0eor1g. o To mRNA nopouord(ouv eupe[o drorupovon Tov Xpdvtov nut- (tons. o H drorpoptxrl oro0epdrrlro rov ;rop(ov mRNA ennped(et rn ouyxdvrptooq roug, ouven6g ror ro rpool-ro rov nptoreivtitv nou nopdyovror o' 6vo rUrropo. T' oyyelrog6po RNA e[vot q4enrd ooro0rl p5pto, oe ow(0eoq pre to DNA Kor, oe plxp6repo Bo096, to r- RNA r<or IRNA. E(vor o).r1Oero 6l or <pootpodteoteptro( 6eopro( nou ouvdSouv to prBovour<treot[6to e[vqt t<6- nog oo0ev6orepor oe o16or; Ue rouq 6eopo0g nou ouv6Souv ro 6eo(uprBovoux.l.eor(6ro L6yro rnq no- pouo(og rnq 2' udpo[u].opr66og orr'1 ptp6En. H oord- 0ero rou mRNA 6ev oqefteror 6prog rup(tog oe outq rq drogopd oMd oro yeyov6g 6rr ro rfrropo nept6- Xouv 1luprd6o 6v(uprov ono66;rqoqg rou RNA, to ono(o ovoprd(owor pr povouxtredoeg (ri bonucleases), prepx6g on6 rrg ono(eg oroleUouv er6xd"ro mRNA. Or prBovouxtredoeg e(vor 6v(upro nou r6Bouv toug groogodemepxorig 6eo;ro(g prero[ri rrov ptBovouxtre- on6(olv oro RNA. Eprgov((ouv erepoydvao enedrlnd- l,5g 6rogoperx6g nporeivtr6g neptoX6g (domains) od- Krnoov evepy6qto ptBovouxtredorlg r<otd rnv eqffuqrl. Tq ondvto nopodefu;roro yvoordlv prBoev(fpttov (xcro- Luru<d RNA) nepil.ogBdvouv noLL6g ptBovouxhedoeg, Svder(r1 yro rrlv opXdyovr'1 npoSLeuoq oufiq rqg or1- prowrrqq 6poorqpr6rrlrog (pL6ne KegdLoto 23). Ot pt- Bovouxl,edoeg 61ouv nollo0g p6l,oug oro x0r.opo, roOdrq ouprper6louv srnv owtypo<p4 rot qv enr6r6p- Ocoor1 rou DNA, mqv enefepyoo(o v5tov geroypdgtov (pre-mRNA, IRNA, rRNA, snRNA t<ot miRNA) Kqt ornv onod6gqoq tou mRNA. Or ptBovouxl.edoeg efuot efue evEoprpovouxtre6oeg efue e[oprBovouxtredoeg, 6nog rpo(veror or4v EIK0NA 22.2 (Blene eil(onq Ev6tqta 3.2). Or ev6ovouxtredoeg r6Bouv 6vo g6pto RNA oe pto eoo- repxq 05oq ror prnope( vo npouno06rouv r1 vo nport- profv ouyrexprprSvrl 6opn n ol.L4l,ou1(o. Ot e[tr:- vour<l,e6oeg orpoporiv vouxleot(dto on6 to 5vo 6xpo rou RNA ror 61ouv ro0optopr6vq roteriOuvoq (notrrr6- rqro) en(Oeoqg - efue 5'-+3'e(te 3'-+5'. Optol-tdveq e{co- vourtredoeg nopo;r6vouv npooxotrl4p6veg oto un6mptotr-ro evc0 orpotpo6v 6to6oXtxd vourtreot(6to, evrir 6Meg rorotrriouv t4v ogo(peoq ev6q { },fuov vou- xtreorr6[rov ror 6riorowor on6 to un6mpol;ro. To neproo6repo mRNA qnoouvt(0evtot rulo(o (6noq q onooUv0eoll rov po6revepyriv toot6ncov), on6te 11 oro0ep6rlro rou mRNA ouvrlOcog sr<gpd(etot rog 1p6vog nyr(oliq (half-time, t1z). O 6pog onooriv- 0eoq rou mRNA (mRNA decay) Xpnorpoilo€fuor ouvrl- 0og evoL..ortrrd rou 6pou ono66prlorl rou mRNA (mRNA degradation). l-ltrr1pogop(eq yro rrl oroOep&r1- ro ev6g ouyrexprprdvou mRNA e(vor r<to6txonotqpr6veg orrq crs-oM4l,ou{eg (BI6ne Ev6qra 22.7) ru dpo 1o- porcrqprorr<6q tou r<dOe mRNA. Arogopertrd mRNA epr- Eov((ouv o(roorlpre[oto 6rogopetu<4 oro0ep6tnro, U' erjpog dtor<riprovorlg 100 eop6q 11 rot nopondvco. Eto E. coli o runtx6q 1p6vog ng(oiq ev6g mRNA e(vot nep[- nou 3 tremd, 6pr<oq optoprdvo mRNA prnope( vo 61ouv Xp6vo qpu(toriq U6fuq 20 6eutep6tremo ri or6trro r<ot 90 Lemd. Irrl (0pr4, xotd tq rpdoq erBldornonq, o Xp6- vog rlpu(<or1g rov mRNA xupto[vetot on6 3 6tog 100 tre- 'e^* e^4 3 4ro4a s4+/a Prpovourtreooeg. 0r e(tovourtreooeg treuoupyouv npog ;lro rore0Ouvorl (unidirectional). flenrouv ro RNA e[re ono ro 5' oxpo e[re ono ro 3'orpo, onetreu0ep6vovroq Eextoproro vourtreor(dro. 0r evdovourtreooeg ropouv ro RNA o' eotoreprro0g rptrrotpodteore- prrouq deopoUq. Mro evdovourtreool ouvrlOolq oroXeuer etdrreg otrtrr1trouXieq ro/l deureporoyelg 6o;t69. Ev6ovouxtredorl tt.? To AyyetrLorpopo RNA Ervor Aoro0l Mopro 629 Boorreg dworeg E' Q' EItrlvouxtredoq EIovourtredorl
  • 121.
    m6, ev6 cnopetd(too, on6lemd 6crlg cirpeq xot, cmd- vto, or6po xot ;r6peg. T' ovritrrotro mRNA oroXeUowot yro notr0 roXe(o xoroorpogrl @6ne Ev&ryeq 22.8 rot 22.9). fevv<o, o Xp6vog nUt(onq npoo6top(etqt Ue na- poMoySg rrlg pe066ou nou rpo(vetot or4v EIKONA 22.3. H ogOov[o ouyr<er<prpdvov mRNA mo r<Urropo e(- vor ouvroropdvq qg rol0qroq o6v0eo4g (petoypo- <pr1 rr ene(epyoo[o) Kot rrlq roXrjqtoq ono66p4or1g. 'Orov out6g or nopdpetpor nopop6vouv oroOepdg, ro en(ne6o rou mRNA grdvouv oe oro0epri xordoroo4 (steady state). To <p6opro tov nproreivc0v nou ouw(- Oewor on5 ro rfuopo xup(og owtKoroTlrp((er qv oE0ov(o trov ow(ororlov mRNA (prolov6tt rdnoto p5- tro no((ouv en[or1g ror ot 6totpop6q qrnv onoreeopo- rrr6rqro 14g prer6qpoonq). H oqproo(o rnq ono- oUv0eoqq rou mRNA ovo6etrvUetot on6 pretr5teg pe- VdInS rL(goxog mrg ono(eg eletdmqr<e 11 oXettrrl ou- verorpopd rrlg rolrirqrog onoofvOeor1g Kot rnq rolrlrrlrog getoypocpqg orq 6rorpoptxti og0ov(o tou mRNA r<ot Bp50r1xe 6rr xupropXe( 11 np6q. To preydLo n).eov6rcr1po trlv ooroOc0v mRNA e(vot 4 trov6tqto roug v' oLLd(ouv ypiyopo qv on66ooq qg trretd- qpoonq ;16oo oLl,oyrilv orrl o0v0eor1 tou mRNA. Ilpogovrrrg, our6 ro nleov5xtqpo e(vot opretd or1- powrr6 r6ore vo toooro0p(oet rrl qorvopevtrrl onord- trn qq 6rlpuoupy(og Kqt 16oo ypliyopnq r<otoarpoEtlg rov mRNA. Avc0polog 6LeyXog tqg oro0ep6qroq rou mRNA lopor<rr1p((er no0oloyrxSg t<otomdoetg, 6ntrlg o xopK(vog, or Xp6vreq <pl,eypov5g Kot n oreQo- vro(o v6oog. 22.3 To Europuorrtrd mRNA Yndpxouv un6 Mopcpr'r mRNP on6 Tn [dvvnon dog rot ro @dvor6 roug e To mRNA otrtrr1trenrdpo g' 6vo peropotrtropevo ntrn0uouo nporeivdrv rord qv trlpipovon Tou orov nuptlvo Kot TIlv tt0p0- govrl rou oro rurropdnl,oolto. o Meprrdq nporetveg tov nupr;vlxtitv mRNP 6xouv pdtro rot oro rurropdntroo;to. * Yndpxouv nopo notrtreq nporeiveg nou npoodevouv RNA. 0t neprooorepeg 6ev 6Xouv Xoporqptorei o16po. E Arorpoperuo mRNA oxett(ovrot ge [eXoptot6q, otrtro 0trtr1tr0- enrxotrunr6peveq, oUd6eq pu0gtonr6v nporeivdtv, 6lUtoup- yrirvrog ro RNA peyroutrtivto. An6 q rn peroypogq ru.lv npo-mRNA orov nup{vo ;r5Xpr ror rrlv Korqorpog4 roug oro rutop6nLo- opq, ro euKopuorrKd mRNA o).).rllent6potiv pr' 5vo pretoBoM6 prevo penept6 p to n porteiv6v. To oti prn).o- ro RNA-nporelvc0v ovoprd(owot ptBovouxtreoTrpo- reivrrd oopori6ro (ribonucleoprotein particles, RNP). I-lo).i.69 on6lg npcuretveg nou npooSSvowot oe npo-mRNA ouprprer6louv oE owt6pdoetg ouppo- eriq Kr ene[epyoo(oq (B].6ne Kegdtroto 21), evcir d].- Leg e;lnSxowor orov 5Ley1o not6trlrog (ou(r1te(tot otrlv Evdtrlra 22.8). H op(;rovorl ev6g mRNA mov nuprlvo nepil.otrrBdveL notrtrd Bqpoto ovodtopr6pgco- oqg (remodeling) nou enqpped(ouv t6oo qv oMq- LouX[o rou RNA 6oo rot rtg oletr(6Ueveq npo- refveg, To drprpo mRNA e(vot xotdtrXq].o yto e(oyco- yn U6vo 6rov 6Xet otroxl.qpo0ef 11 ene[epyoo(o xot 11 oL).qLen(6poo4 rou pe to orr:md npcoteivtxd oUpr- nloro, preto(ri dtrtrcov ro TREX (transcription export, e(oyoyrl petoypo<pqq), ro ono(o preooloBe( ottl oriv6eorl pe rov uno6o15o e(oytoyrlg on6 tov nupn- vrr6 n5po, To cirprpro mRNA 6totqporiv nollonL6g 06oerg np6odeo4g (cls-orotXe(o) yto dtd<popeg puO- ;rrmrrdg nporeiveg, nto ouXvd prSoo ortg 5'n 3' UTR rouq. Evcb noLLdg nupqvu<5q npotetveg onoBdtrLowor nptv 11 rord tr1 6tdpxeto rqq e(oytoyrlg rou mRNA mo ruttop6nLoopro, d}'),eg ouvo6e[ouv to mRNA xt 6Xouv p6louq oro Kurrop6n).oopro. l-to nopd6etypto, pr6).tg Bpe0e( mo rurrop6nLoo;ro, to nuprlvtr6 o0pt- nloxo nou npoodSverot orrl rotr0nrpo ou;rper6Xet oro npoto oupBdv qg pretd<ppoorlq rou veou mRNA, tov rotroril.revo (nporon6po r0rl.o" (pioneering round) trlq prerdqpqolq. Autn q npurtrl Svop(r; qq pretd<ppoorlg e(vor r<p(otpr1 yto to v6o mRNA: ov ro eK- proye(o Bpe0e( eLorropotu<6, 0o xotompo<pe( dpeoo qn6 6vo oforrlpo entr;prloqg (B].5ne Ev6r4to 22.9). KdOe mRNA nou 6Xer nepdoet rov 5ley1o qg prerd- epqonq 0o (o65r,Uer tr1v un6troLn4 (ot1 tou ouvdede- p6vo pre norr<(treg npotetveg nou eL5ylouv rt'l t-- --------::- I trz=zumtn I 75 z. E. E >= 35 >E .oo b* o o E a a a a0 /10 ,/ Aroxonrl ueroYpoQrlq RNA Polll 20 40 60 B0 Aenro pero Tn 0roKonn 100 Ell(0NA 22.3 Mro U60o6oq yro Tov npoodtoptop6 tou Xpdvou nu(onq rtrlv mRNA. H petoypotptl on6 qv RNA notruuepoon ll dror6meror, e[re ge xdnoro rpopporeutLrrl ouo[o e(re otrtrd(ovroq q Oepporpoolo oe oretr6xrl Ue 0eppoeuo(001r4 peto]rtro[1 oro yov(6ro qg Pol ll. l-lpoodrop[(ovrot to en[ne6o ouyxerptpt6vov mRNA g' erruntrlo4 Northern Tl RT-PCR oe dLotpopetxd Xpovtxd olyproruno pero rr1 6Loxonq. H onoddpnon rou RNA, otpou [ert- vrloer, ouvrl0org e[voL rrioo ypnyopn 6me to evdtogeoo npoi6vro rnq 0repyoo(uq dev ovrXve0ovrot. H Xpdvoq lut(onq e(vot o Xpti- voq nou onque(rqr tiore ro mRNA vo n6oet oro Uto6 rr1q opXxfg nunq. 630 KEOAAAIO 22 Iro0ep6r4ro rot Xtopo06r4oq rou mRNA Boorxeg eworeg
  • 122.
    UerdQpoon, rn cnoeep6rnroKot, UeptK6q Qop6q' rnv ;;,i;n i;, oro r0rropo' H "nuprlut''{ torop(o" ev6g *nNrn eivot rp(otprrl rot r<oOop((et ro nenpou6vo tou crro rurroP6ntrooPto. 'Hdn yvop((ou;re nol).6q 6to<popettx6g nproreiVeg npOoieons'-b nnn (RNA'binding. proteins' RBP) r.rt, t ; Bdil qv ovdl'uor1 tou rlovtdttiruoto3',ovog6ve- io,' *- .i"apxouv nol0 neptoo6tepeg' l'1 po BL6nerot 6tt io VourOfouo rou S. cerevisiae rodtronote( nep(nou 600'6togopettr6g nporetveq np6o6eoqg oro RNA' ne- pino, .o 6vo 6riroto tou ouvol'tt<o[ opt0pori rov Yovt- bi,ru -or. Me B6o11 nop6proteg petpqoetq' 0o.nep(pmve xove(g 6tt to ovOpc0ntvo yovtd(topo vo Tleptexet Tlqvo) on6 zboo t5toreg npote"weg' AwSq ot errrprioetg Boo(- io*o, *nu nopouo(o xopoKrnptop6vtov neptoxtiv ipOoJtons mo RNA roi qo(verot 6tt peLl'owrxd 0o Bpegofv xot dltreg neptoX5q' To RNA-or61ot xot ot tr'et- i5;;Gs;;" neptoo6tepcov RBP efvot 6Yvooro' coor6- oo' n,ott,it-oi 6., nolL6g o)'l'r1l'ent6poriv Ue npo-mRNA n mRNA. Aut6 to e(5og ov6}'uor1g 6ev nept- LoprBdvet to peydtro opt0p6 npcote'twilv nou 6ev npoo- 6rivowot oneuOe(oq mo RNA, otrLd oup;ret6Xouv oe o6pnl,oro nP6o6eorlq oro RNA' - -'inuo*,^gg ev6e(ietq yto to tr6yo [-nop(r1g t6oo pe- VoLou opt0poti 6toEopetrr<6v npoteivdlv np6o6eo4q *o ,nrtfn'nporj).0ov on6 to eUpq;ro orl ro mRNA ouvd6owot pe (eXo ptor6q, o)}'6 otr'rlLoenu<otrum6 ge- veq, oUaOeS nee.- Vtel,etag ortg ono(eg 6ywe owtoro(- Xrol etdtr<rirv RBP pe ro mRNA-or61ouq onordLuqUov 6tr owd to mRNA rtodrxonoro(v nptoretvsg pe xowd XqpoKrrlptfixd, n.x' ouprpretopi oe nop6Uoteq Kurro- ii'lgi o"pvoo(eq 11' nop6ptoto ew6ntorl' ..ruvendrq' to penept6pto trov npoodedepr5vtov npolteivrilv KornYo- bil"J,tito mRNA. l-to nopd6etygo, erotowd6eq mR- NA tqg (ri;rqq e(vot 'or6xot" yto np6o6eo- ::: * : " "tptLbitptg on6 ttg 6 ouyyeve(g npcltarvac = -- : Puft xot Puf2 npoodevowot rup(o:g oe mRNA :3- '-- 6rxonototiv trreprBpovtr<6q npultetveg' evrrl 11 Puf3 oo -:- NA yto 1-tttolov6ptot<6g npotetveq K'o'K IUptcptrlvo * ero iiJ*,p"io pow6tro o ouwovtopevoq 6eyXoq rov Ueroperoypotprxov dtepyoottilv tou mR- r.rn ir"ooLoBe(toL on6 rn ouYxpovtog6vq 6pdoq noMo- niiu nap rot ;-rotd(et notr0 pre ro ouwovtou5vo eleyxo -ni ur-ovpoqrlq trov yovt6(trrv nou geoo)'oBekot on6 -ouq or-o,ig ouv6uool.toriq tov ;ietoypocptr<dlv nopo- V6wrov (B).6ne Keqdtroro 28 H opd6o trov mRNA nou uotoo(owot dvo ouyxerptp5vo tUno RBP ovo<p5petot Kor'coq' RNA peyxoulovro (RNA regulon)' 22.4 H Ano66pnon T(DV l'lpoKclpuoTlKdv mRNA I'lePrAoUBdv€l noAAo'Ev(uPo * H onodourlorl rolv porrrlptor(rv mRNA opx((et pe oqoipeorl evog nuporpolotpopLro0 on6 ro 5'orpo' * To Uoro,pro,poputrttrl;t6vo mRNA onodogo0vrot xoro rn 6top- Keto Tlq UeroQpoonq o' tlvo xurtro 6uo Bnportov nou nept- ioUpdr" ev6ovourtreotrurLr6g evio;t6g, rtg onoieg orolou0el neqtn rov rprlgdtirlv nou nporunrouv oe roteu0uvon 3'npoq tr a H 3'notruodevutr(toot1 pnope( vo Dteuro)'0vet rrlv onoOoprlorl rtrooporulv mRNA nou nepteXouv deureporoYrl 6oUI' * Tu x0pto rlv(upo rrlg onodounonq )'etroupyouv toq ouptntroro nou ovoprd(erot otr:1tot[Dto ono6oUnorlq' , H yevtrl rorio svoq RNA peyroutroviou (RNA regulon). rro europu0rtro mRNA npooorlvovtot notrteq npoTeiveq 0t oflottq .i.eyxouv rr1 perorppoorl, ,nu r*in,on roLq oro'eporrro rouq. To unoouvo),0 rov mRNA oro ono[o npoob.veror 11 rbto nptrlrelvq 'eo- : eiror rplpo rou lbtou peyroutroviou. Iro DLoYpoUUo, ro mRNA o rot 0 elvot Uepr] rou peyrou]'ov(ou 1 ' ro mRNA 0' Y K.t 0 rou peyroutrovi- - r 2 r.o.r. 2?.4 H Anooounon ttrlv l-lpoxopucorrxtilv mRNA t]eprtrouBoveL [lo},6.Ev(uuo 631
  • 123.
    Or yvdroerg Uqqyro qv ono66prnorlrov npoKopuon- rc0v mRNA npodplowor r<up(rog on6 preA6reg oro E. coli. MSXpt onygriq, or yevtrdg opldg roXriouv Kor fi' dLLo Borrrlproxd e(64 nou 61ouv ;relerq0e[. Iroug nporopuri:teg, 11 onoS6gnon rou mRNA oupBo(vet rord r1 drdpr<ero qg prerd<ppoonq. To nporopurotrxd prBooro;rdrro ferwofv q pretd<ppoorl nporo0 oLox).q- ptoOe( 11 preroypo<pri: npooSSvowor oro mRNA oe pro 05o4 5vopfig xowd oro 5'drpo ror ouveX[(ouv npoq to 3' drpo. lloMonld ptBooroprdtro pnope( vo [,exwq- oouv rn perdgpoorl rou r6(ou mRNA, 6ro6oXrrd, q44- pori(owog 6vo notruprBootopdno (polyribosome) q notruoop6rro (polysome), 6qlo6r1 5vo mRNA pre noMd ptBoocopdrto. To mRNA rou E coli onodoprofwor on6 5vov ouv- 6uoopr6 evepyor{rolv evdovoux},edorlq Kor 3'-+5' elto- voudiedo4g. To rUpro 1rovondn onod6prqorlg mo E. coll efuor puo drepyoo(o noA.trrirv mq6(ov nou rpo(veror orqv EIKONA 22.5. To opXrr6 Brlpo e(vot q ogo(peor1 rou nupogooqoprroU on6 to 5'drpo, pe rordLorno puo provodtrfl grrlogopu<rl pi(o. H Uovogooqopu].uo;r5v4 popgn dteye(per rqv rorol.unrrl 6pdoq prog ev6ovou- xledoqq (RNAorl E), 11 ono(o npqyporonoo( 6vo opXr- 16 r<6rprpo rovrd mo 5'drpo rou mRNA. Aur6 ro r<6t!rpo oqrlver 6vo 3'-OH oro ovodrr6 ri.dogo r<ot 5' Uovog@oqopr6 oro rq0odtK6 xLdopro ror xoroorpS- rper Lemoupyrxd 5vo povoororpovlx6 mRNA, eq6oov to prBoocopdrro 6e pnopo0v nL5ov vo (erwrloouv rrl prerdrppooq. To ovodrx6 xopprdn ono6opefror on6 pro 3'-+5' e(ovouxtredorl (PNPdoq = gooeoputr6o4 tov nol,uvoux.A,eott6(o:v, PNPase = polynucleotide phos- phorylase). KoOdlq neptoo6repo RNA err(Oeror perd q Suileuorl pgootopro{<ov nou erlov [exw4oer rn Ue- tdtppoo4, our6g o r<Ur<Log 6ui: Brltrrdrtov qg prBovou- xLedorlg enovoLoprBdveror rord pr4r<og rou mRNA oe roteriOuvorl 3'-+5'. H 6repyoo(o efeL(ooeror ro10roro, 6nog cpovepcbvet to yeyov6g 6tr BpoX6o rLdoproro nou nopdyowqr on6 qv RNAoq E ovrlveUowor pr6vo oe perotrtroypSvo r<rirropo nou epqov((ouv e).orrcopro- nrq evepy6rrpo e[olvoux],edo4g. H PNPdoq, 6ncog r<or df,l,eg yvom6g 3'-+5' e[ro- vour<Ledoeg rou E. coli, o6uvote[ vo npolrrlprloer 6ro- prSoou 6(rlovtov neprolrriv. Iuvenci:9, n 6oUn ;r(oXou-0qLrdg mo 3'drpo nolLriv Bor.rqprordlv mR- NA ro npomore0er on6 r4v dpeoq 3'en(Oeor1. En(- oqg, optopr5vo eooreprKd x}do;roto nou nopdyowor pe ndQ4 on6 r4v RNAoq E 61ouv neproX6g 6ewepo- royorig 6oUiq nou 0o pnopo[oe vo noporco],0oouv q 6pdoq trlg e[olvouxLedoqg. Om6oo, r1 PNPdorl ei- vor rrovrl vo n6nrer 6(rloveg neproldg ov undpXer 5vo rprrlpro pov6x.Lrovou RNA prrlroug rouLdlrorov 7- 10 voux.treorr6(ov r<o0o6rrd (3') rou pr(o1ou-04l,rdg. H ptov6xLcr;vr1 ol,l,r1l,ouX(o qo(veror vo Lenoupye( oov rxp(<o;ro yro ro 6v(u;ro. O Rho-ove[dprnroq repUqn- op6g o<prlvet pto ;rov6x.,ovq neptoXrj q ono(o e[vor noLti ptrp4 yro vo l,enoupyrloer oov u<p(ropro. l-to r4v enil.uoq quroU rou npoBl,ri;rorog, puo Borcrrlpror<ri no- Iu(A)- notrupep6o4 (PAP) npoo0Srer oupSg noAu(A) Merdqpooq + I v 5'pp p Y- r.,e rY 'r- 3' 3' - 5'E[tovourtredorl - Ell(0NA 22.5 Anodoglorl rolv porqpLoxtirv mRNA. H onodoplorl (rlv porqpror6v mRNA opX((er ge ro xdtlllro tou rprqooqopxou 5'orpou, ono6f6ovrog Uro UovoqooQopxq opd6o.'Eneuo, ro m- RNA onoDogorivror o' rivov rfrtro 6uo prlpdrrov: ev6ovour],eo)u- rur1 drdonooq, qv ono(o orotrouOe[ 3'-+5'n6Qq tou r],doporog nou qnetreu0epdlveror. 0r evdovourtreotrulr6g dLoondoerg oulr- po[vouv oe rote00uvorl 5'-+3' novo oro mRNA, oro],ou0rirwoq rr1 d r6treu or1 ro u te,Jeuro(o u p rpootrrpor(o u. 10-40 vour<i.eotr6(rrrv oro 3' drpo, rdvowdg ro euo(- o0qro oe 3'-5' ono66trlqoq. To rldo;roro RNA nou xorol,qyouv oe 16rokepo moOep6g 6eureporoye(g 6o;t5g (otog ononoUv enovoloprBov6;revo Brlproro no- i.uqdevu^,[oo4q Kor n5qnq on6 efcovour],edor1. Aev e(vor yvom6 ov 11 noAuo6evul(oor1 e(vor ndwq ro evoprcr{pro B{po yro qv ono65pr4orl rou mRNA 11 ov Xpqorpoilore(ror ;r6vo yrq vo Borl0rloer qv nopoyoyr] t<Loogdrrrlv, prero[rl dtrtrtov Kclr. ro 3'rs}.rr<6 r<].do;ro. Meprrd nerpdproro unodqLrilvouv 611 4 n6tlq evdg m- RNA qn6 qv RNAoq E [oroq e(vor onopo(rqt4 npo- un60eoq yrq rnv evepyono(r1on rnq poly(A) nolu- pepdo4q. Aur6 nr0ov6v eppqve[er yror( r' ordporo mRNA 6ev ono6oprorjwor on6 ro 3'dr<po. H RNAo4 E ror q PNPdo4, po(i pe pro eLrrdoq rr I k$d&' l^N ---&---&---# 632 KEOAIIAiO 22 lro0epdqro ror Xropo06qorl rou mRNA
  • 124.
    5vo entnp6o0eto entrouptr<66v(upo, oXqpor((ouv evo rroir"pr-tlvr16 oti prnl'o ro yvoord cog -ot'l porf6 lo """iOirq"nq (degradosome)' H RNAorl E 6Xet 6trr6 ;4il il otunlo'<o. H N-te)'tr<( neptoxq qg nop5xet i.lu euepyO-qro ev6ovour<Ledor1g, evtir 11 C-reLtrq ne- propj 6vo txp(opo nou ouYKpore( to un6l'otno ouoro- ,,^&. ru,i, n RNAon r<ot 11 PNPdorl e(vot ot nipreg 6v6o- xot eqovo'uxLedoeq otqv onod6prlon rou mRNA' undplouv en(onq rot 6trtre9, p6LLov pe nto nepropt- ;uJvous p6Loui. o p6Loq tov dtr'I'tov vourLeoocbv mrlv ono66prnon rou mRNA drepeuvrl0qr<e o(toLoytirv- roq rouq qotvot6noug pretoMoylrdvov rurtdptov yto nodSuo onb ro 6v(ugo. l-to nopddetygo, 11 o6povono(- non rnq RNAoqq E enrBpo60vet tqv.ono66pnon rou mniln'Xop(q vo rrl moptot6 eweAdrq' Metol'Ld[etg "ou qapouonototiv ttlv PNPdorl q onotodrlnote on6 rq 66o yvtlordq 3'-+5'e[covourtredoeq.6ev 51ouv ou- oromrr6' rog(o en(ntoon crrn ouvotrrx{ mo0ep6qto rou mRNA. Aur6 onorotrUmet 6tt 6Uo onoteo6{note e[c,lvour],e6oeg pnope( vo Sternepotdloouv prdtrtrov *ouou,oO t4v ono66prlorl rou mRNA' Motrororho' pr6- vo 6uo on6 t,g tpe(q e(ovour<)re6oeg (PNP6or1 rot RNAoq R) e(vot tt<ov5g vo nSntouv t<)'1o"y1to pe otq- 0epdq bewepotoye(q 6opdg' Aut6 ono6effinKe oe pe- L6ieg 6tnl,d perol'troypr5vov rundporv, oto ono(o e(Xov o6povonotrl0e( t6oo 11 PNPdorl 6oo t<ot 11 RNAon R. Ie outd to xtittopo, 11 ouyr<6wptoorl rov x).oop&tov mRNA nou nept6louv Seutepotoye(q 6o- p6g ou(dvet. ' 'noila epcorriproto nopop6vouv ovondwrlro oxett- rd pre qv ono66prr1or1 tou mRNA mo E' coli' O yp6- vog rlpr(oriq tcov 6to<p6pov mRNA eprgov((et eupe(o 6toxtil-rovor1, 100 rpop6q rotnl'6ov' H Bdoq outcirvttov orpo(tov 6to<popdrv 5ev SXet rotovoqOe( ntrr1pu:g' oL- trd <po(vetot 6tt orpefietot r<up(<og oe 6uo nopdyoweg' i euotoeqo(o ttov 6toEopettr6v mRNA oe n6qr1 on6 evdovour<Iedoeg notxftet: Svov 9q0p6 npomoo(og npoo<p6pet 11 nopouo(o 6ewepotoyo0q 6oUnS oro 5' a^po.' Op,o1r6vo mRNA ;retocppd(owot qno6ottK6te- Jo o"O 6trLo, o6rlydrwqq oe nurv6tepq ouoxeuoo(o icov ptBootopdrtov nou rcl npomore[ouv' Aev e(vot yrtoorA ov undplouv entnp6o0eto ptovondrto ono66- Uron, tou mnttlA. Ito E. coll 6ev 6Xet Bpe0e( 3'+5' lqt:vouxLedo4, nop6tt rivo t6toto 5v(upro 5xet touto- notrlOe( oto Baciltus subtl/is. l-lt0ov6v or 6totpopettr<69 evdovour)e6oeq Kt e(ovouxLedoeg 61ouv 6torpno0g p6l,ouq. Mto trreL6tr1 UtKpoouorotxtriv o' en(ne6o Yovt- Sui,lroioq npoodr6ptoe to en(ne6o 4000 xot nLtiov mnfln oe o0.ropo pretoLl'oyprdvo Yto rllv RNAoq E' qv PNPdorl n d).Lo oumottrd tou otol'tott6(ou ono- O3prlorlq. To en(ne6o noMrirv mRNA ou[r1Or1rov mo freioLLovpdvo r6rropo, rot6 to ov.opev6pevo (pre(- oon rrlq' ono66prr1or1q). AI'l'o, om6oo, nopSpetvov sro'(6tq'enine6q 11 ox6pro xot getrilOlrov' O Xp6vog rlptEonq erdtrriv mRNA prnope( v' oAl'd(et ond 6totpo- petir6g' rotomdoetg qg rpuototroy(og rou rurrdpou' n.1. ootr(o ri dL}'o e(6r1 stress. Ot uneU0uvot Urlxovt- opro( yt' ow6g rtg oMoydq nopog6vouv dyvtlorot' 22,5 To lleproo6tepo EuKopuoTlK6 mRNA Ano6opo0vTol M6oo Auo MovonoTl6v EEoprdlUevov on6 tloAuo6evuAitrlon r 0t tpononotfoetq xot mo 660 6rpo tou mRNA ro rpoorq- reuouv ond rn 6p6on rtov e[ovour],eoodtv' r To 6uo rupto l.tovondtto onoouvOeoqg tou mRNA opxi(ouv pe onoodevu)(trtorl 11 onoio rorotrUerot on6 nolu(A) vourtred- oeq. " MeiO rqv onoodevul(tlon Unopei v' orotrou0rloer e[te oqo(- peon nlq xoh6ntpog rot 5'-+3'n6r'[Jrl ond evoovourledol' e(- ie n6qn ono e[otvouxtreooqq o roreuOuvorl 3'-+5'' r to dviuUo orpo(peonq Uq rohrjntpoq ovroyrovi(etot ge ro orjpntroro evopEnq Uq UerdQpoonq yto np6o0eor1 oqv 5' rol6mPo. r To e[ootogono, ro ono(o rorotrrjet tr1v 3'-+5' n6$1 rou mR- NA, eivot 6vo UeYoho, e[drtxttxd ouvqprlp6vo otjpdroro' I H ono66prlon ouupo(vet O' eudtoxptto xurroponhooportxd otopor(Oto nou ovopd(ovrot otlltotfdto eneEepYooioq' To euxoputottrd mRNA npoorote0owot on6 ttg e(co- vour<ledoeg gSoto ttov rpononotnp6vtov drpov toug (Err6vo 22.1). H xol'rimpo 7-geOul'oyouovoofuqq no- p61", t po*qo(o on6 5' en(0eoq, evri: q oup6 poly(A)' lroil p," rtg npoo6e6e;r5veq npcoteiveq, nop6Xet npo- *.ioio on6 3' en(Oeor1. E[o(peor1 onote]'oUv to mR- NA tcov torov6v oro 0r1Loonr6, to ono(o rototrrlyouv oe Uro 6opri ;r(o1ou-0q}'r6q ow( trlq ou.pdq poly(A)' Irou g eu r<opudlreg, en(0eoq evdovou rtreootirv ove- (dpqq on5 tr1v oMqAoul(o, evoprcrrlptoq Pnxovt- o;r6q nou XpnotUonote(rot on6 to Bort(pto' ou;rBo(vet ondrto 11 not6. Molov6tt 11 onoo0vOeorl tou mRNA (mRNA decay) loporcrqp(orqre tremope- pSorepo orov (uproprfKnro, ro neptoo6tepo euprlpo- to tcrxtiouv Kol, Yto to r6rtopo ttov 0qtromtrtirv' H' ono66;rqorl rrlq ouwptmtrrlg netov6trltog ttov mRNA e[,opt6tot on6 onoo6evu]'(coo4, 611o6{ opX((et ond(owoq rnv npoororeurxrl oupd poy.(A) H veooxrl- Uoi,oUgun'ou'pd poly(A) (r1 ono(o o-rn.(0Un e(vot nep(- Iou io-so vouxleoidto o6ev(vqg, evci oro Oflr'oortr<d nepinou 200) entroLtirrerot on6 nporeiVeq np6o6e' onq oe polyin) [poly(A) binding proteins, PABPI' H orpa pofv(A)-u,p[oro.o, oro6toxrl BpdXuvorl ;r6Ltq ne- paot,' *o' rurrop6ntr'oopro, dtepyoo(o no-u t<ototr0erot on6 erarx6q vouxtredoeg poly(A) [ovogd(owor enioqq xor onoo6evuldoeq (deadenylases)l' T6o'o orn E[Un 6oo ror oro OqLoonr<d, 11 oupd poly(A) opltxd 9poX6- verot on6 to oUtrtntrot<o PAN2/3' Arol'ou0e( tolUrepq n6rlrlrotv 60-80 r<orol'ofnov A on6 ro orlprntror<o CCR4- ruOf, to ono(o nept6xet rrlv e[rovour]'edoq Ccr4 rot routrdXtorov 8 enrnp6o0eteg unogovddeg- A(tooqpre(- ,.o e'fro, to yeyov6g 6tt nop6;roto otiprnloxo CCR4- NOT eprnl'6r<ovrqt oe norr<(treg 6tepyoo(eg trlg 6r- ti.I To l]eptooorepo Euxopuorrro mRNA AnoDououvrot M6oto AuO Movonort6v E[opttirgevov on6 tloluodevutritool 633
  • 125.
    epoonq rov yovrd(ov,Uero(ri dtrLrrtv 11 evepyono(r1oq rnq peroypqgnq. To oripnLoro Oeo:pekor orroutrlevtr6g puOprorrlg qg 6r<gpoonq rov yovrS(tov, svonot6wog rn Ueroypogri ;re qv ono66prqo4 rou mRNA. T6oo orrl ([pq 6oo Kor. oro 0q],ooru<d undpXouv r<or dMeg vou- xi.edoeg poly(A). O L6yog our4g rqq norrd.6rqrog 6ev eivor or6go oqgnq. A0o Srogoperrxd piovondrro ono66gqorlg tou mR- NA nupodotoUvtor on6 rrlv ogo(peor1 tr.1g oupdg poly(A), 6ncog go(veror orrlv EIKONA 22.6. Zto np6ro pro- vondrr (ErrSvo 22.6A), n n6{n rqg oupdg poty(A) oe ollyo(A) prrlroug *10-12 A nupodore( rrlv orpo[peoq qg xolfmpog on6 ro 5' drpo tou mRNA nou rqto].Uerqr on6 ro ev(uprx6 oripnloxo ogoipeoqq rrlg xoAri- rrrpoq (decapping enzyme complex). Irrl ((9r1, ro oUpnl,or<o onoretre(ror on6 5uo npcorelveg (Dcpl r<or Dcp2), evc0 oro O4l,oonrd nepil,opBdver rrg opr6l,oyeg nporeiveg 6ncrrg en(oqg xor oprotrlSveg enrnp6o0eteg nporeiVeg. H o<po(peo4 qg xoL0nrpog ono6(6er 6vo RNA pe 5' povogoogopuLuopr6vo drpo, un6orprrr;ro yro trlv 5'-+3' e[<ovoux].edoq XRN 1, 4 ono(o n6met ro- lriroro ro mRNA. Ir4v npoygorx6t4ro, ouq 4 n5rfrl efuot 16oo ypnyopn ciore ro ev6tdpreoo nopdy<oyo qg frov odtivoro v' ovoyvoproro6v ;r51pr nou or epeuvrl- tdg ovor6Luquov 6rr ;uo oerpd vour).eol6(cov youov(- vqg (poly-G) pnoporjoe vo moprorloer r4v npoS).oor1 qg Xrnl orn EUUq. 'Onog qo(veror qrrlv ElK0llA 22.7 ot epeuvrlrSg roroore0ooov ;r6pro mRNA c6me vo nepr6- Xouv eooroprr6g neproXdg poly-G ror Bprjr<ov 6rt ouo- oopeu6rov ro ollyoodevu)lo1.ulvo 3' drpo rov mRNA. Aw6 66er[e 5tr (1) q 5'-+3'n6r,Uq on6 r4v ev6ovouxAed- oq qrov to rfpto 1-rovondtr qg onooUv0eo4g r<or (2) 4 ogo(peorlqg roLrimpog ouv6Bowe npw onS qv nlri- pr1 orpo[peorlrrlg oupdg poly(A). Kovovu<d, q r<oLimpo e[vor ovOercru<rj orqv ocpo(pe- orl tnq rotd r4v evepy6 perd<ppooq enerdq oLtqLenr- 6pd pe rrlv Kurropontrooponx{ nporefrv4 np6o6eo4g orq xotr0nrpo (cytoplasmic cap-binding protein), 5vo ouororx6 tou ougnl,6r<ou rou euKopucolKo0 nopdyov- ro 6vop[r1g 4F (elF4F) nou e(vot onopo(rrlrog yro rrl petd<ppooq (BL5ne KeqdLoro 24).'Eror, or prlXovropro( qg prerdqpoonq Kor qg ogo(peorlg rqg r<oi,rimpoq ov- royov((ovror. l-lrig 6poq n onoodevul.[coor1 rou 3' dxpou rou mRNA roOrord qv xoLtjnrpo euno0rl oe ogo(peor1; H prerdqpoo4 nepil.oprBdver gro rpuorxrl oL- ),qtren(6pooq pero[ri qg PABP nou e(vqt npoo6ede;r6- vrl oro 3' dr<po Kqr rou oupnl6r<ou elF4F-xoLfnrpog mo 5' dr<po. H one).eu06poon rnq PABP prSo<rl rqg qnoodevu^,fooqg nroreUeror 6rr onomo0eponore( qv oMqLenbpoo4 el F4F-r<ol,rimpog, orprjvowog ouyy6re- po erre0erpSvrlrrlv roi.rimpo. O ;r4Xovropr6g 6ev e(vor 16oo onL6g, xo0rig e(vor yvcom6 6rr orqv ogo(peoq qg roLUmpog epn).dxowot enrnpdoOereg nptorelVeg. 'Evo o0prnLoro emd oXetr(6prevcov np<oreivcilv, Lsm 1-7, npoo66veror orrlv neproXri oligo(A) ;rerd rr;v on6l.ero qg PABP xor e(vor onopo(rqro yrq rnv orpo(peoq rrlg rol[rnpoq. Enrn]*iov, 5Xouv ovor<o],urp0e( Srdrpopeq npcoteiveg nou onou[dvouv rnv ogo(peor1 qg r<o],ri- rnpoq. Or pullovropro( pre roug ono(oug ourSg ot npro- tetveg dreye(pouv rqv oqo(peo4 5ev e(vor nl.rlpog r<otovoqto(, nop6tr golveror 6n 6pouv eire enrmpo- te0owog ro Unxovrop6 ogo[peorlg qq r<otrUmpog e(re ovoor6Mowog q prerdqpoo4. Iro deUrepo trrovondrr (Err6vo 22.68), q onoo6evu- Nourtredorl poly(A) nno,oFl I [m?c F ". /r'--: it it -, A10 (A) 3'-5'Arrcanvooy/ - r'onooortro, (B) EIrootogolo ^,rS 4+/a t4+4a : , .. . To r0pto povonorto onoouvOeolg nou e[oprtivrol ono qv onoodevu],iroor1 otoug euxoputr.rreg, Auo povonolo opXi(ouv ge onoodevutr(too1. Kot oro duo, t'l oupo poly(A) rovro(ver ono ;.rro vour),eoo1 poly(A) eotg orou qrooeL oe plrog 10 A,'Eneuo 5vo mRNA pnope( v' onodoprl0eI p6oto rou 5'-+3'11 rou 3'-'>5'ptovonolou, To 5-+3' ;rovonon nepiloppover orpoLpeol qg rotrunrpog onrl ro ev(u;ro Dcp rot ndQl on6 r1v e[ovour],eooq Xrn1, To 3'-+5' povonon nepiloppover n5fi ono ro oupn],oxo rou e[trlotrlpor[ou. 634 KEOAAAI0 22 Iro0epoqro ror Xtopo0erlol rou mRNA
  • 126.
    Xprlot;ronoilorl ptoq otrtrr1trouXiogno- Iu(G) yto vo roOoptore[ 11 roreu0uvol rrlg onoofv- 0eonq. Irl (0U1, ov ltto ohtrrltrouXio nou(G) erooX0ei p6oo o' 6vo mRNA, 0' ovoorefieL Trlv npo- titrooq rov e[tlvour],eoodtv. To 5'11 ro 3' rtroopo rou mRNA nou ov0(ororot oqv onodoprlorl 0o ouootrl- peuOet oro rutopo rot pnopei v' ovtXveu0e[ p' erru- nool Nofthern. 3' -5' Anootxty/ 5'- 3'Anooljveton L(oor1 npoq o)rtyo(A) orohouOe(tot on6 3'-+5'ndqr1 on6 pro e(ovour<tredor1 rou r6ptou otirprotog tou mRNA' Au- iJto'Briuo ono56pr1or1q t<ototrUetot on6 to e[tootog6' rro lexoiome), 6vo 6orruhtoet66q orjgnl'or<o on6 5vqv ewe.opepTi nuprlvo, ndvo oto ono(o npoo6Svowot 1'tto ri neptoo5tepeq entnp6o0eteg npotelveq' Mto np6- o,poin ovogopd 56et[e 6tt to e[,<oocrlpot(6to 61et en[- onq'apdo'n ev6ovout<tre6o11g: 11 or1;roo(o trlq ouy'rerptpevrlq ev(u gtr<r1 g evepy6qtoq orrlv-onooUv0e- on'rou tnnrun nopog6vet dyvomrl' To e(ulotoptdtto undpXet oe nop6proro Uopqn oro Aplo(o r<ot en(oqq e[- ,o. oudtroyo rou otopor(ou ono66ptqonq rov Borrrl- p(cov on6 t4v dno$r1 6tr or unogovd6eg tou nuprivo oXeil(owot 6oprtr<d Ue rnv PNPdon' Iuvenrilg, to e(o- or,:1.tot(6to e(vot 6vo opX5yovo ouorottr6 Utoq popto- xnq UnXovnq. To e(ooopdtto no((et en(or1g orlprowtr6 p6Lo crrov nuprivo (BL6ne Ev6rrya 22'2)' H olettrrl orlpooio tou t<dOe pr4lovtopro0 6ev e[vot yvoorn;nop6rt orn E6pn <pofuetot vo uneptolUet to 1rouor,6-, otpo(peorlg qq rotrfntpqq il9u e[optdtot on6 onoo6evulrdron. To govondno ev ;r6pet roLOmov- rot. Erotowd6eg mRNA rnq (6pnq ;retr'eu10r6ov pe gt- Kpoouorotx(eg oe rrirtopo otq ono(q e(Xe onevepYonotrlOe( efue to 5'+3' e(te to 3'-+5' provondl' >e xd6e nep(molor1, pr6vo 5vo prtxp6 noooor6 preroypd- rprov ouEri0nre oe cr16or1 pe ro quotoloyxd r0nopo' Aur6 to erlprlpro uno64lrivet 6tt nol,0 L(yo pr6pto mR- NA trlq (riUnq eEopttirwot on6 to 6vo ( to 6Mo ;tovo- ndtt. "Ltot, npotdOrlr<e 6tt qutd to ptovondtto nou e(opttirwot on6 onoodevul'(oor1 e(vot ro npoenrLeypS- vo lrovondtto ono66prqonq Yto 6Lo to notruo6svutr'to- pr6vo mRNA, ptotrov6tt unoo0votro mRNA pnopef vo e(vot or61ot dMtov et6trrilv l.tovonotttilv nou neprypd- eowot orqv Ev6tqto 22.6. Ar6pro Kot ro. mRNA nou xpqotp,onoiotiv to npoenil,eypr6vo povondtto, ono6o- Uo.i*o, pre 6togoperxrl toX0t4to et6trci yto r60e mR- NA. Ndeg 5peuveg npore(vouv 6t 11 onoS6ptrloq rou m- RNA oulrBo(vet o' eudtdrptro ooltrro{6to oe 6o ro KUr- top6nLoopro, yvomd og oopori6ro enefepyooiog (piocessing bodies, PB). Aw6q or 6op6q, opot6q ge io ,pr-or,*6 pttt<poor6nto, e(vot ouvo0po(oporo Un Ue- rogpo(6prevov mRNA Kqt notK(Iov nproteivcbv nou oxe- t((ovtor UE rotootoLq rnq pretdqpootlq Kor onoo0v0eorltou mRNA, preto(0 dltrtov 11 prr11ovr1 oqo(- peonq rnq rotr0mpog rot q e(tovourledorl Xrnl' l-evt- rd, orq pg Oev undplouv npotetveg np6o6eorlg oe poly(A), yeyov5q nou uno6qL6vet 6tt npw on6 rnv eY- no**oon oe our5g ttg dopr6q oupBofver onoo6evu- iiron. To oto;ror(6to ene(epyoo[og.e(vot 6uvo1-ttt<d' ou[operdlvowot oe g6ye0og ror opt0;r6. ri or6pro ru e[o- qov((owor un6 6roqoperr5g rur-optx6q rot nepo;tott- r<6q ouv0rlreq TIou enrlped(ouv tr1 petoqp-ooq Kot rnv onoo0v0eoq. l-to nopd6elYUo, n onetreuO6ptoorl rov mRNA on6 to noLuoto;-tdtto pr' 6vo qdpptqKo nou ovo- or6Uer rqv 6vop(r1 qg perdqpoor'1q nporol'e( peyolq o[[r1or1 tou opr0ptoti Kot rou prey60ouq tov PB To 6to onot6f,eqto 5Xet rot n Ueptr<( o6povonofioq ouorort- r<c0v t1q onooriv0eorlg. Om6oo, 6tro to mRNA nou e6pe0ouv oro PB 6ev efvot xotodroopt6vo yt' onoofv- 0eor1: optol.t6vo prnope( v' onetreu0epo0oUv yto pet6- epoorl,'Xop(q 6pog vo 6Xet qnoooqnvtore( otte ndrq enil6yowot npoq onel,eu06ptooq ofre Kot ytor(' E(vot 6yv<omo rotd n6oo 11 ono66pr1or1 6}'crlv ttov mRNA ou;rBo(vet oe owd to ocopot(6to 11 o161lo rot tt tret- roupy(eq entteLo(v. Mto npoEovrlq t66o e(vot 6tt 11 ory^3*pror1 rolu ptiv Koroorpenrtr6v ev(tipttov oe onopovcoptdveq neptol6g ro0tord rrlv ono66prlon rou mRNA nto oo<Pohi rot ono6ottt<ti. !e et6xo0q runoprorig ttlnoug undplouv otopot(- 6ro nou nepr6louv mRNA t<ot gotd(ouv pe to PB' ro- 0cirg nept5Xouv rtq neptoo6tepeq on6 ttg nptoteiVeq nou'eptnl6t<owot qrnv r<otoorotrrl qq ;retdqpoorlq Kot rrlv onooriv0eo4. M4rptx6 xoxx[q mRNA (maternal ,hrun granulesl undpXouv mo coor6rtopo noL)'riv opyor,o-1rt'i,u. Aurd to xoxr(o onopt((owor on6 pr6pto niirrrn nou 6roulpo6wot oe pretoqpocrnrrl r<otoorotrq 6cog 6tou evepyonotrl0ofv rffid tqv enox6tr'ou0q ov6- rnuEn. H rotooroLrl ennuyl6vetot pr' ercretopr6vrl qTIo- odevutr(tooq Kot n peroYpoqn Ue notruodevutr(coot1' En(or1g, outd to rorr(o (oog pretoqdpouv mRNA oe ouyxer<prg6veq neptol5g rou oorurdpou (pl'5ne Evd- qia 22.70). To veuptovtx6 xoxxio (neuronal gran' uiesl ovoyvtop[mrlxov oe veuprilveg qq Drosophila' 'Onoq xot to l.rqtptrd rorx(o mRNA, q Lemoupy(o ou- ttilv ttov rorr(rov epntrdretot ornv Koroorol'ri qq petd- qpoonq Kor rn peroQopd er6trrirv mRNA' Mto t6roptrl ^o-nVopio otol-tott6(ov nep0.oprBdvet to xoxxio stress (stress granuiesl. To rorr[o stress 61ouv opretd 6to- ,poptt,o{ orioroorl on6 toug npoqyoigevoug tpe(q li- noug. Oot6oo, en(or1g nept6Xouv peto<ppootxdrg ovevepyd mRNA to ono(o ouvoOpo(owot oe ondvrn- on npoq gto yevrxrl ovoorol,ti r1q 5vop(r1g trlg petd- Qpoonq. *t"S To tleptooorepo Europuolro mRNA AnodouoUvror M6otrl Auo Movonorttilv E[oprdrpevov ono [lc]'Lorle' -r ^l- 635
  • 127.
    22.6 AAAI MovondrroAno66pnonq IroxeUouv tuyrexprpdvo M6pro mRNA * Teooepo enrnprlo0ero povonono ono6o;u1or1g enuetrofv rrl pu0pr(opevq onoddprlol er6trov mRNA. * Arpo[peon qg r<otrunrpog ove(oprrlq ono rrlv onoodevu],uo_ or1 e[etrlooeror orov unopXeL pro porpto notru(A) oupo. * H onoboplol Tov Un notruoDevutrur.rlrevov mRNA Tov r0To_ vtlv nupodore[rot pe npoo0rlrrl ptoq notru(U) oupog oro 3. orpo. qn H onooounorl optopevov mRNA prnopeL v, opXioet Ue neurl ono evdovourtreooq erdrrd yro qv otrtrr1trouX(o r.1 U 6oprl. e 'EvoS oyvtoorog opt0p6q popur;v mRNA oroXeuovrot yr, 0n0_ 6opr1o1 1 gerorppoonrf roroorotrrl on6 m i cro R NA. 'E1ouv neprypoge( r6.ooepo enrnp6o0ero provondrto ono66pqoqg rou mRNA to ono(o ouvotf((owor 6ro_ ypopl.rorrxd ouq :;iljiil:i :j,:..ii r<or ,til :i pio(( pe ro 6rjo r<ripro l.rovondrLo. E(vqr er6u<d yro unoo0vo).o pop(cov mRNA ror nepil.o1_rBdvouv ouprBdwo pu0pr(6pev4g ono66;rr1o4g. 'Evo provondtt nepiloprBover o<po(peo4 u1g xoLrj- mpog ove(dptrlq ond t4v onoo6evu).luloq, 64],o64 11 oqo(peor1 e[e].(ooeror or<6tr1r1 r<or drov undpXer Uro Ue- yd).r1 oupd poly(A). H orpo(peo4 rrlg xo).Unrpoq oKo- Lou0e(ror on6 n5q4 pe Xrn1. H nopdxopqrn rou Brlporog rqg onoo6evul,(rrroqg npouno06rer 6vov ;r41o- vrop6 yro qv enrorpdreuon rnq Unxovnq o<po(peoqg rqg rotrUmpoq Kor u1v ovooroL4 qq np6o6eorlg tou elF4F Xropig q Boq0ero rou ougn}6xou Lsm 1-9. 'Evo on6 ro mRNA nou ono6oproUwor qn6 our6 ro govon6- u e(vqt ro mRNA RPS28B, to ono(o xto6rxonoref rr1 p- Booroporrx4 npcoreivq S28 ror e(vor 6vog ev6tog6pov 1.r41ovropr6g ouro-pU0puoqg. Mro 6oprq p(o1ou-0r1Ar6g m4v neproX4 3'UTR eprnLSKeror orrlv enrmpdreuorl evdg evrq4u'r4 ogoipeoqg trlg r<oi,0mpog. Enrorpfrerr oq ouprBofuer pr6vo 6tov q nporetvq S2g npooddvsror orrl 6opr4 pr(oXou-0q).rdg. Iuvenrilg, 11 nep(ooero eleri Oepqg S28 oro xfrropo 0o nporoAdoer enrdluvoq r4g onooUv0eoqg rou ow(ororlou mRNA. 'Evo derirepo er6trd povondrt lp4orpronorehor oro 0qLooru<d yro rrlv ono66grlor; rou mRNA rrrlv rqrovdnr. Awd ro mRNA e(vor une00uvo yro rr1 ofvOeoq rou rE- pdonou opr0pro0 rov rorovcilv nou lperd(owor xmd rnv owrypoe{ rou DNA. IuoocopeUowor pr6vo rcnd 14 qdon S xor onodo;rotiwot rolfroro oro t6l,og rnq. To ;tr1 noluo6evuAur:1.rdvo mRNA rcov rmovd:v reppcrri(or ror Ue pro 6opr1 pr(oXou-0r11,(og nop6proto ge r4v oupri noMri:v Borcrqproxrirv mRNA. H ono66pr1o4 roug $1a o(rooqpre(coreg opror6r4req pe rnv onooUv0eo4 rou ln- RNA oro Borcrrlpro. Mto no).upep6o4 6ogu<d nop6poo trre q Borcrrlpror4 poly(A) nol,uprepdoq npoo06rer +rlo prxprl oupd noLu(U) owf qg oupdg poty(A). H pxpfl oupd ),er-roupye( oov nl,otg6ppro yrq ro oripdoxo Lsml-7 xqr/n ro e[<oocoprdrto, evepyonouilwoq ro Kt> 0repopr6vo ;rovondrro onoorivOeoqg. Aut6q o rp6nog ono66pr1or1g nopSXer Uro onUowrKri e(e).rrcnx4 ow6e- AqoLpeorl (A) rrlg xo),[nrpog oveIriptrlrrl onrl r4v onoobevutr(roor1 AAAAAMAA Poly(U) notrupepdorlnpooodKn olryo (B) (U) mo mRNA rov totovov, poly(A)notrupepdorl ,rrrr1-) Ev.ovourtreotrurLxn _ Ev0ovouxtreoorl (f) *nrn' pz6'*----------[ft'.-'.'|'iJ*- ev6ovouxtreoorl -Anoouiqq peootropotjpevrl ond miRNA ii{*l*il l? t Atrtro povonorto onoouvOeolg or0 tuKopuoTtKo rurropo. Anerrovr(eror ro opxrxo oulrpov oe roge povonol. (A) Ie oc :-i.vo mRNA 11 rotrunipo otpope(roL nprv vo ouppei onoodevu),irrlo1. (B) To mRNA rov rorovdrv onorrouv pro notru(U) oupo 11 ono(o 0c , ,. ro unoorpogo rrlg onoouv0eols (r) H onodoploq optoprlvtrtv mRNA ;rnopet v' opXLoet pe pro evdovourleotrurLrfl xonrl e(oprtirpev- --,(1v oqou1h. (A) 0progwu mRNA oroler.tovrotlt unobolrqoq roruororqq peroqpuoqq un6 oupnrlpoporyo mRNA-i:-l - 636 KEOAAAI0 22 Iro0epor1ro rorXorpo0eqorl rou mRNA (a) Buorx6g eworeg AMAAAAAA
  • 128.
    Movondrr Evoprurlprooup$fv MeroyevdmepopnUq(m)YnompriIots 5 - 3'ndqrn Anoo6evutr[ooq e{optogevrl on6 npoq otriYo(A) qv onoodevutr(trlorl 3'- 5'ndt}ln Anoodevutr[trlorl e[oprdgevl ond npoq oItYo(A) rrlv onoodevutr[oor1 Aqoipeorl rrlq Aqo(peoq rqq xotrunrpogove(dpqrrl xotrunrpog ond onoo0evutritrlorl Ev6ovourtreolulxo Ev6ovourleotrurtxil govonol xonrl lMovondl rorv Otrtyoouptduvttr[oor1 mRNA rov totovcitv Movonol rolv Zeuyoptrlgo Pooeov miRNA ue to miRNA oro o[Untroro RISC on qvdpeoo oro ouorrluorq onooUveeonq rov npoKo- puordv Kot Tov euKopuor6v. 'Evo tpko Uovondfl opxi(et Ut pto ev6ovouKleolu- nKn Konn e(optri:prevq on6 rrlv oMqlouX(o n q 6oUti. H xonrl oKolouee[ror on6 5'+3'4 3'-+5'n6rprltov r]'o- opdrov. 'Eneno, dvo erro0optoru<6 (scavenging) 6v(u- Uo ogo[peonq rnq Ko^Unrpoq, 6togopertr<6 on6 ro o[pnloKo Dcp, prnope( v' ogotpSoet rnv xo].Untpo. 'E1ouv ovoyvoptore( 6rd<popeq ev6ovouKledoeq nou droonoUv er6K5q 06oetq-or6Xouq oro mRNA. Mto ev- 6toq6pouoo ilep(nroon e(vot 11 oroXeupr6,v4 dtdonoorl rou mRNA CLB2 orrl(6pn, n ono(o ou;tBo(vet g6vo oro 16l,o9 r4g pr(roo4g. H evdovoux).edoq nou rotoLUet rqv ronri, yvtoorq cog RNAo4 MRP, r<otd to preyoLfrc- po ;rdpog rou KurroptKoU r0xi.ou neptop((etot orov nu- prlv(oro Kor ro ptroX6vdpto, 6nou egntrdretot m4v ene[epyoo(o tou RNA, oMd pretotp6perot oro rurrop6- nloollo oro t6Log tqg prfucootlg. To r6ropro trrovondrt e(vot to govondtt ttov microR' NA (miRNA), nou odrlye( r<oteu0e[ov o' evdovoux],eo- Lutrxrl ronrl tou mRNA r1 oe rotootoLq tqg per6gpoorlg tou. Itq ouyrerpqrdvq nepfmtrloq, to m- RNA oroXefetot on6 to (euydptopro pru<pd.tv (19-21 bp) ouprnl4poprortrriv RNA (miRNA-o64yo(, guide miR- NAs) oro ntro(oto ev6g nporeivtro[ oupn].6rou nou ovoprd(eror RISC. To miRNA-o6qyo( npo6plowot on6 preroyporp6prevo yov(6to miRNA ror 64ploupyotiwot on6 q 6r6onoor1 preyol.Uteprov np66popttov RNA. Iuve- ilci)q, n onooroOepono(qoq tov mRNA-ot61ov el.6yXe- ror on6 q pu0pl(Sgevrl Ueroypo<pr1 tov yovtdi<ov ttov miRNA. Ot l,emoprdpeteq rou prrllovtoprori 0o neptypo- qoriv oro Ketpdtroto 30. H oqpoo(o ouro[ tou vSou ;to- vonqrroU otrl ouvotrtr<q onooUv0eon rou mRNA nopop6ver odreurp(vtorrl, oMd 0o ;rnoporioe vo efuot Ueydln. 'EXet 6totun<o0e( 11 np6Bler.frl 5n orov dvOptrt- no Lemoupyoriv routrdXtorov 1 000 miRNA. Iiprgcovo trr' 5vo olor<trqpolpr6vo pow5tro Ytq rnv ono66pqoq tou mRNA, to l.tovondtto onooUv0eoqg nou e[oprrilwor on6 rrlv onoo6evu],(toorl owmpooconeUouv ro npoenil,eyp6vo ouorr]prorq Yto rrlv onoo0vOeoq 6}'tov rcov notruodevutrtcop5vtov mRNA. O Fo0p6q rrlg onoo6e- vuL(o:oqg ro/n dMo Brlporo r4g ono66prqo4g pt6oto ou- rdlv rrov l.tovonotuilv etr6yXovrot on6 cls-dpoottrd ororle(o rou rd0e mRNA t<ot trans-6pooru<otig nopdyov- req nou undpXouv oro r(nropo. I-lopdMql,o Ue ro npo- enAey;r6vo ptovondtto, trettoupyotiv Kot ro 1-tovondno onootivOeorlg nou otoXeUouv etdrd mRNA. npooOson tou oupntrdxou Lsm oro otrryo(A). Arpo(peol rnq rotrdnrpoq 5' - 3' eEorvourtreotruttrn ndQn ond rnv XRNl 3' - 5' e[tovouxtreotruTtKn ndq]r'l onri ro e(oroopdrto 5' - 3' elovouxtreotrulrrl n6Qq 5 -3 xor3 -5 e[ovourtreotrurtxn ndqln on6 ro e[cootogorto lpdobeon rou oupntrdrou Lsm oro 0kyo(U). AQoipeon trlg xotrunrpoq roL 5' - 3' e[orvouxtreotrurtrrl ndQrl ond tqv XRNl 3'- 5'n6Qr1 ond to e[tooogdrto Ev0ovourtreotrurtxrl 0toonqon n roroorotrq qq petogpoolq ftOovriv to neproootepo notruodevutruog6vo mRNA nr0ovrlv ro neprooorepo notruodevutrurr;ievo mRNA 0prog6vo erdtro mRNA 0propdvo erdlKo mRNA To mRNA rolv Lorovtitv oro 0qtroortro l-lotrtro mRNA (dyvooroq opr0pog) EIK0NA 22.9 Kprotpo ouoronro rtrlv l.rovonoldrv onoouv0eorlq rou mRNA oT0 euKopuoTtKd r0topo. 22.7 O Er6rrdq Xp6voq HUt(tufiq Kd0e mRNA EAdyxeror ond AAAnAouxieq rl AoUdq MCsq OTO MRNA * Erdrro cls-ororye(o ev6q mRNA enlpeo(ouv rrlv roXurrlro qg ono6opqoqg rou. s Irorxe(o 0n00T0etpon0tnonq Unopei vo entroXuvouv rriv onoo0veeorl Tou mRNA, sv6 oTotxe(o oroOeponofr1or1q vo qv nepropfoouv. * T0 nLo00r0 0e AU oTotxslo elvot rotvo 0T0tXe[0 onooro0epo- noinonq oro 0r1troortxo, oro ono[o npoodtlvovrot notr[treg nporeiveg. e Opoudveq npoTelveg nou npoo66vowot oe moryelo onomoOeponoi rlqq ol.ltrenLDporiv Ue oumorxo qq $Xovnq onoouv0eorlg rot gol.ov ro enorporeuouv yo qv onodoUnol. * Irorxe[o oroOeponol1or1g unopXouv oe optogtlvo notrf oro- Oepo mRNA. s H ToX0TrlTo onodounolq rou mRNA pnope( v' otrtroEer oe onovrqorl npoq notxftr0 or]Uoro. :] ;r 0 Erdrroq Xpovoq Hpt(orrlg Ko0e mRNA Etr6yXerotono Atrtrrltrouxreq rl AoUeq Meoo oro mRiA 637 Boorx6g eworeg
  • 129.
    Ie rr oqeil.erotq preydA4 6roxriprovoq ro:v lp6vcov npr(ofq 6rogoperxc0v mRNA rou r6(ou xurrdpou; Er- 6u<d crs-morXe(o puloo mo mRNA enqped(ouv 14 oro0ep6rqrd rou. Iuvrl0og ewon(owor ornv 3' UTR, nop6rr undplouv ror oMori. Meldreg o'eninedo o.1,6- xLqpou rou yovr6rcbprotog onord),urpov noMd t6tofue- po ouwnpnpdvo pror(Bo 3'UTR, trov ono[rov 6porg o p6).og nopopSver dyvororog. Optotruivo e(vot 06oerg- o161or yro np6o6eo4 miRNA. A).Lo e[vor 06oerg np6odeorlg yro rrq RBP, r<dnoreg on6 rrg ono(eg 61ouv yvcoorSg Lenoupy(eg orov 6Ley1o rqg oroOe- p6qroq. H roXriqro onoo6evul,(roorlg drogoperu<cbv mRNA pnope[ vo noxfi.er noAU rr 6Xouv neptypo<pe[ oMqloul(eg nou rrlv enqped(ouv. KoLUtepo pel,errltrrdvo e(vor rq oror;1eio qnooroOe. ponoirloqg (destabilizing elements, DE). To rprrlpro yro vo oprore( pro oM4LouX(o og ororle(o onooroOepo- nofiorlg e(vor vo nporole( enrdXuvoq rrlq ono66prqoqg ov eroolOe( o' 6vo oro0ep6 mRNA. H orpo[peor1 ev6g rtltorou orotle(ou on6 ro mRNA 6ev ou(dva ovoyKclorr- rd rq oroOep6rrlrd tou, 6v6er[4 6rt prnope( vo nepdler neproo6tepo on6 6vo orotXe(o onomoOepono(qor1g, 416;ro noLunLox6repo, n nopouo(o ev6g DE 6ev roo6u- vope( pe o6wopro Xp6vo qpl(oqg un6 6Leg trg ouvOr1- req, ener64 dMeg oMql,oul(eg rou mRNA pnope( vo tpononor{oouv r4v onore}eoproru<6rr1td rou. O r<ol,Urepo preLerrlprSvog r0nog DE e(vor ro ntroU- oro oe AU ororyeio (AU-rich element, ARE) nou Bp(- oKeror orrlv 3' UTR nep(nou tou 8% rcov mRNA r<ov 0ql.ooru<rirv. To ARE e(vot erepoyewi r<or 6roxp(vowot oe oprerorjg unotUnoug. 'Evog ttinog onoreLefuot on6 rrlv newqpep4 oL).r1Lou1io AUUUA, o' rivo 4 enovo- LoprBov6gevo ow(yporpo. 'Evog dtrLog rUnog 6ev ne- pr6Xer qv oMr1trou1(o AUUUA oMd e(vor nlo0orog oe U. 'EXouv rouronor4Oe( noM6g nporeiveg np6o6eoqq oe ARE pe erdrr6rrlro yto erbrrofg trinoug RRE r<o/n rUnoug rurrdpov. l-lc0g 6prog l,enoupyo0v ro orotle(o ARE yro vo 6reye(pouv 14 yp4yoprl ono65pr4o.n; l-lol,- l,6q nporeiveg np6o6eorlg oe ARE oLLq).enrdporjv pr' 6vo r1 neptoo6tepo ouororrrd rou trrqlovro;ro0 ono66- Unonq, prero(0 dMov ro e[o:oroprdrro, or onoo6evu].d- oeq Kor ro 6v(upro ogo(peoqg tqg r<oLrimpog, yeyov6g nou uno6qL6ver 611 6pouv enrorporetiowoq ro Unxovr- opr6 ono66gqo4g. To e(cooolprdno pnope( vo npoo6d- veror oileuOe(og oe oproprdvo ororle(o ARE. To morXe(o ARE oproprivcov mRNA enrol0vouv ro Brjpo onoodevuL(oonq rnq onooUv0eoqg. En(or1g (oog 6pouv deurol0vowoq rnv onoteLeogoru<r1 np6o6eor1 rou mRNA oro ool.rori6ro ene[epyoo(og. l-loMd morXe[o DE nAoUoro oe AU ror dMo e(6r1 onomoOepono r4rrr<cilv mo r1e(tov rowono 14 0r1 Kclv oro mRNA rnq (0UnS xor dMov np6runov opyovtopr6v. t-ro nopdderypro, oTn ([pn or npcoretveg Puf npoo66vowot oe erdmd ororXe(o ntrorjoro oe UG ror enmoltivouv rqv ono66prr1or1r<ov mRNA-or6Xov. Ie ouq qv nep(mo- on, o Unxovropr6g onooroOepono(r1or1g enmoX0vet rrlv onoodevuL(<oon eTtorporstiowog r4v onoodevuLdorl CCR4-NOT. Mro ovdl,uorlrov 3'UTR qg (Uprrlg touro- no(r1oe 53 morXefo oMrll,ouX[og ro ono(o oXer(owor pe ro 1p6vo nprEons ttov mRNA nou ro nepdlouv, eU- pnUo nou unodrlLrirver 6n o opr0pr6g rrov 6rogoperu<cirv ororle(ov onomoOepono(qoq g (oog e(vor ;reyol,fte- pog. It4v EIKONA 22.10 ouvorp((owor or yvcrror6g 6pd- oerq rov ororle(orv onooro0epono(4or1 g, Irorleio oroOeponoi4o4g (stabitizing etements, SE) touronorq0rlKqv oe Liyo oouv4Ororo moOepd mRNA. Tp(o mRNA nou peLerrl0nKov oe r[rropo Oq].ooru<oiv 5Xouv orqv 3' UTR otri.4LouX(eg oroOepono(4o4q nLori- oeq oe nupqlr6fuq. Ot nporefveg nou npoo6Svowqr oro ouyrexpq.dvo ororle(o rou mRNA qg oqopfuqg o.lr.qle- nr6porjv Ue rnv PABP, 5v6er(q 6tt 0o pnopo[oe vo ler- toupyoriv npomoteriovrqq rrlv poly(A) oupd on6 ono66prqor1. Ie opropr5veg neprmc0oerg, 6vo mRNA prno- pe( vo oroOeponoq0e( ovootdMowoq to DE tou. Flo no- pd6erygo, ouyr<erpq.r6veg nptotelveg np6odeoqg oe ARE epno6(ouv t4v onooro0epono(qoq rou mRNA on6 ro ARE, prdMov ovoordMowoq rr1 06oq np5o6eorlg ARE. 'Evo nopd6ery;ro pu0pr(6;revqg oroOep6rrpoq rou mRNA nop6ler to mRNA rrlq rpovogepiv4g (transferrin) rcov 04- l,oorrxrilv. Iro0eponore(rqt 6tov puo erdu<r; nporetvq EnrorpdTeusn rou grllovrogorj oroddUnonq Iuvopgo)dyqorl rou PB; 06oq m6Xog yro evdovoudeosn Avumotrrl npdodeorlg rnq npoteivnq onooro0eponofioqg PABP-poly(A) Dcp ffi Nouxtreoorl poly(A) _ ")-> I l<_-___)ur M'.&/ MqlovrolroL pe rouq ono[- oug Leuoupyouv ro orotXe[o onooroOepo- norqolq (DE) xor oroOepono[qorlq (SE). 0r enlnroroerg rov DE rot SE oq oro0epoqro rou mRNA ;:eootrogouvroL rup[og ono lq nprrlrefveg pe rrq onofeg otrtrrltrentdpouv E[orpeol onore],e[ 6vo DE ro ono(o ],er- roupyer olg ordXog yro evdovourtreooq. 638 KE0MA|O 22 Iro0epdqro ror Xtopo06rqorl rou mRNA
  • 130.
    H pti0Uton rrlgoroOepoqtog rou mRNA *c rpovogeptvlq on6 ro en[nedo rou otOflpou To IRE :rlv 3' UTR etvot 0eor1 npoodeorlq yto rrlv nporetvl nou :roOeponotel ro mRNA. H nporreivq nou npoo66verot :ro IRE eivut euo(oOr1rr1 oro en[ne6o otdnpou (Fe) tou ..rropou: npoo6everot oto IRE g6vo 6rov ro enlnedo ot- lnpou e[vot Xopntro. En(neDq otonpou XoUrltrd To mRNA tns tpqvoQep(vnq e(vot mo0ep6 llpotreil'1 :: - npoodevero,:.: :i AAAAMAAA @ AAAAAAAAA npoo66vetot oro orolleio qn6wnonq oro o6rlpo (iron' response element, tnel nou nept61et, 6vo orotXe(o to ono(o ewon((etot qrnv g UTR rot onotelsftot on6 notr'- Ionl6q 6op6q ;r(cn1ou-0qNdq, 6nog qofuerot ornv EIK0NA nfi. H ouyy6veto rrlq npcoteivrlq yu to IRE otrl'6(et pe rqv np6o6eor1 otdrlpou: e(vot 1opr1trrl 6tov 11 06oq np6o- deorlq tou ot6rlpou e(vot roteilrlprpr6vq r<ot ro ow[orpo- go. 'ci.ou q ouyr<5wpcoor1 otdripou mo xirropo e(vot Xo1.rnNl, Xpetd(etot neptoo6tepq tpovorpep(vq Yto etoq' vrjv,r oianpou'on6 rrlv r<urlotpop(o rou o(poroq, on5te ro m RNA rqq tpovorpep(vr1 g oro0eponoekot' H nporchnl np6odecrrlq'oro IRE oro0eponoe( to mRNA ovoor6Mov- rqq rn leroupy(o ttov yerovxrilv oMqlouXttilv onooro- 0epono(qo"r1q. E(uor o[toorlpre(ulrg 61] (6to npotetvrl npoo66vetoi en(or1g o' 6vo otorye(o IRE tou mRNA rrlg eeppmvnq rot to puOg((et ;re no}'0 6togopenr<6 rp6no' H qapptt(vn e(vot pto npoltetvrl np6o6eoqg o't6ripou q ono(o eVr<ttopt(errnv nep(ooeto otSripou oro xfrropo' H npcrnetvq np6o6ecrqg oe IRE npooddverot oe g(q4ouq-Oq- tu5q IRE oqu 5 UTR rrlq Eeppr(vqq 6rov 19 ertue6o ot- dqpou e(voi Xoprl}'6 r<ot ovoor5trtret n'1v oMqMn(6poor1 rou ouprnl6r<ou np6odeoqq onlv ro}'rimpo Ue ro mRNA rnq qeppmvng. 'Erot, 4 pretdgpoorl tou-mRNA rnq Qep- pnfis nopelinodi(etot 6tov ro en(ne6o ot6tipou oro L*r"p" e(vot loprqLd, ouvOrir<eq un6 tg ono(eg oro0epo- noeftot rot prerorppd(etot to mRNA tqq tpovorpep(vr1q' I-1oM6g'npr.iiueq np6o6eorlq oe cis-orotle(o utpi orcrwcrt tpononoqoerq ot ono(eg go(verot 6tt enqped- (ouv u1 trettoupy(o rouq, n'X. tptoogopul'(toor1, peOu- )t(oorl, oMoydg Stop6ptpoorlq e[otr(oq qq np6o6eorlg rel,emov Kqt tooUepe(oor1. T6roteg tpononot(oetg (ocrlg euOrjvowot Ytq nq oLLoydq rrlq tolUqtoq qno- 66Unonq tou mRNA nou endyowor on6 runoptrd of- Uoiq. H onoo6v0eorl rou mRNA oMd(et oe ondwrlorl npoq eupU qdopo neptBotrl'owtr6v rot ev6oyevdrv oq- g6ttov, 1.reto[ti dM<ov 11 np6odog rou KurrclptKori 16- x.ou, 4 rurroptrri 6to<popono(r1or1, oppr6veq, n rpo- rpo6oo(o oe 0pemr6g ouo(eg xot toyeve(g Lortrldr[etq' Mdr.fteq Ue UtKpoouototX(eg Sdet[ov 6rt to 50% nep(- nou rov oLloyriv oro en(ne6o tou mRNA nou npoKo- Lo[wot on6 xurroptr<d oqproto oqeilowot oe ouU- fdwo oroeeponohorlq q onooroOepono(r1or1q r<ot 6xt oe oMoydq rnq UeroYpoqnq. To ncirg enmuyXdvowot our6g ot oMoydq oe prey0to 9o0U6 nopog5vet 6yvt'l- oTo. En(fledo otonPou uUntrd 22.8 Io Neoouvre0elPdvo U6Pto RNA EA6yxovrot Vlo EAoTTdLtoTo Mdoo Evoq tlupnvrro0 Euorhporoq Entripnonq @L9--l , o To etrorroUortro nuprlvtro RNA ovoyvtrlpi(ovrot rot r0r0- orpeqovrot ono 6vo ouorrlpo entrnplolq * To nuprlvtxo e[ulotrll.totto tretroupyet rooo roro rnv one(ep- yooio tpuototroyLrtirv RNA unoorptlgorolv ooo rot roto u1v roroorporpl rctlv etrorroportrtirv RNA. * Iu (0un, ro oupntroro TRAMP entotpore0et ro e[ootr-tltotto oe oviilpotro popLo RNA rol dLeurotr0vet qv 3'-+5' e[ovou- rtreotrurtxfl 6Poo1 tou. * IT0 ufl0oTptlgoro yt' ono66p1or1 p6ool rou oupntrorou TRAMP-e[oroopor[ou nepfioptpovovrot Un ouppogU6vo rl ovtilgoho ouppopp6vo npo-mRNA rot tro0oq repportoptlvo gerdypotpo rnq RNA notrupepoorlg ll Xtrlpiq notu(A) oupo e To neptooorepo peroypotpo uq RNA notruuepoorlq ll totr:q ei- vot rpuqo ooroOn UeroYpoQo llou roroorpdqovroL yprlyopo orov nupflvo, Otro to veoouweOet;r6vo RNA unoBdMowot oe nol- ond Bqproto ene[epyoo(oq petd tr1 peroypoqrl rouq (Bi6ne Ke<pdoro 21). Ze r<dOe Brlpo prope( vo y(vouv ).d0r1. Evto to d0q oro DNA dtopOtr'lvowot on6 notrfto ouorqpoto ent6t6p0cooqg (B6ne Ke<pd- Loro 16), ro Ld0r1 nou ovtXveUowot oro RNA owtge- rconf(owor Ue Kqroorpocprl rov e]'orr<optottt<tov RNA' Iuorriporo enlr{pqoqg rou RNA (RNA survellance systems) undpXouv t6oo orov nuprlvo 6oo rot oto r<urrop6noopo Yto vo 6toXetp[(owot 6tocp6pov et- 6ov npoBlrlpoto, H entrrlprloq nepAoprBdvet 6po- orrlpr6tqteq 66o etdtov: Uto Yto v' ovoyvtlp((et Kol' vo or11lo(vet ro etrorro;trtqrLr6 RNA un6otptol-to Kot Uto dM4 yto vo to rotoorP6<Pet' d *o-o*po.p5oq e(vot to nupqvtr6 e(olotop6rro' O nupqvoq (core) tou nuprlvLt<oti e(cuoopot(ou e(vot oXe- 66v 6protoq Ue ro Kurroponoo1.totLx6 e(u'lotr:pdtto' no- p6tt otrtrq).ent6pd pe 6togopettr<o0g npo:teivtr<oUq ouprnopdyoweg. Arpotpei voux..eot(6to on6 tq sroxeu- pSvo mRNA trre 6p6o'r1 3'-+5'e(rovour<)'edoqg To nupq- To mRNA qq tpovorpepivrlq elvot omo05q 21.* To Neoouvreoetptevo pdpto RNA Etr6Yxovrot yto Etrorrdrporo Mtlotrl Evoq l-luprlvtxou Iuorqpotoc EnL:-c';'c 639
  • 131.
    vrK5 e[oooUdrro 6XernoMonLsq Leroupy(eg, peto[rl dMov 11 ene[epyoo(o oprqrdvtrtv 1.u1 r<to6u<onor4urc0v RNA peroypdcpcov (snRNA, snoRNA xor rRNA) ror q nlnpnq ono66pqorl rtov el,orroportxc0v RNA. To e(to- orrrpdno entorporeUeror orcl unompcitl.roro nou np6- Kerror vo ene[epyoore( tri5oo nproreivrrc6v oupn].6r<ov nou ovoyvop(ouv er6u<69 oMqLoul(eg RNA n do;rdg RNA/RNP. l-ro nopd6eryuo, ro Nrdl-Nab3 e(vor rivo nproreivu<6 6rpep6g pe erdu<6r4ro yto ouyKeKprgdv4 oL- tr4LouX(o ro ono(o enrorporcUer ro e[rrloroprdrro oe qu- orol,oyrxd unootpcbgoro snRNA/snoRNA. Aur6 ro (euydpt nporeivrirv npoo66veror oe ororle(o GUATA/GI Kqr UCUU, owrororXcog. O ouprnopdyowog Nrdi-Nab3 epnA6xeror en(o4g orov reppono;r6 qg peroypoqqg rov pn noLuo6evuLurt;r5votv RNA nou preroypdgowor on6 qv RNA noLuprepdo4 ll. Aur6 uno6q).dtver 6rr ro e(ootopdrro foog enrorporeUeror oneuOe(og mq 06o11 qg oUv0eo4q rouq. AvripoLo ene(epyoo;r6vo, rpononorqpuivo q roxo6r- n).copr6vo pr6pto RNA onomo[v d].troug nproreivrxorig oupnopdyoweq yro qv ovoyvcbpron Kr entorpdreuo4 rou e(oorol.rorfou. Ir4 (rjprq, ro rripro nuprlvrr6 o6pr- n).oxo nou errele( ourq q l,enoupy(o ovoprd(eror TRAMP (or<pcovripuo tov nporeivc0v nou ro onoreLoriv) ror undpler rouldXrorov oe 6Uo Uopqdq nou 6rogd- pouv oq npoq rov rtno qg poly(A) no).uprepdoqg nou nepr5louv. To oriprnAoro TRAMP enrlped(er qv ono66- Unon Ue 6rdEopoug rp6noug: 1. AM4),enr6pd d;reoo pe ro e(cooto;rdrro, 6eye(pov- roq rnv evepySqro e(ovour<Ledoqg nou 6ro0drer 2. l-lepil.oprBdvet ;.uo eAu<do4, q ono(o prdL),ov e(vor onopo(rr'1q yro ro {erri}.rypro qg 6eweporoyo0g 6oUnq ro/r1q preror<[vqoq npo:reivriv nou npoo6d- vowor oe RNA on6 RNP unompd:goro xord rrlv onod6prloq. 3. llpoo0Ster puo ;lrpri 3'otrryo(A) oup6 oro uno- orp6poro-or6Xoug. I1toreUerot 5l 11 oLryo(A)oupd rdver ro oroXeuprdvo RNP xolrirepo un6orpopro yro ro pqxouogd ono65;r4or1g, 6no4 orpgdrg Kor oro Borcrrlpro. H EIK0NA 22.12 ouvotp((er rouq p6Loug tou TRAMp Kor rou efootol.rot(ou. E(vor nfuiov oog6g 6tr 4 ono66- Unorl rou RNA oro Borcrrlpro ror r' ApXo(o rot r1 ono66- Unorl rou nupqvrro0 RNA oroug euxopuolreg oXer((owor ovomu(ror<d. H opor5qrd toug uno6r1).ril- ver 6tr opXdyovog p6).og r4g noluo6evul.(oo4q r.1rov vo 6teurotrUver rrlv ono66prr1o4 rou RNA r<or 6tr q poly(A) npoooppr6or4re opy6repq fiouq euxopucbreg yro rrlv nopd6o(o ow(Oerr'1 Lenoupy(o r4g oroOepo- nofiorlg rov mRNA oro rurropdnlool.ro. floro efuot ro unoorpripqro rrlq ono66g4oqg prdoro TRAM P-e(cooopo{ou ; To oti pn}.or<o TRAM p ovoyvcop(- (er peyd,l.q noxil.fo e].ortrr:;.rotxcbv RNA nou peroypd- gowqr xor qn6 nq rperq RNA noluprepdoeg. Aev e(vor yvom6 ncbg enmuyXdveror our6 roOrig ro RNA-or61or 6ev 51ouv rorvd Xoporrrlprorxd. OproprSvor epeuqtdg unomq p((ouv d,vo prow5.tr o r<rv4ru<ori owoyrovrollori, uno06rowog 6l unoorpci:1loro ono66pnonq on6 ro e(cootrtpduo 0o y(vouv RNA ro ono(o dev ug(mowor ene[epyoo(o oUre xot ouvopltoA.oyolwat 1yxatpa oe puo oprorxrl [opAn RNP. Aur6g o;rrlXovro;rdg onoge0- yer rrlv npoun60eo4 er6u<r1g ovoyvroprorlg rrov ovop(S p4tov nt0ovcbv elorrolprdtcov. Tr e[6oug ovopro].(eg xorodu<d(ouv ro npo-mRNA oe Korqsrpotprl mov nup4vo; 'EXouv ovoyvroptme( 6uo e(6q unoorpoprdrcov. O 6vog r0nog neprloprBdva Un ouppotrU6vo 4 ovcbpoLo ouppotrrprdvo npo-mRNA. Tq ouoronrd rou ocol.rot(ou oupporpqg ouyxpororiv rriroro pretdyporpo Scog 6rou e(re ono6oprqOoUv on5 ro e(oocopdno e(re oLor<i.r1pco0e( 4 xordM4).r1 ouppo- An (qv our6 e(vor erpucr6). l1tmeUeror 6rr ro pow6]to rou KrvrlnKo0 owoyrovropro6 prdl,trov tolrjer Kor orrl ouyrerprp6,vrl nep(nroon. 'Evo npo-mRNA ro ono(o 6ev 6Xer ouppo<pe( Kor ouoKeuoore( onoteLeoporxd ep<pov((er ou[r1g5vo rfv6uvo vo y(ver or6Xog rou pn- Xovrol.rori ono66pr;o4g on6'ro elroou:prdrto. O rp6noq ovoyvrilpro4g rov KoKoouppo;r6votv npo-mRNA 6ev ei- vor yvcoor6g. O deUrepog ttinog unoorpoprdrov nepr- LoprBdver npo-mRNA Xop(q oupd poty(A) e[orr(og LdOog repponollori. Evti 4 noLuo6evul(oo4 npooro- EnrmporeUer to nuplvtxri e[ootopdrto vovrot ono npotreivtroug ougnopoyoweg ot ono[ol entorpore0ouv ro nuplvrro e[ootr;polo. Iq (uUl, o oupnopoyovrog ervor ro ourr-. ,. xo TRAIVIP. H poly(A) notrugepool (PAP I Td4) rou TRAMP npoo0erer pro prrpfl oupo poty(A) oro 3'orpo rorv oroxeupevorv RNA. Etronroporrrd RNP . KoKodrntroUdvo RNA . Ad0og ene{epyoo(o 040 KEQA Al0 82 Iro0epdqro xor Xropo06qorl rou mRNA
  • 132.
    re6er ro yvnoromRNA, orrlv npqyporrK6rrlro Unope( v' onomoOeponote( ro xpuqd ooro0{ per6ypogo (cruptic unstable transcripts, CUT). Aurd ro RNA nou 6ev ru:drronororiv nporelveg (B).6ne en(oqg Evri- rrya 30.3) peroypdgowot on6 r4v RNA noAupepdorl II r<or 6ev rodrronoro0v ovoyvtop(orpro yov(6ro. 0o16- oo oulvd enrroLUnrowor (xor (otog puOpri(ouv) yov(- 6ro nou xtodrronorotiv npcoreiveg. To ouyrerpt;r6vo prerdyporpo noluodevuluilvowot on6 5vo ouoronr<6 rou oupnl6rou TRAMP (Trf4). Aror<p(vowor on6 dtrl.o gerdypogo Ue dyvtoon'l Lenoupy(o on6 u1v unrip;re- rprl omd0erd roug: r<ovovrx6 onodoporiwor on6 ro ofprnLoro TRAMP-e[oocopor(ou opr6ocog perd tr1 o6v- Oeoq roug, nt0ov6v ener64 moXe[owor on5 notruqde- vuL[oo4 e[oprriprev4 on6 rov Trf4. H rjnop[r1 r6rorov peroypdrprrlv onodeffiqre nermrrd yro npd:rr1 <popd oe metr61q (UUnS U' e).orrcoproru<q ono66pqoq RNA orov nuprlvo. Fldvtrl on6 ro rp(o t6ropro rov Uero- ypdgo:v qq RNA noLu;repdoqg ll gnope( v' oTrore- Loriv pr1 r<odrronorqru<d RNA Kor vo un6xerwot oe ypnyopn ono66prr1or1 on6 ro e[ooro;rdrro! Oprotrrdvo CUT go(veror 6l npo6plowor on6 r.!eu6ri 5vop[r1rqg peroypoqrlq Kor ro (6rq to BpoX[Bro RNA npoi6wo tunrrd 6ev epgov((ouv rdnoro Lertoupy(o (n.1 6e dpouv in frans). Om6oo, npoorp6pouv 5vo nopd6ery- po yro ro p6Lo ourrlg r<oOournq rnq petoypocprlg orrl priOproq yeuovxriv r1 enrrol,um6pevrov xrrl6rronorrln- xc6v yovr6(rov (B).5ne Ev1tqra 30.3). 22.9 O'EAeyxoq llor6rnroq Tnq Merdgpoonq Tou mRNA npoyporonolsiTol ond KurroponAoopcrrlKd Iuorfrporo Enrrfipnonq * 0 Ulx0vroU6g onoo0v0eorlg NMD oroXeuer mRNA pe npo- opo rodtrdvto reppono;.rou. * H oToxsuon Tov unooTptlUorov rou NMD onorter 6vo Drorri- prlpevo o0votro nporeiv6v UPF ror SMG. * Ie notrtrouq opyovrolroug, urtrer60, yro qv ovoyvtirproq npo- opov rordrxovlrov reppolopou e[vor 11 oouvq0roq 6opq r; ro oouvr'10Loro g1rog qg 3'UTR. Iro 0r1troolro, o U1xovropoq ovoyvrirptolg eniorlg o(totroyel ror rqv nopouo[o ro0odtrtov oupntrorov ouU0otrnq e[tovltrlv. e 0 Unxovropog onoouv0eorlg NSD oroXeuer mRNA Xtoprg ev- rog ntrotolou rordrxovlo repporropou ror poo((eroL o' evo ouvqplpevo ouvotro npotrelVdrv SKl. * 0 plXovLo;rog onoouvOeolg NGD oroXeuer mRNA pe 0T0orU0 ptpooopolo orqv rodrronotlrtrl neproXrl. Optoprdvo e(6q BoBc6v tou mRNA o[to].oyoriwor p6vo r<ord rr1 perdqpoorl. To ouorqproro ennqprlo4g e[e- LfiOrp<ov yro vo ewonf(ouv rp(o e(6q eontoprotu<rirv mRNA nou enoneil.oriv rqv ntor6rrpo rqg prerdgpo- onq Kor vo ro oroxeUouv yro ypnyopn ono66prqor1. H ': .r,i' i r.: i,,nopouord(et ro unofipoproto yt' ourd ro AAAAAM AAMAAA Ynoorp6poro rov rurropon),ooportr6v ouoqpororv enunplolq 0 gqXovro;rog NMD onodoper mRNA nou 6Xouv npotrrpo '-SLxovlo repgortopo0, nptv ono ro rovovtro. H onoouv0eol NSD onodopei mRNA Xtop(g evrog nLoroiou xo6rrovro reppolo;tou. H ono- , - J0001 NGD onodogei mRNA nou 6Xouv evo oroorlro ptBootrrgolo orrlv rodrxonorrlnxrl neproxrl. Jf .i] 0 'EtreYxoq llotorqrog rrlg Meroqpoorlg rou mRNA llpoyporonoreirot ono Kurropontroopolro Iuoqporo Enrqprlol_c 641 Buolxeg dworcg
  • 133.
    rp(o ouor{Uoro. Korro rp(o nepfioppdvouv ouppdwo ovr.i: 1.roLo u repporropori n1 g ;rerd<ppoorl g : ouvenrig, e[- vor or6nrprl pro enrox6nqon rnq depyoo(og rou qu- orotroyu<o0 rcpporrolloU (BMne Ev6qto 24.15). 'Orov 6vo ptBooro;rdno nou prerogpd(er rprdoer oro ro6116- vro repUorrol.ro0, 6vo (eUyog nopoy6wtov qnetreu06- poonq (release factors) (aroug europucbreg, eRFl xor eRF2) ero6pXeror orrl 06o11 A rou prBoocol-rot(ou, 11 ono(o rord rlv enqlfruvoq xovovxd e[vor efvor xoret- InUU6vn on6 eroepl6prevo IRNA. To oripnLoro rov no- poy6vtc,lv onel.eu0Sporo4q peoo).oBe( orrlv onetrsuO6proon rou oLoxLrlpcopSvou notrunemt6(ou rr Sneno rou mRNA, rov IRNA nou on6pewqv Kor roJV unopov66rov rou prBooroproilou. H onoo6v0eorl nou peoo)roBehor on6 perotrtr6lerg lr,rpig v64po (nonsense-mediated decay, NMD) oro- Xe0er mRNA nou nepr6louv 6vo np6opo r<rodu<6vro rspporropori (premature temination codon, PTC). Om6oo, np6copo r<ro6rr5vto repporrol.ro0 pnope( vo npox0rlouv ror pe d..trouq Unxovrogorig. l-ro nopd6ety- 1.ro, yov(6ro Xtop(q ;retoMd(erg pnope( vo 6qproupyq- oouv ovrilprol,o perdypogo nou nepdXouv PTC L6yro (1) ).dOoug qg RNA noLupepdoqq n (2) prrl olorl,rlpo- U6vnq, eogol4r6vqg 11 evoMor,ctxrlg oupporpqg. Erctprd- ror 6n qXe66v ro 50% rov evolloKrxd ouppoprprdvrov npo-mRNA 6qproupyo0v roul.dlrmov Uro Uopqn Ue PTC. l-1ep(nou ro 30% rov yvoordlv voooy6vrrrv oMq- Lo;r6prp<ov pdl),ov r<o6rronoro6v 6vo mRNA ge PTC. 'Evq r6roro mRNA 0o nopdyer nolunembro pe roLo- Bcop6vo C-rcllrd dxpo, ro ono(o 0eorporiwor r6rofuepo ro[trd yo ro rUnopo e(on[og qg rdorlg rouq vo Troyr- 6e6ouv noMd dMo 1.r6pro oe pr1 Lemoupyrrd oUpnl,o- Kq. To govondrt NMD Lerroupye( oe 6Loug roug euxopurilteg. H or6Xeuoq mRNAnou nepdlouv PTC npouno06ret vo ouprBe( perd<ppoorl xr efoprdror on6 6vo ouwqpqpt6- vo orlvoLo nporeivxc0v nopoy6w<ov, nou nepil.optBdvet nq rpe(g nptoreiveg Upf (Upf1, Upf2 ror Upf3) r<ot t6o- oeprg enmp5oOeteg nporeiveg (SMG1, 5, 6 xor 7). An' 6leq trg nporeiveg NMD, npc0rrl 6po 4 Upf1, 11 ono(o npoo6Sveror oro p$oorol.rdrto nou Bp(oreror oro rrodt- r6vro repporrolloU, er6x6repo oro oriprnLot<o rou TIo- pdyowo onetreuO6proorlq. H np6o6eo4 UPF orlproSe0et ro mRNA yto yprlyoprl onooUv0eorl. l-1op5rr or er6u<o( p6Lot rrov nopoy6wrov NMD nopop5vouv o6teuxp(vt- oror, roor6oo tdto(tep4 oqpoo(o 61er q googopul(ooq qg Upf nou e(vot npoo6e6ep6vq oro p$oo<rl;rdtto on6 rrlv Smg1. Or ouvduooll6veg dpdoerq rouq roto6tx6- (ouv ro mRNA yro ono66pr4or1 on6 ro yevrr6 prllovtopr6 onooriv0eo4q xot 6eye(pouv rn ypiyopn onoo6evuA(to- on. To mRNA or6Xot onodo;roriwor 16oo on6 to 5'-+:r" 6oo rqr on6 ro 3'-+5' ;rovondrr. l-ldrg 6prog 6roxp(vowor ro PTC on6 ro rovovu<6 t<t+' 6x6vro tepproloproU nou Bp[oxeror opretd nto rqOodr- r<d; O gqXovropr6g elet pretrerrl0e[ errevcbg 16oo mn ([pq 6oo Kor oro 0ql,oonxd, 6nou brorpdper rdn<rrg {El K0NA 22.14). Iro 0fli.oonxd, to r0pro o(pro nou rouro- nore( 6vo PTC e(vor q nopouo(o grog ouprBolqg oupporp{q, nou orlgo(vmot on6 ro oripnloxo ougpo- tr{g e[ovitov (exon junction complex, EJC) ro0o6x6 rou np6ropou ro6rrov(ou repporro;.ro0. Enerdrl ro ne- ptoo6tepo yov(6ro rrov ovri:repov suKopuordrv 6ev SXouv rwp6vro rou vo 6tox6mer qv 3' UTR, r' ou0ewt- xd xo6rx6vro repproropro6 6ev orotrou0o0wor on6 06- on ouppqqrlg. Kord rov rrporon6po yfpo rqg yer6gpooqq (pioneer round of translation) ev6q xovo- vtxoU mRNA, 6l,o to EJC Bp(orowor p6oo orqv xodrxo norqnrf neptoXr] rr ercron(owot on6 to 6repX6pevo , '',:'' , Auo plxovrolroi yto qv ovoyvri-rproq evog rodrrovlou repponogou og npourpou. (A) Iro O1trootuo, 1 nopouolo evog oulL- ntrorou ou;rpotrnq eltovur.rv roOodrro rou rtrldxov[ou repplonogou oroXeuer ro mRNA yro NMD. (B) Moltrov oe otroug roug europudlrac pro orpuotro Ueyotrn 3'UTR ovoyvrrrpi(eror ono qv onoorool ovopeoo oro roldrrovro repporropou ror ro oupntroro poly(A)-PABP. Ie xoOe nep(nrooq, rl npo.rrelvrl Upfl npoodSveror oro ptpoorrrpolo repgolopou yro vo nupodorloer rrlv onoo0vOeo1. 642 KE0AM|0 22 lro0epoqro rorXtopo0er4oq rou mRNA Avoyvtirproq PTC p6oro ro0odxori EJC Avoyvtilprorl PTC ond r1v no),rl gurprd 3'UTR
  • 134.
    " t-l -'- u[m,oFTffi- .,r' ,: .,: To onoretreopo rrlq npoodeolg ev6q PABP t<ovto o' evo npoopo rtodrrovro reppouolrou. 'Evo yovidro PABP gerorponlxe (rl01t v0 erqpo(et gto neptoXrl npoodeolg oe RNA evoq qoyou H neproXrl npoodeonq evoutpor60lre o' evo doupoorLxo yov[6to-uno- oTpopo yto NMD. To npoodedepevo PABP epnoOroe rr1 ouv101 yplyopl onodoprlorl ourou rou mRNA ono ro prqXovro;.ro NMD. Aurrl q p6- 0oDog eXer notrtreg eqopgoy6q orn goprorrl protroyro. OXr NIVID t,'t . prBoorotrrdrro. Kord rov nporon5po yUpo qg prerdqpo- onq yro 6vo un6orptrrpo NMD, or nprore"[veg Upfl xor Upf3 npoodSvowor oro(o) xo0o6x6 rordlorno(o) EJC, oroXerlowog ro yro ono66prqor1. To neproo6repo yov(6ro rou S. cereyr.srae 6ev 6ro16- firowor ro06],ou on6 twp6vro, on6re o prqXovtotrr6g ev- ronroprori rov PTC np6ner vq e(vor 6rorpoperrr6g. Ie ourn rrlv nep(moor1, orlpro rrv6rivou e(vor ;uo orpriotro peyoln 3' UTR. Aur6 ono6etX0rlre on6 to erjp4pro 6tr r1 entg{ruvo4 rnq 3' UTR ev6g quoroloyrKorl mRNA 0o pnopo0oe vo ro Uerorp5r{.rer oe un6orpo;ro yro NMD. 'Evo rp61ov ;rowSlo npore(ver 6rt o ooor6q repUorr- op6g rqg prerdgpoo4g o' 5vo ro6rx6vro reppronoproU npo0no06rer 6vo orlpo on6 6vo r<oww6 PABP. MoLov6rr or 3'UTR 61ouv noLri 6rorpoperx6 pnKoq, q <puou<r1 on6- oroon peto[ri rou ro6rrov(ou reppotroprorj ror qg poly(A) oupdq 6ev e[oprdror onoxlerorxd on6 ro Un- xog roOc0g 6eureporoye(g 6op6g xor oMqtrsnr6pdoerg pero[ri RBP prnope( vo rrl oupnr6oouv. H ovdyr<4 yro PABP ono6etl0qr<e oe noMorig opyovrogo0g npoo66- vowoq pro PABP r<owd oro PTC (B).6ne EIK0NA 22.15), on6re ro ow(ororlo mRNA 6e oroXeu6rov n).6ov yto NMD. En(orlg, 4 ovoyvrirproq rou PTC ouprBo(ver ove[dp- rqro on6 q oupporpq or4 Drosophila, o"ro C.elegans, oro rputd ror oe optopr6vo mRNA rrov 0qLoonxri:v, yeyo- v5g nou uno6rlLrilver 6rr ro ptixog ror q 6opq rnq 3' UTR e(vor r<p(oq.ro yto rnv quoroLoyrrl 6ro6troo(o repporr- ogoU qg perdqpoorlg oe 6Loug roug opyovroporig. Opropr6vo guoroLoymd mRNA orolefowor on6 ro g4Xovropr6 NMD. Aurd ro mRNA routonor{Oqxov on6 nerpdproro mo ono(o elorc60rlxov ro en(ne6o rqg Upfl, pe onor6l,eopo peydlrl oU(4oq ev6g unoouv6- Aou geroypdg<ov. O xotdLoyoq rov euor.ol,oyxriv unoorpoprdrov yl.o NMD nept}opBdver pr6pro mRNA pe r6rokepo peydLeg 3'UTR, ro mRNA nou ro6rronoroUv oeArlvonporeiveg (or ono(eg Xpnoqonoroilv ro ro6u<6- vto repUorrol.rorl UGA rog xrodx6vro oelqvoxuoretvqg) xt Svov dyvocro opr0pr6 evqMoKrmd ouppogpr6vov m- RNA. Me Bdorl 6oo yvrop((oupre pr61pr oriprepo, 6s pno- po0pre vo npoBL6goupre 6Lo to unomptilUoro nou onorelorlv m6Xoug NMD. To NMD (oog ono6erX0e[ 6vo oqpowrr6 povondrr yptiyopns onooUvOeoqg yro notrfi.o BpoX0Bro mRNA. Teprlrl 06o11 nprjodeonq H onoorjv0eorl non stop (non stop decay, NSD) oroleUeL pr6pro mRNA lrop(g ew6g nLoro[ou xrodtr6vro teppono;.rori (peoo(o oxnpo orqv Eu<6vo 22.13). E[or- r(oq qq onorulfog reppotrol.rori, 6vo prBoootgd-no ou- veX((er vo preroqpd(er ornv oupd no).u(A) r<oL nL0ov6v orol.rord mo 3' drpo. Ynoorpor;roro rqg onoo6v0eorlg non stop 611;;oupyotiwou<up(cog on6 np6copo repUol.- op6 qg peroyporpqg ror noluo6evul(roo4 orov nuprl- vq. T{itoro np6opo no}.uo6evu},rcop6vo prerdyporpo e(vor nopd6o{o rotvd. H ovd}.uor1 tulo(otv n}.r10uo;rd;v oDNA or ono(or npodpxowov on6 mRNA qg (Upr1g r<or rou ov0pc6nou uno6rlLrriver 6n ro 5-10% rrov ouprBdv- rolv no).uo6evu).(roo4g (orog ouprBo(vouv oe ovo6tr<d "KPUtPd o4pe(o" to ono(o prord(ouv pe to ou0ewu<6 or1- po nol.uo6evuL(oor1g. H or6Xeuoq rov unoorprr;prdrrov non stop nepil.o;rBdver 6vo oUvoLo nopoy6wurv nou ovogd(owor nporei'vsg SKl. To prBooroprdrto oneleu- Oepcbveror on6 to mRNA pe q 6pdor1 qg Ski7. H SkiT 6,1e1 pro neptoXrl GTPdo1g nop6;roto Ue rou eEF3 ror nr0ov6v npoo66veror m4 05oq A rou prBoocol-rot(ou yro vo deye(per rrlv onetreu0Spcooq. H enor<6Lou0r1 enr- mpdreuo4 trov un6).orn<ov npoteivriv SKI xor tou e(o- orrrpor(ou 6Xer og onor6Leopo rqv onoo[voeoq rou mRNA pe xoterjOuvorl 3'+5'. H onooJv0eorl rcov uno- otprrlpdtov non stop prnope( en(or1g vo ouprBe( onouo(o r4g Ski7, on6te xor npoyporonore(tor pe ogo(peo4 r4g r<oLrjmpog ror n5r1.rr1 oe rqreUOuvoq 5'-+3'. H euoroOq- o(o mr;v o<po(peor1 qg r<o).rinrpog 0o pnopo0oe vo o<peftetor o' ercr6nroq rorv PABP xo0rig to npcoron6po prBooroprdno 6rorp5Xet qv oupd poly(A). H VpnVopn onoo6v0eoq rov unoorpto;.rdtrov non-stop o6qyei 61r p6vo oe npooroo[o on6 ro[u<d no],unem(6rq oMd r<or oe oneleu 06pooq noyr6eu p6vcov prBooro prot[ov. Ev6ro- g6pov efvqt ro yeyov6g 6rr ro E. coll lprlotponore( 6vo e[er6rxeugSvo pr; r<o6rxonorqtrr6 RNA (tmRNA) ro ono(o 6po 16oo ctg IRNA 6oo Kor oq mRNA yro vo 6to- ocioeL ptBoorrlprdrro moprorqprSvo o' 5vo nonstop mR- NA. To tmRNA xqreu06ver qv npooOrp<q ev6g pu<pori nenl6(ou nou moleUer ro eLorrol;-torLr6 npoteivtx6 npoi6v yr' ono66pu1or1, nop5ler 6vo xro6rx6vro reppol- o1-roU nou ennp6ner r4v ovor<tix),ooq rou prBooropor(- ou Kor oroxeUer ro e),ortco1lolr<6 mRNA yr' ono66pu1orl on6 r1v RNAo4. TeXvrltd PABP ge v6o neproXrl npdoDeolg :I.-q 0 'EtreYXoq l-lotoqrog rrlg Meroqpoolg rou mRNA l-1poy;loronoLeuor ono Kurropon),oogolro Iuorluoro ErLrc-:-l 643 l-lpoo0ebepdvo PABP
  • 135.
    H onoofvOeol no-go(no-go decay, NGD) mo- Xeier pr6pto mRNA nou 61ouv ordoqlo ptBoorrlpdno oe ro:6u<6vrq u1g rro6rronorrlru<r1g neproXrlg toug (rdro oXnpo oqv Ex6vo 22.13).1-1opo6u<r1 r1 noporcropr6vrl oroorg6rqro prnope( vo npor<iqOe( on6 <puou<6 Xopo- rcqprolx6 opropr6vov mRNA, 6no4 roXupSg 6eutepo- toye(g 6opr6g xor ondvro r<rrl6rxdvto (rrov ono(<ov r' owimorXo IRNA Bp(or<owor or XopqLdg ouyr<ewpdr- oerg). Aur6 ro v6o povondrr enrrnpnonq 6Xer pe),eqOef pr6vo orq (r1;r4 r<or e(vor ro trry6repo xorovoqr6 on6 ro rp(o. H m6leuoq rou mRNA nepd.oprBdver rrlv enr- orpdreuoq rov nporeivriv Dom34 rot Hbsl, or ono(eg e(vor o;r6l,oyeq rov eRFl r<or eRF3, owrmo(Xoq. H ono66prqoq rou mRNA op{(er p' rivo ev6ovoux.,eol.ul- 16 r6rptpo r<or ro 5' r<or 3' xldoproro ndmowor on6 ro e(coorrlprdrro Kot rIV Xrn1. H Dom34 0o pnopoUoe vo e[vor 11 ev6ovoux.l,edoq, erp6oov truo on6 ng neproXSg ng ;rord(er pre vour<Ledo4. l-ror( 5vo rovovu<6 mRNA vo riXer oMql,ou{eg nou e(vor 60oxo},o vo Uerqgpooro6v ror pnope( vq ro rorodxdoouv oe yprlyopq ono66prr1- o4; Trftoeg oMqLouX(eg 0o gnoporioe vo 0eopq0oUv rrlg 6vo dLLo e(dog onomoOeponor4-nro[ morle(ou. E[eLurtrd, n 6roqpnon epnoS(tov yt' ono6orrrrl perd- gpoon uno64Lrilver 6tr e[unrlpetoUv rdnoto o41.lowrxr1 l,enoupy[o mov 5trey1o rou 1p6vou npr(ofq owdrv rtov mRNA. 22.1O Meprrd Evronl(ovror oe Kurrdpou Europuorrrd mRNA Er6rrdq lleproxdq rou " 0 evronrogog rov mRNA e[unlperet drorpopelreg treuoup- yieg oe pegovtripevo rurropo rol ovonruooolrevo 6pppuo. * 'Exouv roroyporpeI Tpsrq Urlxovropoi ylo rov evronropo rov mRNA, + 0 evronlop6g onotreI cls-ororXe(o oro mRNA-oroXo ror Irans-nopoyovreg. * 0 rupLoq plXovropoq evepyou perorpopog neprtroppover qv roreu0uvopevl r[v1or1 rcrlv mRNP roro plroq oddrv Tou KUT- ropooretrerou. To rurrop6ntroopo e(vor rivog noLuo[lvooroq 16pog nou roto).oprBdveror on6 uQq).q ouyr<6wpolon npo- reivov. E(vor dyvr.ooro nog prnope( vo droXSowor to aLerjOepo noluoopdrto Kor ro neproo6repo mRNA prdMov prerogpd(owor oe tulo(eg 06oerg or ono(eg xoOop[(owor on6 ro oqpreio ero66ou rouq oro Kurro- p6ni.oo;ro Kor rrlv on6oroo4 r;v onofo SXouv 6rovri- oeL on6 ow6. Opropr6vo mRNA piero<ppd(owor pr6vo oe ouyKeKprtr-i6vo oqpre(o, xo06g 11 pretdrppoo4 roug r<oromSLLetor ;rdXpt vo rprdoouv mov npoopLo;.r6 toug. Pu0pr(6pievog ewonroprdg dXer nepLypoge( yto ndvol on6 100 erdrrd mRNA, opr0p6g nou oorpo).rog owrnpoooileUer 6vo pirrp6 trr6vo r<),do;ro tou ouv6- 644 KEOAAAI0 22 Iro0eporrlro rot Xtrrpo0eqol rou mRNA Aou. O ewonrql6g tou mRNA e(unrlpere[ drdgopeg oqprowu<5q l,eroupy(eg o' euKoputorrr<orig opyqvr- o1-rorig 6l,rov rrov r0nrav. Tperg xp(orpreg Lenoupy[eg nopouord(owor ornv ElK0tlA 22.16 r<or ou(qrofwor oq ouvSXero. 1. O ewonropr6g erdu<cOv mRNA oro ooxUfiqpo notr- Lrirv (tirrov e(un4pere( t4v eyro0(6puor'1 peMowu<rilv nporUnov oro 6prBpuo (6nrog o d[ovog no].rr6rq- tog) ror op(er ro ovomuftor6 nenpop6vo rurr6- pcov nou edpe(ouv oe drorpoperrr<d o4pe[o. To ewonrotrrdvo prqrprrd mRNA rro6ronoro[v peroypo- gu<o0g nopdyoweg 4 d),Leg nproreiveg or ono[eg puOpr((ouv rqv 6r<gpoo4 rrov yovr6i<ov. Ito rrroxUrro- po rnq Droshophila, ro mRNA rrov yovrd[rov bicoid Kor. nanos ewon((owor mov np6o0ro ror on(o0ro n6lo, owroro(1oq, xor 4 prerdgpoorj rouq nou oKo- l,ou0e( rq yovtporro(r1or1 SXet cog onordl,eopo q 64- l.uoupy(o 6roBo0p(oeov rcov npo:teivrx6v npoi6wrov roug. Or 6roBo0p(oerq Xpnorponorodwor on6 ro xrirropo oe opXrxd md6ro rrlg ovdmu(r1q yro rov npoodroprol.r6 qg npoo0ron(oOrog 05oqg oro dp- Bpuo. To bicoid r<ro6rronoe( 6vov petoypogrr6 no- pdyovro, evcil to nanos Svov xorooroL6o tqg perdgpooqg. Opropr6vo mRNA nou ewonf(owor oE er6u<dg O5oerg rro6tronoroUv roOoprmdq rou ne- npoprdvou rou rurrdpou. l-to nopdderypo, ro mRf.lA rou yovr6(ou oskar ewon((eror oro on(oOro pr5poq rtov rooxurrdpov Kor nupodore( 14 6repyoo(o nou o64ye( m4v ovdnruEn rcov opX6yovrov BLommdrv xurrdpov cro 6prBpuo. 2. O ewonrop6g rou mRNA en(o4g no((et pSLo mrg ooril-rperpeg 6rorp6oerg tou rundpou, 6r1Lo6r1 cng trrm<olr5g 6rorp6oerg ot ono(eg ono6(6ouv 0uyorpud rUrropo nou 6rocp6pouv ro 6vq on5 ro dMo. Atn6 enmuyldveror pe oofprpetpo droXropropr6 rcov xo0e prorrilv rou nenpopSvou rrrlv xurrdprov, ot ono(or pnopef vo efvot nptoteiveg xo/ri ro mRNA nou nq r<co6rronororjv. Iro dprBpuo qg Droshophila, rc mR- NA rou yovt6(ou prospero ror ro ow[ororXo npoi6v (5vog geroypo<prr6g nopdyowog) ewon((owor oe pro neproXrj rou neprqeperqroU rplotori rou ep- Bpriou. Ie preroyevSorepo ord6ro r4g ovdmulnq, n npooovoroltop6vrl xurropx4 6to(peoq rov veupe BLoorrilv 6rooqo).((er 6n pr6vo to e(c(lroro Ouycnpr- 16 rUrropo 0o nepr5ler prospero,6eopreriowdg to vo yfuer rivo pr4rpu<6 rUrropo yoyyl(otv. H oo0p4re- rpq rurropxri 6ro[peoq Xcnoqonore(ror en(oqg on6 rn (UUn yro q 6rlpuoupy(o ev6g Ouyorpxotl rurr6- pou dro<poperxo6 ou(eurcnrotj rUnou on6 to grppr- 16 rrjrropo. 3. O ewonropr6g rou mRNA orouq 6rorpoponorr;pr6- vouq Kurroprxorig t6noug tcov eql,(r<o:v e(vor Svog prlXovropr6g yro rn 6ropeproprorono[qor1 tou rurr6- pou. O ewonrog6g [orog Xpqorpronore(ror rilore vo 6toogol.rme( 5tr ro oumonxd trrrv nol,unpcureivr- rc0v ouprnL6rtov 0o ouw(Oewor xowd ro 5vo oro dMo ror 6rr or npotetveq nou oroXeUowot oe opyo- v(6ro ri e[erdu<euprSveg neprol6g rou rurrdpou eni orlg 0o ouvr(0evror or Kovrrv6g O5oerg. O Boorxeg evvoreg
  • 136.
    EIK0NA 22.18 Tpetgr0pteq tretroupy(eg Tou evTontopou rou mRNA. IXnUottoUriq nporunov r<ot npodtoypoqrl rou nenpogdvou toorur-roPulv ror egBpriov AnUroupy[o 6totpoPettrtlv 0uyorprxtilv runopotv roro rrlv oouggerpq 6ro[peoq rou runoPou Arogeproporono(qor1 rou rurropou o' e[erdrxeuPdveq neproydq ./ @ @ ewonroU6q rou mRNA e[vot tdto(tepo orluowtK6q yro r0rropo pe noltK6rnro, 6noq ot veup6veq. Evd) ro neproo6repo mRNA Ueroqpd(owol' oro Kurrqpt- x6 ocirpro rou veup6vo, flo^.Id mRNA ewon(owot qro veupdqovo Kot rouq 6ev6pkeq. >e ourd flept- LoprBdvetor xot to mRNA rnq p-oKrivnq, rou ono(ou ro npoi6v ouprpret6Xet orrlv ovdmu[q tolv 6ev6ptttirv Kor rov veupo(5vttv. To mRNA rnq p-oKrtvnq ewo- n((erot oe O5oerg evepyo6 petor<ffioqg oe UeY6l'n norxil(o rwqrdlv run6pov. O ewontopt6g tou mR- NA oe veupu<5g getoouvomtr6g O5oetq qo[vetot 6tt e(vot onopo(trlroq Yto rtq rpononoqoetg nou oXetL (owot pre q prd04or1. Iro rfrropo qq ytro(og, ro mRNA rrlg Booxriq nporetvnq qq pruel'ivr1g (myelin basic protein, MBP), ro ono(o t<todtronop( dvo ou- orotrr6 rou el[rpou qq pueL(vr1q' ewon((etot o' 6vo ouyxerptpr6vo 6topr5ptopo 6nou ouw(Oerot pue- L(vr;. Ito <purd, to mRNA ewon((owot orq 9).otdr6r1 nepropj rou rurrdpou. Meptxdq eopdq, o ewont4t6g tou mRNA nepilopt- Bdvet pretoqopd on6 6vo rUrropo o' 6vo 6I'o. Er1 Drosophila, to pqtptxd mRNP ouw(Oewot Kot ouYKev- rpd:vowot oe neptB6trowo tporpodorxd r0nopo rot preroq6powot oro ovonruoo6 gevo olor0nopo pr6oul xurroponlool.tottrc0v dto[],ov. To tputd e[dyouv RNA droprSoou ttov nl.ool.todeopdttov Kot ro Uoroq6pouv oe proxpw6g onoordoetg pt6oto tou oyyetororj ouoni;rotog rou gLouiprotog, Meptr6q qopdq, ro mRNA ;reto96- powot pro(rd oe xoxxio mRNP. H oUorooq outtiv rov xorr(ov nopopdvet o6teurp(vtorq. 'EXouv r<otoypocpe( rpetq Unxovtqro( yto rov ewoTlL- opr6 tov mRNA: o To mRNA t<otov6getot o;rot6prop<po oMd ono6oge(- ror oe 6treg ng 06oetg erc6g on6 rn e6oq qg pretd- Qpoonq. o To mRNA gnope(vo dtolSetot etre0Oepo otrtrdnoyt- 6e0eror orn 06on trlg pretdrppootlg. o To mRNA petoq6perot evepYd mrq 06oetg 6nou 0o geroEpome(. H evepy6q pretorpopd e(vot o r<riptog prrllovtqr6g yto rov ewontop6. H trreroqopd enmuyXdverot Ue rrl pero- r6ntorl rrvqrrlpttov npoteivr6v xotd prrlxog o6tilv rou xunopooretrerori. Itrg olettr5g drepyoofeg, o(tonot- o6wot Kot ot rpetq t0not ptoptot<drv r<wryriprov: 6uvetveg ror rwrlo(veg, ot ono(eg ro[t6etiouv rord ptirog rtov Ut- r<poocotrqv(oxov npoq ow(0eteq roteu0Uvoetg, Kot UUo- o(veg ot ono(eg to[t6eUouv r<otd prir<oq ttov wt6(rov rqg qlctvnq. Aut6q o rp6nog ewontopoti qnore( routrdlt- mov 4 ouorotu<6: (1) crs-motXe(o cno mRNA-or61o, (2) trans-nopdyoweq nou dpeoo ri 6prpreoo npooddvouv ro mRNA orrl oolorri r<wqtrlpto npoteifi, (3) trans-no- pdyoweq nou xotoot6trLouv tq ;retdqpoorl rot (4) 5vo oU arrl pro oy ru po B6Lr1 onq ornv ent0 u prrl( tonoOeo(o' M6Xpr oqgepo 61ouv XopoKnlptfie( pr6vo Liyo cis- orotle(o nou peptr6g Qop6q nopoprotd(owot Ue roXU' 6polirxorig xdrdlxeg (zipcodes)' To motXe(o outd no,r<ilouv rot, nepil,opBdvouv t6oo tdto(tepeq 5op6q 6oo t<ot tdto(tepeg ollrll'ouX(eg RNA nou pnope( vo Bp(orowot onou6rlnote oro mRNA, nop6tt ouvriOcoq ewon((owot orrlv 3' UTR. H ovoyvtilproq toug r;rov 5U- or<o1,4, prdMov enetdrl noMd onotel'o0wot on6 nept- nLordg dewepotoye(g rot rpnotoye(g 6op6g. l-lotr'71'o( frans-nopdyoweg 6Xouv ouoletore( pe tonrrrl UerqQo- p6 mRNA xot xoroorol{ qg prer6qpoorlg. Optopr6vot e[vot ouwrlprlpr6vor oe dtotpopettxoUg opyovtoprofg' l-ro nop66etygo, 11 nporeiwl sfaufen nou npoo66verot oe 6(rtrovo RNA eprnL6retot orov ewontop6 mRNA sro coor6rropo rrlg Drosophila rot tou Xenopus, r<oOdrq en(- orlq Kor oro veuprr6 o0orrlgo qq Drosophlla, tov 0r1- tromtr6v ror (oog rov oxoulrlrttilv t<ot tou rfopto6- ii t?."1S Meprro Euxopuortxo mRNA Ewoni(ovTqt 0t Etdtxriq l-leptoyec rou Kuncoou 645 ^^iA
  • 137.
    (5ppq.Aw6g o notrurdl,qwognopdyowog 61er noMo- nldq neproldg or ono(eg ouv66ouv oritrlntror<o oe Uovo- ndtto petogopdg nou e[optrirwor 16oo on6 orr(vr1 6oo xor on6 pu<poorrrL4v(or<oug. IXe66v ilnoro 6ev eG vot yvroor6 yrcr ro r6topro onopo(rr1ro ouororx5, 611- Lo64 rouq prrllovroproUg oyrupoB6Lrloqg. A0o nopo- 6efyporo pr4lovrop6v ewonropro6 nopouord(owor orrl ouv6Xero. O ewonropr6g rou mRNA qg B-orcr(vqg 6Xer ;releq- 0e( oe tvoBldfieq Kor veupriveg oe xoMdpyero. O ,,rq- lu6poptr6g rc06rrog" e(vor 1.rro ol),qLou1(o 54 vour.treold(rov ornv 3' UTR. H np6odeoq qg npo:rei- v4g ZBPI oe ourl rrlv oMrl).ou1(o oe ouvepyoo(o pre rn Ueroypoqrl e(vot onopofurlreg yro rov ewonropr6, ye- yov6g nou unodrllc0vet 6n to mRNA r4g B-orrr(vr1g 6e- ope0eror oe 6repyoo(eg evtonroprori nporori rov unoBl,q0e( o' enel,epyoo(o r<t e[o10e( on6 rov nuprlvo. E(vor ev6rocpdpov 6t o ewontopr6g rou mRNA rnq B- omfuqg e[optdror on6 or6poro rv(6to orcr(vqg oflq rvo- Bl,doreg ror ox6porouq UrKpoooL4v(oxoug orouq veupc0veg. H yevenrf ovdLuorl rou ewonrogorj rou mRNA rou yovtS(ou Ash'l qg (0p49 npooSeepe rnv nro otrox).q- prop6vq dnotfq yro ro prrlXovropr6 rou ewonroproti xor nopouor6(eril ornv ElKOilA 22.1?. Kord rrlv exBLdorq- orl, ro mRNA rou yovtd(ou Ashl ewon((eror orov ovo- muoo6pevo Bloo16, on6re nporeivq Ashl ouw(0erot pr6vo mo veooxnporropr6vo 0uyorptr6 rUnopo. H Ashl e(vor rorooroL6og rnq Ueroypogrlg nou onoyo- peUer rrlv 5r<gpoo4 r1g evSovouxledoqg HO, puog r Evronroptog rou mRNA qg Ash1. To veoe[oyope- vo mRNA Ashl npoorotrtroror oqv uvlrlpro Uuoo[vn Myo4 Ueoto evog oupntr6r0u Ue Trq nporetveg She2 ror Sh3. H xrvqrlpro nporreivrl peroqeper To mRNA roro pdrog rov rvrdlrrlv orr(v1g rou ovonruooopevou ptroorou. 646 KEOMA|0 22 Iro0epoqro rorXtopo06rrlorl rou mRNA npordvqg nou onome(tor yro rrlv evoMoy4 rou ou(eu- rrrrof rUnou (B].6ne Ev6t4ro 15.20).'Eror, evoMoyrl ouprBo(ver p6vo mo p4rprr6 xUnopo. To mRNA rrlg Ashl tpdpet mqv rodu<onotrlur{ neptol( r6ooepo ororXe(o ewontotrrorj pe doprj p(oXou-0q),r69 oro ono(o npoo66veror q npotetvrl She2, prdMov orov nuprlvo. H npcoreivrl She3 Lerroupye( <og npooqppoorqg nou npoo65veror 16oo mq She2 6oo ror orrlv xrvqrrlpro npu:tefvq Myo4 (en(or1q yvoorri Kqt oq Shel). H npo- reftrr1 Puf6 npoo66verot oro mRNA, roroorSMowog rq perdqpoorl tou. H xrvrlrrlpro np<ote[vq perocp6per ro mRNP qg Ashl xord prp<og rov notrxriv wr6(cov orr[- vnq nou o6qyoUv on6 to ;r4rprr6 rfrropo orov ovo- nruoo6pevo Bl,oot6. Enrnp6o0ereg nporetveg onoro6wor yro ro otoqr6 ewontopr6 Kqr rnv Sxgpoorl tou mRNA qq Ash1. l-ldvrrr on6 20 mRNA rnq (6gnq Xprlorponororjv ro (6ro ;rovondn ewonrotrroti. MrlXovrol.ro[ ewonropo0 or ono(or dev nepd.oprBd- vouv evepy5 petogopd 61ouv onoderX0e( oogcig pr6vo yto L(yo ewonrop6vo mRNA mo roorUrropq Kqr rq npcbtpro SpBpuo. O pr4lovropr6g qq ronrrqq noy(6eu- o4g droxe6prevcov mRNA Boo[(erot or! oupperofn oy- rupc0v nou 51ouv ronoOerq0e( er tov npo-r5polv, or ono[eg 6puog 6ev 61ouv touronorrlOe(. Iro coorrirtopo qg Drosophlla, ro 6roXe6prevo mRNA rou nanos nqyr- 6eUovrot oro on(o0ro uBl,oorrrd ntrdotrro, (germ plasm), pto e[er6treupr6vq nepropi rou rurropon],6- oUoroq xdro on6 ro gLor6. Iro <r:orUrropo rou Xeno- pus, ro mRNA nou evron((ovrqr oro gurrr6 n6lo npriro noyrdeUowor oe xdntog pruor4prc66q, pregBpovo- Bpr0rl 6opn yvomt| org purolovdpror5 vdEoq (mito- chondrial cloud, MC), q ono(o opy6tepo perovoore6er oro qurtr6 nSLo ouprnopooripowog ro mRNA. O UnXo- vrop6q tonrrrlg oro0epono(r1onq rou mRNA 51er nept- ypoge( yro 6vo mRNA ro ono(o en(or1q ewon((eror orov on(o0ro n6lo tou epBpriou qg Drosophila. Zto nprilrpro orddro rqq ovdmu[r1q, ro mRNA rou yovr6(ou hsp38 r<otov6;reror opror6propgo oro e;rBpuir<6 rurro- p6ntroopro, otri.d opy6repo onodopefuot nowo0 err6g qn6 rov n6Lo. H nporeivrl smaug epnl5reror mqv onooroOeponofqor1 rov neproo6repov hsp38 mRNA, rup(og enrorporeUowoq ro otipntror<o CCR4/NOT. Aev e(vor yvoor6 nrig 6rorpe0youv ro mRNA nou ewon((ov- tor orov n6tro. 22.11 [IepiAn$n To rurropxd RNA e[vor q4errrd ooro0rl p6pro ],6y<o rnq nopouo(og runoprrc6v prBovoux).eoo6v. Or prBo- vouxl,edoeg 6to<p6pouv rrlq npoq rov rp6no en(0eor1g ror e(erdrreUowot yto 6rocpoperrr<d unompdrgoto. Aurd ro dv(u;ro ono6Sprqorlg rou RNA 61ouv nol- Lotiq p6loug oro rUrropo, prero(0 dl,l,cov Kor rnv onooUvOeoq tou mRNA. To yeyov6g 6rt ro mRNA e[- vor BpoldBro enrpdner rn ypnyopn npooopproyrl rou <pdoprorog r<,r:v nporeivcbv nou ouw(Oewor on6 ro rUrropo, puOprl(owog qv roXUrrlto rnq Ueroypoe]iq H nporetvrl She2 npood6verot mn doUn U(oxou-0ntudq rou mRNA H nporeivrl She3 ouvd6er qv She2 ge q puoo[vr1 H Myo4 npoo66veror o'5vo w(6to onfuqg lv(Dto orr[vqg
  • 138.
    rov yovr6(ov. Ayyelroq6poRNA pe drorpopettr( oi,- LqlouX(o eprgov((ouv notrU 6rogoperrrri euoto0rlo(o or4 6pdoq rov vour.treqod)v, pre lp6voug nptEorliq nou norril.ouv xqrd 100 gopdg r<or neproo6repo. To mRNA ouv66etot ;r' 6vov petoBoLi.6pevo nLr10u- op6 npu:teivriv xord rqv cop(provo( rou mov nup{vo r<ot q (o( rou oro rurrop6nl,oopro. Yndplouv ndpo notr)69 nporeiveg np6o6eoqg oro RNA, or neproo6re- peq on6 trg ono(eg 6ev 6Xouv or6pro Xoporrqptme(. l-loM6g npcoretveg ;re p6Loug orov nuptlvo onoBdMov- ror nprv ri rord qv e(oyoyrl rou mRNA oro rurrop6- ntroo1.ro. AMeg ouvo6eUouv ro c0prpro mRNA t<ot 6pouv Kor oro rurrop6nloogo. To mRNA ouv66owor g'eu- 6r6rpro, oMd oM4lenu<ol.um5pevq, oUvo^.o np<otei- vdrv (RNA-binding-proteins, RBPs) nou trerroupyo0v orrl gerdqpoon, tn oro0ep6rqro Kor rov ewonropr6. H oprd6o tov mRNA nou protpd(owor 6vov r6ro(repo t6no RBP ovorpdperor Kor. cog RNA peyroutr6vto. H ono66prqoq rou mRNA opX(er Ue rnv orpo(peoq ev6g nupogroogoprrori on6 ro 5' dr<po. Aur6 to Brlgo nupodore( 6vqv xUKIo ewopuilv on6 ev6ovourtredoeg, nou orotrouOe(ror on6 3'+5' e[ovour<],eoAunr{ n6rpr1 r<ov el,e60epcov xtrooprdrcov. H 6o911 g(oXou-Oqhdq oro 3' dxpo nol,trrilv mRNA rq npoororeUet on6 3' ent06- oetg. H 3'-+5' evepy6rqro e[ovoux].edoqg 6reuro].Uve- rot qn6 rqv nouo6evul,(ooq rrov 3' 6xptov, q44;ror((owoq Uro nLorrp6ppro yro ro 6v(u;ro. Or r<0- preg npotetveq nou epnL6rowor mqv ono66ptrlorl rou mRNA trenoupyoriv rog o6prntroxo yvroor6 pe tov 6po oropdrro ono66prrlo'r19. H ono66grlon rov neptoo6te- prov mRNA orn (rjpn, nt0ov6roro Kor ficl Oqtrooru<d, onone( cog npri:to Brlpo illv qnoo6evutr(@o4. H ercre- vrlg Bpdpvorl qg oupdg poly(A) ennp5net vo e[e].t1- 0e( 6vo on6 ro 6rio govondrto ono66prqor1g. To povondrr 5'+3' onoorivOeorlg nepil.optBdvet oqo(peoq rrlg ro).Umpoq Kor 5+3'n6{q on6 e(ovour<}.edoq. To govondrr 3'-+5' onoo0v0eonq roro.l.Uerot on6 to e[co- orr:pdrro, 5vo peydLo orj;rnLor<o e(covouxLedonq. H pe- rdrppoorl Kor rl onooUv0eoq p6oco rou 5'-+3' povonorrori e(vor owoytovtmrr6g 6repyoo(eg ener6ri t6oo ro oritrrnLor<o 5vop[qg qg prerdgpooqg 600 Kor to 6v(utrro ogo(peoqg rrlg r<otrUnrpog npood6vowor cr4v ro).0mpo. To oroprdrro ene[epyoo(og neptSXouv mRNA r<or nporefveg nou eprn].6Kowor orqv onoo0v0e- orl Kor rrlv Koroorol{ qq perdqpooqg rot nrme0erot 6l e(vor oqpre[o onod6gqorlq rou mRNA. 'E1ouv neprypoge( dMo rdooepo provon6rro ono- dSUnonq nou oroXe6ouv er6xd mRNA. Ko0dvo Xpnor- ponore( rov (6to prrlXovropr6 ono66grloqq Ue ro povondrro nou e(oprdrwot on6 rqv onoodevuL[oor1 oMd opX((er drorpopertrd. Iuyrexprpr6vo, opffiouv pre (1) ogo(peor1 qg rol,0mpog ove[dpqrq on6 rrlv qTro- odevuL(u:oq, (2) npoo0rjrq puog oupdq noLu(U) (3) ev- 6ovour<Leol.uttx5 r<6Qr;ro er6tr<6 yro ouyxexpr;r6v4 olrlLouX(o/6oUn (4) (euydpropro Bdoeov pre pr6pro mi- croRNA. Or 6togop6q orouq Xp6voug npr(oiq ttrlv mRNA ogeil,owor oe er6xd cis-morXe(o pr6oo oro mRNA. 'E1ouv neptyporpe( ororle(o onomoOepono(r1or1 g ror oro0eponofionq. Tq neproo6repo ewoni(owot ornv g UTR xor Spo6v toq oqpe(o np6o6eoqg yto npoxefveq 4 microRNA. To ntroriotq oe AU motXe(o (AU-rich ele- ments, AREs) onooroOeponototiv peydlo opt0p6 po- p(ov mRNA oto xrltropo rov OqLoorrxc0v. Ot npu:teiveq nou npoo66vowor oro orotle(o onooroOe- ponoqorlg (ocog 6pouv rup(og entorpoteUowoq opt- opr6vo ouorolrd rou pnxovtoprorj ono66gnonq. H oro0ep6qto rou mRNA prnope( vo puOpr((etot onov- tc0woq oe KurrqprKd oqproro pe rponono(qoq tcov n poo6ev6 prevov n potetvcilv. YndpXouv ouorrlporo ennrjpqoqg rot etrdylou not- 614ro9 nou lemoupyo[v otov nuprlvo rot to xuttop6- nl.oo1.ro r<or oroXeUouv elonopottrd pr6pto RNA yto ono66prr1oq. Itov nuptlvo, ro ef,orotoprdtro 61et 6trr6 p6tro: ene[epyd(eror to <puototroyrxd RNA Ko] Koro- orp5qer to etrort<r:1-rorrxd. To elonopottxd RNA ovo- yvcop((ovror on6 notr(Loug oupnopdyovteg rou e(coorll1.ror(ou nou 6nero entorpote0ouv to e(cootogd- tro. Irq (UUn, o r<0prog oupnopdyowog e(vot ro oripr- nloro TRAMP, ro ono(o 61er opr6troycl Kcrr oe dMouq euxopucineg. An6 ro pretdypogo qg RNA Pol ll uno- crrpcir;roro yr' ono66prqon orov nuprlvo e[vot 6oo 6ev 61ouv ouppoge( oromd rj 6e q6pouv rpuototroytt<6q poly(A) oup6g. To neproo6repo prerdypo<po qg RNA Pol ll (orog e(vor r<pugd ooro0f prerdyporpo (cryptic un- stable transcripts, CUTs). l-lorx0.o mRNA molefowor on6 ro rurroponLo- ol.rorrd ouor{proro enrnpnonq. Kor oro tpto g4etu<d ouor{ goto enmq pqoqq epn},6xowor ou pBdwo ovcOpro- Lou tepporropro6 qg prerdgpoo4g. H onooUv0eor; N- MD oroXe0er g6pro mRNA pre np6opo rodtt<6vto tepporroproU.'Evo ouwrlpqprdvo oUvotro nopoy6wtov (npcoreiveq UPF ror SMG) eprnL6rerot qrnv ovoyvcirptoq Kor rrlv nopddooq ev6g unoorpc0ptotoq NMD oro pqlo- vropr6 yevrrrig onooUv0eoqg. 'Evo np6olpo t<to6tr6vto repporropoU ovoyvrrrp((erot xotd tov npoton6po ytipo qg prerdgpoorlg on6 rnv nqpouo(o ev6g xo0o6rroti oupntr6r<ou oupBoLqg e[ov(rov r1 ev6g oouvrlOtoro pe- yd).ou 3'dxpou. En(or1g, 4 NMD epntrSreror orrlv qno- 66prqoq optop6vrov guototroyu<drv ooroOc0v mRNA. H onoorlvOeol NSD oroXe[et mRNA Xcop(g ew6g ntrotol- ou xto6x6vro tepprortopori rot onore( 6vo ouw4pqpr6- vo orivoLo nporeivc0v SKI ciore vo e[co0rloet tqv oneleu0Spoorl rou noy6eupr6vou prBoorol.tot(ou rot vo enrorpore0oer ro pnxovropr6 ono66prnonq. H onoo6v- Oeo'q NGD orole0er pr6pro mRNA nou 51ouv ordotpto prBooroprdrro oe rro6rr6vto rrlg ror6xonotrpu<r1g nepto- Xrig xor npoxoLe( oneLeuO5ptoorl rou ptBootol.tot(ou rot ono66prqoq. Oproprdvo mRNA ewon((owor oe et6u<69 neptoX6g rou rundpou xor 6e prerogpd(owot pr6Xpr vo qtdoouv orov npoopropr6 roug 96oo oro rrlrropo. O ewontopt6g qnore( crs-orotXe[o ndvo qro mRNA-or6Xo Kot trans- nopdyoweg. E[unrl pere[ rperg rU preg ].enou py(eg: 1. Iro cooxrirrqpo ouvetog6per orrlv eyro0(6puor1 peMowu<cilv npor0nov rou eprBpriou Kot ornv npo- 6royporpri rou ovonruftor<oU nenpopr6vou r<urt6- pov nou e6peUouv oe 6rogoperx5g nepro16q. 3I 11 l]eoltrrun 647
  • 139.
    2. Ie rUnoponou 6torporiwor ooriprprerpo, e(vor rivog prrlXovro;r6 g r<orovo prrl g n p oreivt rciv n opoy6wrov p6vo oro 5vq on6 ro 6rio 0uyotpu<d r(rrropo. 3. Ie oproprdvo r0rropo, t6[rog oroug notropr6voug ri- noug rurrdpov, e(vor rivog pqXovrog5g yro rrlv Ko- 0r6pooq unoKurroprxrilv 6ropeptopdrov. E(vor yvooro( rperg prqlovropro( ewontoprori: o. ono66prqorlrou mRNA oe 5l.eq trg O5oerg er,r6q on6 q 06oq-or51o B. enilerctrf oyxupoB6Lrloq 6roXe6prevcov mRNA orq 06oq-or6Xo y. roreu0uv6pevq perqqopd rou mRNA ndvu: oe o6o09 rou KurropooreLerori. H teLeuro(o e(vor 4 nro rowri pr50o6og xor ofronoe( rouq UoproKotig r<rvrlr4peg nou Boo((owor orrlv oKr(vn Kor rouq trlrxpooro),qv(or<oug. BrBAroypoeio Ievrxd Houseley J., and Tollervey, D. (2009). The many pathways of RNA degradation. Cell 136, 7 63-77 6. 22.2 To Ayyetrtorpopo RNA Eivor Aoro0rl Mdpro 'Lgeuvo Dolken, 1., Ruzsics 2., Rddle B., Friedel C. C., Zimmer R., Mages J., Hoffmann R., Dickinson P., Forster T., Ghaza P., and Koszi- nowski, U. H. (2008). High-resolution gene expression profiling for simultaneous kinetic parameter analysis of RNA synthesis and decay. RNA 14, 1959-72. Foat, B. C., Houshmandi, S. S., 0livas, W. M., and Bussemaker, H. J. (2005). Profiling conditionspecific, genome-wide regulation of mRNA stability in yeast. Proc. Natl. Acad. Sci. (USA) 102P, 1 7675-80. 22.3 To Eur<opurrrrtxo mRNA YnopXouv uno Mopqn mRNP on6 q [ew1oq rltog ror T0 00v0r0 Touq 'Epeuvo Hogan, D. J. et al. (2008). Diverse RNA-binding proteins interact with functionally related sets of RNAs, suggesting an extensive regulatory system, PLoS Biology 6(10), e255. Avorxonnsrl Keene, J. D. (2007). RNA regulons: coordination of post-transcrip- tional events. Nature Reviewsl Genetics 8, 533-543. Moore, M. J. (2005). From bifth to death:the complex lives of eu- karyotic mRNAs. Science 309, 1514-1518. 22.4 H Ano06p1o1 rov Iporoputolrdrv mRNA ilepilogpovet l-lotrtro'Ev(upo Bernstein, J. A., Khodursky A. B., Lin P. H. Lin-Chao, S. and Co- hen, S. N. (2002) Global analysis of mRNA decay and abun- dance in Escherichia coll at single-gene resolution using 048 KEOAAAI0 22 lro0epoqro rorXtrlpo0rlqol rou mRNA two-color fluorescent DNA microarrays. Proc. Natl. Acad. Sci. (us/) 99, 9697-9702. Celesnik, H., Deana, A., and Belasco, J. G. (2007). lnitiation of R- NA decay in Escherichia coli by 5 pyrophosphate removal. MoL Cell27,79-90. Mohanty, B. K. and Kushner, S. R. (2006). The majority of Escherichia coli mRNAs undergo post-transcriptional modification in exponen- tially growing cells. Nucleic Acrds Bes. 34(19), 5695-5704. Avaoxdqoq Carpousis, A. J. (2007), The RNA degradosome of Escherichia coli: an mRNA-degrading machine assembled on RNase E. An- nu. Rev. MicrobioL 61 ,71-87 . Condon, C. (2007), Maturation and degradation of RNA in bacteria. Current 1pinion in Microbiology 10,271-278. Deana, A. and Belasco, J, G. (2005). Lost in translation: the influ- ence of ribosomes on bacterial mRNA decay. Genes Dev. 19, 2526-2533. 22.5 To ileproodrepo Europuolro mRNA Ano6opouvror Mriotrr Auo Movonolrov E[oprtipevov ono [1otruodevutrltoor1 Avorxonqoq Franks, T. M. and Lykke-Andersen, J. (2008). The control of mR- NA decapping and P-body formation. Mol. Cell32, 605-615. Parker, R. and Song, H. (2004). The enzymes and control of eu- karyotic mRNA turnover. Nat. Struct. Mol. Biol. 11,121-127 . Parker, R, and Sheth, U. (2007). P Bodies and the control of mRNA translation and degradation. Mol. Cell25, 635-646. 'Ipeuva Sheth, U. and Parker, R. (2003). Decapping and decay of messen- ger RNA occur in cytoplasmic processing bodies, Scrence 300, 805-808. Zheng, D., Ezzeddine, N., Chen, C. Y., Zhu, W., He, X., and Shyu, A. B. (2008). Deadenylation is prerequisite for P-body formation and mRNA decay in mammalian cells. J. Cell Biol. 182, 89-101 . 22.6 Atrtro Movonol0 Anod0Un0nq lroleuouv Iuyrerprp6vo Mdpro mRNA Avuordnqorl Filipowicz, W,, Bhattacharyya, S. N., and Sonenberg, N. (2008). Mechanisms of posttranscriptional regulation by microRNAs: are the answers in sight? Nature Rev. GeneL 9,102-114. Garneau, N. 1., Wilusz, J., and Wilusz, C. J. (2007). The highways and byways of mRNA decay. Nature Bev. Mol. Cell Biol. 8, 1 13-l 26, 'Epeuvo Mullen, T. E. and Marzluff, W. F. (2008). Degradation of histone m- RNA requires oligouridylation followed by decapping and simul- taneous degradation of the mRNA both 5 to 3 and 3 to 5. Genes Dev. 22,50-65. 22.7 0 Erdudg Xpdvoq HU(onS Kq0e mRNA EtrdyXeroL ond AtrtrntrouXieq n AoUeq M6oo oto mRNA irvlijll ilti:t,:I Chen, C Y. A and Shyu, A. B. (1995). AU-rich elements: charac- terization and importance in mRNA degradation. Irends Biochem. Scl. 20, 465-47 0.
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    von Roretz, C.and Galtouzi, l. E, (2008). Decoding ARE-mediated decay: is microRNA part o{ the equation? J. Ceil Biot. 181, 1g9- 1 94. 22.8 Tq Neosuwe0etyrivo Mripro RNA EtreyXovror yro Etrotrtoporo M6oto Evrl g lluprlvuou luorrlporo g Enrinpnonq 'Epeuva Arigo, J. T., Eyler, D. E,, Carroll, K. 1., and Corden, J. L. (2006). Termination of cryptic unstable transcripts is directed by yeast RNA-binding proteins Nrdl and Nab3. Mol. Cett24,7g5-746. Davis, C. A. and Ares, M. (2006). Accumulation of unstable pro- moter-associated transcripts upon loss of the nuclear exosome subunit Rrp6p in Saccharomyces cerevisiae. proc. Nail. Acad. Sci. (U SA) 103, 3262-3261 . Kadaba, S., Wang, X., and Anserson, J. T. (2006). Nuctear RNA surveil lance in Sacc h aro myce s c e revi s iae; Trf4p-dependent polyadenylation of nascent hypomethylated IRNA and an aber- rant form of 55 RNA. RNA 12.508-521 . Auco'xcnqaq Houseley, J., LaCava, J., and Tollervey, D. (2006). RNA-quatity control by the exosome. Nature Rev. Mol. Cett Biot.7, S2g- 539. Houseley, J. and Tollervey, D. (2008). The nuclear RNA surveil, lance machinery: the link between ncRNAs and genome struc- ture in budding yeast? Biochem. Biophys. Acta 1779, 239-246. Villa, T., Rougemaille, M., and Libri, D. (2008). Nuctear quatity control of RNA polymerase ll ribonucleoproteins in yeast: tilting the balance to shape the transcriptome. Biochem. Biophys. Acta 1779,524-531. 22.9 0'Etrey1og [lordrrirog rrlg Merorppoorlg rou mRNA llpoyporonore[ror ono Kurropontroopouro Iuorrlporo Enrnlprlolg - ,:iii.llliliiil lsken, 0, and Maquat, L. E. (2007). 0uatity controt of eukaryotic mRNA: safeguarding cells f rom abnormal mR- =,-:: :- Genes Dev. 2l , 1 833-1 856. McGlincy, N. J. and Smith, C. W. J, (200S). Alternative spticin_c .;- sulting in nonsensemediated mRNA decay: what is the meanrr; of nonsense? Trends Biochem. Scr. 33, 385-393. Shyu, A. B., Wilkinson, M. F., and van Hoof, A. (2008). Messenger RNA regulation: to translate or to degrade. EMB} J. 27,471- 481. Stalder, L. and Mohlemann, 0. (2008). The meaning of nonsense. Trends Cell Biol. 18(7),315-32i. 'Ipe uvtr Wilson, M. A., Meaux, S., and van Hoof, A. (2008). Diverse aber- rancies target yeast mRNAs to cytoplasmic mRNA surveillance pathways. Biochem. Biophys Acta 1770,5S0-5S7. 22iA Mepuo Europuoruo mRNA Evronl(ovror oe EL6uriq l-leproy6q rou Kuropou 'Epsuvo Blower, M. D., Feric, E., Weis, K., and Heatd, R. (2007). Genome- wlde analysis demonstrates conserved localization of messen- ger RNAs to mitotic microtubutes, J. Cell Biot.179, i 365-1373. Lecuyer, E., Yoshida, H., Parthasarathy, N., Atm, C., Babak, T., Cerovina, T., Hughes, T. R., Tomancak, p., and Krause, H. M. (2007). Global analysis.:of rnRNA localization reveals a promi- nent role in organizing .cellular architecture and function. Cel/ 131,174-187. Avooxtinrloq Bullock, S. L. (2007). Translocation of mRNAs by molecular mo- tors: think complex? Seminars Cell Devel. Biol. 18,194-201. Du, T. G., Schmid, M., and Jansen, R. P. (2007). Why ceils move messages: the biologicalfunctions of mRNA localization. Semrn. Cell Dev. Biol. 18,171-177. Giorgi, C. and Moore, M. J. (2007). The nuctear nurture and cyto- plasmic nature of localized mRNPs. Semin. Cett Devel. Biot. 1g, 1 86-1 93. Martin, K. C. and Ephrussi, A. (2009). mRNA tocalization: gene ex- pression in the spatial dimension. Cell 136,719-30. i.::.-,:,-: 649
  • 141.
    @ Nucleus MedicalArt, lnc./Phototake/Alamy lmages ffiffs ffi l(oroAunK6 RNA Erooyoyrl To lvrpovro rqg 0podog I AuroouppdnrovTor Ue trans- eoreponofiorl * 0t U6vor nopoyovreg nou xpeto(ovror ro rvrprivro rrlg ogo6og I yro Trlv ouToouppoqrl l/? vitro etvot 6uo perotr- trrxo r6vro rr 6vo vourtreor[dro youovoo(v1q. * H ouppoQrl npoyporonore(ror pe duo trans-eoTepoflor- loetq Xorpig ovoyq evepyerog. * To 3'-0H orpo rou oupnopoyovro qg youovoo[vr;g enu(0eror oro S'oxpo rou rvrpovlou orrlv nptirq trans- eoreponoqo4. * To 3 -0H oxpo nou dlgroupyrlOqxe oro r6trog rou nptir- rou e[oviou enu(Oeror oU ouUBotrrt pero(U rou rvrpo- v[ou xol rou de0repou e{ovrou roro rr1 detireprl frans- eoreponolrlorl, * To rvrpovro onetreuOeprirverot un6 poptprl eu00ypoUuou pop[ou nou oX4por((er rur],0 orov ro 3 -0H oxpo rou enueOe[ o' 6vov 6eogo oe pro on6 600 eooreprr6g 06oerg. * Zto Tetrahymena, 6vog eoo:repuog 6eop6g rou tvrpo- viou nou orpoLp60qre eniolg 06Xeror en(0eoq on6 dtr- tro vourtreor[dro oe Uro neproXrl trans-oupporpflq. ffim To lvrprlvto qq 0po6oq I lXlpori(ouv Mro Xopoxqprotr- ru1 Aeureporoyrl Aopq * To rvrpdvro rqg opodog I oXrlpori(ouv 1uo deureporoyl 6op1 U' evv6o d(rtroveg neptoX5g. * To rdwpo rov neptoXrirv P3, P4, PO ror P7 tlXouv ro- rotrunr1 dpoorl. * 0L neprox6q P4 ror P7 oXqgori(ovroL pe (euydpopo ouvtrlplg6vtrlv ouvorvrtrd:v otrtrntrouXLrov. * Mro otrtrrltrouX(o 6intro oqv neproXrl P7 oXqpori(er (eU- yn pdoeorv pe pto otrtrqtrouXlo nou nepr6Xer rq dpoort- Krl G. To Prpo5v(ugo'EXouv fl orr[treg Koro],ulr6g Apooerq * MeroBdtrtrovroq U 06on np6odeonq rou unoorp6po- rog o' 6vo rvrp6vro rqg opodoq l, elvor nr0ovrl 11 eroo- ytrrytl evotrtrorildrv otrtrr1trouXulv nou otrtrntrenrdpot.rv pe q dpoortrrl G. * 0r ovl6pdoerg oroLou0o0v rtroorul rrvlnrrl ev(upov Ue XoUntrrl rorotrurtr1 roXuqro. * Avrtdpooetq nou XprlorUonototioov 2 -0H deououq 0o pnopouoe vo e(vor 11 poorl yro rqv e(etrt[4 rtrrv ou0evl- rtlv xorotrunrdlv evepyorrlrtov rou RNA. ffi 650 KEOMAIO 23 Korotrurtxo RNA Enry€leta KeEaAa{oa: Douglas l. Briant
  • 142.
    @ OptoUevo lvrpovtorrlq Opodog I Krrrbrxonorouv Ev6ovou- rtredoeg nou llpooyouv qv Krvqnrriqro a To rtvnro tvrpdvro rqg opodoq I rotdrronorouv pro ev- Dovourtreooq, 4 ono[o npoygoronorel pro dirtrovq ev_ rop4 oe pro 0601-orrlXo. * To rvrpdvro perorlOeror orrl 06011 rrlg d(rtrorvr1g ewo- Urtq Ueoo ev6g ovnyporpuoU p4Xovro1.ro0 nou 6ropeoo- trope(ror on6 ro DNA. W fo lvrprivro rrlg 0p66og ll 'lotog KtodrxonoLouv flol,utrer- roupytrdg l-lptoreiVeg c To rvrp6vto rqg opodog ll ourooupponrovtot in vitro, otrtro ouvrlOog unopo4Oouvror ond nporelveg rou KoJ- 6rronorouvrot ond to rvrpovro. * 'Evo Uovodlrri rtoDtronorlrtrtj ntro(oro rodxonore( gro nporeiVrl p' evepydrrlro ovr(orpoqrlg peroyporpoor;g, gro evepyrlrrlro poroupoorig, evo por(po nprlodeorlg DNA rot pro DNA evDovourtreoorl. s H evdovourtreoorl dtoono ro DNA-orriXo enup6novrog rqv evoolrotooq rou rvrpoviou oe gro v6o 0e04. * H ovr(orporpq peroyporpdoq dqgroupye( 6vo DNA ovri_ yp090 Tnq otrtrqtrouXiog RNA rou evoogorop6vou tv_ rpov[ou. @ Opropdvo Aurooupponrripevo lvrpdvro Anououv Morou- pooeg e T' ouTooupponrdpevro rvrpdvto pnope( vo Xpero(ovror evepy6rrlieg poroupdo4g nou rodxonorouvror onrj ro rvrprivro ror poqOouv oro dintropo rou o, evepyo roro_ trurtrn doUrt. && tt Korotrururl Evepydrrlro rrlg RNAdorlg p Ogefieror oro RNA c H prpovourtreooq p (RNAooq p) eivor pro prpovouxtreo- npalreiVrl orrlv ono(o ro RNA eXer xorotrunrrl evepy6- TrlT0. * H RNAooq P e(vor onopoirrlTrl 0T0 porrrlpro, r, opXo(o Kor Touq euxoputireg. * H RNAoon MRP rov europuotrdrv oXer((eroL pe qv RNAoorl P xot epnL6rerot orr;v ene[epyoo(o rou rRNA rolrqv ono66prlon Tou mRNA qg rurtr(vr1g B. ffi fo loerdrl'EXouv Korotrunxrl Apdorl * To roerdrl ror ro roooerdq oXqpor(ouv pto orpupoxritpo- In 6oprl p' evepy6qro ourodrdonooqg. o floprjpoteq 6oUdq Unopei vo drlptoupyl0ouv (euyopto- vovrog 6vov rtr6vo-undorpopo nou Droonoror ono 6vo ev(uprxd rtrovo. & Orov o ev(uptrog rtrrovog erodyeror oro rrjrropo, (eu- yoprlvet p' 6vo rtrdrvo-ortiXo, o ono(og oq ouv6Xero 6roonoror. ffi fneqepyoo(o rou RNA luppo(ver oe Megovropeveg Bri- 0erq c 0r unodoxe(q qg onoLrnonptoreivlg-B ror rou ytrouro- pxou urp(orowot e16Lxeg onoprvtiroerg nou rorotruovror onrl Oeoprvdoeq qg ruldrvrlq xor rrlg o6evoo(v4g ror otrtro(ouv qv rodrronorqnrl otrtrr1trouXio. ffi n Ene(epyooio rou RNA Mnopei vo Ko0o6r;yeiror on6 06qyd RNA e Iro Uuoxdvdpro Tou Tpunovoodrporog ouppq(vet erre_ topdvq ene{epyoo(o rou RNA p' evOdoerg 11 onotrouprlg ouptdLv4g. * To RNA unrlorpopo (euyoptiver p' evo odqyri RNA ror onri lg 6uo ntreup6g r4g neproXr;g 11 onoro npdxeuor vo unooreI ene[epyoo(0. * To odlyo RNA nopeXet ro expoye(o yro npoo0rlu1 (r1 onovrorepo onotrotqrl) oup16rvtirv. * H ene[epyoo(o rorotrueror ono ro oopor(dro ene[ep- yoo[og, 6vo o0pntroro p' evepy6r4ro ev6ouvourtreo- o1g, e[ovourtredo1g, retrtr1q ouprdutrorpovoqepoolg ffi xor RNA trtyrooqg. H Iupporprl llporeivdrv E(vor Auroxoroi,urxrl o Mro rvrelvrl 6Xet rrlv xov6rrlro vo rorotruer r4v otpo(pe- o4 qg ond pro npotelvr1 roro rdroto rptlno 6ore vo ouvdriovrot or e[relveg nou Trlv neprpotrtrouv. * H ouppoqrl rov nporeivtirv rorotruerol on6 rqv rvrelVrl. * 0t neproodrepeg tvre(veg eXouv duo ove[oprrlreg evepydrrlreg: ouppoqn nptrlreiv(rv rr evdovourtredoq enotrtopou. l-iepft1Qq To np6to nopo6eryproro r4q r<ordAuoqg nou Boo(- (eror oro RNA ovoyvrop(m4r<ov o' dvo Borrcrlptor<6 6v- (ugo ene(epyoo(og rou IRNA, 14 prBovouxtredo.rl p (RNAdo4 P), ror or' ouroouppom6prevo rwp6vto rqg o;rd6og I oro HNA tou Tetrahymena thermophita. fs rrlv nporonopror<rj Speuvd rouq ornv r<ordl,uorl on6 ro RNA, or Sidney Altman xor Thomas Cech BpoBeU- rrlKov Ue ro BpoBe(o Noprn6L X4pre(og ro 19g9. An6 r4v oplrrq ovor<di.uQq rou xoroLuru<oU RNA, dXouv ovoyvoptore( notrio( dtrLor rrjnor rorolurrxcbv owr- 6pdoeov nou peooloBoUwor on6 ro RNA. Entoqpro(- vetor r6ro(tepo ro yeyov6g 6rr tq prBooropdrro, 6qf.o6r1 ro oripnl,or<o RNA-npotreivc0v nou nopdyouv nenr[6rq (B].6ne Ke<pdl,oro 24, Merdqpoorl), 61ouv 6B 2 3.1 Erooyoyrl H t66o 6rr pr6vo or np<oreiVeg 5Xouv ev(u;rotxrl 6pd- oq frov Bo0td pt(topr6vq or4 Broplpre(o. To oremrr6 yro rrlv rouronoqo4 rcov ev(Upov <og npcoretveg e(1e rtg Bdoerg rou ornv dnorp4 6rr g6vo or nprorefveg, pe rrg norxil,eg tpro6rdororeg 6opr5q Kor rnv norxrl(o r<ov ni.eupu<c6v o;rd6ov toug, erlov rqv eueLr((o vo 6r1pu- oupyrjoouv evepyd x6wpo nou Kqrql,Uouv Brop1l-u- x5g ovrrdpdoetg. Om6oo, rpforgeg geAdteg xov ouorqpdrov nou egnl,6r<owor ornv ene[epyoo(o tou RNA 66er[ov 6rr owri 11 dnorfq e(vor 1.uo unepon].ori- qreuon. f "i;.-! Etooytrtyrl 6b1
  • 143.
    rouronorleei otg prBo5v(upro(ribozymes), 6nou ro R- NA evepye[ og ro roto].utrr6 ouororx6 xor 4 nprotet- vr; cog rxp[copro. O yevu<69 5pog plpo6v(upo 1prlor;ronoefuor yro vo neptypd{.rer 6vo RNA pe roto},uflr<ri 6pdor1. H ev(utrru<rj evepy6r4ro prnope( vo neprypoge( 5nrog 11 ow(ororXq evdg nro oupBorrro0 ev(Uprou. Opro;r6veg xorol.urrr6g Dpdoerg tou RNA roreu0fvowor 5vowr [eXopror6v unoorpo;rdrolv (6topro p tor<69), evril dl.l.eg e(vot ev6opro- ptordg, yeyov6g nou neptop((er rrlv rotol,uur<r1 evepy6- rrlro o' 6vo pr6vo xrir<Lo. To 5v(upro RNAdoq P efvor pro prBovour].eonpco- refvq nou nepr6ler 5vo provodLr6 pr6pro RNA ouv6e6e- p6vo oe pto nporetv4. H RNAdoq P Aerroupye( 6roproproxd ror onorel.e( nop66etypo prBoev(Upou no u roroL(er owr6 pdoerg nolLon).rj g ovor<ri r.tr oor1 g (multiple turnover). Evc0 oplrrd ovoyvrop(orqre oro E col/, ori;repo yvcop((oupre 6rr 11 RNAdo4 p e(vor ovo- yxo(o yro rq Brootpr6qto 16oo rov npoKqpuor6v 6oo Kor rov europuror<i:v. 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To rotv6 orlpre(o tov owrdpdoeov nou 6rexnepot- cbvowor on6 ro roroluru<6 RNA e(vot 6l ro RNA ovo- LopBdver pto ev6opropror4 11 6roproptorrl ow[6poor1 4 ono(o nepil.ogBdver 6rdonoo4 rl Sqproupy(o gooqo- 6eorepx6v 6eoprcirv in vitro. l-1op6r 11 er6rr6rrlro qg ow(6poo4g ror 4 roto).urrrrl evepy6rrlro nopdlowor on6 ro RNA, or npalreiveg nou oLLrllenrdpo6v pe ro RNA (oog e[vor ovoyro(eg yro rqv onore],eoprorrrrl npoypqrono(qo4 qg ow6poorlg in vivo. H ouppogri tou RNA 6ev eivor ro p6vo ;r5oo pre ro ono(o ero5plowor oMoy6g oro n),4pogopror<6 nepre- 16;revo rou RNA. Kord tqv ene(epyoo(o rou RNA (R- NA editing), oMoySg etodyowor oe (eXopror5q Bdoerg r1 Bdoerg npoor(Oewor oe ouyKeKprprdveg O5oerg o' 6vo mRNA. 'Ev0eoq Bdoeov (ouvrlOrrlg rordlotno oupr6(- u1g) npoypotonoefuor oe noMd yov(6ro oro pmoX6v- 6pro opropr6vov l.rovorUttopov/o}.tyoKrjrropov europucorcbv. 'Onrog q ouppo<prj, 6ror ru q ene(epyoo(o tou RNA nepiloprBdver dtdonoorl rr enovdvoorl 6eoprcbv pero(0 vouxleorrd(ov, coor6oo en(onq onore( 6vo ex- poye(o yro rnv rto6rrono(r1o4 tov nlqporpopuriv rrlq v6- og oMqLoul(og. 23.2 To lvrp6vlo rnq AuroouppdnTovrot l.Jr Opd6oq I frons-eorcponoinon * 0r Uovot nopoyovreg nou Xpero(ovror ro lvrpovro rrlg opodoq I yro qv ouroouppogrl in vitro e{uot drjo gerotrtrrro rovro rr rivo vourtreor[6to youovoo[v1 g. " H ouppoQn npoyporonote[ror pe 6uo trans-eoreponouloetg Xop(g ovoyrq evdpyerog. * To 3'-0H orpo rou ouprnopoyovro qg youovoo(vqg enul0eror oro S'orpo rou lvrpov[ou or4v npdrrrl trans-eoreponoqoq. e To 3'-0H orpo nou d1;rLoupyrl0rlre oro rdtrog rou np6rou e(ovrou enu(0eror oq ougpotrq pero[u rou rvrpov[ou ror rou 6eurepou e[ovrou orq 6eureprl trans-eoreponoqorl. e To rvrpovro onetreu0eptiveror uno poprprl euOuypoglrou lLopL- 0u nou oxrlpoTr(et xurtro 6rov ro 3 -0H orpo rou enlreOel o' 6vov 6eopo oe pto ono 6uo eoorepLr6g Oeoetg. * Zro Tetrahynena, iuoc, eooreprrog 6eop6q rou lvrpov[ou nou otpoLp60rlre en(or1g 0SXeror enr0eorl on6 otrtro vour),eo- r(6ro oe pro neproXq frans-ouppoqrlg. lwp6vro rrlg oprddog I Bpfor<owor oe notx[].eg 05,oerg xor oprOporlv ndvco on6 6Uo 1illddeg pr6Lr1. Awi0ero ond r1v RNAdorl P, 6av e[vor onopo(rrlro yto r; Burr- otp6qto. To rwp6vro qq o;rd6og I Bp(or<owor oro yov(6ro nou rtodrronoroUv rRNA orov nupqvo rov Uo- vo rUrropo-rv/oryoxUttopov e u Kop u an rhv T etr a h y - mena thermophila (Blecpopr6og6po) xor Physarum polycephalum (pu(oprUrrlrog). Efvor xorvd oro yov(6ro puxrlrcov xor nptor[ortllv, evo onov((ouv orouq npo- r<opud;reg ror ro (c6o. To rwp6vto r1g opddog I 6Xouv pto ev6oyevq rxov6rrlro vo npoyporonoroUv ouro- ouppog{ (sellf-splicing, autosplicing). (Tr;v (6to rdr6r4ro en(oqg 6ro05rouv Kor ro rwp6vto rrlg otrro- 6o9 ll, nou 0o nopouotooroUv orqv Evotrlru 25.6). H ouroouppog4 ovor<o).0g0qxe og t6r6qro rotv pieroypdrpcov rov yovr6(<ov rou rRNA oto T. thermophi- /a. To yov(6to ttov 6tjo r<[prov rRNA d,Xouv rq ouv1Orl opydvoroq r<oOrrtg Kor ro 60o er<<ppd(owor rr.rg pr6pog lrtog r<oLvrlg preroypoqrr<qg provd6og. To npoidv e(voL dvo np66popro 35S RNA Ue rnv oMrllouX(o tou 1-rLxpori (17S) rRNA oro S'dr<po Kor rnv oM4).ouX(o rou preyd- ).ou (265) RNA npog ro 3'drpo. Ie opropr6vo orel6pl T. Thermophlla, q oMq),ouX(o nou rodrxonore( ro 265 rRNA, 6ror6metot on6 dvo pro- vo6Lr<6 BpoXri Lwp6vto. Orov ro 35S npddpopro RNA snald(eror in vitro, q oupporprl ougBo(ver og our6votru1 ow(6pooq. To rwpdvro onox6meror on6 ro np66popo p6pro r<ot ouooope[erot iog eu0rjypol.lpo r<],do;ro 400 Bdoeorv, ro ono(o 6nero perorp6nerot oe xurLrr6 R- NA. Aurd ro oupBdwo ouvotU((owor crrlv ti,r,'ii,,,. ,_.. H ow(6pooq onorre( 6rjo prero).).rxd L6wo Kr evo vouxLeor(6Lo youovoo(v4g rog oupinopdyowo. Ko;rrd dM4 Bdon 6e pnope( vo unoKoroor(oer rr1 G, evc0 6ev 652 KEOA Al0 23 Kototrunro RNA Boorx6g eworeg
  • 144.
    EE6vto 2 tr- :uPPoQn I I ow(6pqon,sfl1v oTlo(q rl veoouweeetp6vn 3'-OH ol.e ;;;" 6ripo tou eqov(ou 1 enn(oerot oro 6etnepo r(,,i,*o Or 'Orlo trretoqopdq. ouvddowot' xo0tilg 6ev no- A;n;;,i*", el'e0oepo'e[ti''to, q ouvdeoq touq trr6Mov lruBlttt, coq p6poq qg i6roq.ow(6p1on1lo.y onetreu- e"f,i,ut, to twp6vto' To twp6vto onetr'e-u0epoverot coq rr'eJvpoUUo p6pto, oL)'d 11 tp(tq ow(6poorl uoroqo- pdg to PetorP6net oe rrixi'o' ' kae, orddto t4g ow[dpoonq ourooupporpfg oup- Bo(vet ;re trans-ecnepono(nqn, rotd.qv onoio 6voq <po- L,oootr6c eor6poq petotp6netot 6;reoo o' 6vov 6ltro xipis t"i,outon uopOxuon' ot 6eopo( evotrdooowqt ad"o rot rl ev5pyeto 6totrlpe(tot, ouvenriq 11 ow(5po- oi-otu ono,.e( nopoxrl evepyetoq on6 qv u6p6uor1 iou nrp q tou GTP.'kdee enor<6ou0q ow(6poor1 t r.-r*rpono(qor1q 6ev neprLopBdvet r<oOopr1 peto- il;iti -ns ev5pyeioq. rto x0rropo, 11 ouyr<6wptoorlrou bip'tlro, uqnlrl ouyt<pttxd pe q ouyr<6wpoorl rorr RNA r<r 6tot npoo0e( qv ow(6poon npoq-rnv Kortu- gruon xotd tqv ono(o ;tto pretoB-oM trlg 6ewepoto- v"li a"uns tou RNA 0o nopepno6(oet rrlv ow(orporPn ow(Opoo'r1. nw5 enttp6net cnrlv ow(6po9n vlle:I1- pqoei p6xpt vo ol,ortrr1poOe( owivo nepooet oe Korct- L'"tn ,oippor,tos ou&p"oo oro ouppoupr6vo npoi6v Kot rq pn ouppopg5vo np66po;ro g6pto' Metoyporpn I HleKrpoQdPnon oe rtnmn 35S RNA --# +!€!# KuKlorornon | * t Kurtrtxo rvrPovto -,A U EuotlYpouuowrpovto- ' - - H ouppoQrt rou 35S np66popou rRNA rou Tetrahy- mena nopoKolou0e[rot pe qerrpotpoprlorl oe nrlrq H otpoipeorl iou ,uiporrou onoxol'unrerot ono rrlv epqovtorl ;rtoq ptrprlq (6- ;r* ".; rtve(rot yprlyopo, '0rov ro tvrpovto ylvet rurtrtro' peroxt- veirot ppo50repo, 6ntlq tpo(verot ono rrlv utllltroTepl (6W' onome(tot tptqroorpopu<{ op66o: Xpnot- trlonoto6wot ot GTP, GDP, GMP rot 11 i6,o ,1 yououoo(vq, ouventilg 6ev onot- te(toi ev5pyeto. To vourtreor(6to u1g youovoo(vqg npSnet vo nept6Xet pto op66o 3'-OH. To nenPoPdvo tou vourtreott6(ou youovoo(vqq tXvrltrote(rot pe podtoo(- Uqvon. H podevrlpyero oplrrd eto5ple- ror oro onoroppSvo eu06ypoppo twp6vto. To t<otd,tr'otno qq G ouv66etot ue to 5' drpo rou eu00ypo1-t1'tou twpovt- ou g' 6vov t<ovovtr6 <ptooqodteoreptx6 6eopr6. tf ftXOtU 23.2 6efyet 6tt ouprBofuouv rpetg ovrt6p6oerg petotpopdq' Etqv npOq getoqoP6, to voudreot[6to r1q youovoo(vr1g oupneptq5perot oq. ouU- nopdyowog nou nopSXet qv el'er3Oepr1 3'-OH oPrddo q ono[o enr[0etot mo 5 6rpo tou twPov(ou. Aw( q ow(6Pooq OnUrupVti 5vo 5eopr6 getofiti G-twpovl- ou'Kot 6r1;rtoupye( pro 3'-OH ogddo oT o 6t<po tou 5' e(ov(ou' H de6teprl peto- tpopd Boo(etoi oe tr to nop6proto xnptKn G-OH EEtJvto A V.- c '- -- EEovto B P,.<3 tttaa..rrt' I '@*P# + (Itl G- p J (c*Qll ; tta. tt' , !ijt :, l. i i:' .,:,1':.!l'r: To 3'-0H rrlq G entriOerot oro S'orPo rou tvtPov[ou 3.' PG-0H Y pNpNPNPNPNPN t:::.1:: ii. : ;: ;:'':: ;/'": i' I I Y PNPNPNPNPNPN-OH + PG i i ll r'.i:li i'r:'i "li i"r'':' 'r* AE[TEprl geroqoPd To 3'-0H tou e[olv(ou A entr(Oerot oro 5'6rPo tou e[olviou B Tpfu peroqoPd To 3 -0H rou tvtPov(ou entriOerot oTo 6sogo nou on61et 1 5 Pdoetq ono'ro S'orPo : H ouroouPPotP1 nPoYPoronotet- ror pte ovrt6po00tq trans-soTepono(qolg ortg onoieq oupBoiver opeorl ovrotrhoyrl 6eog6v 0t Oeopto( nou DrlpLoupyouvrot oe roOe orobto or1- polvovror pre ro oxtoopevo no(oto' I c-P 8-0H + ?* ? To lvrpovto rnq 0go6oq I AurooupponrovTot Ue Irans-eoreponoiqorl 653 . ,. .. ldffiit& Er.'[i
  • 145.
    t Avostpoqn rurtronofqorl ,ro I 6"s" llpoidwo I"O eu0uypdpprorlq oG4140H AvrfDpoorltrans To onorop;.ulvo lvrpovro ;lnopel vo oXqgorloer r<u- rtroug Xplorponouovrog Uro ond 0uo eoorepirdq e60erq drore v' ovlDpooer pe ro 5'orpo ror [ovovolyer roug rur],ouq pe ovr[6po- o1 pe vepo 1 otrryovour),eor(6r0. H rxov6rrlto rnq ouppoqrlg e(vor ev6oyev4g yro ro RNA xor ro o6orrlgo e(vqr Kov6 vo npoxope( in vitro Xopfg npoo0rl<r'1 rovev5g npcoreivxori ouorfirKo0. To RNA oXqpor[(et pro er6rxrl deuteporoyrl/rpnoroyrj 6o- pn, ornv ono(o or oXenr<5q opddeg rpdpowor owKpu- ord 6ror riore 6vo vour<Leor(6ro youovoo(vqq vo npoo6e0e( oe Uro er6trrl 05oq ru 6neno vo pnope( vo ouprBoriv ot owrdpdoerg 6t6onoo4g xt enovdv<ooqg rou 6eopro0 nou ropouord(owor ar4v Er6vo 23.2. MoLo- v5tt onoreLe( r6r6qro rou (6rou rou RNA, 11 ow(6poor1 e[vot notrri Bpode(o in vitro. Aur5 e[qyehor enerdf rn ur'- vo n ouppoq4 rov rwpov(ov r1g oprddog I unoBorl0e[- ror on6 npcorer*veg, t<ov ono(cov 11 l,emoupy(o 6yrenor or4 oroOeponornon qg 6opr1g rou RNA oe Uro 6rog6p- qcooq euvoir<4 yro 11 ouppogrl. H rxov5rrlro ouggeropig oe t6roreg owrdpdoerg gerogopdg Syxenot orqv otr).qLou1(o rou twpov(ou, ro ono(o e[oro].ou0e( vo efvot 6poonr6 prerd qv onoKo- nrl rou un6 popgrl eu0Uypo;r;rou prop(ou. H Ell(0l'lA 23.3 ouvor.!((er rrg xoroLutrrdg 6pdoerg rou onoxol.rpdvou rwpov[ou tou Tetrahymena. To 3'-0H rnq G41 4 enufOeror onlv pAl 6 q qv pU20 S'exoG.. UUUpAI 6CCUpU20UG To rwp6vro rurLonore(ror 6rov n G oio 3' dr<po (OG) entte0e( oe puo eooreprr4 06o11 rovrd mo 5 'dr<po. O eooreprr6g 6eopr6g droondror r<or ro v6o 5' dxpo pe ro oplu<6 e[<oyevdg voux].eor(6ro youovoo(vr1g (exoG) oneLeu0ep6veror oq eu0Uypoprpo xldo;ro, To ruri.onorqpdvo twp6vto prnope( vo eu0uypopprore[ Ue erdu<ri u6p6).uon rou 6eo;ro6 ovdpeoo orrlv OG r<or ro eoorepx6 rordl,orno nou erxe rl.e(oet rov xrixl,o. Awrl 4 ow[6poo4 ovogd(etor avdotpoE4 xuxAonotqoq (e- verse cyclizafion). Avdloyo Ue rn 06oq qg npororo- yo0g r<uxLono(noqq, ro euOrjypoprpro g6pro nou npoS- ruqe on5 qv u6p6Luor1 nopopSver 6pooru<5 Kor Uno- pe( vo npoyporonotqoer puo Sewepoyev4 r<urLono(- non. To reLrr6 npoi6v rrov ou06ppr4rov owrdpdoeov nou oxoAou0orjv r4v oneleu06proon rou rwpov(ou ulg o;rd6og I ao Tetrahymena e(vot ro L-19 RNA, 6vo euOrj- ypogpo pr6pro ro ono(o nopdye'ror ovoorpdeowoq rn BpoX[rep4 xur].rxr1 6o;r{. Aur6 ro pr6pro 5Xer ev(uplxrl evepy6qro, Xdprl or4v ono(o rffioltier r4v en6moorl Bpo16c'lv oLryovou rLeolS(rov. H owr6poortr6qto rrrlv onorop;r6vrov rwpov(rov enerrefveror n6po on6 qv onLrl ovoorpoe{ rnq ow(6poor1g r<ux}.ono(qo4g. H npoo0fr<q rou oLryovoux).eon6(ou UUU (ovovoryer rov nporotoy4 rUx),o owrdpc0woq pre ro 6eopr6 gero[ri OG-eoorepmoU vouxl,eotrd(ou. To UUU (ro ono(o pror- d(er ge ro 3' dxpo rou 1Sprepo0g nou oneleu0epc00rl<e on6 rrlv npcoroyevrl rurlono[r1o4) y(veror ro 5' dxpo rou oxnporr(6prevou eu0riypo;rpro u prop[ou. l-'l p6remor yro drol.roproxri ow(6pooq nou ovo6exv0er r1v xov6- rqro orivdeorl g 6ti o drogopenr<iv ;ro p (cov R NA. Awrl4 oerpd owrdpdoeov <povepri:ver 6n q owoxo- roi.unrrl 6poorq pr6r4ro owrKoromp((er puo yevmeu ;16- v4 rrov6rrlto rou prop(ou rou RNA vo oX4pror[(er 6vo evepy6 rd,wpo rrov6 vo npoo66ver ouprnopdyoweg youovoo(v4q, v' ovoyvop((er ollyovouxleot(dro Kor vo g6pver rrg owr6pc6oeq oprd6eg oe pro 6topr6peoorl nou entrpdnet q 6rdonooq rr enqv{ivoorl 6e4rcirv. 14..trq rv- rp6vro qq ogdSog I 6ev 61ouv 6repeuvrl0e( 16oo tre- mopeprirg 6oo ro twp6vro tou Tetrahymena, uror6oo or r6r6qr5g roug yevu<d e[vor nop6poreg. H ow(5poo4 ouroouppoerig e(vor evdoyevrlg rdr6- rrlro rou RNA ln vrtro, oMd o p6loq rcov npolteivciv ln vivo 6ev SXer xoOoprcrre( nl;1pog. Oproprdveg evde([erg yro rn oupUeroXrl nporeivdrv nopSlowor on6 Urroxov- dproxd oumrlporo, 5nou 4 oupporpq tcov rwpov(trlv rrlg oprd6og I qnone( ro frans-6poorrrd npoiSwo dl,- l,rov yovr6(rov. Mtq ewunooroxrl nep(nroorl e(vor to cyf78 geroMoypr6vo m6l,e1og rou Neurospora crassa, ro ono(o eprqov((er el,orrogonrc] ouppoqrl noMdrv gr- rolov6prorcbv rwpov(<ov oprdSog l. To npoi5v our6 rou yovr6(ou Bpd0nxe 6rt e(vor q pmoXov6proxrl ouv0etdorl rou rupooulo-RNA. Aw6 e[4yefuor on6 ro yeyov5g 6rr ro rwp6vto prnope( vo npooLdBer Ulo rprroroVn SoUri ninou IRNA, 4 ono(o oroOeponotefuor qn6 q ouvOerd- oq npodyowog 6ror r4v roroluru<r1 owbpooq. Aurrl 11 oXSoq ovdpeoo orq ouv0erdorl Kor rn ouppoqrl e(vor o0prpovrl Uo rnv r55o 6rr 11 oupporprj [er(vqoe og ow(- UUUpAtSCCUpUzoUC UUU ,1 -> oG414oH j -/ ,S'exoG.. U UUpAI6CCUpU20UG 1.15 RNA L-1 9 RNA 654 KEOAAAIO 23 Korotrulro RNA Kurtronohon
  • 146.
    6poor1 dtotr-reooloBo6prevrl on6to RNA, q ono(o mq ouv6Xeto unoBor1O40qre on6 npo:teiveq nou npoo65' ,our-iNA, ot ono(eg opXtrd etXov dtrsg leroupy(eq' H xov6rlro ouroouppogrlq in vitro (otog owtnpoooneilet rn Pooirn BtoXrlpurl olJr4Len(6poon. H 6oUn tou RNA }qtrttoupyei rc evepy6 xdwpo, oMd treroupyef onore- r'.#Uoixa in vivo g6vo 6tov unoBoq0e(tot on6 6vo nptrrte[vtr6 o6Pn].oro. 23.3 To lvrp6vlo Inq Opd6oq I Ixnpori(ouv Mro XoporrnploTlKrl Aeureporoyr'r AoUf mena. Evrit ot 6oprtr6g ovolfoerg xcn6qepw vu 6a qort(oouv u1 6eutepotoVi 6opn rou twpov(ou 116 oprd6og l, 11 tpttotoynq 6oUn rou onorotuiggnKs Ps to, np6o,poro npoo6toptotrr6 qg xpumoMucqg 6e prdq. l-1oli.5q rpuoroMtrsg doprdg tcov twpov(cov rrlg opddog I 6youv 6to)teuxov0el xot enrBeBotr'tlvouv no- l,or6rejo prow6lo 6eurepotoyoriq 6ognq' A0o orn6 r,q.,ep,oX6q, tcov ono(cov or Bdoetg e(vot (euyopro;r6- ,ri p"-oti iouq, 6r1;rtoupyo0wot on6 to (euydpopo u"id,ir ouwqprll.rSvcov oMrlloulttilv nou e(vot row6g oro twpOvro ini oudOoq I' H P4 xotooreu6(etot on6 rrq onn*rlLouX(eq P rot Q, ev6 11 P7 on6 ng oMrltrou- X(eg R xqt S. Ot 6)ieq neptol6g notr<fi'ouv. oq npoq irlv'oMrlLouX(o oe 6togopetu<6 twp6vto' H ovdLuorl preroMd[eov ovoyvrop((er 6vov 'nuprivo' rou twpovi ou nou nepdXet trg P3, P4, P6 rot P7 xot nop6let tr1v eLdltorrl neptoXq 11 ono(o prnopef v' ovo]'dBet Uto Ko- roLutrrf ow(6poor1. To prrlrog rov twpov[ov rnq oprd6oq i notr<fiet notr'ri rot ot ouvotvertrSq oMql'ou- X(eg ewon(owot opretd proxprd on6 ttg npoyprotrr<69 ouprBoLSg ouPPo<Pfg''6prqr6veq owt6p6oerg ori(eu(r1q <p6pvouv nq ouU- BoLeq ouppoqnq ot gto 6rop6ptpoorlnou unoorqp((et inu euEuU,r<ri o*iOpoon. H Pl neprl'opBdvel to 3'dr<po rou S'e(ov(ou. H otrr1l'ouX(o oro eocotepu<6 rou tvrpo- v(ou, nou (euyoprirvet pe to e[,c0vto, t<otrefuot eotoreptrq o).trr1loufo-odnV6q (internal guide sequence, IGS)' H ovoproo(o IGS owovot<Ld to yeyov6q 61 rl neptoXq 3' npoq rnv oMqLou{o lGS, nou nopouotd(erot or4v Et- xovo 23.4, opltxd 0eopri0rp<e 6tt (euydptove Ue rrlv 3'ou;rBo).ri ouppoqrlq, rp5pvowog 6tot oe enorpd ttg 6tio'ou;rBotr5g. Autri 11 oL}'r1)ren(6poon Unope( 6wcog vo ou;rioiver, oMd 6e go(vetot onopo(tr1tq' Mto notrti BpoXeio olnqtrouX(o -rdnoteg qopdq, ;rriroug p6l'tg bOo'[dotrr- preto[ri tov P7 t<ot P9 (euyopdrva Ue gto otrql,ouX(o nou Bpioxetor opr6ocog nptv-rn Spoonr<{ G (OG; e6on 414 oro Tetrahymena) oro 3'dr<po rou twpo- v(ou. H oqproo(o tou (euyopri:poroq rov Bdoeov Yto rn aqgroupy(o tou onopoknrou (nupdvoD rou RNA rov[- qeiot on6 ttg t6t6rqteq rtov cis-dpooru<drv (cis-acting) ;retoMd[eolv nou nopepno6((ouv q ouppoqli roJV tv- tpov(tov tqq ogdSog l. T6toteg preroMd(etq 6Xouv ono- provo0e( oe ptttolovdptord tvtp6vto metre1<ilv nou o6uvoto6v v''ogotp5oouv 5vo twp6vo in vivo' En(oqq' 6,Xouv onopovo0e( oro twp6vto rou Tetrahymena Uero- ,p6po*oq trlv owbpooq ouppo<p{q 9- 9Yq Borcnlptor6 neprBdl,trov. H xotooxeurl rr'1q'EIK0NAI 23'5 enttpSnet t1v noporo)ro0Or1oq qg ow(6poor1q ouppoqrlq oro E' coli. To ouroouppom6gevo rwp6vto tono0etefuqt oe pto 06o11 nou 6tor6met to d6roto t<todu<6vto rrlg olr'r1- ioux(oq Yto rrl B-yoLorro(tddor1. Euventirg, 11 npoteffi gnoie( vo peroqpocne( enttuXtirq on6 to, RNA pt6vo o<p6rou ogopeOe( to twp6vto. H oUvOeoT l F-Votrot<co(t- O&onS on6 to E. colioe out6 to o0orqpo uno6rltrrirvet 6tt 4'ouppocpri prnope( vo npqYporonotrl0e( un6 ouvOtl- req opxetd Stocpopenr6g on6 ere(veg nou enu<poto6v arc Tetrahymena 4 or<6trro Kot in vitro' Evtil to twp6vto * To rvrpovro rnq ouoOoq I oXrlgor(ouv uto deuteporoYl 6oun p' evv6o 0ixtrtlveq nePtoX6g. a To x6vrpo rov neptoXtilv P3, P4, P6 rot P7 tlXouv rorotrurtrrl 6pdor1. e 0r neptox6q P4 xor P7 oxrlpor(ovrot ue (euYoptrlUo ouvupn- gdvtov ouvotverLrdrv otrtrltrouXttitv" * Mro otrtrrltroux(o dintro oqv neptoxrl P7 oxrlpori(et (euyn F0- oeov Ue pro otrtrr1trouX(o nou nept6Xet rrl dpoortr1 G' Otro to twp6vto qg opr66oq I opyovcovowot oe Uto XopoKrnptortr<r1 6ewepotoyrl doprrl.g' ewSo 5)'txeq tiri-pzl. H !:Kcl;& ;:.,i nqpouotd(et 5vo ;row6}'o qq 6eutepotoyotig 6og(q rou tvtpov(ou rou Tetrahy' rqovto r fo-oH P 5,CUCUCd flpdrrn 5'UGCGGG 3 i,eOenee UetuQopd 3'ACGCCC 5' _IGS O P8 s 5' UAGUC 3' 3'AUCAG 5' R 2bp oXrlgot((owot mo 3'drpo rou rwpovlou, , To tvrpovto Uq oUodoq I 6Xouv rotvrl deurepotoytl dogrl nou oXrlgori(etot ono evv6o (euyuptrrpeveq nepLoX6g 0t otrtrqtrouXieq rtov neptoXtitv P4 rot P7 eivot ouvqpqgtveq Kt 0v0- yvolp(ouv ro ororXe[o P, Q, R rot S. H Pl nponinret ono ro (eu- yopopo ovopeoo oro drpo rou optorepo0 e(oviou rot rrlv IGS rou rvrpov[ou: 11 neptoXrl Uero[u Uq P7 rot rrlg P9 (euYoptlvet Ue ro 3'orpo rou tvrPov[ou. ?*:.* To lvrpovLo rnq 0Uo6oq I lXrlpoti(ouv Mto Xoporqptortq Aeureporovl AoL'r 655 4 P6
  • 147.
    Ynorrvrlnlq lwprivto evotoltottitvetot orortodtxdvto 10 Ktodrxdvro Ktrl6xovro Kol0x6vro 23.4 lo PrBodv(uuq 'Exouv llorriAeq KorqAurrrdq Apdoetq * l4eroBotrtrovrog rr1 06011 npoobeolq lou unoorptitltoroq o' evo lvrpovto rrlq oUo6oq l, elvot nt0ovtl 1 etooyoyq evotrtro- rlrtirv otrtr1trouXulv nou otrtrrltrentdpouv pe rr1 DpoorLrq G, i 0r ovndpooeLg orotrouOouv r).oorxi1 uvllrq ev(uptrtv pe Xo- UItr1 rorotrulrul roXu11ro. " Avldpooerq nou Xpnotuonotouoov 2'-0H deopoug 0o pno- pouoe vo e(vor 1 pooq yro Uv e[etrt€n rotv ou0evrtrdv roro- trurutlv evepyorrlrov tou RNA. Mol.ov6tt 11 rorolu-ru<q evepy6qrd roug ovot<otrri<p01- r<e Xdprl orrlv rxov6tlrd toug v' ouroouppdmowot, to rwp6vto rrlq oprd6og I e(vot Lt<ovd v' ovoloprBdvouv Kt dMeg xoro).utu<69 owr6pdoetg rn vitro. OLeg out6g ot owrdpdoeLg Boo((owot oe trans-eoreponotrloetg. Oo rrg ovoAriooupe on6 rr1 oxontd rqg oXSorlg rouq Ue qv [6ro rqv ow[6poor1 ouppo<pr1g. H r<oro).uru<r1 evepydrqro ev6g Lwpov(ou rqq opo- 6o9 I otpeiLetor otrlv u<ov6qtd rou vo nopdyet ouyKs- r<prpr6veg Eeureporoye(g r<or tpttotoye(g 6opr6g nou drlprLoupyo0v evepyd r<6wpo toodrjvopto pie to evepyd r<6wpo rcov ou;tBotu<trrv (npoteivtxt-lv) ev(0ptulv. H ': , ,;i, : '' nopouotd(er rrlv ow(6pooq ouppog4q oe o16 or1 pre ourd to r<6wpo (np6retrot yto rrlv (6to oetpd owtdpdoeov rrlg Etrdvog 23.2) To x6wpo np6odeoqg tou unoorpu.rgoroq oxnpo- r((erot on6 qv 5l.Lro P1, ornv ono(o ro 3'dr<po tol nporrou rwpov[ou (euyopc0veL Ue l'lv lGS. 'Evo x6wpo np6odeoqq youovoo(vr1g oXrlpot[(erot on6 oMrl].ou- Xieq orqv P7. Aur6 ro rdwpo nt0ov6v r<ototroptBdvetot eke on6 dvo ele60epo e(oyevdg vout<l.eot(6to youo- voo(vr1g (exoG) eke on6 to rord).otno OG (06oq 414 oro Tetrahymena). h'1v nptrrrrl ow(6poor1 petogopdq, ro rdwpo np6o6eorlq youovoo(v1g t<otolopBdvetor on6 ro etrerJ0epo vouxtreot(dLo youovoo(vr1q. Metd r1v one).eu06ptooq rou tvrpov(ou t<otoloptBdveroL on6 ro OG. H 6e0repr1 preto<popd onel.eu0eputveL ro evo:prdvo e(ovto. H tpitrl pretoqopd dqpLoupye( to xu- r<Lrr6 rwpdvto. H np6o6eor1 mo un6orpopro nepLl.oprBdvet ;rto oL- l.oyr1 6ropr6peoonq. l-1ptv on6 tqv np6odeorl rou uno- mpcoprorog, ro S'drpo qq IGS Bp(or<etot r<owd mtg P2 xor P8, evc,r prerd t1v np6o6eoq, 6tov oXrlpot((et rqv 6tru<o Pl, Bp[or<etoL xowd oe ouwlprlprdveq Bdoetg pe- ro[rj torv P4 r<ot P5. H owi6poon omtKonote(tot on6 enogdg nou ovrXve6owot fin deutepotoytl 6opt4 tqg :':r'.r:' : ' . :rrlv tpttoroyrl 6opr1, to 6Uo r<6,wpo pre to ono(o Spletot evoMorrLrd o' enogrl q P1 on6louv 37 A, yeyov6q nou unodqtrulvet onpowtKrl petox[vr1oq orrl 05or1 tqg P1. Opropr6veq neporr6por ev(uprtr6g owt6pdoetg nou I rr,teroypo,prl I ruppo,pn Mntre ntrdreg dqgroupyorjvrot pe Xptloq yto p-yolono(6dor1 I Mero,oooon V B-yotrorro(r0oon I I ,r'- ,-.a "' .: EIK0NA 23.5 H rono06qoq rou rvrpov(ou tot Tetrahymena oqv rtodrronoultxf dJrlloux(o rnq B-Ydroxto(tddonq drlptoup- ye[5vo npooDtoptolto yto TI'lv ouToouppoqn oro E. col1. H ouvOe- on tnq F-yqtrono(tdoonq eEerd(etot Ue npoo0nrn ptoq Xlgxrlg dvoorlg nou y[veror pn]re on6 to 6v(upo. H otrtr1trouXio peroq6- peror ono 6vo porrqpLorpdyo, ouven6g 1 nopouo[o gntre ntroxtitv (nou nepr6Xouv potruop6vo BoxrnpLo) unodl)rtlvet entruXlg6vrl ouppoQn. qq oprd6og I rou Tetrahymena o<potpeftot p6vo rou on6 ro mRNA rqg B-yoLorcro(tddoqg orc E. coli,6ev eL vot ooq6g ov 4 ow[6poor1 unoBoqOefuot on6 Bomqpto- 169 nporeiveg. Ie out6 tov npoo6toptopt6, petoL- l,d[erg orrq ouvorvetu<5q otrtrqLou1(eg qq oprd6og I nou droropdooouv ro (euydpoprd toug oropototiv 11 ouppo<pr'1 ror dpo nopeprnod[(ouv rqv 6rEpoorl rnq B- yoLorcro(t6dor1g. Ot petotrl.d[erg prnope( v' ovoorpo- cpo0v pre owrpponrorrx6g oMoydq nou onoxo0toroUv ro (euydpopo rov Bdoecov. Or preroMd[erq onq ow(orotleg ouvolettr5g oM4- LouX[eg rov prroxovdproxrilv twpov(tov rrlg optddog I 6Xouv nopSpoteg entntrioetq Ue ttg ow(orotleg nou nopor4poriwor oro Tetrahymena. Mto prerdtrLo[q oe Uro ouvorvermrl oll.rll.ouXio ;rnopef v' ovqorpoqe( on6 gro gerdLtro(q oe pto oupntrqprr:prottt<ti ouvowenKn oI- Lrll,ouX(o onoro0torc0wog 6tot ro (euydprr;pro: Yto no- pddety;ro, preroMd(etq orn ouvotvertrci oLLrlLouX(o R prnope( v' owrppoTlrl0o6v on6 petotrI.d[etg orrl ouvot- verrcj dtrqtrouX(o S. Iuvotrrxd, ourd t' onorctr6opoto uno6flrcirvouv 6tt q ow(6poo4 ouppog(g qq o;rd6og I efoprdtot on6 to q4r1;rotropr6 6ewepotoyo0q 6oUnS ovdpeoo oe (e[y4 rov ouvorvrrrrdlv oMqLoulrc0v oro eorotepm6 rou tv- tpov(ou. H opfn nou roOtepdrOqre on6 ouq rqv epeu- vryxr] epyoo(o e(vor 6tr aMqLouybq nou aniyouv an6 nq oupBoLiq ouppaertg anarcolwotyo va oyqpauorc( rc evepy6 xiwpo to ono[o xalotd eELxtr'1 rnv aurooup- paEi. 656 KE0AAAI0 23 KorotrurLrd RNA Boorr6g 6worcg
  • 148.
    To rorotrulro RNAdxsi Uro 060n np006s0!g rnq youovooivnq Kor Uio gSon rpo06s0nq rou unootprirporoc 0don npoodeolq iilg youovoo[vr1g -3' d414 :-.'&a.'- ''- 0don nodoOeonc -ffiffi rou unomotiruotoq GGGAGG UUUACCU I fleprdXet Al 6 xor U20 iis:;;z:t:t:;:"t::+**i H G-OH rorotropBovet rn 06on nprlo0eolq G. To 5' e(6vro roro)'ogpoveL q 0do4 npoooeonq Tou un0oTp6U0T0q. -3' 8414 ,tw,6H ':' ,*J;A.GGGAGG#:. -3' 8414 ---> w G*l 5',-@cu . GGGAGG ..' :ii::1r;:.i ,.i;-',:t:.*.:: H G414 Bpioxeror rq 06o11 npoobeorlg G. To 5' e(civro pp[oreror q 0eon npoodeoqq rou unoorpoUoToq + : l-',.j(-ij:' i:i:.:'; ;$:ao;p: H G4l4 Bp[oKeroi oq 06on npoobeoqg G. T0 5' 0Kp0 rou rvrpov[ou pp[oreroi rr1 06011 nprlodeorlq Tou uttooTpopoTo Al: -t-1i"'- / + G,-,S[EffiffiCU.r, GGGAGG..,i; G _j,-i.,:a..:.;{:'.; H e rrop1 Tou ivTpov[ou qq opo0og I oro rRNA rou Tetrahymena 0uUpo[ver Us 0rodoxrKeq 0vTr0p00erg 0v0Ur00 oTrg 0v0l0srq nou KoTo),ouBovouv Trl 0eo1 npooDeor.lq Tnq y0u0v00i vlq Kor ul e60l rpooosqg Tou urooTpidUoToq. T0 oprorepo e[6vro eLrov[(eror p0( ev6 To 6eqrd Uop. 6Lexneportrlvovror on6 LVrpdvLo rlq oUd6oq I rou Tetrahymena ovoertpowor orrlv i ": . r . To prBo- dv(upro l.erroupye( o.rg ev6oprBovour<l.edoq er6rr<q yro ouyr<er<p r pdveq o^.InIo u X(eq Xp! or Uono Lovrog rrlv rKov6rnro rnq IGS vo npoo66ver ouprnLqptrrprotrx6g oLlrl).ouXieg. Ie our6 ro nopdderypo, ilpoo66ver 5vo e[rr;teprr<6 un6orporl.ro ilou nepr6xer rqv o]..qtrouX(o CUCU, ow( rnv ovd).oyq ol,l.r1trouX(o nou ouvneoq nepLSleror mo dr<po rou S'elov[ou. 'Evo vour<].eor[6ro nou neprdler youovoo(vq Bp(oxetor mo x5wpo np6o- 6eor1g youovoo[vlg xt enrr(0eror orrlv o].].r;].ouX(o CUCU r<ord rov (6Lo or<ptBtrlg rp6no Ue rov ono(o 66- leror en(Oeoq ro eEolvro mqv npcoq ow(6pooq ;rero- eopdg. Aurq q ovr(6poo4 6roond rlv oLLqLouX[o-or6Xo o' 6vo 5';-r6pro nou prord(er pe ro lr (f(l(lALTLr EIK0NA 23.7 H 06oq qg IGS otrlv Tprroroyn DoUrl otrtrd(er 6rov oxngonoTe[ n Pl p6oro rpdodeorlq Tou unoorp6U0T0q. S'e[c0vto rr rivo 3'pr6pro, nou <pdpet 5vo re].rx5 ro16- Lorno G. MeroMdooowoq ro ororxe(o lGS, oLLd(er Kor n er- 6rr6rrpo rou prpoev(fpou ro ono(o ovoyvop((er nl.6ov ol,t1).ouX(eq ouprntrr1popronr<5q pre q v5o otrtrr1lou{o orrlv neproX4 lGS. Autrl n rponono[non rqq IGS rirore v' o).d(er q er6tr6rr;ro rou x5wpou np6odeo4q rou unoorp6gorog ennpSner oe dl,i.q RNA-cr61oug vo ugiorowor ene[epyoo(o on6 ro prBo6v(u;ro, to onofo Unope( en(ong vo enrrelSoer owrdpdoerg RNA-tuyrd- o4g. 'Evo RNA nou Korolnyer oe 3'-OH npoo6Sveror oro raiwpo rou uTroorp6poroq rt 5vo RNA nou Koro- )rrlyer oe 5'-G, npoo65vrror fio K6wpo np6odeoqq G. H en(Oeor1rnq u6po(ulopddoq oro gooeopur6 6eopr6 ouvdSer ro 66o pr6pro RNA ;.re onrirLero rou Korolo(- nou G. H ow6poo4 rnq eooqqrdo4g 6e q(er(Eeror dpeoo pe ng owr6p6oerg ;rero<popdg rord rq ouppogrj. Mro ol,ty ovo u rl,eond u<r1 otrLqtro uX(o ou prnLrl p ro prot u<ri pre qv IGS nou roro).r1yer oe pro 3'-groocpopu<q opd6o 66- Xeror en(0eor1 on6 r4v QG. H rpoorpopu<r1 oprddo ;rero- qdpetor orrlv OG xr 6nerro oneLeuOeprirverot 5vo olryovouri.eot(6ro pr' 5vo el.efOepo 3'-OH. Iq ouvS- Xero, n gooEoprrui oprddo prerog6peror efue oe 5vo ofu- yovoux.treor(6to nou rorotrrlyer oe 3'-OH (ouorootrrd, ovoorp6qowoq rrlv ow(6pooq) efue oro vep6 onel.eu- Oepd:vowog ov6pyovo rpd:orpopo r<r ol,or<Lr1 privowoq puo ouOewrrrl ow[6poo4 groorpordoqq. Ot owt6pdoerq nou rorol,riowor on6 ro RNA nepr- ypdrpowor 5nog or r<i.oox6g ev(upru<69 owtdpdoerg ;re 6poug rrvrlrxqg Michaelis-Menten. H EIK0NA 23.9 ovo- tr(er rrg owr6pdoetg nou rorolfowor on6 ro RNA. Or np5q qg Kn,r Tu)v owt6pdoeov nou roroLtlowot on6 ro L: GGGAGG Enoqdg gero qv npdo0eorl rou unoorptiporog Iii.4 To Ptpo6v(upo'EXouv llorx[],eg Korotrunxec Apooerg 657 w Enog6g nprv on6 qv npdodeoq rou unoorpolcio( s', @@s,cu GGGAGG
  • 149.
    'Ev(u;ro Yndmporyo KM (mM) Avur6xtrosn flmin) t i5'ffi*3' GGGAGG5' * S,*CUCU-0H + loooet6Sq 1 9 pdoeov lwprlvLo L-19 RNA RNAdsnq P nlripnS RNAdon P RNAdoq T1 p yotrorro(tddorl RNA 24 pooetov 0.0006 0,5 cccccc 0.04 1 7 npotRNA 0.00003 0.4 pretRNA 0.00003 29 GpA 0 05 5,700 trorro(q 4,0 1 2,500 5'_Y@3' +u + 5'UCU-oH 3 ' EIK0NA 23.8 0r KoTotrurKdq qvTrdpdoerq rou p0oev(6[0u rspr- trogpdvouv trans-eoTeponoqoerg ovdgeoo oe Urq oUd60 Tnq ee- onq npdoDeonq Tou uflooTpouqroq Kor Urq oud6o TIq e6onq rpdooeonq rnq G. RNA efuor XoUnMq, yeyov6g nou uno6rlLri:ver 6rr to R- NA npoo66var ro un6orpo;r6 rou Ue uqrnln er6u<5r1ro. Qor6oo, or oprOpro( ovor<UrLoorlq yro nq owr6pdoerg nou roroLriowor on6 ro RNA e(vor 1.uxpo[, yeyov6g nou owtxorompi(er q Xoprllll rorol,uurrl roXriqro. ['lpo- mxd, ro pr6pro RNA ou;rnepup6powor Uetov 6ro yevr- 16 rp6no 6nog xor ro nopo6oorord 6v(upro, ov Kot e[vor oletrxd opyd ouyrpnrrd pe rouq nporeivrro(g rorol,Ureg (6nou ro runw6 e0pog tov oprOpcirv ovorU- rtroonq efuor 103 dcog 106 minl). Ot 6poor4pt6qreg r<ov prBoev((pulv enerctdO4rov Ue rnv ovor<dl,urlq 6n prnope( vo pu0;uoroUv on6 ouv- 66teg (BL5ne Ev6r4ta 30.3). Aw6g or pu0pronr6g cis- 6poorrx6g neprol6g xol,otjvtor prpo6ro16nreq (riboswitches). Ee 6l,oug q(e66v rouq prBo6ror6meg, puo geroBolrj dropr6pqooqq r<o0op(et rlv xotdoroorl on/otf. A[rooqpe(corq e[o(peor1 e(vor ro yov(6ro g/mS rtov Gram-0erx6v Borcrrlp(ov, ro ono(o g4rlpor(er 5vo ouro6roonrirpevo ppo6v(ugo nopouo(o 6-rproogopm{g yluro(opr[vqq (GlcN6P). H tlK0NA 23.10 ouvorf((er 11 p[0pror1tou prBodror6mq g/mS. (l-ro neproo6tepeg ].e- mop6peeg, Bl$ne Ev&1ra 30.2). Av ro evepy6 r<5wpo e(vor 1.uo enrgdvero nou er06- rer pro oerpd evepyolv opddrov oe ndyo prero[rl rouq ox6on, y(veror rorovoqr6 ndlg ro RNA e(vor rxov6 vo nopdoXer 6vo rorol,utrr6 r6wpo. Ee pro nporelw1, or 658 KEOA AI0 23 Korotrurrxd RNA EIK0NA 23.9 0t ovn6pdoerg nou rorofujovror on6 ro RNA 6Xouv ro Oro XoporrrlpLolxd pe lq ovrL0pdoerq fiou rorotruowor on6 nptoretveg, gotrov6r 4 roXUqrd roug e[vor BpodUrepn. H Krvr 6(ver q ouyx6vrptoorl rou unoorptirgqroq flou qnquefror yro vo enr- reuX0e(ro pro6 qg Udyromq roXUrrlrog: e(vor 6vo ovr(orpoqo p6- Tpo Tnq ouyy6verog rou ev(Ugou yro ro undorproUd rou. 0 optOpdg rov ovorurtrtiroeorv e[vor o optOpdg rtrlv lrop(trrv rou u[o- orp6gorog nou gerogoprpt0vowor oT1 povddo rou Xprlvou o' 6vo p6vo rordruilxri x6vrpo. 6poorr6g opddeg nop6lowor on6 ng nAeupu<69 otru- o6eg ru:v o1.uvo(6rov, or ono(eg epr<pov((ouv r6ro(repr1 nomil.io ror nepr).oprBdvouv Oenr5g r<ot opwlnx6q rowr- 169 oprddeq, r<o0rilg r<or u6p6<poBeg oprddag. Iro RNA, or 6ro06orpeg p((eg e(vor neptoproprdveg ror onotetrotiv- tot t<up(tog on6 rrg er,ee0eqdveg oprd6eg rcov Bdoecov. Bpole(eq neproX6g rou RNA ouyr<poro6wor oe ouyKe- rp rp6vr1 6eweporoyrl/tpnoroyrj 6ro 96 pgoorl, nop6- Xowoq 6ror puo evepy6 entgdveto r<ot 6roqpriwog 6vo nep$dMov oro ono[o ;rnope[ vo droonooroUv Kqr vq oXrlporroro0v 6eopo(. Oo(verqr ovon5rpeurcro 6n q o).- LrlLen(6poo4 pero(U rou xorol,uru<od RNA Kor rou unoorpdlprot6g tou Boo(etor oro (euydpropro tcov Bd- oeov npoKerp6vou vo 6qproupyq0e( 6vo evepy6 nepr- BdLl,ov. Arnotru<d Kqldwo (ouvriO<og Mg2*) no[(ouv orlpowrr5 p6Lo mr1 6oprl ror runxd Bpiorowor mo evepy6 r<6wpo, 6nou ouwov((ouv rtg 06oerg tov drd- eopov oprd6tov. To 6rno).txd gerotrtru<d Kffir6wo en(- oqg nof(ouv dpreoo p6].o mrlv evdovourl.eo],ulrr1 6pdoq tov prBoev(ripov tcov roet6cbv (BL6ne Ev6rrrya 23.9). Or e[efurcrx6q npoerrdoerg our6v rcov ovor<otrri{re- <ov e[vor o[roo4pre(orreg. H "6tXoo;r6v4 npooonu<6upo" rou yeverrKori prrlXovtoproU -orov ono(o ro RNA Bp(oxe- ror oe 6l,o ro ouoronrd, oLld ot nporeiVeg ovotroprBd- vouv r<oro.truru<69 owr6pdoetg- onoteloUoe ndwo 6vo yp(go. Oo[veror on(0ovo ro ev6eX6prevo ro npdrro ou- oniproro owrypoqqg vo nepre(1ov r<or voux.trsivurd o[6o xor nporeiVeq. Yno06ore, to01600, 6rr to npoto ou- orqporo nepe(Xov g6vo 6vo ouroqwrypog6gevo vou- rLetvu<6 o[U pe nprrlr6yoveg roro].wrr6g evepy6qreg -g6vo 6oeq Xperd(owor yto rq 6rlproupy(o r<or rq 6td- onoon rpoogo6teoreprrciv deoprdrv. Yno0riore en(or1g 6l q ouprpreroXrlrov 2'-OH 6eopc0v onq onpepvdq ov- ndpdoerg ouppogqg ovdyeror oe our6g rrg opXdyoveg r<oroLuttr6g evepy6rqrcg. Me ourrig rtg nopo6oXdg, 0tooqordorl RNA trtyxdorl
  • 150.
    G-C P3 G -e) o-o o-o a- o- o- o- a- o- o o OP3,1 o o o a U .0r0xl rou mRNA To oIIo[0 KoorKonotel T0 ev(uuo nou nop0Yet :-:oorpoprril ytruro(opivn (GlcN6P). '0rov 1 GlcN6P npoodevetot ' , pLpo6v(upo, onoono ro 5'orpo rou mRNA, odpovonot6vroq : <L epno6[(ovT0q rrlv nepou6po nopoYoYrl rou ev(rjpou. 0t ne- : :Xeg, nou e(vot orlpovrLr6q Yto fll Dtorrlplon rrlg evepyou rptro- :','ouq doUnq, errov((erot pntre, evdt I0eol Dtoonooqg i .ol1polveror ;t' 6vo roruvo petroq. (Avorunoorl ono Curr. Opin. :.'uct. Biol., vol. 17 T. E. Edwards, D. J. Klein, and A. R. Ferr6-D' --rar6, Riboswitches: small-molecule recognition . . ., pp. 273- --9 Copyright 2007, pe qv odeto rou Elsevier [http://www.sci- . -.r e d irect. com/science/jou rnal/0959440X) . -:ope( vo 6rorunto0e( to entXe[prlpro 6tt ro ou0ewtr6 .cur<.treivrr5 o[U rlrov to RNA, r<o0cog ro DNA 6e 6to05- iL 2'-OH u6poluLopd6o xot ouvenc,.rg 6e 0o prnopo[oe . : drexneporooer tdtoteg owrdpdoetg. Ot npu:tefveg -:,,ov npoor50rp<ov Xdpn orrlv tt<ov6tqtd toug vo -oOeponotoriv q 6opr1 tou RNA. Xdprl orq preyotr0te- :- eue).r((o rouq, ot nptoteiveg orrl ouv6leto ov6LoBov trg r<ototrulx6q owLdpdoetg, odrlyo.lwog teAtxo c:: ' nolrjn).or<o oriyXpovo prllovtopr6 trlg 6xgpoorlQ rt"; yovt6(rr.lv. 23.5 AptapCvo tvrp6vro Tnq Opd6oq I Kor6rronoro0v Ev6ovourAedoeq nou tlpodyouv Tnv KrvnrrK6rnro * To rtvlro rvrpdvro e[vot trovo vo eloXtopouv oe v6eq Oeoetq. * To rtvlro tvrpovto Uq oUodos I rtodtronotouv gtu ev6ovou- rtreoo1, q ono[o npoyporonote( pto 0trtrtovr1 evro;lr1 oe pto 0doq-oroXo. * To tvrpdvto perorlOeror oq 06orl rr]q dLrtrovr]q evroplg p5- ou.r evoq ovlypoqtrof glXovtopou nou 6togeootrope(rot ono ro DNA. Optopdvo rwp6vto t6oo trlg opddog I 6oo r<ot ttlg oprd6og ll nepr5Xouv ovotXrd nLo(oto ovdyvtr;or1g nou prerogpd(owor oe npora(veg. H 6xgpooq rov npco- teivo-lv enrrp5net oro twp6vto (e(te mrlv ou0ewtr<q Uopen rou tl:q RNA eke cog 6vo DNA owtypogo rou RNA) vo r<Lve(rot, on6re e(vot txov6 vo eroXtope( oe Uto r<otvoripro yevoprxrl 06oq. To twp6vto t4g oprddog I ror ll e(vot 6ro6e6oprdvo t<ot undpXouv t6oo os npo- ropuorreg 6oo rot o' europutrtteq. To Lwp6vto trlg o;rd6og I petovooreUouv ;t5oto prrllovLopttrtv nou 6to- preootroBorjvrot on6 ro DNA, evot ro twp6vto tqg oprddoq ll petovooreUouv prSoo prqlovtol.ttov nou dto- ;reootroBotiwor on6 ro RNA. H r<rvqttr6rrlro rcov twpov(ov npotoovtlver-i01re on6 6tomouprrroetq ottg ono(eg t' oMrlL6propqo rou olelro6 yovr6(ou 616<pepov oq npoq qv r<otoXtl rou tv- tpov(ou. l-lotru;toptptopol ovo<poptrd pre qv nopouo(o 11 qnouo(o rwpov(rov e[vot t<otvo( oro pirol6v6pto rov pu- rr[cov. Aut6 e(vot ouprBot6 Ue rnv dnoQrl 6tt ro ouyre- r<prprdvo twp6vLo npoqL0ov on6 eLolopqon oro yov(6to. Mto ovdluoq ovoouv6uooproU ttDv 6tomquptooerr.rv mqv ono(o Xprlotponotrl0r;re to yov(6to tou peydtrou r- RNA tou puroXov6p(ou rnq (UUnq 6pt[e qcog oqv ntOo- v4 6tepyoo(o etolrrrpqorlg ttrtv Lwpovlo.lv. To yov(6to tou peydtrou rRNA tou prttoldv6ptou qg (Upr1q 6XeL 6vo twp6vro oprd6oq I nou nept6Xet Uto Ko- 6u<onotqrLr( oMrl).ouX[o. To twp6vto Bp(or<erot ot opt- opdvo oreLdxn (0UnS (xootiwot to*), evo onouotd(et on6 dMo (to ). Ot on6yovot t<ov yevettr<lv drooroupto- oeov petofri rou o+ Kor rou ro 6ev 66otoov rov ovope- v6pevo yovotuntr6 l.6yo, xoOrig ouvqOotg rltov co*. Av Oeoprlooupre ro ordLeXog ot oq 66q r<ot to ordLeloq to rog 65r<111, odrlyotiprome ornv dnoqrn 6tt oe 6to- oroup6oerg o+ x 0)- nopdyetot 5vq v5o owtypo<po tou rwpoviou oro o- yovt6(topro. 'EtoL, 6l.ot ot on6yovot e(- vor o+. MeroMdletg prnope( vo ouprBoUv oe onotodqno- I Ktodrronororjv Evdovourtreooeg nou llpooyouv qv KLvnrLxo:-.: 659 aa o-o o-o a-o G -C o-o o-o G -CDOO lJ -tJ c -c G G."y' o-o o-oP2 o-o o-o a-o orffiou' ox- O' UA o-o o-o o-o o-o o-o O-O A.A A.O AA Oa O'A t.o-o o-o o-o o-o o-o var a oA t -G - c P2,1 a U GA Go oo A- i :<-'t- 1oo.o-t?O-O-l 23.S 0prop6vo lvrpdvro rnq OUd0oq Boorxdg dworcg AC G
  • 151.
    RNA I Ev6ovourtreoorl Fa A'*'qOeor1-oroyo I*u**u7.*R;{& To rvrpovro ovlypdtperot rr 6nerro evoorpor6veror > &6u{{ .{Xb{,& 'Evo rvrpovto xui6tronoLe( ;:ro evdovouxtreoorl nou npoyporonorel pro 6frtrtov1 eviop4 DNA. H otrtr1trouXio Tou rvTpo- vlou ovrtypoqeror rr eneuo evoopordtveror oqv evroprl. 660 KE0AAAI0 23 Korotrulro RNA re norprr6 ordl,eXog pe onor5l.eopo n Ul Mev6SLeto yovotunrrrl r<orovoprrl vo Xdvetor. OptoprSvo meL5X4 epgov((ouv rovovtr6 drolropropr6 pre ioo opr0;r6 o+ ror co- onoy6vrov. 'Otov Xoproypoeo[wor, ot preroMd(erg rou ore).5Xoug ro- Bp(orowor r<owd orqv neproXrl Snou 0o pnoporioe vo eroxoprloer ro rwp6vro. Or trreroMd- [erg ev6g ore].ri1oug co+ Bp(orowor oro ovoyvtoonr6 nLo(oro rou rwpov(ou xor eprno6((ouv tqv nopoyruyq npcoreivqg. To yeyov6g out6 unoorrlp((er ro prow6).o qg EIKONAI 23.11, orqv ono(o n npotretvrl nou ro6u<o- noefuor on6 ro rwp6vro o' 6vo or6Lexoq o*, ovoyvop(- (et 14 06o4 6nou 0o 6npene vo eroXcopqoer to rwp6vro o' 6vo mSl,eXog o- Kor euvoe( rqv rlrlpovoprrr6tqrd TOU. OproprSvo rwp6vro r4g oprddog I r<o6xonoroUv ev- 6ovouxledoeq nou to xq0rmoUv rrvrlrd. YndpXouv roul.dXrmov n6we oxoy6vereg yovrd(iov yro ev6ovou- r).edoeg enorrtoprori (homing endonuclease genes, HEGs). Ario ouvrl0ropr6veg ou<oy6veeg HEG eivor r1 ev- 6ovoux),edoq LAGL-IDADG ror q ev6ovoux.l.edon rou rourro6 His-Cys. To rwp6vro r4g opd6oq I nou nep6- Xouv HEG owrnpoooneUouv 6vo ;urp6 noooord rou ouvo).xo6 oprO;rof rov nuprlvKcbv rwpovirrrv qg opd- 6o9 l. Ev6 51ouv rouronot40e( nep(nou 1200 nup4vu<d twp6vro opddog l, Lry6repo on6 30 nepdXouv HEG. To rwp6vto rrr nepr6ler 6vo yov(6to HEG nou rodxo- noei "rr1v ev6ovouxl,edorl l-Scel. H l-Scel avoyvap((et rc yov(6Lo u-uq m6yo yo 6kAuv4 0pa0or7. H l-Scel ovo- yvrop(er pro oMqLou{o-or61o trrrlxoug 18 bp, nou ne- prSXer 14 05oq ero1c6pr1o4g tou rvrpov(ou. H otri.rlLouX(o-o,6Xog 6roondror oe rdOe xLdrvo rou DNA oe on6orooq 2 B6oerg on6 rrlv 3'neproXrlqg 06o119 er- ox6pnonq. 'Eror, ot 06oerg 6rdonoo4g onSlouv 4 bp t<or 64 puo u pyo6v enxpeprdgevouq provori g r<Lrirvo u g. Au- rn n Uopqn drdonooqg oXet((eror Lre ro XqpoKrqptonrd rov rpovoflo(ovfcov 6rov perovooreUouv oe vrleg 05- Eq6vro lvrpovro E[rrtvro &h..6kfir"/&&t& lrrrrrarrrr.rr.rrrra 060r1-oroXog *b#&ift{g-&ffi oerg (Btr5ne KeqdLoro 17). H 6(xi.rovq ewop4 pdLtrov nupodore[ pto 6epyoo(o yovtdroxrlg perotponrlg, oqv ono(o 4 oMqLouX(o rou yovr6(ou o+ owrypdeeror yro v' owrroroorqoet qv oMq).ouX(o rou yovr6(ou co-. H owbpooq ouv(orqrot oe prerd0eoq prSoot ev6g Unxovr- opori owryporprlg r<or npoyporonoekor ;r6vo oro en[ne- 6o tou DNA. H erq4ripqoq tou rwpov(ou dror6met rqv oMrll,ouX(o flou clvoyvopf(eror on6 rqv ev6ovoux.tre6- o4, 6roogo).(owog Stor rr1 oroeep6rnro rou 1lop(ou. l-1op6poro rwp6vro ouprd ;reroq6pouv opr<erd 6ro- <popenrSg ev6ovour.ledoeg. Yndplouv 6rorpop6g ong l,emopSpeeg qg erqXd:pnonq: yro nopd6erypo, q ev- 6ovouxl,edoq nou rco6tronore(tor qn6 ro rwp6vto td rou gdyou T4 droond puo 06or;-or6Xo 11 ono(o Bpiore- tot 24 bp ovo6rrd tqg 05oqg etoXdrpqoqg rou rwpov! ou. H drdoroon rnq otrL4),ou1(og rou twpovlou on6 qv oMrl).ouX(o r4g ev6ovoux.tredoqg rov((eror on6 ro yeyov6g 6rt or (dreg oMqloufeg ev6ovour<].eoorilv Bpi- oKowor ong rweiVeg (oMr1Lou1(eg nou rco6rronororjv ourooupponr6preveg npcotetveg - BL5ne Ev6r4ra 2s.12). H norril.(o rov ev6ovourl,eoocirv oqpo(ver 61 6ev undpXer opoloy(o prero(0 rcov oMlLouXuirv rrov ow! ororxov 05oeov-m6Xov. Or oXerrrdg 0rioerg-m6XoL 6Xouv ;reyd).o pqrog (oe r<L(proro 14-40 bp), dpo ror Ueyoln er6x6rr1to. H erdu<6rrpo e(oogoL((er 6tr ro w- tp6vto 0o noMonl,oord(etor pr6vo prloto eroXdrpqoqg oe pro 1.rovo6rxr1 05or1-m6Xo xor nou0evd oMoti oro yovr6(opro. Aur6 r<oLe[ror enolxrop6g rtov lwpovirov (intron homing). lwp6vto nou peroE6pouv ollqLouX(eg or ono(eg ro6tronoroUv ev6ovour<Ledoeg Bp[oxowor oe notx0.o Bomrl pro rot ;rovo rUrropo u g/ollyo x6rropou g eu xo- purilreg. Aurd t' onore).64roro evroXriouv rrlv dnor[r1 6n ro twp6vro nou prerorpSpouv rtobxonorqnrSg oMrl- Loul[eg [ex(vrloov trtg ove[dpqro ororle(o. 23,6 To lvrp6vro Tnq Opd6oq ll 1ooq Ko6rronoro0y l-loAuAelToupytKdq fl poretveq * To rvrpovro r1q ogodoq ll ourooupponrovror ln yltro, otrtro ou- vrl0trl g unopol0ouvrot on6 nptoretve g no u rtrld txono lo uvror ono ro rvrpdvro. * 'Evo U0v06rKo rodrronotllr6 n),0(oro rodrronore( Uro npul- relvrl p' evepyorqro ovlorpoql g peroyporpoorlg, ;uo evepyo- qro poroupooqg, 6vo gor(Bo npoodeolq DNA rot ;uo DNA ev6ovour),eoo1. s H evdovourLeooq droono ro DNA-oro1o enupenovroq rrlv evotrrportool rou lvrpovlou oe gLo veo 06o1. I H ovr(oTpoq1 peroypotpoorl d1;uoupyei evo DNA ovr[yporpo rrlg otrtr1trouX(og RNA rou evoogoropevou rvrpov(ou. To ro).rirepo XopoKrrlpro;-r6vo r<rvq-rd rwp6vro rrlg Buorxfg evvoreg
  • 152.
    cud6oq ll rotdtronoto0vprto povodtr<r1 npoltetvq oe -Lo neptoltl tou twpov(ou nSpo on6 tov t<otol'uttx6 .upqvo -or. U tuntrrl npcotetVq nept5XeL prto evepy6- :'tto N-teLtxrlq ow(orpogrlq UeroYpoq6onq, pto Kev- .crxr1 neptolq nou oXeti(etot Ue Uto entxouptt<t1 :,,,eoy6tqto q ono(o unoBorl0d to 6(nLotrro rou tvrpo- , rou orrlv evepy6 6opr1 tou (xoeitot protoupdorl' -aturase - BLSne Ev6tqra 23'7), ptoneptoXll np6ode- rtg oro DNA r<ot prto C-retxr1 neptoX( p' evepy6tqto : ,,5ovouxtredorlq. H evdovouxl'edor1 (erwd qv ow(6poor1 qg pret60e- :-"q r<L 61et tov [6to p6tro mov enotrtop6 6no:q q ow(- -cLp1 evdovouxledoq tou twpoviou qq oprd6oq l' H :.rio-rpo<Pn pieroypotpoorl nopdyet 6vo DNA owtypotpo :; rwpov[ou nou evooUoro0qr<e orrl 06o11 enotrtopo0' - evdovout<tredor1 en(oqq droond oe notr'0 loprql'6repq ;-;ydqto 05oetg-or61ouq nou protd(ouv, 1op[q vo ei .:.'novoprot6tuneq, Ue q 05on enotrLopori, oSqyowog ; ivor-opdrool rou twpov(ou oe v6eg 06oetq' H ,,.,, '. :i. nopoUotd(et rqv ow(6poor1 petdoe- :-c ev6q tunrrorl twpov(ou rqq oprd6oq ll' flptrrtov, 11 ., lovout<tredon npoYUoronote( prto 6(xl'ovr1 ewoprrl r-3'"' owtouv0ettr<6 (owcilvul-to, antisense) xtr<rlvo' H - :rroo1 tou ouvOetrrori x.I.o-rvou enttuyXdvetot Ue Uto :...5poor1 owiorpoqrlq ouppo<pqq, rord qv ono(o ro ="A Lwp6vto evotrll.tottlverot ovdpeoo oro efrovto tou l'.A Aur6 to 6v0eto RNA twp6vto evepye( nL5ov tr:g .-*cye(o Yto rrlv ow(mpo<pq preroypoqdoq lXe66v - r rc twp6vto rrlg opd6oq ll dXouv evepy6qto ow(- - :cof g pretoyporpdoqq nou e(vot et6txrl yto to twp6- : tt ow(orpoQrl UeroYpoqdoq drlprtoupye( 6vo DNA -. .!'poqDo tou twpov(ou pe tefur6 onot5Leopto rqv ev- - --oroon tou twpov(ou orq 06o11-o161o un6 Uopqtl - 't.-rvou DNA. 23.7 Optap6vo AurooupponT6pEvo lvTpovlo Anotro0v MorouPdoeq * T' ouTooupponrogevro tvtpdvto pnopei vo Xpeto(ovrot evep- yoqreq poroupoorlq nou rodtronotouvrot on6 ro tvrpovto rot porl0otiv oro Dintrorpo rou o' evepyo xorotrurtrtl 6opfl' l-1op6tt ro twp6vto ttov opddtov I xot ll 6Xouv rrlv tKo- v6rqro ouroouppogtlq in vitro, un6 rovo.vtt<5g ouvOr1- xeg ouvr'10toq Xpetd(owor BorlOeto on6 npo:tefveq' Koi ot 60o rUnot twpov[ov (ocog t<trl6tronoto[v evtp- y6trlteq poroup6oqg, nou unoBoqOo0v rqv ow(6po- on rnq ouppoqrlq. ' H'evepy6tlto protoupdoqg e[vot rgnpo tou 1'tovo6t- xori ovotXrof n].oto(ou ovdyvtootlg nou t<<o6tronote(tot on6 to Lwp6vto. Ito nopd6erYpo rov twpov(u:v nou ror6tronotoriv evdovoux}'edoeg enotrtoptoti, to ;-tovo6t- 16 npo.lteivtt<6 npoi6v 51et evepy6trlto t6oo ev6ovou- rledoqg doo xot potoupdorlg. H ovdtruoq getotr'- Ld[eotv 6ei;yet 6tt ot 60o evepy6tqteg e(vor ove[dprq- ."g. U 6oprtr<r1 ovdtruoq entBeBorc6vet to 5e6opt6vo tcov pretoMd(eolv t<ot 6e(Xvet 6rt ot evepy6tqteq qg ev6o- vouxtredoqg Kot rnq protoupdorlg nop6Xowot on6 6to- qopettrd evepyo rdwpo tqg npo:relvr1g,.r<o06vo on6 to'ono(o xtodtxonotekot ond 6tocpopettxq neptoXrl H ouvOnop(r; tov 6[o evepyorlto-lv oqv (6to nptoteivrl unodqtrutvet qv e(etrtrcrtrrl nope[o tou twpov(ou' H ": , :' ', uno6ntrovet 6tt to twp6vto e[e]'(10r1re on6 6vo ove[dptrlro ouroouppon-r6prevo orotXe(o' Etr<d(e- ,'n?Vr0don-oroxoq ,72rru, I IMeroypoQn Evdovourltedon / Awi'rporpn UetoYPotP0on ev6ovoure6ori 5'& 3', EvorLlgortrloq "rou RNA tvtpoviou nou orotrou0elrot ond ovr(orPoql UeroYpoQn Yto ro oXrlPonogo cDNA . H .vTioTp.qrl ueroyporpoon/evbovourtreoorl nou rootronote[rot ono 6vo tvrpovLo enrrpdnet o'6vo ovr[ypotpo rou RNA vo , , *:oro.l0e[ oe Pto 050r1-otoXo. ?t.: 0proU6vo Aurooupponropevo lvrp6vto Anotro0v Moroupdoeg 661
  • 153.
    *6.&{4 *$ffiW,t 6{, I,-ll.q*L;* I - rwoo*o*#%,-l ITo yovidto rnq w ; EvDovouxtreoorl evdovourtredorlq I evotouur6veror I oro tvrprivto * I, Torwpdvrorrrr, JLill-Aqvevoovourieoqff-l To yov[6to rrlg goroupdo4g evour gordvetor oro rvrpdvto W I I * To rvrpovro rpeper ev6ovourtreoorl K0r uoToup0orl '' , . ) To tvrpovlo lerlv1oe otg ove[opqu1 otrtr1trouXio, nou xodrronorouoe ovo ourooupponrogevo RNA. H evotoporool qq otrtr1trouXLoq qg evdovourtreoolg drlproupyloe 6vo rvrp6vro enorxropou nou 1rov rrvqro. Iq ouv6Xelo, 1 evourgdrrrlorl qg otr- trltroux(oq rrlg poroupoorlg evloXuoe qv rxovoqro rov o],trqtrou- Xr6v rov rvrpovlov vo 6rnl6vovror orrlv evepyo doUrl yLo rn ouppoqrl roug. tor 6l nop6goro e(vor r; 6tepyoo(o Kor yro to rwp6vro qg oprd6og ll, H evoopdtoorl oe our6 ro ororXe(o puog oL).r1LouX(og nou rol6rr<onore[ ev6ovour].edoq rou npoo6(6er xrvrlrtx6t4ro. Oo16oo, 4 evotrrpdro.ron Uno- pe( vo droropd[er qv lrov6rqro qg RNA oMq],ou1(og vo 6tnLrrtveror mrlv evepyq 6opr1. Aur6 0' ooro0oe n(e- orl yro Boq0eLo on6 nporetveq nou 0o pnopoUoe v' onororoorrloouv rnv rrovdtrlro ovod(n).oor1g. H evoot- p6roor1 truog tS,rotog oMrllouX(og oro rwp6vro 0o dro- rqpotoe rqv ove(oprqo(o rou. Opro;r6vo twp6vto qq ogddoq ll, oro16oo, dev rco- dLronororjv potoupdoeg, To ouyrerprprSvo rwp6vLo [oo:q Xpqorpronotoriv nporeiveq (roo6rivopreg Ue rrq Uo- roupdoeq nou xr.odrronororjwor on6 to rwp6vto) nou r<ro6 tronoro tjwor on6 o).).qlo u1(eg ro u yovLdtco proto g rou [evtorr1. To yeyov6g ow6 uno6r;].rrtver pro nrOovrq e[e].trrrxr1 nope(o yro rouq yevrr<oig nopdyoweg oup- poqnS. O nopdyowog (or,-rg [er<[vr1oe otg proroupdorl nou Boq0oUoe eLdLxd orn ouppoerl ev6g ouyxexprpd- vou rwpov(ou. H xro6rr<onorqtrrq o).trr1lou1[o onoUo- voOrp<e on6 ro rwp6vro oro yovLd[ropro rou (evtorq r<or 662 KE0MA|0 23 Korotrunro RNA orrl ouvdlero e[e].iX0qr<e dlore vo ].enoupye[ o' eupG repq xl,(prorq unoorpoUdrtov oe oX6on Ue rlv ouOev- rrr4 oLl,qloul(o rou rvrpovfou. O roro].urrr6g nuprlvog rou rwpov(ou 0o prnoporioe vo 6Xer e[eh1Oe( o'6vo snRNA. 23.8 H KoroAurrrfi Evepy6rnro Tnq RNAdonq P OgeiAeror oTo RNA * H prpovoux),eoo1 P (RNAooq P) eivor pto ptpovour),eonpot- refuq orrlv ono(o ro RNA 6Xer rorotrurtxl evepydrrlro. e H RNAoon P e[vor onopo(rrlTr] 0To porrrlpto, r' opyolo xo. roug europudreg. a H RNAoon MRP rtlrv europuor6v oXer((eror pe qv RNAoor P ror epntr6reror orqv ene[epyoolo rou rRNA ror qv ono6o- Urlorl Tou mRNA qg rurtrrvqq B. Mto on6 tg np6reg ono6e[[erg trrtv roro].unr<lrv 6u- vororqtorv rou RNA 6d04re Ue rrlv ovdl,uorl rnq pr- Bovour<i.edoqg P (RNAdor; P) rou E. coli. Evc6 opXrxd ovoyvrop[orrlKr oro Bor<rrlpro, q RNAdoq P rouronoL- qOrlxe <og Boorr<4 evdovour<Ledol nou epnLdreror orqv ene[epyoo[o tou IRNA oroug neptoo6repoug, ov 6Xr 6Loug, rouq opyovro;rotig, Borcrqpro, oplo(o xt euKopuoreg. Iqv oni.oUorepn UopAn qg, 11 Borcrrlproxrl RNAdoq P 6r[orotor oe 66o ouorortrd: dvo RNA 350-400 voux.tre- ottS[ov Kor l-uo npcoreivrxrl unopovd6o. H RNA unopovd- 6o trov Borcrqpfrr;v, 6rov onopov6verot in vitro, etrrgovl(er r<oro).urtrr1 evepy6qto. H RNAdon P ttrtv opXo(ov Kor rov euKopuorotv onoreekot on6 6,vo provodrr<d RNA 6opur<d ouyyevSg pre ro RNA rtov Borcr4p(rov, oMd 61er geyoL0repq neprernr6r4ro oe nporelvr1. To RNA 61er e).d1rorr1, ov 61r roOd).ou, r<oro).ulxr1 evepy6qto, 6rov pe).erdror in vitro. Tunrxd, n RNAdon P tcov oplo(ov o.tr- LrlLenrdpd pre rSooeprg nptorelveg evo tr;g (6p49 pre ev- v6o ror rou ovOpolnou trle 66ro. Ie 6Leg rrq nepr- m6oerg, to npcoreivu<6 ouorolx6 e(vor ovoyro(o yro r1v evepy6qro qq RNAdorlq P in vivo. MeroMd[erg e(te rou yovr6fou rou RNA eke rou yovrdfou rqg npcorefurlg odpovonororjv qv RNAdoq P in vivo, enoprSvtoq yv<r-lpi (oupe 6tr rot ro 66o ouororu<d e(vor ovoyr<o(o yto rn,i evepy6rqto rou ev(6prou. ApXu<d e[Xe unora3:. :; - npo.lrelvq nope(1e rqv xoro].urrrr1 evepyorn:c . , - -: RNA eiXe rdnoto entxouprr6 pdlo - n,X, ponOc-:: --np6o6eoq rou unoorpc,-rUoroq, enetdq neotoit. -a:..:: BpoXe(eq oMrlLouX(eg oupnr1po-lpolKrq ur :.1 i.-.i. p6veg nepLoXSq rou DNA. Auro( or p6lor, rr.rr::: :,-. orpdgr;xov r<oorrtg orn npoyUortxdqro ro B,1 I ::-: rlv xoro.trulr4 evepy6qro, evol or nporervoc rtJt, t - 6optxrlornpr(n. Avo),riowog r' onoteL5oporo [-le qv nop:-]:- t- ro RNA e(vor 5v(upro, Kdee "6v(upo" roro.trua. :- : : Boorxeg ewong
  • 154.
    onoon notr 6vunoorpoudrov. nop6rr n KorolurtKn evopy6rrlro Bp[oretor oro RNA, ro nproteivu<5 ouorort- r<6 ou(dver nol0 r4v roXfqro qg ow[6poor1q, 6nolg go(vetot on6 tqv ori[4oq tou optO;toti ovorrix].rooqg (BL6ne Ex6vo 23.9). Err6g on6 rqv RNAdoq P, or europurirreg nep6- Xouv Uro dltrq onopokrlnl ev6ovouxLedorl Booropr6vrl mo RNA, qv RNAdoq MRP (mitochondrial HNA pro- cessing, ene[epyoo(o puroXov6proro0 RNA). Auti q ev- douvouxl.edoq onoteLefuor on6 6vo 6opmd ouyyev6q rqrolulr6 RNA ror ;rorpd(eror nol.tr5q npo:relvrr6g unoprovddeg Ue rrlv RNAdorl P. Evd: opXrxd ovoyvop(- orrp<e on6 ro p5l,o rnq ornv ene(epyoo(o rou prroxov- Eptororj RNA, 4 RNAdorl MRP rup(rog tretroupye[ orov nuprlvo, 6nou ene[epyd(eror ro np66po;ro prBooropur6 RNA. H RNAdoq MRP (orog en(or1g no((er o4;rowu<6 p6Lo orq pU0prorl rou KurrqprKo0 xUr.trou, ro069 ou1.r- ger6Xa orrlv onoS5prnon rou mRNA tr'19 r<urd.fuqg B. H ovoyvriprorl r4g RNAdoqg MRP e(vor npox.trr1nrf, ro- Oci:g go(verot 6tr ro npoteivrr6 oumonr6 ourrqpe(tor oe preydlo Bo0p6 pero[0 RNAdortg P r<ot r1v RNAdorl MRP, evri q peroBoLrl r4g er6x5qroq oq npoq ro un6- fipogo ogeftreror mqv owotr].oyr1 rou roroLurtroU RNA. 2 3,9 To loerOfi 'Exouv KoroAurrrfi Evepy6rnro * To LoeLdrl ror ro roooetd4 oXrlpot(ouv 1uo orpupor5qoin 6oUn p' evepyoqro ourodroonoorlg. o l-lop6uoLeq dou6q unopei vo 6q;uoupyq0o0v (euyoprirvovrog 6vov rLdrvo-undorptrllro nou 6toonoror qnd 6vo ev(ugrrd rtr6vo. a '0r0v o ev(uprr6g xkovog erooyeror oro rUrropo, (euyopdrver p' 6vo rtr6vo-ordXo, o ono(og oul ouveXero droondtor. 'Evo or6prq nopddery;ro qg u<ov6rqroq rou RNA vo Lemoupye( og evdovouxledo4 nopSXeror on6 trreprxd grrpd (-350 nt) <pulrd RNAs nou drexneporcbvouv pto ow(6poor1 ouro6r6onqo4g. 'Onog Kor orrlv nep(mat- orl rou rwpov(ou opddog I rou Tetrahymena, e(vor nt- Oovrl q rotooreu( Soprc0v nou prnope[ vo ],erroup- yrloouv o' e[coreprrd unoorpd:poro. Aurd ro prxpd gutrxd RNAs rorordooowor oe 6Uo yevu<69 oprddeg: ro roer6f Kor ro toooer6{. To roer6ri (viroids) e[vor ].ot1.roy6vo pr6pro RNA nou Lenoupyo0v ove(dptrlto Xcopfg vo neprBdMowot on6 rdnoro npo:- reivx6 5).urpo. To roooer6{ (virusoids - ;repu<69 rpo- p6g roloriwor dopuEripo RNA, satellite RNA) rilouv nop6poto opydvrrloq, oMd neprBdtrI.owqr on6 nprore(- veg <purrrdrv urlv r<o0clq ouoreud(owor pro(( pr' 5vo rx6 yovr6(orpro. To roooer64 6e pnoporiv v' owLyporporjv ove[dpqro, xo06g Xperd(owot t4 Bori0ero trr:v rcilv. To roerdq Kor ro toooerdq owrypdgowot 1.uloo ru- l,r6pev<ov r0xLrov (BI6ne Err6vo 12.6). O xAtiryoq rul RNA nou ouoxeud(eror orov 16 roleko xkitwg qr (plus strand). O ouprntrqpcol.torr6q xLtivog nou nop& yeror xord rlv owrypogrl tou RNA rol,ekot xArbvog ilqv (minus strand). Yndplouv nouprepri rot ttov 60o r<trc[vrov, ouv Kor n].4v. Kot or 5Uo t0not provol.teporiq 6qpuoupyoriwor pe drdonoon rnq oupdg ev6q xuht6pre- vou r0x,trou Q. To rurlrrd povopepf rou xl.<itvou ouv nopdyowor ouvddowog t' dr<po ev6g eu0UypopUou provopreporig, T6oo ot xLrivor ouv 6oo rot ot xtrrivot ntrr1v rov toet- dc6v xor rtov toooer66v ug(orowot outo6tdonoorl rn vlf- ro. Opto;r6vo RNA droonoUv in vitro un6 rovovtx6g ouvOrjr<eg. l.trLo 6roonoriv pr6vo prerd 6vo r0rl,o 06p- Uovonq xor rfri(r'1q: our6 unodrlAdlver 6rr ro onopovo- ;r6vo RNA 6Xer oxordtrtrr1lr1 6ropr6pqrrloq, oMd ;rnope( v' onorcrloer evepy6 6ropr6pqtooq ov ono6rqroX0e( xt enovo6rorolOe(. To roer6ri xor roooerdti nou ug(orowor outo6rdono- or1 qlqpot(ouv Uro "oqupoKdeclln" 6ewepotoyn 6o- Uri mn 06o11 drdonooqg, 6nog qo(vetor mo ndvo U6poq rnq ElKOllAI23.14. To o<pupor<5goLo prBodv(ugo ouirouv oe Uro ou<oy6vero prBoev(Uptov nou nepilo1.r- Bdvouv tov 16 qg qnor(n5og 5 (HDV), ro prBo6v(u;ro goupr<drog Kor ro 6opurpoprr<6 ptBo5v(upro Varkud (VS). Aenoupyu<d, o HDV onone( 6ro0evr1 geroMu<d ron6wo yro vo npooOfoer q drdonoorl, evcb to ogu- pox6rpo).o prBodv(upro Kor ro ogupordgol,o gouprdrog 6e Xperd(owor p6rotr.o. H orlproo(o rov perdMorv orrl 0r ouv0werrr6g xegol6q dxouv tperq p(oXoug-filtrdg xor ouwrlprlg6veq pdoerg M(oXoS 3 et E, Arfmoon cG / ^f u^,y' GNCGA IINNNNN cNGA ..cNNNNN Mtoxoq 2 u*ou' vrioxos r Korotrurtrd x6vrpo 0r rerpotr6g drlptoupyorjvror $ o}.$trenibposrl ovdpeoo oe drjo oupnhrlpol-ronrd p6pro RNA Ktrovoq-unoorpouo 8tg ,auonooQ I C6 / aal Y"( PPP G,..G"C"CGA "U.E-GAGCOH H0 CG"6Ga ^AGCUCGGppp^G.- ^utI -u"G Ev(ugutiq xtrrrlvog n fl(0f{A 2*.14 0r neproX5g ourodtdonoolg rorv roerddrv Kor Tov toooetddrv dXouv 1uo ouvorveruq otrtr4trouX(o ror oXrlpor[(ouv gro oQupordQotrn beureporoyrl doUrl U6oro evdopoproro0 (euyoprir- poroq. 0r reqotr6g enioqq drlptoupyouvror Ueoo (euyopdrgoroq ovdgeoo orov rtrdlvo-unoorptrllto Kor Tov (ov(uprro, rtrdrvo. H t*.S T0 loerdq'EXouv Korotrulrq Evepvo:-.: 663 Boorxeg evvorcq
  • 155.
    6r6onoor1 rou prBoev([prouVS e(vot or<dprq ooorp4g. 'Oi.' ourd ro prBo6v(upo, oo16oo, npoyUororroto0v gro 6rdonooq nou rorol,o(ner 6vo 5'-OH dxpo rc 6,vo 2'-O'- r<uxAr6 goorpodteorepr6 dr<po. Ze ow(Oeoq pe 6l.o ro un6lorno prBoriv(upro nou riXouv ovoyvrrlprme( pr61pr o4prepo, ro oeupor<dgo).o prBodv(upro rr dLl.o pr6l4 qg ou<oySvetdg roug 6e Xper- d(owor nporeivx6 ouqrour6 yro vo ).eroupyrloouv rn rzrVo, xoOrilg q oMqLouX(o oufig rqg doUnq enopre( yro rq 6rdonoo4. l-ro ro ocpupor6cpoLo prBo6v(upro ro evepy5 r<6wpo e[vor 1.uo oMrl].ouX(o p6].rq 5g vour<].eo- n6(ov. H rerpo).r1 neprSler rpetg neproldg p(oXou-O4- Ltdg, rcov ono(olv n eSon r<or to prdye0og e(vor oroOepd, roOrilg xor 13 ouwqp4pr5vo vouxleor(dro, r<up(og ong neproXSg nou ouv66ouv ro rulwpo r4g 6opqg. Or ouv- rqpqg6veg Bdoerg ror or 6rnlo( g(oXor ouv0drouv 6,vo RNA p' ev6oyevrl n<ov6r1ro 6rdonoong. Mro evepy6g regol4 en(onq dqptoupye(ror on6 ro (euydpopro ev6g RNA nou owrilpooone0er puo nleupd qg doprig trr' 6vo RNA nou owrnpooonerier qv dl,Lq nLeupd qg. To rdrrrr pr6pog t4g Err6vog 20.14 nopou- otd(et rivo nopddery;ro r;g r<ego).rjg nou drlpuoupyefuor on6 rov uBpt6ropr6 ev6g propfou 19 nt g'5vo pr6pro 24 nt. To uBp(6ro trlpe(ror q ogupor<6goLq 6oprr1 pre rrl Srogopd 6n Ae(nouv or oyxriLeg I r<or lll. 'Orov ro RNA 19 nt npom(0eror oro RNA 24 nt, 6rdonoorl ouprBoiver orqv rordMrl).4 05or1qg re<pol,r]g. Mnoporipre vo Oeco- pr;oouge 5rt o ndvco (24 nl) xLcbvog ourotj rou uBpr6(- ou e(vor ro 'un6orpopro' evci o rdro (19 nt) xlrivog ro '5v(upro'. 'Orqv ro RNA 19 nt ovoperyvrjeror Ue nep(o- oero RNA 24 nt, 6roonc6wor noMon).d ow(ypoeo rou 6eUrepou. Aur6 uno6ql,c6ver 6rr undpXet dvoq xrlxl,oq (euyopriprorog 19 nt-24 nt, 6rdonoorlg, 6rdorooqg rov 6roonoopuivrov xLoo;rorov on6 ro RNA 19 nt ror (euyo- p6poroq tou RNA 19 nt p'5vo v6o un6orpopro 24 nt. Iuvencbg, ro RNA 19 nt e(vor vo prBoSv(upo pr' evepy6- t4ro evdovouxLedoqg. Or nopdprerpor ulg ow(6poor1q efuor nop6proreq Ue lq ow(ororleg dtrtrcov owrdpdoeov nou KotqAUowor on6 ro RNA (Fl,5ne Erx6vo 23.9). 'EXer 11611 ovo).u0e( 11 r<puoroMu<ri dopq ev6g eLdXr- orou oeupor<5<po),ou pgoev(riprou. Irrlv eLdlrmq 6o- p4, coor6oo, rl opxffeKrovxrl rou evepyorj r5wpou 6ev ro0toroUoe oocp5g ncirg 0o pnoporioe vo npolrop{oer 11 rcordLuoq. l-l p6ogoro, ovolri04 r<e 4 rpuoro..f.rxr1 6oprl rou nLrlpoug o<puporSgol,ou prBo6v(uprou on6 ro Schlsfosorna mansoni (6vo nopdotro), yeyov5g nou 6rotpdrnoe rlv rordl,uoq. Aur4 q 6oprr1, nou nopouord- (etor opprorrxd mqv EIK0NA 23.15, onoro).Untet pro r<p(otprq rptroroyq oAlrl).enidpoorl ovdgeoo oe pro npoe[oXrl rou pr(oXou I ror tr1g 0r1]ldg rou ;r(o1ou ll. H oMq).en[6pooq oroOeponoe( ro evepy6 xdwpo oe rd- roro dtopr6ptpooq d:ore q G12 v' orporpe( npor5vro on6 ro 2'-OH qg C17, 6n^.o6n qn6 ro 6eopr6 nou 6roond- ror eUKoLq, rot vo 6qproupye(ro enr0eur6 2'-o[uy6vo. Me q oepd rnq, n G8 nop5Xer u6poy6vo yro tr1 oro0e- pono[4oq rou veooxnponopuivou 5'-OH drpou mo 3' npoi6v drdonooqg. Elvor 6uvor6g o oXedroqr6g ouvduool.uirv ev(riprou- unoorpdlporog xovciv vo oXrlpor(oouv eLdXroreq oeu- 664 KE0AAAI0 23 Korohulxri RNA IVlioXoq I [4(oxog Ill Ell(0NA 23.15 H dop4 rou orpuporeqotrou prpoev(rjgou ouyKpo- re[ror o' evepy6 rpuoroy1 dLop6prpoo4 pSoto otrtrqtrentdpooeov pero[u trtv luoXrltirv oyrutrtlv, nou enrorlgo(vovror ond ro petr1. H 06orl qg dLoonoo4g enroqpo[veror pe ro r6xxrvo petroq. l-lpo- ooppoog6vo on6 M. Maftick and W. G. Scott, Cel/ (126): 309-320. pordqotreq 6op6g. Aur6g ot 6opr6g rilouv Xpnoqonor- r10e[ yro vo 6e([ouv 6n r; etooytoyrl rov rordl.q],cov ;rop[cov RNA o' 6vo r<rirropo prnope( vo ennp6rper tr1v ilpoyporonofio4 ev(uptxc0v owr6pdoecov rn vivo. Ztrlv npoyponr6trpo, dvo prBodv(upo oXedtooprdvo Kor' ou- r6v rov tp6no e(vor 1.uo e[orperxd er6u<q ev6ovoux].ed- on ileproprollof nou roreu0[veror evovr(ov ev6g RNA-m6Xou. Av ro pgoSv(upro reOe( un6 rov 6).ey1o ev6g pu0pr(6Uevou unoKrvrlrrl, gnope[ vo Xp4orpronor- q0e( 5ncoq n.X. ot antisense rorooxeu6g yro v' o6povo- nouloet erdxd tqv 6r<ppoorl evdg yovtd(ou-or61ou un6 ro0opro;.r6veg ouvOrlxeg. 2 3.'l O Ene{epyoofo rou RNA lupBoiver oe Mepovtop6veq Bdo€tq * 0r uno0oxelq qg onotrtnonporelv1g-B xor rou ytrouroprrou utp(orovror erdrreg onogrvooerq nou roro),uovrol ono 0eopr- vooeg qg ruld[vr1g ror qg odevoolv1g xor otrtro(ouv rlv Ko- Drronoullq otrtr1trouXio. 'Evo Boou<6 o[(iilpro qg propror<qg BLo].oy(og e(vot 6l 11 oLAq).ou1(o ev6g mRNA owrnpooonerier p6vo 6,rr e(vor rro6rr<onorqprSvo oro DNA. To r<ewprr6 66ypo npodBLene puo ypopprrxrl oX5on, onlv ono(o pro ouve- plg oMrlouX(o DNA preroypdgeror oe Uro oMrlLou- X(o mRNA nop Ue rq oerpd rnq Llerogpd(eror oe no).unem[61o. H Unop(r1 Sroxexoppr6vov yovrdicov r<oL 11 oqo(peo4 rcov rwpov(tov pr6oir.t rrlq ouppoerlg rou M(oXoq ll < )'--+ c'.G 1 n- G,"A t ^"n:]*C.;--"Ai/s A-J G.---"U G A JC FI QI Boolxeq eworeg
  • 156.
    RNA eroqyoyov 6voentnL6ov p(Uo orn drepyoo(o rqg Srgpoo4g tcov yovtd(tov (Bh6ne Ke<pd).oto 21). Iuvo- mr6, 11 ouppoqq npoYporonote(tot oto en(ne6o tou RNA r<r 6Xer og onor5leopo trlv o<po(peoq pr1 rodtro- norqlrrilv oLlqAouXrdrv (twpov(<ov) nou 6tox6nrouv -ng ro6rronorqtrr6g oMql.ouX(eg (e[c0vto) ndvo orqv oMrlLoufo tou DNA. H 6tepyoo(o efvot 1.tto UeroQo- pd nlr;po<poptcilv, oqv ono(o q npoyprottrrl r<r:6tro- notrlrrrrl ollrlloulio tou DNA nopogdvet opetd- 9rnq. AL).oy6q ttr;v ntr4 porpo p tcirv no u rcodtrono toUwot oro DNA npoyporonotofwot oe optoprSveg e(otpenr<5q neprmdoerg, xup(tog t<otd t4v nopoycoyrl v5rov oMq- l.ouXuilv nou rcodtronotoUv ovoooo<porp[veg oro 0qtro- onrd ror ro m4vd. Tdtoreg oli.oy6q ouprBo(vouv et6xd oro ooponKd rrlrropo (B Leg<por<r3rropo) aro ono(o ouw(0ewot ot ovooooeotp(veq (B)r5ne Kerpdtroto 18). N5eq ntrqpogop(eg nopdyowot oro DNA ev6q ot6ptou rot6 11 dtdpreto rnq ovoKqroor<eurig ev6g yovt6(ou ovooooqorpfvqg rot ot n].r1po<pop(eg nou efuot t<codxo- noqp5veg oro DNA preroBdMowqt on6 oroprotx6g pre- toLl,d[etq. Or nLrlpotpop(eg tou DNA ouveX((ouv vo pretoypdgowot ntord oro RNA. H enefepyoo(o tou RNA (RNA editing) e(vot 1.tto 6tepyoo(o mrlv ono(o ot nArlpoEopbq aMd(ouv oro en[ne6o rcu mRNA. Aw6 onoro],0me-rot on6 xotqord- oerq orrq onoteq 11 rodtronotqrtr<ri oLlrll'ou1[o ev6q RNA 6toq6pet on6 qv otrtrql,ou1(o tou DNA on6 qv ono(o peroypdqrp<e. Ene[epyoo(o tou RNA ouptBo(vet oe 6rio Drorpopetrr6g rotoordoetq, 11 r<oOepr(o pre 6to- gopetrrd ofttq. Iro x0nopo ttov orproortrdrv, undp- Xouv neprmciloetg 6nou owtrordmoorl ouprBo(vet oe ;uo Bdo4 rou mRNA pretoBdl,Aowoq rrlv otr).r'1}'ou1(o rou ro:srronoroUpevou nol,unemt6(ou. Aurrl n owtKo- rdmoorl e(vot ro onotStreopo t4g onop(vcoorlg e(te qg o6evoo(vnq npoq tvoo(vq efue qq r<un6fvqq npoq oupr6(v4. Iro puroX6vdpto tou tpunovootitl-toroq, eu- prirepeg oMoy6g ouprBo(vouv oro pretdypoqo noMdlv yovr6[trlv, 6rov oum4pottrd npoor(0ewor 11 onotro(- qowor B6oerg. H EIK0NA 23.16 ouvor!((er ttg oMr;Lou1(eg rou Yovt- 6(ou r<or rou mRNA u1q onol.tnonpotetvqg-B (apo-B) oro 6wepo xor ro qnop trov Or1troortxdlv. To yovtd(copto nepr6ler 6vo provoSxd dtor<eropptdvo yov(6to, tou ono(- ou 4 ol"qtrouX(o e(vot novopot6runq oe 6Loug toug roroig, Ue pro rodrronorqtrxq neptolq 4563 t<todtro- v(o:v. Aur6 to yov(6to preroypdqetot oe mRNA nou Ue- rorppd(eror oe Uro npcoretvq 512 kDa, q ono(o owrnpooonef er qv nln pn xo.t6txonotqtu<r1 oM4Lou- 1(o mo 4nop. Mto Bpolftepq Uopqn qg npoteivrlg (-250 kDa) ouw(0erot mo 6wepo. Auq q nptoteivrl onoretrefror on6 ro N-te].tr6 plo6 rrlg n]rlpoug npo- re[vr;q. Meroqpd(erot on6 6vo mRNA, tou ono(ou t'1 oM4l,ouX(o e[vot novopor6tunrl Ue rrlv ow(orot1r1 tou qnotog g' e(o(peor1 puo oMoyrl on6 C oe U oro rtrl6t- r6vro 2153. Autrl r1 owxotdoroo4 oMd(et to ro6u<5- vro CAA qg ylouropr[v4g oe ro6tr6vto tepprortoprof UAA. Enerdrl 6ev undpXet evoMorrtr6 yov(6ro { eE6- vro oro yovt6(co;ro yro vo rto6xonotqoet q v5o otrtrq- To yov(6to qg onotrtnonporeivqq B 61et 29 e(drvro .!!.ffi ro xtrldtrovto 2153 rto6tronote(q ytroutopfurl t c AA Ene{ePlooto uAA :: I o&rororyP To ouppopdvo mRNA mo rlnop xtrloxonote( pto nept5XeL ro xtldtxdvto UAA npote'tvn 4563 rotdointov nou repuotl(et rn o0v0eoq qg nporetvnq oto rotdtrotno 21 53 EIK0NA 23.16 H oltrn)ouxio tou yovt0[ou qg apo-B eivot [6ro mo 6vrepo roL to r1nop, oilro n otrtrnhou1fo tou mRNA Tporottot- e[ror pre otrtroyn Urog Foonq, n onoio OlptoupYei 6vo rol0txovto reppolopoti oro 6vrepo. Loul(o ror de Bp60ry<e r6noto oMoyri mo np6tuno rnq ouppoqrlg, ovoyroortrd ouptnepo[voupe 6tt rd- noro oMoyrl o'uv5Bq d;reoo orrlv oMrlLoul(o rou RNA petoypd<pou. 'Evo or6prq nopddetypo e(vot ot uno6ole(g tou ytrouropur<o6 orov eyr<6goAo rov opoupofrov. H ene- (epyoo[o oe pto 0do4 ;retorpdnet ro rro6tr6vto t4g yl.owop(vqg oro DNA oe rodtr6vto rrlq opyrvfuqg oro mRNA. H oMoyrl on6 y)routoprtvrl oe opyw(vq enrlped- (er t4v oycoyrp6rqro tou StoULou n enop6vtog 51et o4- prowu<r; en(nroorl mq pri0;uoq rnq po{q rrov t6wtov dropSoou tou 6rqUlou. Ie dtrq 05oq tou uno6oX5o, 6vo ro6rx6vto opytv[vqg perorp6netot oe rodtr6vto ylur<rvr;9. To ou;rBdwo nou neprypdenKov yto rouq uno6o- 1e(g qg apo-B ru rou yl.ouropKoU e[vqt to onorStre- opo anastwritoe@v onq ono(eg oqotpe[tot q optvoptd6o rou vour<Leorr6rro0 SoxruL(ou. H ene[epyoo(o qg apo-B nporo),e( oMoy4 rrlS Czrse oe U, evci xot ot 6tio olloy6g orov uno6oX6o tou yloutoptro6 e(vot A npoq l(voolvq). Ot onogwcioetg tqg onoltnonporetvqg B ro- tol0owor on6 q deopuvdoq t4g rutt6(vqg APOBEC (apolipoprotein B mRNA editing enzyme complex, otil-t- nlor<o ev(Upro:v ene(epyoo[og tou mRNA t4g ono].t- nportefvrlg B), evdr ot onopwrioetq orov uno6oX5o tou yAouropuxorl on6 ng 5eo;uvdoeg qg odevooivqg nou 6pouv oro RNA (roLotiwot ADAR, adenosine deami- nases acting on RNA). Aut6q o tUnog ene(epyoo(og go(vetor vo ouprBo(ver r<up(tog oro veuptr6 o0mqpo. Yndplouv '16 (duvqrtr<o() or6Xot rcov ADAR otq Drosophila melanogaster ror 6lor e(vot yov(6to nou eU- nL6Kowqr mr1 veupo6roB(Boo4. Ie noltr6g neptmdl- 23.10 Ene[epyoo[o rou RNA Iuppo(vet oeMegovtop6veq Bdoetq 665 I wTo mRNA tou evr6pou
  • 157.
    EIK0NA 23.'17 Hene[spyoo(o Tou mRNA npoypoTorore[Tur 6T0v pro deopLvdorl snrdpd oe pro odeviv4 grog drilootoop6vlg nepLo- Xrlg ev6g ore)drg (euyoprop6vou RNA. oerq, n ene[epyoo(o o].].d(er 5vo optvo[ri oe pro ].er- roupyu<d oqprowrxrl 06oq qg np<otetvrlg. To 6v(upro nou TrpoyUoronoroUv rrlv onopr(vooq ou- yyd 61ouv eupe(o er6rr5rpo - n.X. n r<oLwepo preLeq- ;r6vr1 6eo;rrv6o4 rqg o6evoo(vqg enrdpd oe rdOe rordLorno A ;uog 6rnLooroql5vqg neproXqg rou DNA. Om6oo, 11 onopr(vooq qg o6evoo(vnq Kor rqg ruu6(- vnq oro RNA ep<pov(er er6r6r1ro. To dv(u;ro ene[ep- yoo(og oXer[(owor Ue rrq yevrKSg 6eopuvdoeg, o].].d nepdXouv dLl,eg neproX6g q enrnp6oOereg unopovd- 6eg nou puOpr(ouv t4v et6u<6qrd roug. Irrlv nep[mro- on rnq ene(spyoo(oq rnq apo-8, 11 rorolurrrrl unoprovdSo ev6q ouprnL6r<ou ene[epyoo[og qXeti(eror pe rr1 Bomqpror<4 6eopuvdoq qg xurtd(vqg otrld 6Xet pro enrnp6o0eq neproXrl np6o6eoqg rou RNA nou Bo40d orqv ovoyvrilpron rnq erdu<rjg neptoXqg-or61ou npog ene(epyoo(o. Mro er6rrci 6eo;rwdoq qg odevoo(- vrlq ovqyvop((er rrq neproldg-or6Xoug mo RNA rou uno6o16o rou ytrouropu<oti, evc0 nop6proro ouprBdwo Trpoyporonoto0wor oro RNA rou uno5o16o rnq oepo- rovfuqg. To orjpnl,oxo (orrlg ovoyvrop(er puo et6xr1 ne- proXq qg 6ewepotoyorig 6oprig xord rp6no ovdl,oyo ge ro 6v(upro rponono(r1oqg rou IRNA 11 0o prnopo0oe v' ovoyvrop((er oneu0e(og pro vourl,eorr6rxrj oM4l,ou- X(o. H ovdmu[r1 ev6g in vitro ouorriprorog yto rnv ene- [epyoo[o rqg apo-B uno6r1].6ver 6rr rord].].4].og or6Xog e(vor ;uo qXeru<d purp4 oMqLou{o (-26 vou- rtreotbto) nou neprBdLl,er rnv neproXrl ene[epyoo(og, H EIK0NA 23.17 6efiyer 6rr orqv nepfmooq tou RNA yro tov uno6o15o rou ylouroprro0, ovdpeoo ornv e[epyo- op6vr1 neproXq rou e(ov(ou Kot Uro ou;rn).4pcogotrrrl oM4LouX(o rou xo0odrro0 rwpov(ou q4rlgod(eror pro 6(xl,ovq neproXq, yvomrl og GluR-B, nou e(vor ovq- yxo[o yro qv ovoyvr6prorl rou or6Xou. 'Evo ].dOog (eu- ydprol"ro pr6oo orq 6(xlrovrl neproXri e(vor ovoyro(o yro er6u<ri ovoyvcbpron. Iuven6g, drorpopeurd ouonjproro ene[epyoo(og prnope[ vo 6Xouv Srorpoperu<69 onorq- oerq ovqgoprrd pe qv erdx6rrlro yro qv oMrll,ouXio rov ufloorpoprdtcov roug. 23.1 I H Enefepyooio Tou RNA Mnopei vo Ko0o6nyeiror on6 O6nyd RNA * Iro Utrox6vdpr0 Tou Tpunovoorogorog ouppolver eKTeTopsvrl ene[epyoo(o rou RNA p'ev0eoeLg r1 onotrorqeg oupLdivrlg. e To RNA unoorporpo (euyoprirvet U' 6vo odnyo RNA xot ono ttg 6uo ntreupeg qq nepioXqg 1 ono(o nporeuor vo unoorel eneIepyooro. * To od1y6 RNA nopdXerro er;loyefo yro Tlv npoo0rtrn (n onovrorepo qv onotrouprl) ouprdrvrirv, * H ene[epyoolo rorotruerot ono ro otopor[dro ene[epyoolog, evo oupntroro p' evepyorrlro evdouvourtreoolg, e[ovouxtre- ooqq, retrurlg ouptdu),orpovotpepoorlq xot RNA trtyrooqg. 'Evog 6togopelr<69 rlinog ene(epyoo(og onoro).Ume- tor on6 lq ewunoorox6g o).).oydq qq oMrlouX(og oro npoi6wo noMdrv yovr6f<ov rrrlv Urroxov6p(olv rou tpunovoocitllorog. Irqv npotq nep(moloq nou ovoKo- ).Urp0r1re, q oMq).ouX(o rnq unopovd6og ll rrlg o[er6o- orlq rou r<uroXpcbporoq 5Xer Uro eooreprK4 prerot6nrorl noro(ou nou 6ev npoBAdneror pe Bdorl q voux.treort- drr<r1 oMr1l.ou1(o rou yovr6(ou coxll. Ot oMrl).ou1(eq rou yovL6(ou Kor rnq npolre'ivr1g nou 6(vowor orqv ::....i, ..:.: , " ouwr]poUwor oe noMd e(6q rpunovooropro_ rog, ouvenolg 11 p60o6og ene[epyoo(og tou RNA 6ev e(vor onor.treronxrl yLo 6vo ;r6vo opyovropr6. H ooullgrov(o ovdpeoo orqv oMrlLoul(o rou yovt6(- ou coxll Kor ro nporeivrx6 npoi6v ogeil.etot orrlv ene- (epyoo(o rou RNA. To mRNA tou coxll neptSler ;uo Adevoolvr1 lvoo(vr1 NHI C I-rl * -c -6 -N, HN -C .c -N.. I tr cH-> I lt cH HC --n, -C -g/ HC --n, -C -g/ 'r | ,, L Ktodrronotl96vo oqv otrLr1trouX(o rou yevtogworl DNA Metor6nrorl tou ntroto[ou ovdyvoorlg Atr)trftouXio RNA /UtrqlouXio nporeivrl g ouloro ntro[oto ovoyvtoolg nou noporr;pe[Tot oTrlv npolreiVl drlptoupyeuor ono rqv evoorporooq reoooptov vourtreoldlrrrv ouprdrvrlq. 666 KE0A Al0 23 Korotrulxo RNA I S S L G I K V E N i:L€ V C AUA UCA AGU UUA GGU AUA AnR GUA GAG MC eUGJGUA GGU AUA UCA AGU UUA GGU AUA AAA GUA GAU UGiJ AUA CCU GGU AGG UGU MU ISSLGIKVDCIPGRCN Boorxeg eworcg
  • 158.
    U UAUAUGUUUUGUUGUUUAUUAUGUGAUUAUGGUUUUGUUUUUUAUUGGUAUUUUUUAGAUUUAUUUAAUUUGUUGAU ru @9AAUACAUUUUAUUUG UUU-GUUAAUUUUUUUGUUUUGUGUUUUGGUUUAGGUI @ UUUUUUGUUGUUGUUGUUUUGUAUUA TUnUo qgotrtrr1trouXrog mRNA coxlll rou T. brucei.l-lotrtr6q oupL6rveg Dev rodtronorouvrot oro DNA (rorrtvo) n oQotpouv- ror ono ro RNA (olUo(vovrot pe T). 6v0eor1 rcoodpov vour.l.eott6(orv (ouptd(veg) y0po on6 qv neproXri rqg petot6ntorlg nLoto(ou. H 5v0eor1 ro- Oop((er ro rotdMqlo ntro(oto ovdyvtloqg Yto rnv npo- retv4. Aev 61er ovorolug0e( dtrLo yov(6to coxll nou vo q6per q petot6ntoq nl.oto(ou: 6rot ouvdyetqt 6tt ot enrnp6oOereg Bdoerg etq4opo0v rotd rr1 6tdpreto rqg preroypogr1q rj petd on5 owrl. Mto nop6ptoto ooupqo- v(o ovdgeoo oro mRNA Kot rrq yevulptr6g oMrltrouX(eq Bp(oreror oe yov[6to to6 nopo;ru[oioU SVS xqt tou toti t4g rLopdg rot o<peil,erot oe npooOrlrrl xorolofutov G oro mRNA. l-'lop6proto ene[epyoo(o oM4].ouXtcilv tou RNA oupt- Bo(ver ror oe dMo yov(dto xot nepil,oprBdvet t6oo ono- ).o1969 6oo xot npooOrireg oupr6(vqg. H oouvti0tmq nep(nrrooq rou yovr6(ou qq o[etddoqg lll tou xuto- Xpc0prorog c (coxlll) :rou Trypanosoma bruceiouvotf ((e- ror crrnv EIK0NA 23.19. fdvu an6 ra ptod xatd^olfta rou nRNA e[vu oupt[veq nou 6ev xadxonoolwat oro Yo- Iovt6[togo fl poeneqepyoou6vo AAAGoGGAGAGAAMGAAA RNA mRNA Guide RNA v[61o. H o(yr<pro'q ovdpeoo oro yevoptK6 DNA rot ro mRNA deftver 611 6ev undpXet rpnpo rou mRNA ndvo on6 7 vouxl,eodSto nou vo prqv 6Xet unosre( oLLoyti ror 6u erodyowot ouorotl(eg oupr6lvqg pr{rouq 5og 7 roroLo(nov. Or ntrlpogop(eg yto rqv et5xrl 6v0eoq tov oupr6tvri:v nopSXowot qn6 6vo o6qy6 RNA (guide R- NA). To odr1y6 RNA nepdXer gto oLqLoufo ouprntrrlpo- prorrr{ pe ro otrlord ene[epyool.tdvo mRNA. H EIK0NA 23.20 6e(tyet 6vo prow6lo yto t1 6pdoq rou oro yov[6to rou r<uroXpcilgotog b tqg Leishmania (6vo dMo nopdor- to). H oltrqLoufo oro ndvo prdpog qg ex6vog defiyet to qu0ewu(6 prerdypogo 11 ro getdypo<po 6ncog e(vot npw on6 qv ene[epyoo(o rou RNA. Tq revd defiyouv nori ero6pXowor Bdoerg rord t1v ene[epyoo(o, Orcrc0 ouprd(veg np6ner vo etq(oprioouv oe ouri qv neptoXti 6ore vo 6rlpuoupyq0s( puo dyrup4 oL}'4l.ou{o mRNA. To odqy5 RNA e(vor ouprntrqpoporxS pe to mRNA yto AAAGCGGAGAGAAAAGAAA A G G CTTTAACTTCAGGTTGTfiATTACGAGTATATGG !t,,;t1,';g'pr * A G G C UUUMCUUCAGGUUGUUUAUUACGAGUAUAUGG I Zeuvupr,lucvo pe ro o0qy6 RNA t llpoene[epYoo[6vo Rpvq66664qAGAMAGAM AG GcUUUM6UU6AGGUUGUUUAUUA6GAGUAUAUGG RNAmm*83ilXmm$$m$XXXX OOny6 RNA AUAUUCAAUAAUAAAU UUUAAAUAUAAUAGAAAAUUGMGU UCAGUAUACACUAUMUMUMU I rvor,rpotrooq ouptdwrlv v AAAGCGGAGAGAAAAGPAA UUUAUGUUGUCUUUUAACUUCAGGUUGUUUAUUACGAGUAUAUGG il illllil lHII}I ltil Ilil lIAUAU UCAAUAAUAAAU UUUAAAI.'AUAAUAGAAAAUUGAAGU U CAG UAUACACUAUMUMUMU I t--', ;-.1;; :' .. .:j ; - I mRNA AAAGCGGAGAGAAAAGAAA UUUAUGUUGUCUUUUAACUUCAGGUUGUUUAUUACGAGUAUAUGG ,;:rr. .' ,r:', lr ,r.r To npoene[epyoop5vo RNA (euyoptirver p' rlvo o6qyo RNA ror on6 rtg 6uo ntreup6g qg neptoXqq nou nporettot v0 uro- orei ene[epyooio. To odnyo RNA nopdXer dvu erpoye[o yro qv evourlroroorl Tov ouptdtvtirv. To mRNA nou nporuntet ono qv evotlporol- o1 e[vot oupn],r1pogolr6 tou o61you RNA. 3;.'!i H Ene[e pyoo(o rou RNA Mnope(vo Ko0o6lyeuot ono 06qyo Rri,+ 667
  • 159.
    MURF3(5') CyB-2 O6ry6RNA MURF2-1 Coll-FS MURFII-2 CyB-1 l:.:.;i7:,r::125 95 MURF3 CoillCytsMURF4 tMURFl NDlCoilMURF2Cot ND4 ND5 :r,..:i,::: 'l i .': To yovLd[oU0 Tnq lelshrnarla nepr€Xri y0v[6r0 nou KodrKonorouv np0ensqspy0og6v0 RNA ovoperKTo Ue UOV06eq, n6u K0]- dtronotouv t' onopo[qro odrlyo RNA yto q onuroupy[o rov ooortirv mRNA otrtrr1trouxrdv. 0prourvo yovroro eXouv notrtrontrd o61yo RNA. CyB e[vot T0 nporeEepY0opevo yovisLo tou ruroXptirlrorog b, evdr CyB-1 roL CyB-2 ro yov[6r0 ro]v 06ly6v RNA nou epn],6rovror orrlv sneqepyoo[o Tou. o41-rowm{ 5rcrooq, nou nepil.o1lBdver xor nepgdtrtrer rqv eile(epyooUdv4 neproXrl. Tunu<d, q ou prnLq po proru6r4- rq e(vor nro ercrevrlg ornv 3'n).eupd tqg ene[epyooprd- vnq neproxnq, Kor. ilro BpoXe[o mqv 5'n].eupd. To (euydpo;ro ovdpeoo oro o64yd RNA ror ro npoene[ep- yoopr6vo RNA og4ver revd 5nou o(euydpcoreg A rou o6rlyoti RNA 6e Bp(orouv oupn).qproporu<6g Bdoerg oro RNA nprv on6 qv ene[epyoo(o. To o6r1y6 RNA nopdXer 5vo erpoye(o nou enrp6ner r4v evocoprdroon rov Koro- Aohcov U oe our5g ng O5oerg pe puo 6epyoo(o nou 0o neptypoqe( orq ouv61ero. Otov oloxL4pto0e( r; owi dpoon, ro o64y6 RNA onoondrot on6 ro mRNA nou e(- vqr nL5ov 6ro06orpo yro ;retdrppoorl. H e[eL6[r<euo4 qq re].rxr1g ene[epyoop6vqg oMq- ).ouX(og e(vor opxerd nol,rin),oxr1. Iro nopddetypo rou rwolp6prorog b rqg Leishmania, pto prorprd neproXrl tou perdyporpou urp(ororor ene[epyoo(o pr' 6v0eoq ou- voLrrd 39 roroi.o(ntov U, 4 ono(o go(veror 6u onone( 6Uo o6qyd RNA nou 6pouv oe yenovtr5g 06oerg. To np6ro odr1y6 RNA (euyop6ver pe rrlv mo 3' 06o4 ru 6nemo 11 ene[epyoop6vq o].].q].ouX(o yfverot ro un6- orpopo yro nepon6po ene[epyoo(o on5 ro end;levo o6qy6 RNA. To o64yd RNA xrodrxonoroUwot oq ove- [dprrlreg preroyporprxdg povd6eg. H E!K0NA 20.21 no- pouord(er 5vo 1dprr1 rrlg oXerrxrlg neprol4g rou 1-utolov6proxod DNA qg Lershmanra. O Xdprrlq ilepr- troprBdvet ro yov(6ro rou rurolpriprorog b, to onoio r<co- drronore( qv o).).4Aou1(o nprv on6 rrlv ene[epyoo(o, ror 6Uo neprolSg nou r<o0op((ouv ro o64yd RNA. To yov(6ro trrlv r<tiprrov r<todrronotrlnr6v neprolcbv e(vor ovdperrcro pre ro yov(6to trov ow(ororXcov o6qyc0v RNA. Oe<op4nrd, pro ;rerdMo[r1 e(te rou yovt6(ou e(te ev6g on6 to 6rio ow(ororlo o6r1yd RNA 0o ;rnopoUoe v' oLL6[er rrlv npororoy4 oM4IouX(o rou mRNA dpo Kqr rnq npolrefv4g. Me yeverrrd r<pn4pro, xoOep(o on6 ourrig ng provd6eg 0o prnoporioe vo 0ecop40e( 6tr ono- reLe( rpqpo rou yovr6(ou. Or provddeg ergpd(owor ove[6prr1ro, on6re oogoA6g 0o dnpene vo vo e(vor ougnLrlpcolrorx5g in frans. Iuvenc0g, ov unqplov 6ro- 06orpreg preroMd(erg, 0o dronrm6vopre 6rr yro vq Ko- 6rrono140e( 4 npororoyrlg o).l.r1LouXio tlroq 668 KEOAAAI0 23 Korotrurud RNA provodu<4g nporeiv4g Xperd(owor tperg ou;rnLqpoJUo- ttr6g oprddeg. O loporrqpro;r6g tolv ev6ropuiocov nou u<p(orowor pepr4 ene(epyoo(o uno6r1).cirver 6l q ow(6poon ilpo- lrope( rotd ;rrlrog rou npoene[epyoopr6vou RNA oe r<o- re[Ouvor1 3'-5'. To o64y6 RNA r<o0op((er rrlv er6x6rrlto torv ev06oeov oupt6ivrlg prSoot tou (euyopdrprot6g rou Ue ro npoefle(epyoopSvo RNA. H ene[epyoo(o rrov ouprdwcbv xoro].Ueror on6 6vo 20S ev(upur<6 ofpn).oro nou xol.efuor oapailo eneep- yoolaq (editosome), onore].efuor on6 nep(nou 20 nprrl- -retveg rcor neptSler pro ev6ovouxl.edoq, pro re),rr4 o u p 16 uLotpovogepdorl [f UTdorl), puo 3'-5'U-er6u<r1 e[o- vour<Ledorl (eEcoUdofl Kor Uro RNA Lryr<do4. 'Oncog <po(- veror. orrlv EIK0NA 23.22, to oropor(6to ene(epyoo(og npoo66ver ro o6qy6 RNA r<or ro Xprlorpoilore( yro vo (euyopc0oet Ue ro npoene(epyoo;r6vo mRNA. To RNA un6orpopro 6roondrot oe Uto nepropl nou (nr0ov6v) ovoyvtop[(eror on6 rlv onouo(o (euyopuiproroq Ue ro EI 3', *"q&ff I rvoovourreoon ry&ff . TetrLrrl I f oupr0utrorpovoqepoorl '(TUTdon) :ry J **o ^,y*ooqY .. .,, , l1poo01r1 :l onolouprl roro),0[nov U npoyporo- nole[ror pe Droonool rou RNA, otporpeoq 11 npooOr1u1 U ror ofv- deol riov orptov. 0t ovlDpooerg roro),0ovror ono evo oupntroro ev(uprrrv uno qv ro0odrlyloq rou odrivou RNA. 5',
  • 160.
    o64y6 RNA: truooupr6ivrl evool.rotriveror 11 onolo(rperot yro vo (euyopcioer pre ro o6qy6 RNA r<t 6nerq ouv6Serqr ro RNA un6orpo;ro. H tpupargopuq oupr6fur1 (UTP) ei- vor 11 n4yrl rorv r<orol.o(nov oupr6fur1g ror npoor(Oeror pe q 5p6o4 rrlg TUTdorlg. H onoi,orqri rov rorolo(nrov U 6ropreooLoBefuor on6 pro e[oUdoq, q ono(o ouvepyd- (eror pre pto 3'gtoogor6oq enmp5nowoq oro RNA nou unoBtrrlOqr<e o' ene[epyoo(o vo enovoouv6e0e(. Or 6opdg ;rop(cov nou 6Xouv unoBLq0e( oe prepu<r1 ene[epyoo(o unodrltrdrvouv 6l ro r<otd].orno U npoor(- Oewqr 6vq q rpopd nopd r<ord o;rddeg. l-10ov6v 11 ow(- 6poon npoXrrrpe( 6roprSoou 6ro6o1rrd:v rfrtrcov oroug ono(oug npoor(Oewor r<otdtrorno U, 6ort;rd(owot trlg npoq rn ouprntrrlpoprorx6rrltd rouq pe to odqy6 RNA ror 6ror4po0wor ov e(vor ono6errd evdl orpotpotiwot or4v owi0erq nep(mcooq: ouven6g, 4 rotooreur] rqg ooord ene(epyoopr6qg oMqtr ou(og ouptBoiva orodto- xd. Ae yvop((oupe ov or (6tor t6nor owt6pdoerrrv eprntr5- Kowor. orr;v ene[epyoo(o nou npoo06rer rord.trorno C. 23,12 H tuppogfi llporeivdlv Elvor AuroxoroAurr16 r Mro rvretvrl 6Xer rr;v rrovdqro vo rorotruer rrlv orporpeon rnq on6 pro npolre[vr1 rord r6roro rpono tiore vo ouv06ovror or e[reiveg rou Trlv neptpotrtrouv. r H ouppoQn rov nptoreivdrv roroLferor ond qv twelVrl. t 0r reproodTepeg rwetVeg 6Xouv 6rio ove[opqreg evepyoq- req: ouppoqn nporciVtiv rt evdovourtredorl enotuqrou. H ouppogri rov nporeiVdrv (protein splicing) SXer to (6ro onordLeopo Ue rrl ouppogrlrou RNA: puo o).- L4l,ouX(o nou owrnpooonerjeror fio eooreptr6 ev6g yovr6(ou 6ev owrnpooorne6eror orrlv nporeivq. To r;rq;roro rqg nprore(vqg ovoprd(owqr Kor' ovoLoy(o pe rn ouppogq tou RNA: elreiVeg (exteins) e(vor or o),).4)rou1(eq Trou clwrnpoo(l)neJowor mnv drptprl nporetvrl, evri rweiVeg (inteins) or oMrll.ouX(eg nou ocporporiwor. O prqXovropr6g yro qv o<po(peon rrlq tv- retvqg e(vor eweLtirg 6torpoperu<69 on6 rov ow(oror- Xo rnq ouppo<pqg rou RNA. H EIKONA 23.23 6e(yyet 6tt ro yov(6to ;rerorppd(eror oe Uro np66poprr; np<oretvrl nou neprdXer trlv rwelvq xr 5nero q rwei4 orporpefuor on6 r4v nproref,rr1. fldvo on6 350 nopodetyporo oup- pogns npoteivdlv efvor yvoord ror 6rodedopt5vo oe 6l,eg ttq r<oqyop(eg rov opyovroprc0v. To tuntx6 yov(- 6ro rou ono[ou ro npoi6v ucp(ororor ouppogti 6Xer 1.uo l.rovodrq rweiwl. H npcirq rwetvq ovor<olriq0qKe oro yov(6to rnq D- NA noLuprepdoqg ev6q opXo(ou un6 poprp{ Uroq no- pepBoM6pevrlg oMrll,ouX(og oro yovbro q ono(o 6ev or<oLouOe[ roug r<ovdveq yro ro rwp6vro. Irrl ouvdlero onodeffiqre 6n 11 roOop4 nporeffi prnopef v' onoMo- nporeivnefu ; Exioprl n1g . * rwervnq ono ' Trlv fipolervn w4EIK0NA 23.23 Koro tr1 ouppoqn nporeiVdrv, ot e[retveg ouvD6- ovrot orporptirwug qv tvre[vr1 ond r4v npotelVq, ye( on6 ourrl qv oMql.ouX(o oe Uro outor<oro).uttr4 ow(6poor1. H ow(dpoo4 6ev onone( npoo0rp<r1 ev6p- yeroq Kor npoyUoronoefuor drogSoou rnq oepdg ovo- drord(etov beoprcbv nou nopouotd(etot mrlv EIKONA 2X.24. H ow(6pooq drernepouiveror on6 qv twetvq, prolov6n q onoreleopotrr6ulrd rrlg enqped(eror on6 rrg e(teiveg. H np6tr1 ow(6pooq e(vor 1.uo en[Oeoq on6 pto n].eu- pu<r1 oLuo(6o -OH n-SH tou npritou o;tvo[5og rrlq rv- reivrlg orov nentr6rx6 6eo;r6 nou ro ouvdSer Ue rnv npc0rr1 e(retvq. Aurn q ow(6poor; preroqdper qv e{tei'- vr1 on6 r1v o1.uvotetrrrr1 oprddo qq rweivqq o' 6vov qru- Lrr6 deopr6 N-O { N-S. 'Enerro, our6g o 6eopr6g dSXeror en[Oeor1 on6 tr1v -OH n -SH n].euprr4 qluo(6o rou npdlrou oprwo[6oq rqg 6e[repqg twetv4g. To ono- rSl,eopro efvor 11 prerogopd qg e[teivrlg 1 or4v n].eupt- 14 oluo(6o rou ogrvoreLu<o0 o(6o9 rrlq e[teivrlg 2. T6Log, q C-re}'l<r1 oonopoy[vq qg rwetvrlg xur.l.onore(- ror, evoi 11 rel,u<r1 NH qq e[retvqg 2 enn(Oeror mov or<uLrr6 6eopr6 c6ore vo rov qwrKqroorrloer pr' 5vo ou1.t- Borrr6 nemrdx6 6eopr6. KoOepr(o on6 outSg rrq owr- 6pdoetg prnope( vo ouprBe( ou06p;rqro pre noLri XoUnIn roXrir4ro. l-ro vo npoyproronor4Ooriv ouwovropSvo ror oprerd ypnyopo rirore vo entteulOe( 11 ouppoqrl qg nptotetvqg Xpetd(eror rordluorl on6 rqv rwefvq. Ot tweiVeg 51ouv Xoponrqprcrxd yvop(oproro. Bp(- oKowor cog ev06oerg ew6g nl,oro[ou oro eooreptx6 ro- 6rronor4rrrrilv oMrllou1rc6v. Avoyvrrlpf(owor on6 t4v 0nop[r1 o;r6],oycrlv yovr6(rov nou 6e 6to05touv rn ouy- rer<pr;.r6vr1 6v0eoq.'E1ouv puo N-refuxq oep(vn r1 ruorei- vr1 (nou nop6let Uto -XH nl,eupu<ri oluo(6o) Kor Uro C-relrrq oonopoyfvrl. H tunrxq twetwl 51er pto otrl.q- l,ouX[o -150 oprvo(5ov oro N-re].u<6 dr<po r<or -50 o;u- vo[6cov mo C-retrx6 drpo, or ono(eg epntr6rowor orrlv xordl.uorl rnq ouppoqrlq rov nproreiv6v. H oMqlou- {o oT o r6wpo qg twetvrlg 6Xer dMeg }.emoupy(eg. 'Evo oouvq0roro XopoKrqptoru<6 notrtrriv rweiv6v eL ?* ii H lupporpf l-lporeivorv Elvor Auroxorotrulrl 669 Boorxfg fvuoreg E[retv1 lvTsivn Elrerwl reDNA IBNA
  • 161.
    X=Sor0 vor 611 6Xouvevepy6rqro ev6ovouKledonq enoxrol.rori. Mrq evdovouxledoq enoxrol.rori 6roond ro DNA-or51o 6q;uoupyriwog eror puo Odo'q 6nou prnope( vo eroX<r:pr1- oer q oMql,ouX[o rou DNA nou ro6xonore[ t4v rwefvr1 (BI5ne Eu<5vo 23.11 orqv Ev6trya 23.5). H ouppog4 rurv npcoreiv6v r<r 11 evepy6rqro rqg evdovouxtredo4g enou<to1-rori e(vor ove(dprqreg treroupy(eqrnq rwetvrlg. Aev dXoupe xorovoqoer nLrjpcog ndrg ouv6Sowor or 6rjo our5g evepy6rrlreq p6oo oe pro rweiv4, oor6oo 61ouv nporo0e( 6Uo rUnor prow6lcov. liprgovo Ue ro nprilro, opXu<d unrlple rdnoro e(6og oriv6eo4g ovdpre- oo orrq evepy5rqreq, oMd opy6repo Sytvov ove[dpr4- reg xr 6tor x6noeg rweiVeg 61ooov qv evepy6rrlro rnq ev6ovouxLedonq efior.KtoUo0. IUprgovo pe ro 6e0repo prow6lo, or rwe'fveg (orog [er<(vqoov rog pov66eq oup- pqeiq nporeivriv Kr 6nerq (or nro noMdg) 66X0r1r<ov etoBo).r1 on5 ng ev6ovoux.l,e6oeq enorKroUori. H un60e- or1 ourri e(vor ouprBor4 pre ro yeyov6g 6n or ev6ovou- rledoeg enorrroprotj go(veror 611 6Xouv en(oqg eroBdMer Kor oe dMoug r0noug provd6cov, xup(rog oe rwp6vro qq o;rd6og l. 23.13 llepiAn$n H ouroouppocp{ e(vot 1616111ro 60o oprddov rwpo- 670 KEOA^AI0 23 Korotrurx6 RNA EIK0NA 23.24 0r 6eopo[ ovodrqroooovror p6oo ond pro oxotrou0fo frans-soTeponoqoetov olg ono(eg ougper6- Xouv or -0H ogddeg qq oepiv4g 4 qg Opeov(vr1q Kor rl- SH opo6o rrlg ruoretvrlg 6tog 6rou ouvde0o[v or e[reiVeq pr' 6vo nemrdxd Deogd xr one],eu0epo0e(11 rv- Te(vn U' 6vo rurlonotrlpdvo C-relud 6xpo. v(tov, ro ono(q e(vor 6ro6e6oprdvo oe povor<rit- topoug/o)lyor0rropou g euxopudlreg, npoKo- pucorrrd ouorrlproro xor 1.rrroX6v6pro. Or ovoyr<o(eg nLrlpo<pop(eq yro rnv ow(6poor1 Bp(oxowor orqv oMr1l,ou1[o rou rwpov(ou, 1-ro- l,ov6rr, in vivo, srnv npoypotrx6r4ro q ow(- 6pooq unoBoq0ekor on6 npotrefveg, l-ro ro rwp6vro 16oo qg otrrddog I 6oo rqr qg otrrd- 6o9 ll, 11 ow(6poo4 onorre( ro oXrlprorropr6 grog er6trqg deweporoyorjg/tpnoroyorig do- Unq Ue BpoXeieg ouvorverrr6g oMqLouX(eg. To RNA rov rwpov(cov r4g opd6og I 64trloupye( ;uo 6opul oqv ono[o 4 oM4].ou1(o-un6mpr,r;- Uo ouyKporefuor pr5orr: r4g neptoX(g IGS rou twpov(ou, evol dMeq ouwrlpqprdveg oM4).ou- X(eg 64;rroupyoriv prro neproX4 np6o6eorlg vouxLeorr6(ov youov(vr1g. Aur6 ouprBo(ver pre trans-eorepono(r1oq orqv ono(o ou pprer6Xer co g ouprnopdyowog 6vo xord).orno youvoo(v4g. Aev qnotre(ror nopop; ev6pyerog. H youovoo(- v4 6roond ro 6eo;r6 ornv 5' ouprBo).r1 ;rero[ri e[cov(ou-rwpov[ou. 'Eneno, ro udpo[[].ro mo eLef0epo dr<po tou e(cov(ou entr(Oeror orrlv 3'ouprBo- L4 e[rov(ou-rwpov(ou. To rwp6vro rur].onorefuor ror ldver 14 youovoo(vr1 Kor rrq retrewo(eg 15 Bdoerg. Mro oerpd ouyyevcilv owr6pdoeov xoro,l.riowor ;16oro ent0Soeov qro uq eocoreprro ri g qrrlogo6remepu<oUg deoprorig on6 ro rsfur6 G-OH rordlorTro rou rwpov(- ou. Me qv nopoXrl tcov xordMql,rov unoorpco;rdrr,lv efuor duvorri 4 xorooreu{ prBoev(riprov nou npoytlo- ronoroUv norrfi.eg roroLurrr6q owr6pdoerq, mtg ono(eg neprLoprBdvsror Kor 4 evepy6rqro rnq rpov- o<pepdo4g vou xi.eor6(ov. Opro;rdvo ;utolovdpror<d rwp6vro qq oprddog I xor ll 51ouv ovorXrd ntrq(oro ovdyvo:or1g. Or nporer*veg nou t<to6txonoro0wor qn6 ro rwp6vto rrlg oprddog I e(vor ev- dovouxledoeq nou npqypqronororiv 6(xtrcoveg ewopr6g oe 06oerg-or61oug rou DNA. H ev6ovour<.l,eoLunrri 6rd- onoon nupodore( pto 6repyoo(o yovr6rorqq Uerorpo- m'19, or4v ono(o 11 oMrl),ouX(o rou (6rou rou rwpov(ou owrypdgeror orq 05or1-m61o. Irrg nporetveg nou Ko- 6xonotoriwor on6 ro rwp6vto qq oprd6og ll oupnept- l,oprBdveror pro ev6ovourledorl, 11 ono(o (ar<rvd 14 perdOeoq, Kor Uro ow(orpogq preroyporpdoq, nou eTrr- rp6ner rnv owrypoe{ ev6g RNA owrypdqou rou rwpo- v(ou m4 06oq-or61o. Auto( ot rUnor rwpov(cov (ocoq nepd.oprBdvouv evepy5rrpeg ;roroupdorlg nou ouv6p6- Uouv ornv ogo[peor1 rov twpov(rov oro0eponorcirwog il a _tu I .HX QHz l I H Yn2-u:',; -N -i { rr.rivnz l________J (i,, -In-iY ilHn XH CH, -J;g zHN il L ir;),, ; *, ---=
  • 162.
    ro oxn Uortop6rnq dewepotoyoti g/tproroyo 0S 6o U( S tou evepyori rSwpou. Kotoluttrdg owrdp6oetg 6texnepotrirvowot on6'ro RNA oumottx6 qq RNAdorlg P. Tq RNAs rtov toooet- 6cirv npoyprotonoto(v ouro6tdonooq Utqq oocpuporS- qol.r1g" 6opriq. Igu por6rpoLeq 6opr6g oxq got(owot ovdpeoo oe 6vo RNA un6mpopo Kot to RNA ev6g pt- Boev(Upou, yeyov6q nou xo0odqye( et6tr<6 q 6tdono- orl oe ouyrerptprdveq oLl,rlLoul(eg. Aut6q ot ovrt- 6pdoetg mrlp((ouv tqv dnorlrq 6tt ro RNA oXrlpot((et erdrrd evepyd r<6wpo nou 51ouv roroLwtxrl evepy6- rnro. H ene[epyoo(q rou RNA oMd(et qv oM4].ouX(o ev6g RNA rotd q pretoypotpri 11 rot pretd on6 ouq. Ot olloydg e(vot onopo(tqteg d:ore vo 64prtoupy40e( pto rro6xonotlnrci oMrll,ou{o ;re v6r1 go. Awtt<otoor6oetg gepovcopdvtov Bdoerov ouprBo(vouv oro Oqtroottrd rot no(pvouv rn Uopqf onogtv6oetov, ortq ono(eg eke 11 C perotp6netot oe U e(te 11 A petorp5nerot oe l. Mto t<o- rotrwrrtj unopovddo nou oXet((erot Ue rn deogrvdoq r1g r<wr6(v1q n rnq odevoo(vqg Letroupye( og tprlpo ev6q preyol.0tepou oupnl,6rou to ono(o eggov((et et6t- x6t1ro yto Uto ouyrer<ptpr6vq dLqlouX(o-or6Xo' I-1 poo0rjr<eg rot onotrotg5g (ouly6repo ouptdiv4g) oupBo(vouv oro puol6v6ptq rou tpunovooritrloroq Kot orouq nopq;ru[oioti g. Ercretop6veg owtdpdoetg ene- [epyoo(og npoyporoTloto tiwot qrq rpunovootill.toto, sro ono(o oXe66v ot prtodg Bdoetg ev6g mRNA npo5p- Xovror on6 ene[epyoo(q. H ow6poorl qg ene[epyo- o(oq Xpqorpronote( dvo erproye(o nou onotetre(tot on6 5vo o6qy6 RNA ouprntrqpcoporrr6 qg oLr1l'ouX(og tou mRNA. H ow(dpoorl rfiol0etot on6 ro oopotbto ene- (epyoo(og, 6vo otipnLot<o ev(riprov nou nepfioprBdvet gto ev6ovouxledoq, pto e[ovoux].edoq-te]'u<r1 oupt6u- l,orpovorpepdoq xot qv RNA ).tyr<doq rot lp4otpronoe( el,e[0epo voutd.eot(6to oq flnYn Yto rrlv npooOrlrq ti oneLeu0epdlvet 6toonoop6vo voutd,eot(6to pretd tqv onoLotgrj. H ouppogri rcov npoteivcov e(vot ouroroto)r,wtxr1 ow(6poo4 nou npoyporonote(tot ;16oo owr6pdoeov prero<popdq 6eopd:v, evcil 6ev onome( nopoXri ev6pyet- og. H rwetvrl r<ffiolUet qv onoxonf rnq on5 ng e[tei- veq nou rrlv neptBdll,ouv. IloLL5g tvtetveg 6Xouv evepy6qto ev6ovourtredonq enoKroUo0, 11 ono(o e(vot ove[d6r1q on6 qv evepy6r4to rnq ouppoqnq npco- reivcbv. BrBAroypogio 23.2 To lvrpdvto rnq 0Ud6oq I AutoouppdnrovTot Ue trans-eorepono[qor1 ii,ti.t,iL{:1. +l Cech, T. R. (1985). Self-splicing RNA: implications for evolution. lnt. Rev. Cytol. 93, 3-22. Cech, T. R. (1987). The chemistry of self-splicing RNA and RNA enzymes. Science 236 1532-1539. Vicens,0. and Cech, T. T. (2006). Atomic level architecture:' group I introns revealed, Trends Biochem. Scr. 31 , 41 -51 . 1.r:*::'.,tt Been, M. D. and Cech, T. R. (1986).One binding site determines sequence specif icity of Tetrahymena pre-rRNA self-splicing. transsplicing, and RNA enzyme activity. Cel/ 47 ,207-216. Belfort, M., Pedersen-Lane, J., West, D., Ehrenman, K., Maley, G., Chu, F., and Maley, F. (1985). Processing of the introncontain- ing thymidylate synthase (td) gene of phage T4 is at the RNA level. Cel/ 41 ,375-382. Cech, T. R, Zaug, A. J., and Grabowski, P. J. (1981). invitrosplic' ing of the rRNA precursor of Tetrahymena: involvement of a guanosine nucleotide in the excision of the intervening se- quence. Cell 27, 487 -496. Kruger, K., Grabowski, P. J., Zaug, A. J., Sands, J., Gottschling, D. E., and Cech, T. R. (1982). Self-splicing RNA: autoexcision and autocyclization of the rib0somal RNA intervening sequence of Tetrahymena. Cell 31, 1 47 -157 Myers, C. A., Kuhla, B., Cusack, S., and Lambowitz, A. M. (2002). tRNA-like recognition of group I introns by a tyrosyltRNA syn- thetase. Proc. Natl. Acad. Sci. US/ 99 2630-2635. 'Epeuvc Burke, J. M., lrvine, K. D., Kaneko, K. J., Kerker, B. J., Oettgen, A. 8., Tierney, W. M., Williamson, C. 1., Zaug, A. J., and Cech, T' R. (1986), Role of conserved sequence elements 9L and 2 in self-splicing of lhe Tetrahymena ribosomal RNA precursor. Cel/ 45,167-176. Michel, F. and Wetshof, E. (1990). Modeling of the three-dimen- sional architecture of group I catalytic introns based on compar- ative sequence analysis. J. MoL Biol.216, 585-610. 23.4 To Prgo6v(uUo'EXouv [lorrltreq Korotrurtr6q Apooetg Avaoxdnqol Cech, T. R. (1990). Self-splicing of group I introns. Annu. Rev. Biochem. 59, 543-568. 'Epeuvo Edwards, T. E., Klein, D. J., and Ferre-D'Amare, A. R. (2007). Ri- boswitches: small-molecule recognition by gene regulatory F- NAs. Curr. lpin. Struct. Biol. 17:273'279. Serganov, A, and Patel, D. J. (2007). Ribozymes, riboswitches and beyond: regulation of gene expression without proteins. ffaf. Rev. Genet. 8'. 776-790. Winkler, W. C., Nahvi, A., Roth, A., Collins, J. A., and Breaker, R, R. (2004). Control of gene expression by a natural metabolite- responsive ribozyme. Nature 428, 281 -286. 23,5 0propdvo lvrpovto Tnq 0U060q I Ko:6lronotouv Ev6ovourtredoeq nou npo0Y0uv qv Ktvlrtx6qro i!'i+{-illi:i Belfort, M. and Roberts, R. J. (1997). Homing endonucleases: keeping the house in order. Nucleic, clds Res. 25, 3379- 3388. Haugen, P., Reeb, V., Lutzoni, F., and Bhatacharya, D. (2004). The BrBtrroypotpLo 671
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    MET6q>paon Avonopliyno1 on6: I.L. Mainprize, et al. , Molecular Biolo- gy of the Cell 17 (2006):5063-5074. Copyright 2006, Ame- rican Society of Cell Biology. H (j)W10ypo(j)[o nopoxwp~8T]Ke onci mu~ David W. Andrews , McMaster University KOL lain Mainprize, University of Guelph. E:rttµileta Ke<palaiov: Cheryl Keller Capone 61) Etoaywy!'] flB H Mnaq>paon t.LaKp[v£TOL 0£ Tp[a I1Mta: 'Evap~Tl, Em- µ!']KUVOll KOL Tepµanoµ6c; • To pt~oowµano nepLEXEL 1p£L<; neptoxec; np6o5wric; TOU tRNA. • 'Eva aµtvoaKuAo- tRNA £LOEPX£TOL OTll 8!:'.ori A. • To nrnn5uAo- tRNA npooMv£TOL 011181::011 P. • 'Eva aµtvo~u npoo1f8£TOL mriv no>.unrnn5LK!'] a>.uo[5a µ!:'.ow µnaq>opac; 1ou no>.unemt5[ou an6 TO nrnn5uAo- tRNA 111<; Stone; POTO aµLVOaKUAO-tRNA Tll<; Stone; A. OI) EL5LKO[ Mnxavtoµo[ EAEVXOUV TllV AKp[~£La Tll<; Mnaq>pa- 01']<; • H aKp[~£La TT']<; µ£TOq>paoric; £AEVX£TQL an6 £L5LKOU<; µ11xavtoµou<; 0£ K08£ OTMLO. 8 H 'Evap~T'] om BaKT!']pLa Xp£LO~£TOL 30S YnoµovME<; Kl EmKOUPLKOU<; napayovT£<; • H!:'.vap~n Tll<; µnaq>paonc; anOLT£[ ~£XWPLOT£<; Pl~OOW- µLKE<; unoµovM£<; 30S KOL 50S. • Enforic; anm1ouvrnt ot napayovrnc; !:'.vap~ric; (IF-1, -2 KOL -3), OL ono[OL npo0Mvov10L one; 30S unoµovMec;. • Mta 30S unoµovMa nou µnaq>!:'.p£L napayov1ec; !:'.vap- ~11<; npooMv£TOL 0£ µLa 8£01'] !:'.vap~T']<; OTO mRNA OXll- µm[~ovrnc; tva ouµnAoKo !:'.vap~nc;. • 0 IF-3 np!:'.n£L v' an£A£U8£pw8e[ WOT£ OL 50S unoµo- va5ec; va npoo5e8ouv 010 ouµnAoKo 30S-mRNA. 674 KECl>AAAIO 24 Mnaq>paon - H'Evap~T'] nepLAaµ~OV£l Zrnyapwµa B00£WV Mna~u TOU mRNA KOL TOU rRNA • LTO ~aKTT']pLaK6 mRNA Tl 81::011 !:'.vap~nc; anoT£A£lTOL an6 TO KW5LK6vto !:'.vap~nc; AUG, 10 ono[o aKoAou8£f -10 ~(Joete; µe10 10 e~aµep!:'.c; nouptvcilv Shine-Dalgar- no. • To rRNA 30S Pl~OOWµLK!']<; unoµovMac; TWV ~aKTT1Pl4JV 5ta8!:'.m µLO ouµn>.ripwµanK!'] ann>.ouxra nou ~rnyapw- V£L µ£ TllV aAAT']Aouxra Shine-Dalgarno KQTO TllV !:'.vap~T']. 8 'Eva EL5LK6 EvapKT!']pLO tRNA ::£KLVO TT']V no>.unrnTL5LK!'j AAuo[5a • Hµnaq>paori ~£Ktva µe tva aµtvo~u µ£8£tovrvric;, nou ouv!']Swc; KW5LKonOL£tTOL an6 10 AUG. • t.Laq>oprnKd tRNA µ£8£tov[vric; eµnAEKOVTGL 01riv !:'.vap~n KOL TllV emµ!']Kuvon. • To evapKT!']pLO tRNA 5La8!:'.m µova5LKO 5oµLKO xapa- KTT']PLOTLKO, rn ono[a 10 5taq>oponmouv an6 6Aa rn un6Aoma tRNA. • H oµMa NH2 TT']<; µ£8£tovrvric;, nou npooMvnOL 010 ~aKTllPLaK6 £VapKT!']pLO tRNA, £[VOL q>opµUALWµEVT']. 8 HXp!']on TOU fMet-tRNAf EAEVX£TOL an6 TOV IF-2 KOL TO PL- ~oowµOTLO • 0 IF-2 npooMm TO evapKT!']pLO fMet-tRNAf KOL TOU rnt1p!:'.n£L va £LO£A8£L OT1l µepLK!'] 81::011 P111c; 30S uno- µovMac;.
  • 166.
    &!) Οι ΜικρέςΥπομονάδες Αναζητούν Θέσεις Έναρξης στα Ευκαρυωτικά mRNA • Οι ευκαρυωτικές 40S ριβοσωμικές υπομονάδες προσ­ δένονται στο 5' άκρο του mRNA και το «σαρώνουν» έως ότου εντοπίσουν μια θέση έναρξης. • Μια ευκαρυωτική θέση έναρξης αποτελείται από μια αλληλουχία δέκα νουκλεοτιδίων η οποία περιλαμβάνει ένα κωδικόνιο AUG. • Οι 60S ριβοσωμικές υπομονάδες συνδέονται με το σύμπλοκο στη θέση έναρξης . &11) Οι Ευκαρυώτες Χρησιμοποιούν Ένα Σύμπλοκο Πολλών Παραγόντων έναρξης • Οι παράγοντες έναρξης απαιτούνται σε όλα τα στάδια της έναρξης , μεταξύ άλλων η πρόσδεση του εναρκτή­ ριου tRNA, η πρόσδεση της 40S υπομονάδας στο mRNA, η μετακίνηση κατά μήκος του mRNA και η πρόσδεση της 60S υπομονάδας. • Το ευκαρυωτικό εναρκτήριο tRNA είναι ένα Met-tRNA το οποίο διαφέρει από το Met-tRNA που χρησιμοποιεί­ ται στην επιμήκυνση, ωστόσο η μεθειονίνη του δεν εί­ ναι φορμυλιωμένη . • Ο elF2 προσδένει το εναρκτήριο Met-tRNAi και GTP σχηματίζοντας ένα τριτοταγές σύμπλοκο , το οποίο προσδένεται στην 40S υπομονάδα προτού αυτή αλλη­ λεπιδράσει με το mRNA. • Ένα σύμπλοκο πρόσδεσης της καλύπτρας προσδένε­ ται στο 5' άκρο του mRNA προτού αυτό αλληλεπιδρά­ σει με την 40S υπομονάδα. m Ο Παράγοντας Επιμήκυνσης Tu Φορτώνει Αμινοακυλο­ tRΝΑ στη Θέση Α • Η EF-Tu είναι μια μονομερής πρωτεΊνη G, της οποίας η ενεργός μορφή (προσδεδεμένη σε GTP) προσδένεται στο αμινοακυλο-tRΝΑ. • Το σύμπλοκο EF-Tu-GTP-aμιvoaκuλo-tRNA προσδένε­ ται στη θέση Α του ριβοσωματίου. fIID Η Πολυπεπτιδική Αλυσίδα Μεταφέρεται στο Αμινοακυλο­ tRΝΑ • Η 50S υπομονάδα έχει ενεργότητα πεπτιδυλοτρανσφε­ ράσης που παρέχεται από ένα rRNA ριβοένζυμο . • Η νεοσυντιθέμενη πολυπεπτιδική αλυσίδα μεταφέρεται από το πεπτιδυλο-tRΝΑ της θέσης Ρ στο αμινοακυλο­ tRΝΑ της θέσης Α. • Η σύνθεση των πεπτιδικών δεσμών παράγει αποακυλιω­ μένο tRNA στη θέση Ρ και πεπτιδυλο-tRΝΑ στη θέση Α. &! Η Μετατόπιση Μετακινεί το Ριβοσωμάτιο • Η μετατόπιση του ριβοσωματίου μετακινεί το mRNA διαμέσου του ριβοσωματίου κατά τρεις βάσεις. • Η μετατόπιση μετακινεί το αποακυλιωμένο tRNA στη θέση Ε και το πεπτιδυλο-tRΝΑ στη θέση Ρ, ενώ αδει­ άζει τη θέση Α. • Σύμφωνα με το μοντέλο υβριδικής κατάστασης, η μετα­ τόπιση συμβαίνει σε δύο στάδια, στα οποία η 50S με­ τακινείται σε σχέση με την 30S και στη συνέχεια η 30S μετακινείται κατά μήκος του mRNA ώστε ν' αποκατα­ σταθεί η αρχική διαμόρφωση . fm} Οι Παράγοντες Επιμήκυνσης Προσδένονται Εναλλακτικά στο Ριβοσωμάτιο • Η επιμήκυνση απαιτεί EF-G, που έχει δομή παρόμοια με τη δομή του συμπλόκου αμινοακυλο- tRNA-EF-Tu-GTP. • Η πρόσδεση του EF-Tu στο ριβοσωμάτιο O!eJ·.lΞ~ -: , EF-G και το αντίστροφο. • Η μετατόπιση προϋποθέτει υδρόλυση του GTP, πυροδοτεί μια αλλαγή στον EF-G, που με τη σειρά ~~ πυροδοτεί μια αλλαγή στη δομή του ριβοσωματίου . m Τρία Κωδικόνια Τερματίζουν τη Μετάφραση • Τα κωδικόνια UAA, UAG και UGA τερματίζουν τη μετά­ φραση . • Στα βακτήρια, συνήθως χρησιμοποιούνται με σχετική συχνότητα UAA>UGA>UAG. S Τα Κωδικόνια Τερματισμού Αναγνωρίζονται από Πρωτεϊνι­ κούς Παράγοντες • Τα κωδικόνια τερματισμού αναγνωρίζονται από πρωτεϊ­ νικούς παράγοντες απελευθέρωσης, όχι από αμινοα­ κυλο-tRΝΑ. • Οι δομές των παραγόντων απελευθέρωσης τάξης 1 μοιάζουν με τις αντίστοιχες δομές του συμπλόκου αμι­ νοακυλο-tRΝΑ-ΕF-Τu και του EF-G. • Οι παράγοντες απελευθέρωσης τάξης 1 απαντούν σε ειδικά κωδικόνια τερματισμού και υδρολύουν το δεσμό πολυπεπτιδίου- tRNA. • Οι παράγοντες απελευθέρωσης τάξης 1 υποβοηθούν­ ται από τους παράγοντες απελευθέρωσης τάξης 2, οι οποίοι εξαρτώνται από GTP. • Ο μηχανισμός είναι παρόμοιος στα βακτήρια (τα οποία διαθέτουν δύο τύπους παραγόντων απελευθέρωσης τάξης 1) και τους ευκαρυώτες (οι οποίοι διαθέτουν μόνο ένα παράγοντα απελευθέρωσης τάξης 1). 8 Το Ριβοσωμικό RNA Διαπερνά και τις Δύο Ριβοσωμικές Υπομονάδες - - - - - • Κάθε rRNA φέρει πολλές διακριτές περιοχές που δι­ πλώνονται ανεξάρτητα. • Σχεδόν όλες οι ριβοσωμικές πρωτεΊνες έρχονται σ' επαφή με το rRNA. • Οι περισσότερες επαφές μεταξύ των ριβοσωμικών υπομονάδων πραγματοποιούνται μεταξύ 16S και 23S rRNA. Τα Ριβοσωμάτια Περιέχουν Πολλά Ενεργά Κέντρα • Οι αλληλεπιδράσεις στις οποίες εμπλέκεται το rRNA είναι κρίσιμο μέρος της λειτουργίας του ριβοσωματίου. • Το περιβάλλον των θέσεων πρόσδεσης του tRNA κα­ θορίζεται κυρίως από το rRNA. Το 16S rRNA Διαδραματίζει Ενεργό Ρόλο στη Μετάφραση • Το 16S rRNA διαδραματίζει ενεργό ρόλο στις λειτουρ­ γίες της 30S υπομονάδας. Αλληλεπιδρά άμεσα με το mRNA, την 50S υπομονάδα και τ' αντικωδικόνια των tRNA στις θέσεις Ρ και Α. Το 23S rRNA Έχει Ενεργότητα Πεπτιδυλοτρανσφεράσης • Η ενεργότητα της πεπτιδυλοτρανσφεράσης εντοπίζε­ ται αποκλειστικά στο 23S rRNA. Οι Ριβοσωμικές Δομές Αλλάζουν Όταν Συνδυαστούν οι Υπομονάδες • Όταν σχηματίζονται πλήρη ριβοσωμάτια, η κεφαλή των 30S υπομονάδων περιστρέφεται γύρω από τον αυχένα. • Το ενεργό κέντρο της πεπτιδυλοτρανσφεράσης στην 50S υπομονάδα είναι πιο δραστικό στα πλήρη ριβοσω­ μάτια συγκριτικά με τις μεμονωμένες 50S υπομονάδες . Περίληψη 675
  • 167.
    24. l E1oaywyn 'EvamRNA m:pt8X8L µta 08tp6 KWOLKov(wv nou aMri- A8mopouv µ8 TO OVTLKWOLKOVLQ TWV aµLVOQKU/1.0-tRNA 8TOL WOT8 µLa OVTLOTOLXJ'"] 08Lp6 aµtvosewv va 8Vowµa- TW88l 08 µta no11.un8ITTLOLKfl OA.uo(oa. To pt~oowµano nap8X8L TO n8pl~CI/I./I.OV YLO Tr] pu8µLOJ'"] Tr]c; aMr]A.8n(- opaOr]c; µ8TOSU TOU mRNA KOL TOU aµLVOQKU/1.0-tRNA. To pl~OOWµCITLO ouµn8plcp8p8TOL OQV 8VQ µLKpO µ8Ta- VQOT8UTLKO 8pyoOT60Lo, TO ono(o Tasti58U8L KOTa µfi- Koc; Tou 8Kµaydou ot8s6yoVTac; Taxdc; KUK11.ouc; OUV880r]c; n8ITTLOLKWV 08oµwv. Ta aµLVOQKU/1.0-tRNA 8KTOS8UOVTOL µfoa Kl esw an6 TO µ6pto µ8 an(OT8UTO µ8yC1A.rJ TOXUTrJTO 8VW nap6Mri11.a 8vano8eTOuv aµtvo- sfo. En(Or]c;, napayOVT8c; 8nLµr]KUVOJ'"]c; KUKA.LKQ aMr]- /l.8nLOpOUV µ8 TO pt~oowµano KOL ot'(OTOVTOL an6 auT6. Mai'.;( µ8 TOuc; 8mKouptKouc; nap6yovr8c;, TO pt~oowµ6- TLO nap8X8L TO n11.fip8c; 8Upoc; TWV opOOTr]pLOTr]TWV nou anOLTOUVTOL yta 611.a Ta ~fiµma TJ'"]c; µ8Tacppaoric;. H EIKONA 24.1 napoumai'.;8L nc; oX8TLK8c; OLQOTCI08Lc; TWV OUOTQTLKWV Tr]c; npwTe'Lvoouv88TLKr]c; OUOKWr]c;. To PL~oowµano anOT8/l.8LTQL an6 Mo unoµov6o8c;, Ol ono(8c; otaopaµm(i'.;ouv 80LKouc; p611.ouc; OTrJ µ8Tacppa- OrJ. To ayy8A.LOcp6po RNA OUV088TQL µ8 Tr] µLKpr] uno- µovaoa: 08 Ka88 xpovLKfl OTtyµfi dvm npooo808µev8c; -35 ~CI08Lc; mRNA. To mRNA OLOTP8X8L Tr]V nopda TOU KQTCI µfiKoc; Tr]c; 8mcp6v8LOc; KOVTCI OT!'"] ouµ~0/1.r] TWV unoµovaowv. .6uo µ6pta tRNA dvm n6vroT8 8V8py6 OT!'"] µ8Tacppa0r]: OUVWWc;, r] 8nLµr]KUVOJ'"] TOU nOA.Un8- nTLOLOU n8pL11.aµ~aV8l OVTLopao8Lc; Ol ono(8c; ouµ~a(- vouv 08 µ611.tc; Mo an6 Ta (aop6) i58Ka KWOLK6vta nou KaMmovrm an6 TO pt~oowµano. Ta Mo tRNA 8Loep- xovrm a' WWT8plK8c; 8808Lc;, nou 8KT8LVOVTQL KQTCI µfi- Koc; TWV unoµovaowv. 'Eva TplTO tRNA (owe; napaµev8L navw OTO pt~oowµ6Tto acp6TOu xprimµonotri8d mri µ8- Tacppaori, npOTOU QVQKUKA.W88l. H ~QOLKr] µopcpfi TOU pl~oowµm(ou 8X8L OUVTr]pr]88l KOTa TrJV 8se11.LsrJ, wOT6oo unapxouv oacpdc; napaMa- yec; we; npoc; TO OUV0/1.LKO µey88oc; KOL TLc; QVQAOYL8c; RNA KOL npwTe'Lvwv OTO pl~oowµana TWV ~OKTr]p(wv, TOU 8UKOpUWTLKOU KUTiapon11.6oµmoc; KOL TWV opyavL- o(wv. H EIKONA24.2 OUYKPLV8l TO OUOTQTLKCI TWV pl~OOW- 220 A /VVVVV. mRNA 35 ~GoEwv o<( 0 0 N EIKONA 24.1 IuyKp(ow; µf:y£8wv 0£1XVOUV 6n 10 PL~oowµcmo £(VOL apK£16. µ£yo.Ao wo1£ va npooi'iev£L tRNA KOL mRNA. 676 KE4>AAAIO 24 M£To.(j)paol'j Pljloowµdna rRNAs npw,EivEc; r Bmrn')pta (70S) 50S 23S = 2904 ~GOEi<; 31 MG½a: 4,2 MDa 5S = 120 ~Goe1c; 60% RNA .._30S 16S = 1542 ~GOEi<; 21 28S = 4718 ~ci.OEI<; 49 Bn>..aOTLKG (BOS) '9 60S 5,8S = 160 ~ci.OEI<; MG½a: 4,2 MDa 5S = 120 ~ci.OEI<; 60%RNA .._40S 18S = 1874 ~ci.OEI<; 33 EIKONA 24.2 Ta Pl~oowµo.na £!VOL µ£yo.Ao pl~OVOUKA£0TIPWT£l- VlKO. owµOTlOLa, nou n£pLexouv n£pL0061£po RNA napo. npw1tlvrJ KOL OL0Kpfvov10L 0£ µ£yo.A£~ KOL µLKP£~ unoµovdo£~. µm(wv OTa ~aKTfipta KOL TO 8ri11.aOTLK6. Km TO Mo d- vm pt~ovouKA.eonpwTe'LVLKa owµm(ota, TO ono(a n8pte- xouv n8ptoo6T8po RNA an6 npwTdv8c;. Ot pt~oow- µtKec; npwrnTv8c; dvm yvwOTec; we; npwTdv8c; r (r-pro- teins). K688 pL~oowµLKr] unoµovaoa n8pl8X8L eva KUPLO rRNA KL optoµev8c; µLKpec; npwT8'(v8c;. H µ8ya11.ri unoµo- vaoa µnope( en(oric; va n8pl8X8L µLKPOT8pa RNA. LTO E. coli, ri µtKprJ unoµovaoa (30S) anoT8A.8LTOL an6 TO 16S rRNA Km 21 npwrnfv8c; r. H µ8y611.ri unoµovaoa (50S) n8pl8X8L 23S rRNA, TO µLKp6 5S RNA KOL 31 npwTdv8c;. M' 8S0lp80r] µLa npWTe'fVr], nou ~p(OK8TQL 08 T8008p0 avr(ypacpa ava pt~oowµaTLo, un6pX8L µ6vo eva avr(- ypacpo K688 npwTdvric;. Ta µdi'.;ova RNA ouvtOTouv TO KUplO µepoc; TJ'"]c; µ61'.;ac; TOU ~QKTJ'"]PLOKOU PL~oowµm(- ou. Bp(oKovrm navrou KOL, µ6Mov, OL n8pt006T8p8c; PL~oowµLK8c; npwTdv8c; OTJ'"]V npayµaTLKOTJ'"]TO 8PXOVTQL o' enacpfi µ8 TO rRNA. Iuv8nwc;, TO µ8(1'.;ova rRNA crxri- µm(i'.;ouv au16 nou µ8pLKec; cpopec; 88Wp8LTOL we; OK8A.8- T6c; KCl88 unoµovaoac; - 8VQ OUV8X8c; vfiµa nou KUpl0p- X8l OT!'"] ooµfi KOL Ka8op(i'.;8L nc; 8808Lc; TWV PL~OOWµLKWV npwrn'(vwv. Ta pt~oowµana OTO Kunap6n11.aoµa TWv euKapuw- Twv dvm µ8yaMT8pa an6 T' avr(OTotxa Twv ~aKTrJ- p(wv. H ouvoA.LKfl n8pL8KTLKOTJ'"]T6 TOuc; T6oo 08 RNA 600 KOL 08 npwTdvri dvm µ8yaMT8PrJ: TO µdi'.;ova µ6- pta RNA dvat µaKpuT8pa (Ka11.ouvTOL 18S KOL 28S rRNA) Km unapxouv n8ptoo6T8p8c; npwTdv8c;. To RNA 8SOKOA.ou8d v' anoT8A.8l TO Kup(apxo oumanK6 an6 TJ'"]V anOljJr] TJ'"]c; µ6/'.;ac;. Ta PL~oowµana TWV µLTOXOVop(wv KOL TWV x11.wpo- n11.00TWV OLOKPLVOVTQL an6 TO PL~oowµaTLQ TOU KUTIQ- pon11.6oµmoc; KOL na(pvouv nOLKLA.8c; µopcpec;. Optoµe- V8c; cpopec; 8XOUV TO µey88oc; oX800V TWV ~QKTr]plOKWV pt~oowµm(wv Km n8ptexouv 70% RNA. 'NJ...orn, dvm µ6vo 60S KOL n8pl8XOUV <30% RNA. To pt~oowµano n8pL8X8t noM6 8V8py6 Kevrpa, Ka- 8eva an6 TO ono(a KQTQOK8UCl/'.;8TOL an6 µLO oµaoa npwTe°LVWV nou OA.A.r]/.8nLOpOUV µ8 µta n8pLOXrJ TOU pl- ~OOWµLKOU RNA. Ta 8V8py6 K8VTpa anOLTOUV ciµwri ouµµ8TOXrJ TOU rRNA o' eva ooµLKO Tl QKOµr] KOL KQT0- /1.UTtKO p611.o. Optoµev8c; KOTa11.uTtKec; 11.8LToupy(8c; anm- TOuv s8XWPLOTec; npwTdv8c;, wOT6oo Kaµ(a an6 nc;
  • 168.
    opaow:; 5£ µnop£iv' avanapax8£i an6 µ£µovwµev£c; npWT£°fV£c; rJ oµ6i5£c; npwTE°LVWV: A£lTOupyouv µ6vo OTQ n>-.aima Tou p1l3oowµmiou. H av&uari n1c; A£tToupyiac; Twv 00µ1Kwv oumm1- Kwv Tou p1l3oowµmiou µnop£i va y(v£1 µ£ Mo n£1paµa- T1Kec; µ£86oouc;. H av6>-.uari µ£TaM6~£wv avayvwp(~£1 £ti5tKec; p1l3oowµ1Kec; npwT£°fv£c; 11 l360£1c; TOU rRNA nou ouµµ£T8XOUV 0£ ouyK£Kptµev£c; avnop60£1c;. H ooµtKl1 av6>-.uori, nou ouµn£p1>-.aµl36v£1 6µwri Tponono(riori TWV OUOTQTlKWV TWV p1l3oowµm(wv KQl ouyKpl0£tc; npOK£lµ8VOU Va TQUTOnotr]80UV OUVTf]pr]µeva xapaKTr]- plOTlKO Tou rRNA, rnuTonm£i TrJV £VT6mari Twv ouma- TlKWVTa ono(a £µnMKOVTQl 0£ OUYK£Kptµev£c; A£lTOUp- y(£c;. 24.2 H METOcppaon ~IOKpiVETOI OE Tpia ITOOIO: 'Evap~n. EmµnKuvon KOi TEpµaT10µ6c; BomK&~ &VVOlt:~ • To p1~oawµdno n£p1tx£11p£1c; n£p1oxtc; npoaowric; 1ou tRNA. • 'Eva aµ1voaKuAo-tRNA £1atpx£10t 0111 Stari A. • To nrnTlOUAO-tRNA npoaOEV£TOl OTll 8tari P. • 'Eva aµ1vo~u npOOTf8£TOl OTllV no;l.unrn1101Kn a;l.ua(oa µtow µnaQJopdc; TOU no;l.unrn11o(ou ano TO n£m1ou;l.o-tRNA 111c; Storie; p OTO aµ1voaKUAO-tRNA TllC: Storie; A. 'Eva aµtVO~U µ£Tacpep£TQl OTO ptl300WµOTlO µeow £v6c; aµ1voaKu>-.o-tRNA. H npoo811KrJ mu mriv au~a- v6µ£Vf1 npwT£°ivtKrJ a>-.uo(oa npayµmono1£iTm µfow µlac; aMri>--w(opaaric; µ£ TO tRNA TO ono(o £LX£ µ£Ta- cpep£l TO nporiyouµ£VO aµ1vo~u. Ka8eva an6 aUTO Ta tRNA l3piOK£TQl 0£ µta OlaKplTl1 8eari n6vw OTO p1l30- owµ6TlO. H EIKONA 24.3 0£l)(V£l OTl Ol Mo 8fo£tc; exouv 01acpop£TlKO xapaKTrJplOTlKO: • 'Eva £to£px6µ£vo aµ1voaKu>-.o-tRNA npooi5ev£TQl OTrJ 8£01') A (A site). npiv an6 Tr]V £LOOOO TOU aµt- voaKUAO-tRNA, rJ 8eori £K88T£l TO KWOlKOVlO TO onoio aVTmpoown£U£t TO w6µ£vo aµ1vo~u nou np6K£lTat va npom£8£i OTrJV a>-.uo(oa. • To Kwi51K6v10 nou aVTtnpoown£u£1 TO mo np6ocpma £VOWµmwµevo aµtVO~U OTrJ V£00UVT£8£tµ8Vf1 nOAU- n£ITTlOlKl1 a>-.uo(oa l3p(OK£TQl OTrJ 8£01') P (P site). H 8fori QUTl1 KQTQAaµl36V£TQl an6 TO ntTITl~u>.o- tRNA, eva tRNA nou µ£Tacpep£l Tr] V£00UVT£8£tµevri no>-.un£mti5LKl1 a>-.uo(oa. H EIKONA 24.4 unoi5£LKVU£l 6n TO aµ1voaKUALK6 6Kpo ou tRNA £VTon(~£Tat OTrJ µ£y6>--ri unoµov6oa, £vw OTO 6Mo 6Kpo To aVT1Kwo1K6v10 aMri>--w1op6 µ£ To mRNA OU £!Vat npooi5£i5£µevo OTrJ µtKPl1 unoµov6oa. LUV£- wc;, Ka8£µ(a an6 nc; 8foe1c; p KQl A £KT£LV£TQl KQTO µ11Koc; Kat Twv Mo p1l3oowµ1Kwv unoµov6owv. npoK£Lµ8VOU TO ptl300WµOTLO VQ OUV880£l 8VQV n£- ITTIOLKO owµ6, npen£L va l3piOK£TQl OTrJV KQTOOTQOr] Kw1itK6vm "n' Kw1itK6vm •n+1• Hee011 PouyKpan:( To aµtVOOKUAO-tRNA TO m:nn1iu>.o-tRNA ElOEPXETOl OTTJ 81:'.0T] A _. M&TOKlVTJOT] p1poowµ01(ou 3' 1 nptv an6 10 oXT]µanoµ6 ne:mt1ilKOU 1ie:oµoti TO nEnTLOUAO-tRNA KOTOAOµ~dve:1 TTl 81:'.0T] P, EVW TO aµtVOOKUAO-tRNA TTl 81:'.0T] A NEOOUVTL81:'.µEVT] a>.uo(1ia - ;~::-,1,... ......... 2rxnµanoµoc; nem11i1Koti 1ie:oµou: 10 no>.une:m(o10 µ£Tmpepe:Tm an6 TO nEITTL1iUAO-tRNA TTlC: 81:'.0T]c; POTO aµLVOOKUAO- tRNA TTlC: 81:'.oT]c; A 3HµETOTOnlOT] µe:TOKLVE( TO pt~oowµdno KOTd eva KW1ilKOVLO/ TOn08ETE( TO ne:mtOUAO-tRNA OTTJ ee011 P/ TO anOOKUALWµl:'.vo tRNA EYKOTOAE(ne:1 µ!:'.ow TTlC: Seonc; E/ T] 81:'.0T] AElVOl di5ELO y10 TO w6µe:vo aµLVOOKUAO-tRNA ........ Kwi5tKOVlO "n+1" / Kwi5LK6vto "n+ 2" EIKONA 24.3 To p1~oawµd.no n£p1tx£1ouo 8ta£1c; y1a npoaowri Q)OpT1aµtvou tRNA. nou (j)QLV£TQL OTO 1311µ0 1 Tr]c; EtK6vac; 24.3, OTQV TO n£- ITTl0UAO-tRNA l3ptOK£TQl OTrJ 880T] P Kat TO aµLVOQKUAO- tRNA OTrJ 8fori A. Ixriµanoµ6c; n£nTlOLKOU owµou ouµl3atV£l OTQV TO nOAUn£ITTtOlO, nou cpep£TQl an6 TO n£ITTlOUAO-tRNA, µ£rn<pep£TQl OTO aµtvo~u, TO ono(o cpepnm an6 TO aµtVOQKUAO-tRNA. AUTO TO 1311µ0 npo- Ta aµLVOOKUALKd dKpa TOU tRNA a>.>-n>-emopouv µeoa OTTJ µe:yd>.T] p1~oowµ1Kl'j unoµovdoa Ta OVTlKW1iLK6VLO npo0Mvov101 OE1iml,.avec; TptnAE:TEC: TOU mRNA OTTJ µ1Kpl'j p1~oowµ1Kl'j unoµovdoa EIKONA 24.4 Ot 8ta£1c; PKOi ATOn08£TOUV TO QUO tRNA nou aA- AllA£mopouv KOTO µnKo<:; KOi TWV QUO pl~OOWµLKWV unoµovdowv. 24.2 HM£TdQJpaari t.1aKp(v£10t a£ Tp[a I1do1a: 'Evap~ri, EmµnKuvari KOi T£pµanaµoc; 677
  • 169.
    To aµ1voaKUAO-tRNA El0£PXETOI 011BEOrJ A To no>.unemio10 µernq,tpnm OTO aµlVOOKUAO-tRNA Hµn01dmo11 µETOKIVE[ TO nEmlOUAO- tRNA OTrJ BtOrJ P EIKONA 24.5 To aµtVOQKUAO-tRNA £l0£PXETQl CJ1TJ 8t011 A, napa>..aµ~dvet TTW no>..unrnnOtKf] a>..uoioa an6 TO nrnnou>..o-tRNA KQl µ£TQ(j)£- P£TQl OTTl Stan Py1a 10v rn6µevo KUKAO emµf]Kuvoric; Ono8eu:1 awm~ rnno8en1ari TWV aµLVOOKUALKWV 6.- Kpwv Twv Mo tRNA µfoa mri µ£y6.t..ri unoµov6.<3a. H auyK£Kptµevri OVT(<3poori KOTOAU£TOL ano Tl'l µ£y6.t..ri unoµov6.<3a TOU pt~oowµm(ou. H µ£TOcpop6. TOU nOAUn£mt<3(ou <3riµ1oupy£( TO pl~O- awµ6.no nou napouo16.i'.;£TOL mo ~~µa 2, onou TO anoa- KUA1w1,,1tvo tRNA, xwpic; KOV£VO 0µ1vo~u. ~piOK£TOl OTTl etori P, £vw eva vfo n£mt<3ut..o-tRNA EXEL <3riµ1oupyri8d mri etori A. auTo TO m:m1<3ut..o-tRNA dvm µaKpuT£po KOT6. eva KOT6Aomo 0µ1vo~foc; ano TO m:m1<3ut..o-tRNA nou ~plOKOTOV OTTl etori P KOTO TO ~~µa 1. To p1~oowµ6.T10 µ£TOKLV£LTOL Twpa KOT6. µ10 TpmM- TO KOTO µ~KOC: TOU ayy£ALOcpopou RNA. AUTO TO m6.<3LO KOA£LTOL 1,J&TOTOTIIOI') (translocation). H µ£TOKivriori £~6.y£L TO anOOKUALWµevo tRNA ano Tl'l efori P, onou £to6.y£L TO nem1<3ut..o-tRNA (~Mn£ ~~µa 3 mriv EtK6va 24.3). To £n6µ£VO KW<3LKOVLO y10 µ£T6.cppoori ~pLOK£TOl Twpa mri Sfori A, eTmµo va uno<3£X8£L TTlV doo<3o £voe; vfou 0µ1voaKut..o-tRNA, oriµeio mo onoio ea ena- vat..ricp8d o KUKAoc;. H EIKONA 24.5 ouvoljJii'.;£L TTlV oMri- t..£ni<3poori µ£TO~U TWV tRNA KOL TOU pl~oowµmiou. To anOOKUALWµevo tRNA £YKOTOA£(m:1 TO pl~OOW- µ6.no µfow Tl'lC: Sforic; E, µ1ac; 6.Mric; Sforic; npoo<3£- oric; tRNA. H etori OUT~ KOTOA0µ~6.V£TOL napo<3LK6. an6 TO tRNA, nou ~pioK£TOL mri <3L0<3tKaoia va £YKOTOA£i- ljJ£L Tl'l Sfori P KOL v' an£A£U8£pw8d ano TO p1~oowµ6.- no mo Kunapont..aaµa. 'EToL, ri po~ Tau tRNA £LVOL µfoa OTTl etori A, <31aµfoou TTlC: etoric; P KL e~w ano Tl'l Sfori E (~Mn£ rn(oric; EtKova 24.28 oTriv fv6r17ra 24.12). HEIKONA 24.6 OUYKPLV£l Tl'l µ£TOKLVl'l0Tl TWV tRNA ...• 0EOrJ E .·...... ..... ....0tm, P •• ••••••••• ~ =~ ·- • a-•.-a---.aaaa a a a aaa a aaaaa I EIKONA 24.6 Ta tRNA KQl mRNA µ£TGKLVOUVTQl OLaµtoou TOU PL- ~oowµm[ou npoc; TTlV 101a KGT£tl8uvori. 678 KECl>A/AIO 24 M£Td(j)paori KOL mRNA, nou £LVOL <3uvmo VO 8£wpri8d we; £VO d<3oc; 0<30VTWTOU TPOXOU, ITTOV ono(o Tl OVT(<3p00Tl K080<3Tl- y£(TOL ano TTlV 0AATlA£n(<3poori µ£TO~U KW<3LKOVLOU KOL OVTLKW<3LKOVLOU. H µ£T6.cppoori <310Kp(v£TOL a£ Tp(a m6.<310, nou na- poum6.i'.;oVTm mriv EIKONA 24.7: • H tvap~I') (initiation) n£pLA0µ~6.v£1 nc; OVTt<3p6.0£1c; nou auµ~a(vouv npornu axriµm1m£( o n£m1<31Koc; <3wµoc; µ£Ta~u Twv Mo npwTwv 0µ1vo~ewv Tl'lC: npwTE'fvric;. npouno8eT£L npoo<3£0Tl Tou p1~oowµa- T(ou mo mRNA, WITT£ VO axriµOTLITTEL £VO ouµnt..o- KO £VOP~TlC: nou n£pl£X£L TO npwTO aµLVOOKUAO- tRNA. AuTo £(vm eva oX£TLK6. ~paM ~~µa mri µ£- 'Evap~11: 11 µ1Kpn unoµovaoa OTrJ 8£011 npdoownc; mRNA ouvoua~e1m µe 111 µeya>.11 unoµovaoa Km npoootvnm 10 aµ1voaKuAo-tRNA EmµnKUVOrJ: 10 pl~OOWµ0110 µ£1aKIV£l1Ql K011l µnKo<:; 1ou mRNA, emµ11Kllvovrnc:; 111vnpwn:fvn µe µernqiopa and 10 nrnnouAo-tRNA 010 aµ1VOQKUAO-tRNA Tepµanoµ6c:;: 11 n0Aunrnni'i1Kn a>.uoioa ane>.eu8eptiJV£10l and 10 tRNAKm10 p1~oowµa110 olima1m and 10 mRNA EIKONA 24.7 Hµ£Tdq>pa011unoo1ap£[TQ10£ Tp[a OTdOlQ.
  • 170.
    TC!cppaori KOL ouvfJewc;Kaeopil;el TT']V TOXUTT']TO µe TT'JVonoia ea µeTOcppaOTei TO mRNA. • H &mµ~Kuvo11 (elongation) nept11.aµp6veL 611.ec; nc; OVTtopaoetc; an6 TT'] ouv0eOT'] ,OU npJnou nemtOl- KOU owµou ewe; TT']V npoo0fJKT'J TOU TE:11.WTOLOU aµL- vo~foc;. T' aµtvo~fo npoOTieeVTOL OTT'JV 011.uoioa eva TT'] cpop6: T1 npoo0fJKT'J ev6c; aµtvo~foc; eivOL TO Ta- xurnpo pfJµa TT']c; µeTacppaOT']c;. • 0 T&pµaT10µ6c:; (termination) neplfl.aµpaveL Ta Pri- µma nou anOLTOUVTOL yta TT']V ane11.eueepwof1 TT'Jc; 011.oK11.ripwµevT']c; no11.unemtotKfJc; 011.uoioac;: TOUT6- xpova, TO ptpoowµ6Tto otiOTOTOL an6 To mRNA. ~LacpopenKec; oµaoec; E:nlKOUplKWV napay6VTWV unoPorieouv TO ptpoowµano oe K60e OT6oto. Evepyeta napexeTOL oe otacpopa OT6ota an6 TT'JV uop611.uOT'] TT']c; TptcpwocpoptKrJc; youavivric; (GTP). Km6 TT'JV evap~T'J, ri µLKPrJ ptpoowµLKrJ unoµovaoa pooMveTOL OTO mRNA KOL OTT'] ouvexeta evwveTOL µe µey611.ri unoµovaoa. Km6 TT'JV emµfJKUVOT'], TO mRNA uaKLVeiTOL otaµeoou Tou ptPoowµmiou KOL µeTOcpp6- £TOL oe TpmMTec;. (nap6n ou~ewc; µtMµe yta µeTO- ivriori TOU ptpoowµm(ou KOTC! µfJKOc; TOU mRNA, 8LVOL epLOOOTepo pea11.LOTlKrJ Tl tMa TT']c; e11.~ric; TOU mRNA taµfoou Tou ptpoowµmiou.) Km6 Tov rnpµanoµ6, ri pwTefVT'] KOL TO mRNA ane11.eueepwVOVTOL, evw Ol ~e- wptOTec; ptPoowµtKec; unoµovaoec; OllOTOVTOL npoKet- evou VO ~OVOXPT']otµOnOtT']eouv. 24.3 E101Koi Mnxav1oµoi E.Myxouv mv AKpiBEIO me; METCJcppaonc; 1 BOOIK&I; &VVOl&I; • H aKpf~£ta Tll<; µndcppaoT]<; £A£YX£TQl and £lOlKOU<; µT]xavt- oµouc; 0£ Kd8£ 010010. rvwpil;ouµe 6Tt ri µeT6cppaori eivOL yevtK6 aKptPfJc;, C!pf'] OTT'] ouveneLO TIOU OlOnLOTWVOUµe OTOV K00op(- ouµe TT'JV 0Mri11.ouxia ev6c; no11.unemtoiou. 'Exouv npayµmonotrieei 11.iyec; 11.emoµepdc; µeTpfJoetc; TT']c; ouxv6TrJTOc; M0ouc; in vivo , wOT6oo yevtK6 mOTeue- TOt 6Tt KUµaiveTm oe KMµaKa mu ev6c; 11.68ouc; ova 104 ewe; 105 aµtvo~fo TIOU 8VOWµOTWVOVTOl. /aµpa- VOVTOc; un6ljJT'] 6n ot neptoo6Tepec; npwTdvec; nap6- 0VTOL oe µey611.ec; noo6TT']Tec;, auT6 oriµa(veL 6n ri ouxv6TrJTO M0ouc; eivm noM xoµri11.fJ yta va exet K6- no10 eniopaOT'] OTO cpmv6Tuno TOU Kunapou. ~ev eivot 6µwa npocpavec; nwc; emTuyxaveTOl TOOO xaµri11.11 OUXVOTT']TO M0ouc;. npayµmL, Tl cpuori TWV eµ- n11.eK6µevwv µT']XOVLOµwv OTIOT£11.et Y£VlKO eeµa yt' Op- 8TC! pfJµma OTT']V eKcppaOT'] TWV yovtoiwv: • nwc; TO evc;uµa nou ouveeTOUV RNA avayvwpil;ouv µ6vo TT'] paori nou eivot ouµn11.ripwµmtKrJ µe TO eK- µayeio; • nwc; 01 ouveeTaoec; avayvwpil;ouv µ6vo xa tRNA Kot aµtvo~fo; • nwc; eva ptpoowµano avayvwpil;et µ6vo TO R nou OVTtOTOlX£i OTO KWOLK6vto OTT'] 0eOT'] A; Kaee nepimwori eernt eva nap6µoto np6P11.riµa: nwc; µnope( va otaKpt0ei eva ouyKeKptµevo µe11.oc; an6 oMMrJPTl TT'JV 0µ600, TT'Jc; onoiac; 611.a Ta µe11.ri µrnp6- l;oVTot TO LOLO yevtKC! xopOKTT']plOTlKO. nteav6v OTIOlOOrJTIOTe un60Tpwµa µnope( OPXLKQ VO e11.8et o' enacpfJ µe TO evepy6 KeVTpO KOTC! TUXT'J, aM6 enetTO TO 11.680c; unoOTpwµma anoppimoVTot Kot ano- oeKT6 y(veTQl µ6vo TO KOT0/.1.T']II.O. To KOT0/.1.T']fl.0 un6- 0Tpwµa ppioKE:Tot n6VTa oe µetoljJT']cpia (µta an6 Ttc; Tfooeptc; paoetc;, tva an6 Ta eiKooL aµtvo~fo, tva an6 TO - 30-50 tRNA) , ouvenwc; TO KplTrJpta OlC!KptOT']c; npe- neL VO eivot OUOTT']p6. H EIKONA 24.8 ouvoljJ(l;eL TT'] OUXVOTT']TO M8ouc; OTO pfJµma nou µnope( va enripeaoouv TT'JV aKpiPeta TT']c; µeT6cppaoric;. Ta Meri OTT'J µeTOypacpfJ Tou mRNA eivot on6vta, < 1o·6 . np6KetTm yta oriµaVTtK6 OT6oto yta TrJv aKpi- Peta, Ka8wc; K6ee µ6pto mRNA µeTOcpp6l;eTot oe noM6 npwTe"tvtK6 aVTiypacpa. Ot µrixovtaµoi nou e~aocpaM- c;ouv TT'JV aKpiPeta TT']c; µeTOypacpfJc; e~eT6l;oVTot OTO Ke- cp611.oto 19. To ptPoowµ6no eivOL uneu8uvo yta Mo Tunouc; 11.aewv OTT'] µeT6cppaori. Mnopei va np0Ka11.foet µua- T6mori Tou n11.otoiou avayvwaric; (frameshift) napa11.ei- LUXVO-rT)rn MSou~ I ~ /dSo~ ~dITTl I M£TOTOTILOT) I nAma[ouovdyvwari~ /d9o~ I oµtVOOKUAO-tRNA t /dSo~ 0µ1vo~u -/d9o~tRNA I 10-6 10-s 10-4 EIKONA 24.8 LTO 01acpop£TLKd OTOOLa TT]<; µ£Td(j)paOT]<;, Ad8T] ouµ~aivouv µ£ ouxvdTTJTO and 10·6 twc; 5x 10-4. 24.3 ELOlKO[ MT]XOVtoµo( EMyxouv TT]V AKpf~£la TT]<; M£Td(j)pOOT]<; 679
  • 171.
    nowoq Uro Bdon6rov 6roBd(er ro mRNA (rl 6roBd- (owog pro Bdoq 6Uo gopdq -nptirro og teleuro(o Bdoq ev6q ro6rrov(ou rr 6nero o:g npdrrl Bdor'1 rou en5trlevou xro6rrov(ou). Aurd ro l,d0r1 e(vor ondvrq (oup6rqro -10-5). En(or1g trrnope( vo enrrpdqUer (ro- ro)ro(ptoogo ev6g LdOog opuvoorulo-tRNA pr' 6vo rco6rx6vro, pe ouv6nero vo evoroporc0veror ).60o9 o;u- vo[ti. Aw6 prd].].ov e(vor ro oulv6repo Ad0og orn Ue- r6qpoo4 (ouXv6r4to -5 x 10 4). ELdyXeror on6 rrl 6o;111 tou ptBooroprot(ou Kor rnv r<rvqnrf qg 6rdoro- oqg (BL6ne Ev6trpa 25.1 5). Mro ouvOerdorl rou IRNA prnope( vo nporol*ioet 6uo rrlnoug LoO<irv: vo ronoOerloer Ld0oq oprrvo[0 oro IRNA r4g 11 vo goprcioet ro opuvo[ri qg oe ].d- Ooq IRNA (BI6ne Evotrpa 25.9). H evocopdroo4 Ld- 0og opuvo[dog e[vor nro rorvrl, (oolg enerdq ro IRNA npoorpdper peyolUreprl enrrpdvero [e rnv ono(q to 5v- (upro pnope( vo 6rlproupy{oer neptoo6repeq enogdq yro vo SroorpoL(oer er6rr6rqto (specificity). Or ouv0e- rdoeg rov optvooKuLo-tRNA 6ro0rirouv er6rxorig ;rr1- Xovroporjg yrq rnv enrd16p0ooq Lo0cirv npo'roU oneleu0epto0e( 6vo IRNA pre ).dOog <popr(o (BL6ne E- v6tqra 25.1 1). 24.4 H 'Evop[n oro BoKTtipto Ynopovd6eq Kt EntKouptKo0q Xperd(eror 3OS Ilopdyovreq e H evop[I qq peroqpoo1g onoue[ Eextoproreq prpooorgrr6g unopovodeg 30S ror 50S. * En[onq onouotivror ot nopoyovreg evop[1q (lF-1, -2 rot -3), ot ono(ot npoodrlvovror otrg 30S unopovodeg. e Mro 30S unopovodo nou gerorpeper nopoyovreg evop[qg npoo66veror oe pLo 0eo1 Svop(1q oro mRNA oXlpor(ovrog evo o0pntroxo evop[r19. * 0 lF-3 npener v' one),eu0epot0e( tlore ot 50S unogovodeg vo npoode0ouv oro o0gnLoro 30S-mRNA. To Borrrlpror<d prBooroprdtro nou lprlottrlonotoriwor ornv entprlKuvoq Uroq nolunemLdLr<r1g oLuo(6og ug(- orowor rr;g 70S ooll.ror(6ro. Kord rov repprotropr6, one).euOepovowqr on6 to mRNA cr.lg eeU0epo pLBo- o<r.lprdrro r; prBooroprxdg uno;rovddeg. tre ou(ov6pevo Bor<rqpto, ro neproo6tepo ptBoorrtprdtro ouvO5rouv nporelVeg : 11 de[opevrl rotv e].eU Oepo-lv p rBooropror[cov nr0ov6v nepr5ler -20o/o r/ov prBooropror(ov, To ptBooroprdtro r4g eLe60eprlq 6e[opev4g ;.rnope( vo Srooro0oriv oe leloprm6g unopovd6eg: ow6 oq- ;ro(ver 6rL to 70S prBooopdtro Bp(or<owot oe 6uvopu<q Looppon[o pre ug 30S xor 50S unopovddeg. H |vaplrl tqq pefiEpoo4g 6ev elaptotaL an6 oxipata pLBoortt- paua oMa avalapBavetat an6 tq eyuporcq unopo- vddeg, or ono(eg otrtrrllenr6porjv r<ord qv ow[6poor1 rrig 6vop[49. H ri{*i!]ti !;i.} ouvoil((et rov r<0r<}.o rov pr- 680 KEOAAAI0 24 Meroqpool 30SunopovdDeg :eXoptotdg Ae(opevrl 'Evup{n EnqrKuvoq Teppottogtig EIK0NA 24.9 H evop[1 onoLre[ etreU0epeg pgooopLr6q urotto- vddeg. 0rov ro pgooropdrto onetreu0epdrvovTor oTov reppouoprl, or 30S unopovoDeq npoo66vouv nopoyovreg 6vop[r1g ror 6ri- orovTor dlgroupytowoq etre0Oepeg unopovd6eg.'0rov or unopo- vodeg enovoouvduo(ovror yro v' onodtiroouv rivo treuoupyrxri prpooogdno oqv 6vop[1, onetreu0ep6vouv Touq nopoyoweg. Boocoprrdrv unopovddrov r<ord tr1 pretdgpoorl oro Bo- rcrripro. 'Evop(q ouprBo(vet or puo erdrxrl oLlqLoul[o rou mRNA yvroorrl cog neprol{ np6o6eo4g rou plpooo- poriou (ribosome-binding site) (ouprnepr).oprBdver n1v oMqLouX(o Shine-Dalgarno - BLdne en5prevq Ev5rq- ro). l-1p6r<error yro Uro BpoXe(o oLlqLouX(o Fdoeov npw on6 ulv x<o6rxonorqtu<f neproXrlg (BL5ne Err6vo 24.12),4 ono(o e(vor oupnlqpro;rorxrl ev6g rp4prorog tou 165 rRNA (B).6ne Ev6rqro 24.18). H prxpri Kor n Ue- ydLrl unoprovd6o oMrll,entdpo0v orrlv neproXrl np6o- 6eonq rou prBootopror(ou olqpot((owog 6vo d0ucto prBoocopdrro. H ow(6pooq ougBo(ver oe 6Uo BnUoro: . Avoyvcbprorl rou mRNA npoyUaronore(ror 6rov npoo66veror puo prxpri unopovddo oXqpror((owog 6vo olpnAoxo 6vopl4q (initiation complex) orrlv Tre- ptopl np6o6eorlq rou prBooroprot[ou. o 'Enetto prro preyd),r1 unopovddo ouv66eror ge ro o[ pn].oxo oXq poil(owog 6vo n],r1 peg prBooro prdrro. l-iop6n q 30S unopovddo eprnLdreror ornv Svop[q, 6ev enopre[ on6 g6vr1 rnq yro ng owr6pdoetg np6ode- orlq rov mRNA ror IRNA. Anore( entnp6oOereq npto- refveg yvom5g og nop6yovreg 6vop[49 (initiation factors - lF). Tdtorot nopdyoweg Bp(or<owor ;r6vo nd- vo orrq 30S unoprovddeg rr oneleu0eprivowor 6rov or 30S unopovd6eg otrL4l.enrdpo0v pe ng 50S unopovd- 6eg nopdyowog 70S prBootoprdrro. Aurrl 4 6pdo4 6ro- rp(ver roug nopdyovreg Svop{r1g on6 rrg 6opur6g nporetveg rou prBoorollffi[ou. Or nopdyoweg Svop[r1g ooXol.oriwor onox,.eronrd Ue ro oxrlpotopr6 rou oup- nl6xou Svop[qg: onouord(ouv on6 ro 70S prBoorr:pr6- lo Kqr 6e 5to6po;ror((ouv rdnoro p6lo oro ord6to rqg enrpuiruvorlg. H Elf0NA 24.'10 ouvorf((er ro md6ro rqg 6vop[qg. Boorxeg evvoreg llopdyorneq
  • 172.
    1 H 30Sunouovddo npoo66verot oro mRNA tcail-J G...... A AuvogLrl rooppon[o Ae(ogevrl 70S prpoorilgoriolv EIK0NA 24.10 0r ropoyovTsq 6vop{19 oToeeporototiv nq etreU- 0epeq 30S unopovqdes Kor rrp0o66vouv ro evopKTnpto IRNA oTo oriUrIoKo 30S-mRNA. To Borrrjprq XpnorponotoUv rpetg nopdyovteg 6vop(r19, lF-1,1F-2 Kqr IF-3. Or nopdyoweg outo[ onot- roUwot yro rnv e(oo6o oro oUpnloro 5vop[qg t6oo rou mRNA 6oo rot rou IRNA: . O lF-3 61er noMonl,Sg Letroupy[eg: mo0eponote( rrg (eleri0epeq) 30S unogovdSeg rot ovomSlLet qv np5op4 np6o6eoq qq 50S unoprovddog. Ko0t- md rrg 30S unoprovd6eg trov6g vo npoo6e0o0v ong O5oerg dvop[r1q rou mRNA. En(or1g, tog t1.ui1.to tou oupntr6xou 3OS-mRNA, etr6yXer qv or<p(Bero q q ovoyvri p rorl q ro u n pdlro u ol.uvooxul.o-tR NA. . O lF-2 npoo66ver 6vo erdrr6 evopxuipto IRNA r<t el,6yXer r1v efoodd rou oro prBoocoptdtto. . O lF-1 npooS6vetor oe 30S uno;rovd6eg oq rprlpo rou ntrrlpoug ouprnL6r<ou 6vop[qg. [1poo66vetot rowd orq 06oq A Kor nopeUro6(et qv e(oo6o tou o1lwooxul,o-tRNA. H ew6ntoq tou prnope( eniorlg vo epno6(oer qv np6o6eor1 qg 30S unopovddog pe qv 50S unopovddo. I1o),),59 6opur<59 prel6teg unod4trtilvouv 6tt o lF-3 51er drio 6tor<pm6g, odpd ogotpu<5q neptoX6q, Ue rnv C-reLtrrl neproXrj orrl 06o11 enogrlg yto rrlv 50S ndvo orrlv 30S unopovddo Kor rnv N-teLxri neptoX4 xowd orq 05oq E qg 30S. H npdrr4 l.enoupy(o tou lF-3 eLdylet qv tooppon(o prero(ri tov ptBooco;ru<tilv xoroor6oeov, 6nog Eofuerot orqv El(0t'lA 24.11. O lF-3 npoodSvtrot 01 etre[0epeg 30S unoprovd6eg, or ono(eg oneleu0epdrvowot on6 tr1 Etreu0epeS unopovodeg t- I t I H 30S unopovodo pe rov lF-3 Unopet vo npoode0et I oro mRNA. oxr opoc xot oqv 50S unopovo0o i I 0 tF-3 npener v 'nttrsuospooet yt. v0 unopeost " * . . . ' I vo npooDe0el 11 50S unouovci5o EIK0NA 24.11 H 6vop[r1 onoLtei 30S unopovodeg nou getoq6- pouv lF-3. de(qrevrl tov 70S prBoocopror(ov. H nopouo[o tou lF-3 nope;rno6((er rrlv enovoo0v6eorl qg 30S unopovddog pe Uro 50S uno;rovd6o. O lF-3 Unope( v' oMrlLenrdpd- oer dpeoo pre ro 165 rRNA r<or undpXet orlprowu<r1 ent- rdLur!4 pero[ri rov Bdoerov tou 165 rRNA nou npoorore0owor on6 rov lF-3 Kot rov ow[orotXtov Bd- oeoJv nou npooroteUowot on6 rqv np6o6eor1 qg 50S unogovddog, yeyov6q nou uno6qtrcilvet 5fl nopoxtol,rjet 14 quoxri ouprBoLrl tov unopovddtov. Iuvenrig, o lF-3 ouprnepup5perot cog nopdyowog owt-oltr4lenbpoo4g, nou 6Xer org onot6)reopotnv nopogovrl qg 30S uno- provddog orq de(opevrlrov etreri0eporv unopovddtov. H ow(6poor1 peto[ti lF-3 ror 30S unoprovddog e(vot orot- Xetoprerprx4: 5vo pr6pro lF-3 npoo6dverot ovd unopovd- 6o. Enerdrl 11 noo6rlro tou lF-3 e(vot oXettrd ptrprl, ouvdyeror 6u q 6ro0eotpr6rqrd rou ro0opf(et tov opt0- pr6 ro:v ee0Oepov 30S unopovd6tov. H 6efrepq ].emoupy(o tou lF-3 e].6yXet tqv xov6rl- ro rov 30S unoptovd6rov vo npoode0oUv mo mRNA. Mrxp6g unopovd6eg np6na vo 6to05touv lF-3 nporetp6- vou vo g4rlpot(oouv otipmloxo 6vop(qg pe to mRNA. O lF-3 npdner v' onetreu0epoOe( on6 ro o0pnLor<o 30S- mRNA d:sre vq prnopSoet vo npoo6e0e( 11 50S unopovd- 6o. M6tg oneleu0epo0e(, o lF-3 ovorurl.6vetot opr5ocog Bp(orowog puo dMq 30S unopovd6o. T6Log, o lF-3 el,6yXet t4v ot<p[Beto ovoyvdrptorlg rou nprilrou ol.rloorul.o-tRNA r<ot Fon0d orov npooovoro- Lropr6 rou rpoq rrl 05oq P rrlq 30S unopovd6og. H nprirq on6 owSg rg Lemoupy(eg 6Xer ono6o0e( oqv C-relrnj neproXq tou lF-3 (BL5ne Ev6t4ta 24.7). H N- teltxrj neptopl rou lF-3 e(vot tono0etrlpr6vq drme vo Bor10d fiov npooovotoLtop6 rou optvooKu).o{RNA npoq rn 06oq P qg 30S uno;rovddog onoqpdooowog rq 06o11 E ev6oo o lF-1 onogpdooetrq 05or1 A. O lF-2 dXer 6p6or1 GTPdorlq e(oprc(rprevqg on6 to prBootoprdrto: nopouo(o prBooto;ror(rov npodyet tr1v :i 0 lF-2 geper ro IRNA orn 0don P 3 0r lF oneleu0ep6vovror xor npoo66vetot 11 50S 2&,4 H 'Evop(r1 oru Boxqpro XpeLd(eror 30S Ynopovddeg xt Enxouptro6q flopdyovres 681
  • 173.
    udp6Luorl rou GTPone),euOeprilvowoq qv ev6pyero nou e(vor ono04r<eup6vr1 oro 6eop6 uUnInS evdpyerog. To GTP u6poLtieror ;16119 npoodeOe( 11 50S unopovddo yro 14 drlpuoupy(o rou ntrrlpoug prBoocogot(ou. H 6Ld- oroon rou GTP 0o pnoporioe vo epnl,6reror fin Uero- BoI4 qq 6ropr5prpcooqg rou prBoocopor(ou, Stot 6qre or ouvevopSveg unopovd6eg vo perorponoUv oro evepy6 70S prBooroprdrLo. 24.5 H 'Evop[n lleprAopBdv€t ZEUydpoUo Bdoeulv Mero[ti mRNA Kot rRNA '. Iro gorrrlproro mRNA 11 0eo1 evop[1q onore],e[rot ono ro rodrrovro 6vop(1q AUG ro onolo oroiouOei -10 pooerg ge- ro ro e[opep6g nouplvrilv Shine-Dalgarno. ,, To rRNA qg 30S pr$oooprrnq unopovodog rotv Borqpuov Dro0erer pro oupntrrlpo;rolr1 otrtrr1trouX[o nou (euyoprlver pe qv otrtr1trouXLo Shine-Dalgarno roro qv evopil. To orlpo yro rnv 6vop[q prog no].unemrdrrrlg otruo[6og e(vor 5vo et6tx6 rto6u<6vto Svop(4q, ro ono(o orlporo- 6ore( qv oppl rou n).oro(ou ovdyvotorlg. Iuvrl0otg, r<o- 6rx6vro Svop[qq e[vor qtptn}.6ro AUG, toor6oo oro Borcrrlpro XprlorUonororiwot en(or;g ro GUG xor UUG. 'Evo mRNA nepr6ler nol),69 rprnAdreg AUG: nog dpoye ovoyvrop((eror ro r<ro6rxdvro dvop[qg oq oeern- p(o qg perdgpoorlg; Or 0ooetg tou mRNA dnou opX((er q perdqpooq rouronotofwor on6 qv np6o6eoq rou pr- Booorpror(ou oro mRNA un6 ouvOrp<eg or ono[eg ovo- m5).Louv rqv entpqxuvorl, r<o0r1).rrtvowoq ro ptBootr-l- pdrto mrlv o<peqp[o. Av oro r<o04].cop-rdvo oripn].or<o Svop[qg npooreOe( prBovouxtredorl, 6].eg oL neproxeq rou mRNA nou Bp(orowor 5lo ond ro prBooroprolo 0' ono6opr40oriv, evc': or neprolSg nou 61ouv npoo6e0e[ oro pLBoorrtl-rorro 0o Trpoororeurorjv, 6nog qo(verot oqv . To npoororeupSvo r<Ldo;roro ;rnope( v' ovocrq0oJv Kor vo loporcrqptoroJv. Ot oMq).ouXieg Svop[r1g nou npoorore0owoL on6 ro BorurlpLor<d pLBoooprdlo dlouv prrp<og -30 Bdoeotv. Or nepLoXdg np6odeorlg rou prBoooprorfou 6rorpoperL- rurv BorcrrlpLoxrr;v mRNA eprgov((ouv 6io xorvo Xopo- KI1proflKo: o To r<odrx6vro dvoplrlg AUG (n Lrydrepo oupo GUG n UUG) ndwo oupnepL).opBdveror ornv npocrroreu- pdvq otrI.q).ou1[o. o Ze ondorooq 5og 66ro Bdoeov ovo6Lr<d rou AUG Bp(or<eroL puo oMqLouX(o nou owrororXe( oro e[r1g elopep6g 11 oe rprl;ro rou: 5,..AGGAGG..3' Autq q nolunouprvrr<r1 ol.).r1).ouX(o e(vot yvcrlorrl orq oMqlouX[o S h i ne-D al garno. E(vor ou prn].r1 potl.rortr<r1 ;re 682 KEOAAAIO 24 N{sToQpoorl r^r{F, /t' 0Dnyoq KoDrronorrlrtrui neptoxrl Shine-Dalgarno < 1 0 pdoerg ovodLxd'rou AUG Al-li '0tre9 ot neproxdg oro rdv.rpo rou tivop(1q 6to0erouv npoorortuUtvou 6tloouvotvertro xtrdoporoq orotxe[o EIK0NA 24.12 l-leproX6g npdodeorlg rou prpooogorfou novto mo mRNA pnope( v' onopovtoOouv on6 oupntroro 6v0p[rlq. 0r nepo- X6q 0ur6q nepilogpdvouv qv ovodrrrl otrtrntrouxio Shine-Dalgarno ror ro xtodurjvro evop[nq" pto eforpetrrd ouw4p4pSvq o).).4).ouX(o rowd oro 3'drpo rou 165 rRNA. (To eripog qg ouprnlqpcoprorrr6- rqrog drorp5per ovd).oyo ge to mRNA ror pnope( vo ercre[veror on6 r4v xewpu<r1 oMrl],ouXio reoodpov B6- oeov GAGG 6o9 puo oMr'1),ou1(o ewSo Bdoecov.) l-potrr- pr6vr1 oe ow(mpog4 rotef0uvor1, q o].].4].ouX(o rou rRNA efuot ro e(oprepdg: 3'...UCCUCC...s', Apoye, rord rqv np6o6eoq rou mRNA oro pgooro- pduo q oMrlLouX(o Shine-Dalgarno (euyoprirver Us ro oupnl4prllporu<6 qg rRNA; Meto).)d[etg efue orn Uto eke mqv dtri,q oMqlouX(o uno6rll.rr:vouv rr1 oqproo[o rouq orrlv Svop[q. Irlprerox6g preroMd(etg orrlv oMrl- ouX(o Shine-Dalgarno prnope( vq nopegno6(oouv 14 prerdrppoo4 tou mRNA. EnrnLriov, 11 erooyoyrl ;rerotri.d- (eov orrl oupnl.qpolprotu<ri oMqLoul(o rou rRNA e(vor enp),oBr1g yro ro r[rropo r<or pretoBdMer ro np6runo qg perdrppoonq. H xoOoproru<r1 engeBo(cooq qg ow[- 6pooqg rou (euyoprilgoroq rov Bdoerov e(vor 6rL puo perdLLo[r1 orqv o].].4].ouX[o Shine-Dalgarno evdg mRNA pnope( vo roroorotre( on6 pro prerdtrtro(4 oro rRNA nou onoroOrord ro (euydpopro rtov Bdoeotv. H oMqLou{o oro 3'6xpo rou rRNA e(vor ouw4pr;- prdvq prero[U npoKopuord:v xr europucorcbv, Ue rrlv e(o(peor1 6n oe 5loug rouq euKopucirreg undpXer ono- Lorqrl qq oL).4l,ouX(og nSwe Bdoetr:v C C U C C, n ono(o e(vor ouprnLrlpoprorLrrl qg oM4louX(og Shine- L---T----------l npdo0eon rou prpootopor(ou oqv neprcXn SvopEnq tou mRNA npoo0rlKrl vourtredolq yto 6trou rou onpomdreurou mRNA Anopdvoorl rtrdoporog rou npomoreug6vou mRNA Boorxeg 6vuoreg AACAGGAGSAU UACCCCAUG UCGAAGCAA... npoototeu gSvou rtrdopotoq
  • 174.
    Dalgarno. Iuvendrg, prdMov6e ouprBo(ver (euydporpro Bdoeov preto[ri rou euKopuotrxoU mRNA rot rou 18S rRNA. l-1p6r<error yro o4prowu<4 dtogopd oro prrllovropr6 rqg 6vopl4g. Iro Bor<rqpro, Uro 30S unoprovd6o npooS6veror d;reoo oe pro Odoq rou prBooco;rodou. 'Eror, o111prot(- (eror dvo o[pn],or<o 6vop[qg oe Uro o].trr1].ou1(o nou neprBdMer ro ral6u<5vro Svop[r1q AUG. 'Otov ro mRNA efvot no].uormpovrx6, rdOe rcodrronot4rrrrl neprol4 opX((er pe pto 05o4 np6o6eorlg prBoocopror[ou. H rptio4 qq Srrppoolg rov Borcr4prordlv yovrd(otv orlpo(ver 6n q prerd<ppooq ev6g BorrrlproroU mRNA npolcope( 6ro6o1u<d 6rop5oou rov ororpov[rr.rv rou. M6- fug to ptBoocoprdtro npoo6e0o0v orrlv nptirq rrrrdrronor- rpu<r; nepro1rl, ot en6peveg rcodrronorqru<5q neproXSg 6ev 6Xouv or<6prq preroypoge(. M6rg y(vet 6to06orgr1 4 deurepq 06oq rou prBooroproilou, 11 prerdgpooq Bpiore- ror xo0o66v 6rop6oou rou nprirrou otorpov(ou. To ou;rBdwo olg rro6rronoulnx6g neprol6g norri Aouv ovdLoyo Ue ro mRNA. Ilg neptoo6tepeq ilepr- nrrioetg, ro ptBoocoprdrro prdAov npoo6Svovror ove[dptrpo orrlv opXrl rdOe ororpov(ou. H ouq06ore- pq oerpd rrov ou;rBdwolv nopouord(erot mqv EIK0NA 24.13. Otov repporrore[ 11 o[vOeor1 qg npc0rqg npotrei- vnq, ro prBoocopdno eyrorole(nouv ro mRNA r<or 6ti- orovror orrq unopovddeg toug. 'Enerro, rivo vrlo prBoocopdtto np6neL vo ouvopl,rooyq0e( orqv en6trrev4 rodxonorqtu<r1 neproXq xor vo (errvrloer q perdqpoorl oro en6gevo otorp6vro. Ie opropdvo Borcr4pLord mRNA, 11 pretd<ppoorl rcov 6mtrovrov ororpov(rov e(vor ou(euygdvrl, enet6ri to prBo- ocopdrto onorcrofv np6oBooq oro xco6tr6vto 5vop[r1g rou 6e[tepou ororpov(ou r<o0dlg o].ox].r1p6vouv rrl Ue- rdgpoorl rou nprinou. l-to vo ougBe[ ow6, ro peoo6rd- 6rnUo peto[ri rtov 6tio ro6rronorqrrxciv neptolrirv npdner vo efuor prxp6. Mnope( vo e[oprdtor on6 rlv uqnln rontrti nurv6rrpo rcrlv prBootol-ror(cov. EvoMo- rrtrd, 4 nopdOeo4 O5oeo:v repprolo;ro[ xr Svop[r1g enrpdnet vo nopqKqpq0oUv opropr6vo on6 to ouvr10r1 oupBdwo prero(0 rov ororpov(cov. 'Evo pLBooroprdtro Evop(11 l Tepgortogoq 'Evopqn t' r<oLUmeL -30 Bdoerg rou mRNA, ouvrniue :: * ::: -, oe vo 6pXetor towdlpovo o'enocprl g evo <*5.,:. : repportopori Kor Fe rnv en6pevq 05o1 dvoplqe, :.r. vo prlv on6louv nopd ).tyeq p6vo Bdoerq. 24.6 'Evo Er6rr6 Evoprrrlpro IRNA lerrvd rnv lloAunenr6rrfi AAuoi6o " H peroqpool opXi(er p' evo rorotrorno pe0erov[v1g rou ou- v10og rt.rdtxonore(ror ono io AUG. ,, Arorpopelro tRNA pe0eLovrvrlg egntr6rovror orrlv evop[r1 ror rqv enrgrlruvol. - To evoprqpro IRNA dLo0erer povoDrxo boprro XoporqpLort- ro, ro onofo ro dlotpoponorouv on' o],0 ro unotrorno tRNA. '. H ogo6o NH2 rrlt Usoerovivrlq nou npoodeveror oro poxrq- proro evoprrflpro tRNA e(vor rpopputrulpevl. H orjvOeoq 6lrov rcov npcoteivdlv [exrvd pe ro [6ro oprr- vo(rj, rr1 peOerov(vq. To IRNA nou ovoyvop((ouv ro rodtr6vLo AUG preropdpouv ;re0erov[v4, evo 6rio ttj- not IRNA prero<pSpouv our6 ro otrrtvolrj. O 6vog rJnog XplotponoreLror orqv Svop[q r<r o d].].og orrlv ovcl- yvroptoq rcov rrr;6u<oviti-rv AUG r<ord trlv entgrlruvoq. Iro Bor<tqpro, ro 1-rLtol6vdpro r<or toug g.ropon).d- oreq, ro evopnrqpLo IRNA preropdper dvo rot6otno trle- Oetov(vr1g, rou ono(ou 11 oprrvoprd6o 51er rpoppruLro0er cn1r1 por((owog 5vo pr6 p ro N -rpop p utro- peO er ovuho-t R NA. To IRNA efuor yvoord org tRNAlMet, evcb ro opuvoor<ulo- IRNA ouvrlOog 6r1).rr-rveroL pe rr1 oUw;rqor1 fMet-tRNA. To evoprcrrlpro IRNA onorcrd ro rpononorqpr6vo opr- vo[ri rou p-re puo ow(6poor1 60o plUdrotv. l-lproro <pop- troveror ;-re ro oprrvo(0, olqpror((owog MettRNAt. Irrl ouvdleto 11 ow(6poor1 <pop f-iuL(cr-loqq, nou nopo uoro(e- ror orrlv , onorLe(et qv eLe[Oepr1 opuvopro- 6o: dror, ro evopr<rqpro IRNA 6e pnope( vo o'uprprerdoler ,C7' C,t,^ --- '9ch 'oc/-,/)n. .bn, ,, 2,.[ - ruH' -%..8-s$' I U_U I 0 l^l I l- t -I1poiq rodrronorrllrrl neptoxil I U -U I QDtr o 1ls 3L_-'-_lJ Aetireprl xol6rxonoulrixrl neprox4 'EvopErt ouppolver ove[opqro oe roOe olorpovro ,. -. notruororpovrrou mRNA. 'Orov I neproXrl pero[u duo or- ,-::'iirov e[vor porpurepq ono ro prlrog rou prBootrrporlou, l Dro- , rrn 0T1 0eo1 repporropou oro),ou0efroL ono ove(oprlrq - -. e. opEq oTo tttopsvo oroTpovto, IMet-tRN& N-rpopputro- Ue0srovutrotRNA ru-q,upFu^u-.. Terpoijdporpulitrrd TsTpoU6ooQuAIrKd elK0filA 24.14 To evoprrrlpro N-rpopputro-pe0erovutrolRNA (fMet-tRNAt) oXlgor[(erot Ue QopUutr(toon rou ge0erovutrotRNA, orrlv ono(o trrg oupnopoyovToq Xplorporore(ror ro qop;rutro-re- rpoUdporputrtrrxrS. 2{..6 'Evo Erdrx6 Evoprrflpro IRNA :errvd rqv l1otrunenrt0Lxr1 A-uoi0o 083 1 0-rpopUulto- Boorxeg ewocg
  • 175.
    qnlv enUnKUVon'rnq o).uo(6o9,lcop(g 6prcog ow6 vo no- pepBo(ver or4v xov5r4rd rou vo exruve( puo npotefvr1. Aur6 ro IRNA Xp4orponorefuor ;r6vo or4v Svop[q. Avoyvcop((et ro xro6rr6vro AUG xot GUG (onopodtxd ror ro UUG). Tq KodrK6vro 5ev ovoyvcop((owor e[(oou ocomd: 11 ouXy6r4ro r4g Svop[4g perciveror nep(nou oro pro6 ov ro AUG owrKoroora0e( on6 to GUG 6n<og en(oqg Kor ov Xcrloryonot40e(ro UUG. To IRNA nou e[vor uneU0uvo yro rnv ovoyv6prorl trov xrodrxov[rov AUG o' eooreprK5q O5oerg e(vor ro tRNAmMet. Aur6 ro IRNA owrororXe( trr6vo o' eoorepr- rd rrrl6rx5vro AUG. H peOerovlvrl rou 6e prnope[ vo eop- ;ruLurl0e(. floro Xoporcqprorrr6 drorplvouv ro fMeltRNAf qg Svop[4q on6 ro Met-tRNA, rqg enr;rrlr<uvo4g; I4proo(o 6Xouv oprqr6vo lopor<r4pronrd rrlg oMrllouX[og rou IRNA, 6nroq ouvotp((eror or4v E|(0NA 24.15. Meprxd e(- vor ovoyxo(o yto vo ilopepnod(oouv rn Xpnon rou eKKr- vrn{ m4v enrpr4r<uvor1, ev6 dtrl.o yro rq },eroupy(o rou orqv 6vop[r1: . H gopputr(oor1 5ev e(vot ono).frog ovoyro(o, ener- 6]i to pq qoppru},uoprdvo Met{RNAr prnope( en(or1g vo Lettoupy4oer cog exrrvrlulg. Om6oo, n gopUu- L(oor1 Betrrrciver rrlv onoreAeoportr<6r1ro pe rnv ono(o Xp4orponore('ror ro Met-tRNAr, enerdrl e(vor dvo on6 ro XopoKrrlptorrrd nou ovoyvrrlpf(owor on6 rov nopdyowo lF-2, o ono(og npoo6Sveror oro evoprcr{pro tRNA. o Or pdoerq nou owu<p((ouv n prq rrlv dMq ornv re- l,eurofo 06o4 rou pr(oXou orov ono(o npoo65veror to o1-uvo[rj efvot (euyoproprdveg oe 6l.o rq IRNA ercr6g on6 ro tRNArrret. MeroMd(etg nou 64puoup- yo0v dvo (eUyog Bdoeu:v oe outrj rr1 06oq rou tRNnu"t rori ennp5nouv vo lerroupye( orrlv enrpui- KUVorl. Iuvenc0g, 11 onouo(o ouroU rou (e0youg no- pepnod(et rn Xpnon rou tRNArMet or4v enrprqruvorl. 0opUUtrrorU6vo oUrvoIri Xop(q (euyopotto Pooeurv 3 (eUyq pooeov G-C r:;';1li;&:*.:i To ftVlet-tRNAt 6Xer povodtro XopoKTrlproTrKo nou ro [eXopi(ouv trlg ro evoprrlpro tRNA. 684 KEOA Al0 24 Meroqpoon En(oqg, ro (eUyog e(vor onopolrr1to yro r4v ow[- 6poon rpoppruL(tooqg. o To tRNAtMet, Kor. p6vo our6, 6ro0ritet pro oerp6 3 (euycirv G-C oro pr[oXo nprv q Oq].rd nou nept6la ro qwrKo6rr6vto. Aurd ro (eUy4 Bdoecov ononoriv- ror c0ore ro fMet-tRNAtM"tvo pnope( vo evoopmu> 0e( oneu0e(oq crrn e6on P. Iro Borrqpro Kor ro l.rrroX6vdpro, to xordLorno 1-rupl.rqyrorof (gop1.rutroprd6o) orqv evoprrrlpro pre0er- ov[vq ogotpe(ror on6 truo er6rx4 onogop;ruldon, o]n- ptor((ovrog 6,vo r<ovovrr<6 oprvorel,rr<6 dxpo. Av 11 peOerovfuq npoop((eror oq ro oprvoreLrx6 opuvo[f rqg npondfig, our6 e(vot ro 1lovodrx6 ovoyro(o Bqpo. l-le- p(nou mrg puoSg nprrneiveg, 11 preOerov(vq rou dxpou ogotpe(ror on6 gro ol.rwonemr6doq, q onofo 6qpuoup ye( 5vo v5o dr<po on6 to Rz (opXrrd ro 6eUrepo oprvo- [6 nou e(Xe evooprorro0e( or4v o].uo(do). 'Orov ror ro 6Uo Bqgoro e[vot ovoyro(o, ouprBo(vouv 6ro6oXu<d. Ot owr6pdoerg orpo[peo4g npoypororrotofwor ypnyopo, ;rdMov 6rov 11 veoouwr0Sprev4 nolunemr6u<r1 o).uor6o grdoer oe prrlxog to 15 o;rwo[6o. 24.7 H Xpfion Tou fMet-tRNAr EAdVxeTot on6 Tov lF-2 Kot To PlBooropdflo o 0 lF-2 npoo06ver ro evopxrrlpro fMet-tRN& ror tou ennp6ner vq ero6tr0er or4 gepLr4 0eor1 P r4q 30S unogovoDog. Kotd q ;rerdqpooq mo Boxrrlpro, n ongoo(o rov rr.u6rrov(rov AUG rqt GUG <e[opr6rol on6 ro ouu- gpo(6pevo,> (context-dependent).'Otov ro r<o6rx5- vro AUG Xpnotgonore(ror yro 6vop[4, droBd(eror og <pop1-rulo-pe0erovivq, ev6 6rov Xprlorponoe(ror gSoo orrlv rcodu<onorrlru<r1 neprop1, oro nolunem(6ro npo- m(0eror preOerov(vq. H oqproo(o rou xrodrrov(ou GUG efoprdror or<6prq neproo5repo on6 rlv ew6nto{ rou. 'Orov e[vor ro np6ro rcodrx6vro, npoor[0eror gopUu- l,o-pe0erov(vq, evril 6rov Bp(oreror oro eooreptr6 ev6g yovr6(ou, npoo6Sver Val-tRNA, 5vo on6 ro Kovo- vxd pui).q rou ouv6Lou ro:v IRNA, nop6Xowog BoLi- v4, oripr<pcr:vo pe ro yeverrK6 r<c66xo. l-1uig epprqveUowor ro "ouUqpo(6pevoD rov xto6xo- v(ov AUG ror GUG; H EIK0NA 24.16 qvodeKvtier rov r<p[- or1.ro p6),o rou prBoor.opor[ou, 6rov evepye( oe ouv6uootrl6 U' enxouprKo6g nopdyoweg. I' 5vo orlprnLoxo 5vop[r;q, oro mRNA e(vor npoo6e- 6ep5vr1 pr6vo 11 purprl unoprovd6o. To r<rrr6rx6vro rivop- [49 Bpioxeror p6oo oto rpqpo rqg 05o4g p nou perorp6peror on6 rq ;rxpr1 unopovddo. To pr6vo otruvo- oruLo-tRNA nou trlnope[ vo roroore( p6pog rou oupr- nL6rou 6vop{49 e(vot ro evoprcr4pto, ro ono(o dro06rer q povodu<q rdt6qro vo ero5pXeror oneuOe(og orn Ue- prr4 06o4 P yro v' ovoyvrop(oer ro r<ro6x6vt6 rou. I q*, i &&{3 & & & * Boorx6g eworcq
  • 176.
    M6vo ro fMet-tRNAfetodpletoL orn Uepxl 06oq P n1g 30S Mrjvo ro aa{RNA eto6pletot or! e6on A rou nlrlpouq 70S . : ':,: l/16v0 ro fMettRNAf Unope[ v0 Xpnotuonotrloe( Yto , :-i- ono rtg 30S uno;rovo6egl , A),),0 olttvoorutro{RNA (aa- ': , - rpener v0 Xp1orponotl0ouv yto entpfruvo1 ono ro 70S pt- - , --0Ti0. ','loLrg 11 ;reydr1 unoprovd6o npoooptqOe( oro oO;i- -' :(o, or pepLr<dg 06oetg np6o6eorlg tou IRNA pero- --rrcwor ortg or<Spoteg 0doetq P rot A. To evoprcrrlpto '''::-rRNAf rcoro).oprBdveL q 0dor1 P, evot q 06o11 A 6to- - :.::r yro rlv eioodo rou outvooKuLotRNA nou elvot , --:i.qpo;,-rotLx6 pe to 6e[tepo rco6tr6vto tou yovt6[- : - C npci.rrog nemt6u<6g 6eopr6q crlqprot((etor preto[ri - - - ./opKrrlprou Kor rou en6pevou o1-tLvoot<utro-tRNA. - evop(r1 entrpote( dtov 6vo t<to6tr6vto AUG (n , - 3 Spior<eror gSoo oe Uto 0do1 np6o6eoqg ptBooto- -rr r;, enetorl p6vo to evoprrrlpto IRNA pnope( vo et- , .'iaL or1 preprx4 06orl P nou crlqprot((etot 6tov 11 30S - -: -:vo6o npoodeOe( de novo oro mRNA. H eorotept- ' - r. tryvoJon entrpote( opy6tepo, 6tov to xtrt6tr6vto : - : r ,ror o' enogrl p' 6vo ptBoot,.lporto, ro ono(o ouve- -. .o ;.rerocppd(et dvo mRNA, enet6q pdvo to Kovovt- . r l-.',/ooKUIo- tRNA e(vot trovd vo eto5trOouv srrlv -. - ^- Acnn A! vevrl,. 3. onrroupLro( nopdyoweg e(vot xp(ot;.rot Yto rov : : .-,.i rlq Xpnorlq rov optvooKuLo-tRNA. Otro to o;.tL- - :. -r,c-tRNA oMqLent6porjv pe to pLBootoprdtro prSool -:::-ioqg o' 6vov entrouptrd nopoyowo. O nopd- , ,-1. nou Xprlotponote(tot orrlv 5vop(q eivoL o lF-2 : .-t Evorqta 24.4) rot o ow(motlog nopoyowog - - - '.norponote(tot mrlv entpqr<uvoq e(vot o EF-Tu : .- t Evotrlta 24.10). I :cooyowog 6vop[r1g lF-2 tonoOerc[ to evoprcr4pto : '.: llr 0eor1 P. Iplprot((owog 6vo oUtrrnl'oro etdtrd -. -- ",let-tRNA, o lF-z 6too<po).((et 6tr pr6vo to evoprrrl- : : :l"iA ror rov6vo on6 ro rovovtrd oprtvoor<ulo-tRNA .: :--.rerd1et orrlv ow(6poo1 6vop[4q. Awtorp6goq, o EF-Tu, o ono(og ronoefie[ optwooruhetRltlA orn Atorl A 6e ;rnope( vo npoo66oet fMet-tRNAt, to onoio otrveiliq 6e Xp4orgonotefuor xotd r4v enqlrlxuvoq. 'Evog entnp6oOetog SleyXog Yto rrlv oxpiBeto ilpoyUoronoeftot on6 tov lF-3, o ono(oq oroOeponote( qv np6o6eoq tou evopr,criprou IRNA ovoYvop((owoq ro ooor6 (euydpolpro Bdoeov pre rq 6eriteprl r<ot tpfur1 Bdoq rou ro6xovfou Svop(r1q AUG. H EIK0HA 24.17 nopouotd(er tremoprepdlq rn 6to6oXr1 tov yeyov6rtov, ptSoto tcov ono(ov o lF-2 tono0ete( to evopr,crlpro fMet-tRNAr or4 05or1 P. O lF-2, npoo6e6e;r5- voq pe GTP, oMqtrent6pd pre q 06oq P qq 30S unopo- vd6og. Ie ow6 to o4trle(o, q 30S uno;rovddo ;reto<p6pet 6Loug toug nopdYoweq Svop[4q. To fMet-tRNAt npoo- 65vetot orov lF-2 ndvco orrlv 30S unoprovd6o n Snemo o lF-2 prerorpdper to IRNA pr5oo orq pepu<r1 06on P. 24.8 Or Mxpdq @doerq'Evoplnq Ynouovd6eg Avo(nro0v oTo EuKCrpuoTtKd mRNA r 0r europuortr6q 40S ptFoooutr6q unopovodeq npoo66vov- ror oro 5'orpo rou mRNA rot To (oopdlvouv,, 5oq orou evro- nloouv 1uo 0501 5vop[qq. . Mr0 euKopuonul 0601 evopltlg onoretre(rot onrl pto 0trtrr1- trouX[o d6ro vourtreotLd(trlv 1 ono(o nepiloppovet 6vo rotdt- rovto AUG. o 0r 605 prgooourx6q unopovodeg ouvd6ovrot pe ro ofpntroro oq 06o11 evop[r1q. IUpn)toro 30S-mRNA 0 lF-2-GTP ouvduo(eror ge ro ougntroro IuvdSeroL ro evqpKrnpro tRNA Iuvdriovror or 50S uno- govoDeg rr onetreu0ep6vovrot or nqpoyovreq lFl -3 @lJ. IF-] IF-z IF-3 GDP P; l:iliii"lA il.:l 0 lF-2 e(vot onopo[qrog yro qv npoodeol rou fMet-tRNAt mo oupntroro 30S-mRNA. Mero qv npoodeon rou 50S, otror or nopoyovteg lF onetreuOeptitvovrot rot ro GTP dtoonorot. ,*o. tr a *4.* 0rMrrp6g Yno;rovo6eq Avo(nrouv 06oetg'Evop{lq oro EuropuorLrc -l',: 685 Boorreq 6vvorcg rF-3 ffi rF-l v -.{ I tF-2 rS..,
  • 177.
    H Svop[q qgprerdgpoonq oro xurrop6n].oopo rov europuordlv prord(er pre 14 Srepyoo(o rolv Borcr4p(ov, oMd q oerpd rov yeyov5rov e(vor dro<poperxrl ror o opr0p6 g rov eTr rKo u pxrilv nopoy6wtrlv preyoL0repoq. Oprog6veg 6rogop5g oXe{(owor pe ro drotpopetrx6 rp6no Ue rov ono(o or BorcrrlproxSg 30S rr or euKo- puortr6g 40S unoprovd6eq Bp(orouv ug 06oerg npSo- 6eoqg rouq npoKerpdvou vo (er<rvqoouv rq perd- qpqon rou mRNA. lroug europuc0reg, ot pur<p6g uno- povd6eg npciro ovoyvtop((ouv ro 5' dxpo rou mRNA rr 6,neno perorrvoriwor Trpoq rr1 06o4 5vop(qg, 6nou ouvoworjv trg preyd),eg unoprovddeg. (Iroug TrpoKo- pucbreg, or ptr<p6g unoprov66eg npoodrivowor orreu- Oe(og or4 05or1 5vop(r19.) I1e66v 6tro to europurolxd mRNA e(vor l.rovoor- orpovxd, oor6oo r<d0e mRNA ouvrlOcog e(vor nofui po- rpUrepo ond ro ovoyro(o yro l1v r<ro6rrono(4or1 pr6vo tqg nporeivnq rou. To prSoo mRNA oro europuronr6 r<urop6nloo;ro 51er grlrog 1000-2000 Bdoerg, rpSper 1-rLo pe0uLr<oprdv4 roLUmpo oro 5'dxpo r<or 100-200 Bd- oerg noLu(A) oro 3'dr<po. H gq perogpo(6pev4 5' ol,LqLoul(o-o64y6g e(vot oXenrd BpoXe(o, ouvrlOcog <100 Bdoerg. To grjr<og r4g r<todrronorrlnr<r1 g nep roXq g ro0o p((eror on6 to g6ye0o g qg nporeffiq. H Un preroqpo(5;revq B'oupd ouxvd e[- vor oprerd porpr6, preprxSg qopdq 5og -1000 Bdoerg. To npcbro XopoKrnpronr6 nou ovoyvropf(eror r<ord q perdrppooq evdg europurotu<orj mRNA e(vor 4 peOu- Luopdvq r<o).Umpo, 11 ono(o oqpro[ver ro 5'dr<po. Ayye- )lorp6po RNA tcov ono(rov ot r<ol6nrpeg SXouv o<por- peOe( 5e prerogpd(owor onoreLeoporrrd in vitro. H np6o6eo4 40S unopovd6ov mo mRNA onome( noL- Lo6g nopdyoweg Svop(qg, ;rero[ri dMrov npu.neiveg or ono(eg ovoyvop((ouv q 6opuj qg xoLrJmpog. Tponono(4o4 rou 5' dr<pou npoygoronote(ror oe 6Lo oXe66v to rurroprrd ror ru<d mRNA r<or efvor ono- pokrlq yto q perdgpoorl roug oro euKopuotr6 xur- top5n).oqro (evcb 6ev efvor ovoyxo(o oro opyov(6ro). H provodxri e[o(peoq ourod rou r<ov6vo o<popd oe ].iyo u- rd mRNA (n.X. rou noLrolori)ro ono(o 6e q6pouv ro).ti- mpo: g5vo owd r' oouvr'1Ororo ru<d mRNA gnope( vo Ueroepqoroiv in vitro Xop(g xolfmpo. Xp4or;ronorotiv pro evo.trl.orcrrrr1 o66, nou nopor<dpmer rrlv ovdyrrl yro r;v r<o).rimpo. Mrilpr rri:po e[erdoope 14 6epyoo(o rqg Svop[r1g oo vo rltov ndwo 6ro05orpq q 05oq 6vop(49. H 6ro0e- ot;r6rrp6 rqg, oo16oo, ev651eror vo neprop((eror un6 deureporoye(g 6op6g. H ovoyvclprorl tou mRNA onor- re( noMo0g enrnp6o0eroug nopdyoweg: 5vo onpowr- x6 puipog r4g Leroupy[og roug e(vor 4 orpofpeor1 rov 6norov deureporoyriv doprcbv oro mRNA (BL5ne Err6vo 24.22). MepLr5g qopdq, ro r<to6x6vro Svop[qg AUG ewoni (eror oe on6oroor; ;rxp6repq on6 40 Bdoerg qn6 ro 5' drpo rou mRNA, on6rc 16oo q r<o).Umpo 6oo rot ro AUG Bp(or<owor prSoo orrlv neproxq np6o6eoqg rou pr- Booropo{ou. Ie noMd mRNA, o016o0, 4 r<oAUmpo xot ro AUG on6Xouv neproo6repo -oe orpo[eg neprntdl- oerg, 5og ror 1000 Bdoerg. H nopouo(o qg ro).0mpog 686 KE0MAI0 24 Merdqpoorl e[oro].ouOe( vo e(vqr ovoyxo(o nporerp5vou vo oxnpo- lore( 6vq oro0ep5 oUpnLoro oro ro6u<6vro Svop[qg. l-1c09 pnopei ro prBoorrlprdlo vo Boo((eror oe drjo 05- oerq nou on6louv r5oo noLri; H EIK0NA 24.18 nopouord(er ro pow6lo "odpoorlq" (scanning model), oriprqovo pe ro ono[o 11 40S unopro- vd6o opXu<d ovoyvcop((er r4v 5' r<olUnrpo xr 6netro "perovooreUer,, rord prlxog rou mRNA. H odpcoorl on5 ro 5'dr<po e(vor ypoprpur<4 6epyoo(o. 'Orqv or 40S uno- povddeg ooprivouv rqv neptoX4-o6qy6, pnopofv vo nporol5oouv rnqn rov qoupKerdrv qg 6eureporoyorig 6oUnq pe oro0ep6rqro <-30 kcal. 0o16oo, rpoupxdreg peyoLUrepq g oro0ep6qrog noporo).ri o uv 11 ovoorflt- Louv rq perovdoreuoq. H perovdmeuoq oro;rord trr6).rg 11 40S uno;rovd6o ouvowqoer ro xo6u<6vro 6vop[qq AUG. Iuvq0rog, 11 npdrq rprnl,5ro AUG mr1 oerpd e(vor to r<ro6rr6vro Svop(qg. Aurq roOouorr1 r'1rprn).6ro AUG, coor6oo, 6ev enopxe( yro vo oroporloer u1 petovdoreuon: ovoyvoF p(eror rog ro6rr6vro 6vop(r1g ;r6vo 6rov Bp(oreror oro oorm6 nl,o(oro. Or orlpowtr5repor xoOoprmdg rou nloro(ou e(vot or Bdoetq orrg 06,oerg -4 Kor + 1. 'Evq ro- 6tx6vro 6vop[49 ;rnope( v' ovoyvrrlp((eror orrlv our1- LouX(o NNNPuNNAUGG. Or tperq Bdoerg noup(vr1g (A r1 G) nprv on6 ro rcodx6vro AUG r<or 11 G opr6oog pre-rd prnope( vo enqpedoouv rrlv onoreLeoponr6rqro rqg perd<ppooqg r<ord 10 eopdq. 'Orov 11 oMrlLouX(o-odq- y5g e(vor ;ror<prd, enrnp6o0ereg 40S unoprovddeg prno- pe( v' ovoyv<op(oouv ro 5' drcpo npotori 4 npc0rrl unoprovddo eyr<oro).e(r,Uer q 05o4 Svop[qg, drlpuoup- ycirwog puo oupd unopovddrov nou npoxope( r<ord pq- Koq rnq oM4LouX(og-odqyoU npoq rn 06o11 Svop(r1q. YcccIccauccMe0utruog6vr1 xotrunrpo 060r1 6vop(r1g : H puprl unopovodo npooddveror oq Ueoutrrcou6vn ro),unrpo p**f d- otr..'-or' i.r-a. ./1- r:, G C C A C CAU G G 2 H puprl unopovobd perovooreuet orn 06on ,.*f...- -^ .^ ^-?-^ ccc$)cnuce 3 Av rl 0trtrn),ouxio-odnyoq efvor poxpro, 0r unoUovoDsq gnope( vo oXqgor[oouv oupd 8*'*@_,-=.=^- ,.a:*:..."r;.1-"i1 eccfi ccc:liljt To europuonro prpootrlporto ;lerovooreuouv o , to 5' orpo rou mRNA oqv neproyrl 6vop[19, q ono[o nepLtrop3':. vet 6vo rodlxovro 6vop[1q AUG.
  • 178.
    Iro nto onoteleopottrdpetoqpo(6pevo mRNA prdMov toXUet 6rt ru:6tt<6vto Svop[4g e(vot to npdno AUG. Tt oupBo(vet, <11016oo, 6tov pro rptnLdro AUG ev- ron((erot orqv 5'o;rerdqpoo.rn neptoxn; Yndplouv 60o nrOovo( prqXovtqro( 6toguyqq yo 6vo ptBooopdno nou [errv6 q odprrloq oro 5' 6r<po. O nto rol6g e(vot 6rt 11 odptoorl e;rqov((et dtopporl: our6 orlpo(vet 6tt 6vo pt- Booog6tto pnope( vo ouveXfoet ndpo on5 6vo pr1 evop- rcrqpro AUG enetdq 6e Bp(or<etot mo orom5 ntrq[oto. Irq ondvro nep(mcooq nou ovoYvop(oet to AUG, ev65- Xeror vo (erwrloet q petdEpoorl, oLI.d tep;roil(et npw ro ooor6 rto6tr6vto 6vop[r1q rt 5netto ouveX((et q od- poon. Iroug europud:teg, ro neptoo6tepo oupBdwo 6vop[qq nepil.o;rBdvouv odpooq on6 tqv 5'xo].fmpo' Om6oo undpXet 6vog evoLtrolmtr6g tp5noq Svop{r1q, Trou Xprlotponoteftot t6(oq on6 optqt5vo ttxd RNA, oro ono(o puo 40S unoprovd6o ouv66etot oneu0e(og pe Uto eocDreprrrl 06oq yvomri tog eooreplx{ plpootoplx{ 06oq ero66ou (internal ribosome entry site - IRES). Ie out4 rqv nepfmooq, rdOe rcl6x6vto AUG nou tu- 1o(vet vo Bp(or<erot mqv S'opretdqpoorn neptoXrl no- pordpnretot evteL6g. To yvtrlord ototXe[o IRES epr<pov((ouv neptoptop5vrl otrrotroy(o oq flpoq qv oMrl- LouX(o toug. Atorp(vowot rpetq linot ;le Bdorl qv otr- TrqLenidpoo4 rouq pe rq 40S unoprovddo: o 'EvoS rUnog IRES neprdXet to t<to6tr6vro Svop(4q AUG oro ovo6tr6 6pt6 tou. H 40S unopovddo npoo65vetot oneuOe(og oe out6 Xpqorpronord:wog 5vo unoo6vol,o ttov (6tcov nopoy6wov nou onot- to[wot yto rrlv Svop[r1 oro 5' drcPo. o 'EvoS 6Mog tUnog ewon((etot 6o9 100 vour.tr'eorf- 6to ovodtrd tou AUG rot onqtte( petovdoteuoq truog 40S unopovd6oq, nt0ov6v t<ot nd}'t p6oo ev6g prlxovropori odptoo4g. o 'EvqS ondvrog trinog IRES otov t6 qq qno{ttdog C prnope( vo npoodSoet oneuOeiog prto 40S unopovd- 6o Xrop(q vo Xpetd(etot rov6vo nopdyowo 5vop- EnS. H dtodoXrl tolv oupBdwov e(vot 6to<popetrr<t1 on6 r<dOe dLLrl Svop(r1 orouq euKopucireg. Metd t4v np6o6eor1 40S t<ot mRNA, npoo65vetot 5vo oripnLot<o to ono[o nept6Xer nopdyoweg Svop[4q Kor ro evopKrqpto IRNA. H Xpnon trov orotlefo:v IRES 61er t6tofuepr1 or1;roofo orrl l,o(po[q on6 ntropvoi6, 6nou npolroovor<o],U<p0r1- xe, enet6rl o t6g ovom5Met tq pretd<ppoorl tou [evtorrl xorootpS<powog ttg 6opr6q qg xotr0mpog rt ovoor6L- ).owog toug nopdyoweg 6vopfilq nou npoo65vowot oe out5g. 'Evog t6totog m61oq e(vot 11 unopovd6o elF4G (BL5ne Evorqta 24.9), rl ono(o npoo66vetot oro 5' dxpo rou mRNA. 'Etot, q ).oigo[r1 nopepno6[(et q gerdqpooq tcov mRNA tou fevtot4, evrir enttpdnet oto tuxd mRNA vo UeroqpomoUv enetdrl Xp4otpronotoriv IRES. H np6o6eoq oro0eponote(tot m4 05o4 6vop(r1q. M6).rg q 40S unoprovddo evoOe( Ue Uto 605 unopovd- 6o, to nl.rlpeq ptBootoprdrto ewon[(etot orrl 05oq ne- pro1q nou toutonote(tot pSoco npoo6toproprorj npooroo(oq (proiection assay). H 40S unouovoOo noo- ororerier pLo nepLoltl 6otq 60 Booerg: orov c. 6-3 - -- povddeg npoode0oriv oro oJpnAor<o, n npoorcre - -. .-. neproXrl ouoreMetot xot <ptdvet oro prqroq rolv 3l--- Bdoetov nou XopoKrnp((et roug npoxoputrlteg, 24.9 Or Europudreq XpnoluonolotjY 'Evo I0UnAoro lloAAcbv I'lopoY6vrtov'EvopEnq e 0r nopoYovreq Svop[r1g onotrouvtot oo 6tr0 to otoDto t4g Svop[qq, UeroE0 otrtrtov n npdoDeon tou evoprqptou IRNA, 11 npoo6eon Trlq 40S unopovodog mo mRNA, 11 perox(vr1or1 ro- rd prlxoq tou mRNA xor 11 npdodeol rqq 605 unopovoDog' I To tuKopuolrrl evopxttlpro IRNA e(voL 6vo MeitRNA ro ono(o 6Lorp6pet onri ro Met-tRNA nou XpnotUorote(tot otlv enrpqruvol, toordoo 1 peOetovivrl tou Dev e[vot tpopputrttrlg6- vl. * 0 elF2 npoo66vet ro evoprqpto Met-lRNAi xot GTP oxnuort- (ovrog 6vo tptrotoy6q orlpilroxo, ro ono(o npood6verot onlv 40S unopovoDo nporo0 ourl otrtrr1trenr6pooet ge ro mRNA. r'Evo oUuilroro npoodeorlg qg xohtjntpoq npooD6vetot oro 5' orpo tou mRNA nporou outo olJqtrentDpooet pe qv 40S unogovd6o. H 6vop[r1 orouq euKoputi:rcg 61et to (6to yevu<d 1o- porcrrlptortrd Ue rrlv ow(orotlq Borcrqptor<r1 on6 rrlv 6norlq 6tt Xprlorpronote[ 6vo et6tr6 ro6tr6vto Svop- [r1q r<or to evoprcrqpro IRNA. H Svop[r1 oro rurrop6- nLool-ro rov euKclputotcitv lpqotpronote( cog evoprcr(- pro ro6rx6vro to AUG. To evoprcrrlpro IRNA efuot (e- Xoprm6, o).L6 q preOetov(vr1 tou 6e goppru).ttivetot. Ko).e(tot tRNAfl"t. H droqopd preto(ri tou Met-tRNA qg 6vop[r1g Kot rnq entptqruvorlg 6yretrot qnoKlet- orrr6 oro rUnUo tou IRNA: to Met-tRNAi Xpnotuonot- e(ror orqv 6vop[r1 evri to Met-tRNAm orqv entprlruv- on. TouldXtmov 6tio Xoporrqptmtrd e(vot povo6trd yro ro evopKrrlpto tRNAlM"ttqg (U;rqq: 6to05tet Uto oou- vn0rorn tprotoyrl 6oprl ror tpononoekot Ue qooqo- putr(oor1 rnq 2' 05oqq qq ptB6(49 mq Bdo4 64 (ov nopepnodtme( our4 1 rponono[4on, ro evopr<tqpto IRNA pnope( vo lp4ottrronotqOe( mqv entprrir<uvoq). 'Eror, 11 Boox4 opln rnq 6tdrptoqg ;reto[ti Met-tRNA yro rrlv Svop(q Kor rrlv ent;r4ruvot1 6totqpefuot oroug europurilrcg, toor6oo q 6oprr4 gdon qq 6to96pet on6 rqv ow(orotXrlrcov Borr4p(ov (yro oriyr<pror1, BL6ne Et- x6vo 24.15). To eu roputrlttrd rtirropo 6to05touv neptoo6tepoug nopdyoweg 5vop[r1q on6 to Bormripro -o tp51ov rot6- ),oyoq nepDr.oprBdvet 12 nopdyoweq ot ono(ot onot- ro0wqr dpeoo n SpUeoo Yto rrlv Svop[r1. Or nopd- yoweq 6Xouv nop6proeq ovopoo(eg ge toug qw(mot- Xoug Borcrqptot<otig r<ot d4trtilvowot pe ro np60epo "e" ,.,. :: 0t Europutirreq Xprlotponotouv 'Evo l0pntroro I-lotrtlv [looe', :'. * ' Boorxeg evvoteg
  • 179.
    .-Met .0, J#p , 4E4A,. Ir.-nt.'lt JG 3 t^to l .g'kj.-.:u4 E -i. .: I (eukaryotic: euxopuorx6g). Apouv oe 6Ao rq orddro rrlq 6epyoo(og, prero(ti dMrov: . sxrlpqrrop6g ev6g oupnl,6r<ou dvop[r1g pe to 5' drpo rou mRNA . o)C4porroU6q ev6g oupnl6xou pre ro Met-tRNAi o np6o6eor1 rou ouprnl6r<ou mRNA-nopdyowo mo oUpnl,or<o M ettRNAi-nopdyowo o 6reur6luvorl rou prBooro;ror(ou vo ooprioer ro mRNA on6 ro S'dxpo 6rrrg ro npcbro AUG o ovftveuorl qg np6o6esnq rou evoprcr{prou IRNA oro AUG orq 05oq 6vop[qg r<or o dropeooLdpnon yto rq ouv6vooq qg 605 unopovd- 6qq. H Ell(0NA 24.19 ouvor.p((er ro md6ro qg 6vop[qg r<ot defiyer roug nopdyoweg 6vop[4g nou eprn].6Kowor oe xd0e ord6ro. O elF2 pro(( pe ro Met-tRNAi, rov elFO, tov elFl xor rov elFlA npoo66veror mq 40S prBootopu<ri unopovddo oXrlproil(owog ro 43S oriprnl,oxo npo6vop- Enq. Ot elF4A, elF4B, elF4E xor elF4G npooddvowqt oro S'dr<po rou mRNA oXrl;ror((owog to orJpnAoro npSo6eorlg t4g ro).0mpog. Aur6 ro oUpnLoxo oMrlle- nr6pd pre ro 3'drpo tou mRNA pr6ool rou elF4G, o ono[og pre rr1 oepd rou otr].q].enr6pd ge rqv nporefvr1 np6obeoqg rqg nol,u(A) oupdg (poty(A)-binding protein - PABP). To 43S o0prnlor<o npoo6SveL roug nopdyov- teg Svop[qg oro S'dr<po rou mRNA r<ot ovo(qre( ro Ko- 43S odynloxo npoiuup{4q: elF2, elF3, Met-tRNAi, elFl , elFl A 5 I0ptroro np{oieo6 rqq xulrmpqq + mRNA, eIF4A, B,E,G To 43S oulriloKo npooddveror oro 5'dKpo Tou mRNA To 48S o$griloro oXrlgor[(erot oro roDrr6vto 5vop{49: elF2, elF3, elFl,1A, eIF4A, B, F EIK0NA 24.19 Mepxo[ nopdyovreg evopfig npoodrlvovror orrl 40S prpoooprrq unopovodo oXlpori(ovrog ro 43S oUpntroro npo6vop[4q, ev6 otrtrot npood6vovror oro mRNA. Mritrrq ro 43S oupntroxo npoode0e( oro mRNA, ovo(r1re( ro rordtrdvro 5vop[nq xoL pnope[ v' onogovtoOe( og 48S oupntroro. 688 lcaAitl0 24 Merogpoorl I * IVe0erovIvn brf :::r.!l:;rl .,,.,::l Koro qv evop[1 oroug euxopuotreg, o elF2 oX1- porL(er evo tpLroroy6g oupn),oro Ue T0 Met-tRNAi rot GTp. Auro ro rpuoroyeg oupntroro npoodeverot ong etreuOepeg 40S uno- Uovodeq, or ono[eg npoodevovror oro 5'orpo rou mRNA. Apyo- Tspo oTrlv ovr[6poo1, ro GTP udpo],ueroL ror o elF2 one],eu- 0epriveror uno poprpl elF2-GDP 0 elF2B ovoyevvo qv evepyo u0pal 6rr<6vro Svop(qq. Mnope( v' onopovc,.rOei o:q 4gS or-ip- nLor<o 6vop[r19, H unopovddo elF2 e(vot o r<p(orprog nopdyowog yro qv np6o6eoq rou MettRNAi. Awi0eta on6 ro Bor<rr1- pLor<d lF2, provo;.repr1 nporelvq nou npoo66ver GTp, o elF2 e(voL ereporprUepr1g nporeivq nou npoo66,ver GTP, q ono(o onorele(toL on6 o, B r<oL y unoprovddeg, xotrr(o ond rrg ono(eg 6ev e(vot op6).oyr1 pre ro Borcrrl- pLox6 elF2 (B).6ne Exdvo 24.36 o.irlv Evor7ta 24.15). O elF2 e(vor evepy6g 6rov npoo66vet GTp r<or ovevepydg npoodSver duprrrorpoprxq youov(vq (GDp) . H ;,rr';:-, ,r r iri,t nopouotd(er qv npdo6eoq rou elF2-GTp mo Met-tRNA. To npoi6v prepu<5q eopdq xo.eftoL rprroro- y6g orJprn),or<o (ternary complex) (on6 ro rpfo ouorol- r<6: elF2, GTP r<or MettRNA). H ouvoppro).6yr1oq rou rproroyo0g ougn).dr<ou puOpr((eror on6 tov nopdyov- ro owoMoyrlg voux.l.eorr6(ov youov(vr1g (guanine nu- cleotide exchange factor - GEF) , elF2B, o ono(og owoMdooeL GDP pe GTP entrpdnowoq rlv u6pd).uor1 tou GTP ond rov elF2. H : ii',rilrl iri.: I rtopouoro(et rrlv ronoOdrr;or1 rou Met{RNAi ndv<rl orq 40S unopovo6q ond ro rprroro- y6g oUprn).or<o. Mo(( Ue rouq nopdyoweg elF.l, elFlA rot elF3, to rprroroy6g oUprni.or<o oplprot((er to 43S o0pn).oxo npodvopl4q. H ow(6poor1 6ev e[oprdrot ond qv nopouo(o mRNA. O elF3, nou onor-re[ror yro rr] 6toqpqorl ro.lv 40S unopovd6r.ov oe r<ordmooq 6rd- oroorlq, e(vot rivog notr[ peyd].og nopdyowog pe B-10 unotrrovd6eg. O elFl r<or o elF'lA, nou e[vot opr6],oyog pe to Borrrlproxd lF1, go(verot 6tr evrolJouv rnv svep- y6rrlro 6tdoroo4g rou elF3. H r;'::.'-'lil" *irr i''i nqpggstd(et qv Oprd6O torv nopOy6v- rov nou npoo66vowot oro 5'dr<po rou mRNA. O nopd- yowoq elF4F efvoL 6vo npotreivtx6 orjprn].or<o, ro ono(o nepdler rpeLg on6 toug nopdyoweg dvop(r1g. Aev e(vor oo<p6g ov ouvoplloi.oye[ror otg oripn].or<o nptv npoo6e- 0 elF2 onoretre[ror ono o. B xor y unogovo0eq elF-28 ..+ GDP GTP 0 elF2B nopdyer rnv evepyd [opqn rou elF2
  • 180.
    elF3 6torrlpe( els00epeg40S unogovdOeg &*w0 elF2 npoo06vet Met-tRNA mo 40S -rr@-'Ea* 0 elF2 e(vor Pto GTPoorl 0 elF2B eivot o nopoYowoq ovTotrLoYnq ?GDP??GTP i-qr..i :::i,rr,'r ]]:;.J I 0r nopoyovreq 6vop[r1q npoooevouv 10 svopKTrl- rro MettRNA orn 40S unopovodo oXlpoii(owoq evo 43S otlp- roro. Ie preroyeveorepo orodLo 11g ovri6poonq' ro GTP -,0potrueror rt o elF2 onetreu0epdrverot uno Uopqn elF2-GDP 0 =,F28 ovoyevvo Tlv evepYo UOPQ1. Oei' sro mRNA 11 ov ot unoUovddeg npoor[Oewot [eXolpt- cro yto vo olrlpror(oouv to o0pnLoKo ndvco crro mRNA' O nopdyowog elF4F neptdXer rnv unopovddo np6o6e- orlg rnq r<oLJmpog elF4E, r1v e)'trdoq elF4A t<ot trlv rnopovd6o-*trp(tlpto, elF4G. M6).tq npoo6e0e( orrlv xo- i,ln.rpo o elF4A ietuiyeL onorodrlnote 6ewepo-royr1 5o;u1 Bp(or<erot ortq nprrlteg 15 Booetq. rou mRNA' H :vdpyeto yto ro [et[].ry;ro nopdXetot on6tlv udp6luorl .ou ATP. I-1epott6p<o (et[l'Lypo qq 6oUrtq rotd ptrlxoq :cu mRNA enttuyXdvetot on6 tov elF4A po(i Ue rov no- :oyowo elF4B. O r0ptog p6troq tou elF4G e(vot vo ouv- 5eer oLl.o ouorotLrd tou oupnL6rou 6vop[r1q' H unopovddo elF4E e[voL f-ito eor[o yto ptiOptoq H ivepy6rqto trlg ou(dveL pe Qooqopul(trlon, n ono(o nu- :oborettot on6 epeO(oproro nou ou(ovouv rq pretdqpo- lr-l Kot ovoorpeQerot qn6 epeO(optoto nou rnv Koro- :rdMouv. H unol-tovo6o elF4F 6Xet 6pdon xwdorlg nou :tr-locpopulr<ovtt rov elF4E. H 6to0eotp6trlto tou elF4E reyletot en(oqq on6 npolrelVeg, ot ono(eq npoo66vov- 0 elF4F etvot 6vo steporptUepfq nou qnotele(tot qnd: elF4G, pto nPtrlretvn-trp(oUo elF4E, nou npood6vetot ornv 5'Ue0u)tttoU6vrl rotrtintpo elF4A, [to etrtrdorl nou [etulQet qv 5'0ogr1 0 elF4G npoo66vet 6uo efltnpoo0ttouq nopoyovreq 0 elF4B Dreyetpet irlv elF4A etrtrooq H PABP npood6verot orqv 3'notru(A) oupd ',r,l.i ;4 ,1;: To ereporptpepeq elF4F npoo66verot oro 5' orpo :'l mRNA ro06q rot oe otrhouq nopoyovreg nteovdq oMnlsltt6Pdoetg: 0 elF4G nPoo0dverut mov elF3 To mRNA npooDdvet elF4G, elF3 xot q 40S unoPov66o EIK0NA 24.23 Altrntrent6pdoetg olq ono(eg eptilr6rowot nop6- yoweq 6voplng elvot otlgovttr6q 6tov ro mRNA npoo66ver0t oro 43S otipntroxo. tor oe out6v (rotroriwot 4E-8P1, -2 xot -3), nporeqdvou vo epnod(oouvrq treroupy(qtou ornv 6voplt H nopouo(o t4g nolu(A) oupdq oro 3-6t<po tou mRNA 6ieye(pet ro oxnUqrrop6 tou ou;rntr6xou Svop- inq *o 5'dr<po. H PABP npoo66vetot cnqv nproteivrl- ,nptrUo elF4'G rdvowog ro mRNA vo utoOedloet pto r<ur<).rr<r1 6tdto[4, orqv ono(o ouyrpote(tot t6oo to 5' 6oo Kqt to 3'dxpo (Bl,6ne Etx6vo 24'22)' O o141'tott- o'trr6q owrlg qq rLeror4g OrlLtdq drele(pet rn. peroYpo- en Kot n Fnap onotre(tot yt' outrl qv enidpoorl' H irlitlLtt iapoorl PABP/e|F4G n6vco mo mRNA npo6yet rqv entmpdreuon tou 43S oupnh6rou mo mRNA' 6nog en(or1g Kor rnv np6odeoq qq 605 unoprovd6og' H ElK0]'lA 24.23 6eiXvet 6tt ot oMrlentdpdoetq nou epnLSrowot or4v np6o6eon rou mRNA pe-ro 43S oript- nior<o 6ev 6xouv npoobtoptote(nlrlpog, otrLd tpofvetot vo eprn),.5xouv rouq elF4G r<or elF3, to mRNA Kot rrl 40S unopovddq. H unotrlovddo elF4G npoo'66vetot orov etFi ror peooLoBe( orqv np6odeorl trlq 40S ptBo- oto;ux(g unopov66og mov elF4F, 6po t<ot orrlv eflt- *pa-"rotl rnq fio o6pnLor<o' Ouotomtxd, o elF4F ionogt-ti io, "ir+o Stot tirore vo pnope( vo npooeLrU- oet q prtrpr] ptBootogtr<r1 unopov66o' M6irq'n UtKpn unopovd6o npoodeOe( mo mRNA' 1r"-ouo*rltt flpoq to np6to (ouvrl0coq) rtodtr6vto nUO. H odptrloq unoBoq0efuot on6 roug nopdyoweg elFl rot elF1A. Aurrl 11 6epyoo(o onone( 6ondvq ev6p- yetog un6 Uopgd ATP: ouvendlq, nopdyoweg nou oxe- t((owot Ue rnv u6p6truon tou ATP (elF4A' elF4B rot eti+f1, en[or1g dtodpoprot((ouv rdnoto p6tro oe out6 to B,iUo. H Ell(0NA Za.Za Oefpet 6tt rl.prtr<p-q unoprovddo mogord p6Ltg <pt6oet mq 06o11 Svop[r1.q, on6te to evopr,crlpto IRNA (euyoptivet ge.to rro6x6vto 6vop[r1q AUG oXnpmi(owoq 6vo oro0ep6 48S o0pnloro' l-'lp6o6eoq tov 605 unopovddtov mo otipntroxo Svop(r1q 6e gnope( vo ouprBe( npototi onogorpuvOo0v ot elF2 rot elF3' Aw6 dropeootroBe(ror on6 tov elF5 xot ovoyrd(ettov elF2 vo u6poL0oetro GTP tou' H qv- {6poor1 TlpoYgqronoekot ndvto cnrl prtrpq ptBootopur<q unopovddo t<ot npouno06tet (euy6ptrlgo B6oetrlv tou evoprcrlptou IRNA pre to ro6tr6vto 6vop(qq' 'Otr'ot ot u"iio*b, nopdyoweg nt0ov6v onetreuOepdrvowot pt6- trrg oXrlponore( to nLrlpeg 80S ptBoocopdtro' 689 PABP ?4.S 0t Europu6reg Xprlotponotouv'Evo l0pntroro l-lotrtrdrv [10p0'' o';l'J'
  • 181.
    0 elFs efldyerrnv udpdluon Tou GTP 0fl6 Tov elF2 0r elF2 ror elF3 onetreu0eptirvowor 0 elF5B 0roueootrope( unopovd6oq EIK0NA 24.24 0t elFl Kor elF3 pol0ouv ro 43S ougnhoKo 6v0p- [19 vo oopriroer To mRNA 6tog rlrou qrdoer o' 6vo rodtxrlvto AUG. 0 elF2 udpotruer to GTP rou, on6re pnope[ v' onetreu0epo- Oer po(r pe rov elF3. 0 elFSB dropeootrope( orqv 6voor1 605-405. T5).o9, o nopdyowog elFSB entrpdnet mqv 605 unopovd6o vo evoOe( pe ro oUprnLor<o oXqprot((owog 6vo nlqpeg p$ooropdtro, ro ono(o e(vot 6roqlo vo (ert- vqoer qv entprlruvorl. H unoprovd6o elF5B 51et nop6- proro otrqlou1[o pre rov npoKopur,nu<6 nopdyowo lF2, nou en(or1g udpoLrSet GTP (nop6Mqtro pe ro p6),o tou orqv np6o6eon rou evopr,mqprou IRNA). M6Ltq onel,euOepo0oriv ot nopdyoweg, ;rnope( v' oMqLenrdpdoouv Ue ro evoprcr4pto IRNA rot ttg ptBo- oogu<5q unoprovd6eg o' 6vov dMo r<6r<i.o 5vop(qg. H unopovd6o elF2 51et udpotr(oer ro GTP rqg, ouvenc0g npdner v' ovoyewrlOe(. Aur6 ennuyXdvetot on6 tov elF2B, Svov GEF o ono(og er<ron((er ro GDP 6rot c0ore vo prnope( v' owrKorooro0e( on6 GTP. H unoprovd6o elF2 onorcl.e( or51o pti0puorlq. l-1oL- L6g puO;tortr5g rrvdoeg enr6poriv orrlv o uno;rovd6o rou elF2. H rpcoorpopuL(rooq nopepno6((er qv ovoySv- vnon rou elF2B o' evepy6 Uopgn. Aut6 neptopi(et tn 6pdon rou elF2B o' 5vo r0xl.o Svop[qg xt 6tot ovo- qr5trl.er q prerdgpoorl. 24.10 O l-lopdyovToq EilrUfiKUVonq Tu Ooprdlver AprvooruAo-tRNA oTn O6on A -' H EF-Tu e[vor 1uo uovoueplq nporelvr; G, rrlg onotog rl evsp- yoq UopAn (npoode5egevl oe GTP) npoo66verot oT0 0ptv00- rutro- tRNA. " To ouUntroxo EF-Tu-GTP-oUrvooruLo- IRNA npoo06veT0t 0Trl 0eo1 A rou prpoorogorlou. M6)lg cnlrlpottore( to nLqpeg prBooc,rptdtto oro ro6t- r6vro 6vop(r19, 6o e(vor 5ror1.to yto 6vov r<Ur<Lo 6nou 6vo ol.rrvooxuAotRNA ero6pXetot or4 06o11 A rou pt- oqv npoodeorl rrlq Boorogor(ou, rou onofou 11 06oq P r<ototroprBdvetot on6 5vo nemrdulo-tRNA. Onoto6rlnore optvoorulo- IRNA errc6g rou evoprcrqptou pnope( vo eto6tr0et orrl 05oq A. H e[oo66g rou 6ropeoo].oBekot on6 Svov no- p6yovro enrp{xuvoqg (elongation factor, oro Bo- rcrrlpro o EF-Tu). H drepyoo(o e[vot nop6proto orouq europud:reg. O EF-Tu e(vot 1-uo e[otpettxd ouwnpn- ;r5vr1 npcoretvrl oro Borrrlptcl Kot to trrmoX5v6pto, o96- Loyrl pe rov ow(morxo euKopuotu<6 nopdyowo. Aw[Oetq on6 tov opr6].oyo nopdyowo orqv Svop[4 (lF-2), o EF-Tu ouv65eror Ue ro prBoorrlprdrro pt6vo rotd 14 drepyoo(o ero56ou rou ol-uvooKul,o{RNA. 'Otov ro ol.uvooxul.o-tRNA Bpe0e( orq 05oq rou, o EF-Tu eyro- tole(ner ro prBooo:prdrro yro vo epyoore( [ovd pre dMo o1-rwoor<ulo-tRNA. 'Eror er<6qlcbvet qv xurilr4 oUvde- orl Ue ro prBooroprdrro ror 6tdorooq on6 out6 nou Xo- porcrqp((er rouq entroupu<oIg nopdyoweg. H EIKONA 24.25 nopouord(er to p6Lo tou EF-Tu srrlv rono0ftr'1oq rou ogrvooKutro-tRNA or4 05oq A. O EF-Tu, provopeprlg npoteiq nou npoo65ver GTP, e(vot evepy6g 5rov e(vor npoo6e6e;r6vr1 oe GTP r<ot ovevepy6g 6rov eL vor npoodedeprdvrl oe dr<pcoogoprrrlyouovfir4 (GDP). To drpep5q o{pntroro EF-Tu-GTP npoodSvet optwoot<uo- tRNA g4rlpror((owog 5vo rproroy6q orl;rntroxo optvoo- ruLotRNA-EF-Tu-GTP nou npoo6Svetot pdvo orn e6oq A oe prBoodrporo, rov ono[cov n eSon P e(vot 11611 rcnet- InUU6vn on6 nemr6ulotRNA. Aurq e(vot q xp(oq-rq ow[- 6poon nou drootpotr((et 6tt to ol.twoorulo-tRNA rot to nemr6utrotRNA e(vor ocoqrd rono0erqpr6vo yto ro 96- prorropr6 nemtdxo[ deoprori. To o;.rwooruLo-tRNA <poptdlvetot orq 05oq A oe 66o srddro. l-lprinov, ro drpo tou owtrro6txov(ou npoo66- vfior orrl 05oq A qq 30S unopovddog. 'Enero, n ovo- yvcirprorl ro6tr<oviou-ovrrro6rr<ov(ou nupo6ote[ gto preroBol,rl orrl 6rog6prpoon rou ptBootogot(ou. Aurq moOeponote( qv np6o6eon rou IRNA xor npoxolei u6p5luoq rou GTP tou EF-Tu. To CCA dt<po tou IRNA getoxtvefuor nL6ov orn 05oq A rrlg 50S uno;rovd6og. To 6rtriep5g oti pnl.oro E F-Tu -G D P onetreu Oeprrivetot. Awrl n Uopqn rou EF-Tu e[vor ovevepy6g t<ot 6ev npoo66vet onoreleoporrd ro opwooruLotRNA. O nopdyowog owoMoyrlg vouxtreott6(trtv youov[- vrlg EF-Ts 6ropreooLoBe( orqv ovoy6wnon rnq Xpqot- ponotrlpdv4g troperlq EF-Tu-GDP orrlv evepy6 popgn EF-Tu-GTP. ['lprirtov, o EF-Ts ereoni(et to GDP on6 rov EF-Tu oXqpror((owog to ouvduool-t6vo nopdyowo EF- Tu-EF-Ts.'Eneno, o EF-Ts erron((etot on6 GTP eTlovo- oXrlpror((owog EF-Tu-GTP. To evepy6 dtgepSg o0;rnLo- ro npoo66ver oprwoorutro-tRNA, evci o onel.eu0epop6- vog EF-Ts prnopef v' qvoruKlo0e(. Yndplouv -70,000 p6pro EF-Tu ovd Borcnipto (-5% rou ouv6lou tcov Borrrlptorcilv npcoteivciv), nep(nou todpt0po trre ro pr6pro opwoor<utro{RNA. Aw6 unodq- Lcbver 6tt to neptoo6repo opr.vooKuLotRNA nt0ov6v Bp(or<owot oe rprroroy4 ori;rnLor<o. YndpXouv pr6vo -10,000 pr6pro EF-Ts ovd r(rropo (nepinou todpt0pto pe ro prBooroprduo). H rtvrlttr4 qg oLlqtren(6poor1g prero[ri EF-Tu r<or EF-Ts uno64].rivet 5rt to otipn].oxo EF-Tu-EF-Ts u<p[ororor pr6vo nopodu<d, Sror c6ore o EF- 690 KEoA Al0 24 MetoQpoon 0r elFl ror elFl A enupdnouv q oopoorl + 48S afgni"oxa '1-.: Boorxeg cworeg
  • 182.
    EIK0NA 24.25 ToEF-Tu-GTP ronoOere( To oUrvoo- rutrotRNA oro pgoorogdlo xr 6neuo onel,eu0epo- veror og EF-Tu-GDP.0 EF-Ts drogeooiopei or4v ovrxorooroo4 rou GDP ond GTP. H ovr[6poo4 xoro- votrrlver GTP rr onetreu0ep6ver GDP. To provodrrd ogrvooruhotRNA nou de pnope[ v' ovoyvtoprore[ on6 ro EF-Tu-GTP e[vor ro fMet-tRNAf, tou ono[ou I odu- vop(o vo npoode0er ro egnodr(er v' onovr4oet o' eototepud xtrtdtxtivro AUG ror GUG. Tu yprlyopo vo perorpdneror orn Uopqn nou npoo66- ver GTP r<t 6neno o' 5vo tpnoroydg oripnloxo. O p6Log tou GTP oro rprroroy6g o0prn).oro 61er petrerr10e( pe owu<ordoroo( rou on6 5vq ovdl,oyo nou 6e prnope( vo u6poLu0e(. H 14prrq 6voor1 GMP-PCP 61er pto y5<pupo pre0ul.ev(ou mq 05o4 rou o(uy6vou nou ouv66,a rtg B r<or y eoogoprKdg op66eq qro GTP. I'lopouo(o GMP-PCP, prnope( vo ox1lgorrore( 5vo rpno- roy5g oripnLoKo, nou npoodSver o;.uvoorutro-tRNA oro ptBooroprdno. Om6oo, nentrdrr6g 6eo;r6g 6e ;rnope( vo oxrlpor.ore[, ouvencig 11 nopouo[o rou GTP eivor ovoyr<o(o yro rrlv np6odeoq rou oprvooKuLotRNA orr; 06orl A. Y6p6Luoq 6ev onqrefuor npoq ro nop6v. H xrppopuxiv4 (kirromycin) e(vor 6vo owrBrorrx6 nou ovom6l,Ler rq Lenoupy(o rou EF-Tu. M6Lrg o EF- Tu npoodSoer r<ppoprur<fur1, nopop6ver rrov6g vo npoo- 6dver oprrvoor<uLo-tRNA orq 05oq A. C)o16oo, ro o0pntror<o EF-Tu-GDP 6e prnope( v' oneleu0epo0e( on6 ro ptBooropdrro. H ouvelrlg nopouoio rou nopepno6(- (er ro oX41.rotop6 nemrdrrof 6eopori pero[0 rou ne- rrrr6ulo-tRNA xor rou ol.rrvooxutro-tRNA. 'Eror, ro ptBooroprdtro or<rvrlronotefuor ndvrrl oro mRNA npoKo- )rriwog nofoq rrlg prerdgpoorlq. Auq 11 en(6poor1 rrlq Kppopur<fur1q ovo6errvfer 6rt q ovooroLrl ev6g Bqporog tqg petdEpooqg ovoorStrtret xor ro en6pevo Brlpo. O 6yog e[vor 6n 1 ouvepg no- pouo(o rou EF-Tu nopepno6((er ro opr.vooKu).rr6 drpo rou oLuvooKuLo{RNA vo ero6}.0er o-r4 06oq A qg 50S unoprovd6og (BL6ne Eu<6vo 24.30).Iuvenrig, q oneLeu- 05poor1 tou EF-Tu-GDP e(vor ovoyr<oio riore ro prBo- oalpdtro v' ovoldBet ro oxnporropr6 rrov nenrt6rr<rilv deotrrcbv. H 6ro opXrl nopoqpefuor oe dMo orddro r4g perdqpoorlq: pro ow(6pooq np5net vo oLorlqpo0e[ oomd nporoU ougBe( q en6prevrl. H oll.rllen(6poon Ue rov EF-Tu enronq 6ro6pqroil- (er rdnoro p6tro orov 5Ley1o nor6rltog. Apuvoor<ulo- To ogrvooxutrotRNA erodpxeror mn g6on A rrlg 30S unopovddog To CCE oKpo gerorle[ror u6oo orn 06on A nq 50S unoUovo6oq IRNA erodyowor offl 05or1 A lcop(g vo yvrop((ouv ov t' owu<trl6u<6vrd roug roptd(ouv Ue to xrrrdx6vro. H udp6- uoq rou EF-Tu-GTP efuor oXeru<d Bpo6e[o: 6ropre( ne- proo6tepo on' 6, tr onorrefuor yro rn drdorooq ev6g Ld0og otruvoor<utrotRNA on6 q 06oq A: 5ror, ro nepro- o6repo LdOog optvoorutrotRNA orporporiwor oe our6 to ord6ro. H one).eu0ripcDorl rou EF-Tu-GDP pretd qv udp6Luo4 efuor en(or1g Bpo6e(o, ouvsnrig 6noro ldOog ol.rwoor<ulo-tRNA SXer ru16v enrBrciroer pnope( vo 6ro- oro0e( oe ow6 ro orddro. H Boorx4 opXrl e(vot 6tt or ov- n6pdoerg nou 4rnLdrouv rov EF-Tu ouprBo(vouv opr<erd opyd cbore vo enmpdnouv oe Ld0og ollwoorulotRNA vo 6tooro0orjv nporof noyt6ewoUv orq perdgpoorl. Itoug eur<opurirreg, ro oprvooKuLotRNA gSperor mo prBoorol.rdno en(oqq on6 tov nopdyowo eEFlo oe puo ow(6poor1 11 ono(o neprtroprBdver drdonoorl ev6g 6eopo0 uUnInS ev6pyerog oro GTP. 'Onrog ro ilpoKo- purotx6 opr6l,oy6 rou (EF-Tu), efuot 6g0ovq npcoreivrl. Metd qv u6p6Luo4 rou GTP, 4 evepy6g propqq ovo- yewdror on6 rov nopdyowo eEFlBy, nou e(vot ow(- ororxoq rou EF-Ts. 24.11 H floAunenrr6rrfi AAuoi6o Merorp6peror oTo AUrvooKUAo-IRNA -, H 50S unopovodo 6Xet evepy6qro nenldu),orpovoqepoorlg nou nop6Xeror ono 6vo rRNA prpo5v(u;ro" ,, H veoouvrrOdgevrl notrunenrrdLrri otruof60 peroq6peroL ono ro nenldutrotRNA qg Oeoqq P oro oprvoorutro-tRNA qg 0eoqg A. .:. H ouv0eol ro.lv nenlDrrdv Deopdrv nopoyet onoorutrrtrlgdvo IRNA oq 0eor1 P roL nenlDutro- IRNA oq 05o1 A GTP .'d ?r Tu-Ts -.7 O '....- Ts '. a uot or..t iTpuoroyeq f,S..r otigntroro 5 ;::".".', H fiotrunenldLrrl Atruorbo Meroq6peror 0ro AItv0oKLi,,1-:i',- 691 -W...^ -...r"trr,trt' o.tK' u.. Tu-GTP Boorxdg eworeg
  • 183.
    To prBoorr:;rdrro nopopr6verorn 05crq tou, evri q no- Lunemt6tr<f o].uo(6q entprlr0vetor Ue rn prero<popd rou nol,unenrrd(ou nou e(vor npoo6e6ep6vo oro IRNA orq 06oq P npoq ro ol.rloorutro-tRNA orq 05or1 A. H ow(dpoorl nopouord(eror orrlv EIKONA 24.28. H uneriOuvq eevpy5qro yo q o0vOeon rou nemrdu<ori deoproU rqlefuot nenrdutrorpovogepdo4 (peptidyl- transferase). H nemrdulorpovo<pepdoq e[vor ].emoupy(o qq pre- yo).rtq (50S n 605) prBoocoprrqg unoprovddog. H ow(- 6poon nupodotefuor 6rov o EF-Tu oneLeuOepri:oet ro ollwoor<ulu<5 drpo-rou IRNA rou. Irq ouv61ero, ro opr- vooruLx6 dxpo otopefuor oe Uro rono0eo[o r<owd oro drpo rou nemrSuLo-tRNA. Auri n 06on 6Xet evepy6rq- to nemrdulorpovogepdonq, n ono(o onlv ouo(o 6ro- orpoL((er rqv toXe(o pretorpopd rrlg nentr6rr<49 oLuobog oro oUr.vooKULotRNA. T6oo ro rRNA 6oo r<ot ot nporeiveg rqg 50S unopovd6og e(vot ovoyxoleq yr' out4 rqv evepy5r4ro, <oo16oo 11 (6ro 11 6pdon rrlq Ko- rd).uor1g e(vor t6r6r4to rou prBoorrlpxo0 RNA ror rrlg 50S unoprovddoq (BL5ne Ev6rrya 24.19). H gUoq qg ow6poorlg perogopdg onoroLriqOrl<e on6 tqv rxov6rr;ro rou owrBrortroU noupopux[vr; (pu- romycin) v' qvoor5Mer q ;rer6gpooq. H noupoUuKivn protd(a ;r' 6vo qtvo{0 npoo6e6epr6vo or4v re}.xr1 o6e- voofur1 rou IRNA. H EIKONA 24.27 6efyer 6t 4 noupol.ru- r[vr1 61er 6vo N ow[ rou O nou ouv66er dvo opuvo(ti oro IRNA. To owrBrorx6 owrprerton(etor on5 ro prBo- orr:prdrro og eroepX6;revo outvooKuLotRNA: 6netro, ro noLunem(6ro nou e(vot npoo6e6e;r5vo oro nemt6ulo- IRNA petogSperor orrlv oprddo NH2 rrlg noupoprur<(vr1g. H p((o rnq noupopur<(vr;g 6ev oyrupoBole( orq 06- oq A rou prBooropor(ou r<t 6rot ro npot6v nol,unemr6u- Lo-nou po pu r[vq one].eu0epcbveror on6 ro p $ooro pdrro :: ', IxnUortoUoq nenndrrou deogou ou;rporvrr Ur 0v- r(6poo1 pero(u rou notrunenndlou rou nenrr6utro-tRNA orrl 0eor1 P ror rou olrrvo[dog rou ol.uvooru],otRNA orrl Odort A 692 KE0MA|O 24 lvlsToqpoo1 Agwouxulo-tRNA CHI CH. r,r / lloupoUuxrwl ;:i,1,::r.r, li.::': H noupopur[v1 ptpe[rOt rO OprvOOru],0-tRNA, ro- Orirg lroro(er ge opo.rpotro o;uvo[u ouvbedepevo oe pro p[(o oox- Xopou-pooqq. un6 propcprl noLunentrdu).o-noupol.rux(vqg. Aur6g o np6copog repprotro;r6g qg prerdgpoorlg e(vot uneU0u- voq yro rq BorcqpLorcr6vo dpdorl tou owrBroru<oU. 24.12 H Merordnron MeToKtv€i To PrBoourpdrro r H Ueroronrorl rou prpooropor[ou gerouvel ro mRNA dropeoou tou prpootrlpor[ou roro rperg pooerg. r H Ueror6ntorl peronvel ro onoorutrtop6vo IRNA orrl 06o1 E rot ro nentdutrotRNA oq Oeoq P, evo odero(er q 0eorl A. c luUrptrrvo pe ro povr6tro uppr6rrlg rorooroorlg, 11 peror6ntol oupporver oe 6uo orodto, oro ono(o q 50S perorrve[ror oe oX6oq pe rrlv 30S ror oq ouvdXero q 30S perorrve[ror roro prlrog tou mRNA riore v' onororooro0e( | opxurl droprlptprrt- 0rl. O xUr).og rqg npooOqr4g optvo[5trlv ornv ou(ov6pre- vq nolunemtdrx4 o).uo(6o oAox).rlpcbvrror pe rn 1-te- rat6ntoq (translocation), r<o0rog ro pLBooropdrro t- NHz NHz AgtvoIri Boorreg ivuoreg
  • 184.
    I npoxope[ rp(o vouKleot(droxord UnKoq rou mRNA. H Ell(0l,lA 24.28 napouord(er n6g 4 peror6nrorl ono- BdMa ro ogdpuoro IRNA ond t4 06o11P, 6ror dsare vo pnope( vo eroSL0et ro v6o nemrduLo-tRNA. To pr- Booroprdtro 6ro05rer nL6ov pro d6eto 05oq A 6rorprrl ytq rnv e(oo6o rou o1-uvooKuLotRNA nou owtororXe( oro en6pevo xto6u<6vro. 'On<og 6efiyet q etx6vo, oro Borrripro ro IRNA pero<p6petor on6 rr1 05or1 P orq 05oq E (on6 qv ono(o onoBdtrtreror oneu0e(og oro r<urrop6nloo;ro) . Erou g e u rop u cireg ono Bdtrl,eror oneu0e(og oro rurrop6nl.oollo Xrop[g vo preoo].oBe[ ;uo 06oq E. Or 06oerq A ror P e<prnneUouv 16oo q pre- ydl,r; 6oo r<or rr1 trurp{ unopovddo: r1 06o4 E (oro Bo- rrrlpro) ewon[(eror rup[rog orrlv 50S unopovddo, evcil 61er xdnoreg enog6g Kor ornv 30S unoprovd6o. Or neproo6tepeg on6rgerg qXeurd pe rq peror6nroq ot<olou0orjv to 1tow6,lo uBptixqq xatdoraorlq (hybrid state model), oriprgcovo pe to ono(o 4 peror6nrorl npoy- fu'lrr+ rii pri+trinr:ri. To onooxutrtol.rtlvo tRNA geronve[roL orrl 06o11 E, ro nenrrdutrotRNA;leroxrve[ror on1 06o11 P EIK0NA 24.28 'Evo porqpLoro pgoorrrlrolo 6ro06rer rperg 0ri- oerg npoodeorlq yro To IRNA. To opwoorutrotRNA ero6pXeror orrl 06o11 A ev6g prpootrlgor(ou nou SXer nemrdutro-tRNA oq 06o11 P. H ouv0eol nenndrrou deogou onooxutrr6ver ro IRNA qg Oeor1g P xor ono6(6er nenndulotRNA oq 06oq A. H perorontorl UeroKr- ve[ ro onoorutrLrop6vo IRNA or4 0604 E xor ro nenndutrotRNA arl Oeol P. tRNA EIK0NA 24.29 To povr6)ro qg geror6nroqg npore(vouv 6Uo md- Dro. Ilptilrov, roro ro oX4golopo nenudrrou deopou ro oprvooKu- trrxd dxpo rou IRNA oq 06o1 A enqvoronoOerefror orq 06ori P. Aeurepov, ro dxpo rou ovrxoldrrov(ou rou IRNA enovoronoOe- retuor or4 Odon P. trloronoefuor oe 6rjo ord6ro. H EIK0NA 24.29 6e(1ver 6rr npcbrov ouprBo(ver pro geror6nron rnq 50S uno;rovd- 6o9 oe q(6oq Ue qv 30S unopovd6o, nou oroLouOe[- rot on6 ;lo deftepq petot6ntorl 11 ono(o ouprBo(ver 6rov q 30S unoprovd6o geroxtvr;Oe( r<ord pujrog rou mRNA, onoro0rord:woq rnv qp1Kn 6rop6pcproo4. Bd- oq ouroU tou 1.row6l,ou qrov q nopotrlpqoq 6l ro 1.ro- r(Bo rcov enorpd:v nou npqygqronore( ro IRNA pe to ptBooroprdrro (pretpdrwor pe Xqprcl ono{noor1) pero- BdLLeror oe 6Uo ord6ro. 'O'rov npomeOe( noupogur<(vr1 o' 5vo prBootopdrro pe opwoorufutr:pr5vo IRNA orrl 06- o4 P, or eno<pdg tou IRNA Ue rnv 50S unoprovddo geto- BdLl,owor on6 rq 06oq P orrl 05on E, evdr or enorp6g pe rnv 30S unopovddo nopopr6vouv operdBlqreg. Au- 15 uno6q).drver 6n q 50S unoprovdSo 5Xet perorpone( evri 11 30S unoprovddo 6ev 6Xer peroB).q0e(. H epprrlve(o ourc0v rov onoreLeopdrov e(vor 6rt, nprilrov ro opr.vooKultrd drpo rov IRNA (ewon((owor mqv 50S unopov66o) peronvoriwot oe v5eg 06oerg (evdr ro dxpo rov qwKo6rxov(ov nopopr6vouv npoode- 6e;r6vo or' owrrro6u<6vrd roug orrlv 30S unoprovd6o). Ie out6 to ord6ro, to IRNA npoo6Svowor onoretre- ollotxd oe uBprdrxdg nepto16g, nou onorel,oUwor on6 ng 06oerg 50S E/30S P xor 50S P/30S A. Irq ouv6Xero, q petoxfurloq enenre(veror orrg 30S unopovddeg, Sror cirore 4 neproXrl (euyopdr;roroq preto(ti qwKo6rxov(ou- rcodrxov(ou vo Bp(oreror orq oolorrl 05oq. O nr0ov6re- pog tp6nog yro vq 6qpuoupyl0e( 11 uBprdrrcj rordoro- oq e(vor vo petortvrl0e( 11 puo prBooroptrrl unopovdSo oe oX5oq pernv dM4, r<ord rp6no rilore q peror5nrorl H 50S unogovdOo getorwe(tot oq npoq To IRNA nou onoptrrlfire eyrorohelner Drogdoou qg 06oqg E EroepXdgevo oluvoorutro- I To nenldutrotRNA ppioKeror orn 06on P, To oUrvooKutrotRNA sro6pxer0r mn 06on A I 3.1.'i* H Meror6nLoq Merouvefro Prpooirrporto 693 tlpw q perotdnroq:
  • 185.
    vo nepilqppdver nporcru<d6rio ordbro, Ue rnv rovovu<r1 6opr{ rou prBooroprodou v' onoroOrordror oro 6eUrepo orddto. To ptBoorrlprdno owrperconf(er 5vo evdtoEripov 6(- trnUUo r<ord tq petor6nroq. l-'lp6net vo droondoer noL- L6g on6 rg eno<p5g rou Ue ro IRNA cirore vo enmpone( 11 preror<fur1oq. Tour6Xpovo, roo16oo, np6ner vo 6ro14- pqoer ro (euydpropro prero[ri IRNA rot owrro6rrovfou (6toondlwog ro (euydpro;ro rou onooKuhopr6vou IRNA p6vo rq ooorrl Xpovrrri orryUfl. Mro nt0ov6rqro e(vor to ptBoooprdlo vo peron(nrer prero[rJ evoL].orclrdlv, Stoxptrcirv 6ropropg<6oerov. H evoMoyrl 0o prnopoUoe vo ouv(ororor oe oMoyrig oro (euydpcopo rov Bdoeov rou rRNA. H oxp(Beto r4g gerd<ppoorlq eilnped(eror on6 oproprdveg peroMd[erg or ono[eg enrlped(ouv rrg evoMornr6g 6rord[erg (euyopdr;roroq rov Fdoecov. H nt0ov6reprl eppr4ve[o e(vor 6tr q en(6poo4 6ropeoo],o- Befuor on6 q 6rorpoprrrl roX{ np6o6eorlq rov evoMo- rcnrrilv droprop<pciloeov oro IRNA. 24.13 Or llopdyovT€q EnrUfiKUVonq flpoo6dvovror EvoAAoKTtKd oTo PrBooopdrro .. H entprlruvol onotre[ EF-G, nou 6yer doprl nopolroro pe rr1 6op[ rou oupntrorou ol.uvoorulo- tRNA-EF-Tu-GTP. e H npoodeol rou EF-Tu oro pLpoorolrolo onoxtre(er rov EF-G xor ro ovrlorpoqo, = H Uetoronrorl npouno06rer udpotruorl rou GTP, 1 onolo nu- podorel pLo otrtroyr1 orov EF-G, nou Us Trl oerpcr qg nupodo- rei pro otrtroyq oq doprl rou prpooroporlou. H perordnroq onorte( GTP r<r 6vov enrnp6o0ero no- pdyowo enrprqr<uvoqg, rov EF-G. Aw6g o nopdyov- tog e(vor r<tipro ouoronr6 rou rurrdpou: e(vor noptov oe ouyr<6wprooq '1 owrypdqou ovd ptBooroprdrro (20,000 ;r6pto ovd xrirropo). To ptBootripidlo oduvororiv vo npoo65oouv EF-Tu xor EF-G rout6lpovo, ouvenotg 11 prerdrppoorl orol.ou- 0e( rov r<rjr<l.o nou nopouord(eror ornv 6nou or nopdyoweg evoMorrtrd npoo66vowor Kr one- LeuOepo.lvowor on6 ro prBoocoprdrro, 'Erot, o EF-Tu- GDP npdner vo oneLeu0epo0e[ nporoU npoodeOe( o EF-G r<r Snetro o EF-G np6ner v' oneleuOepo0e[, npo- rorj npoo6e0e( ro orjprnLoKo oUrvooKutro{RNA-EF-Tu- GTP. H rrov6rlro r<dOe nopdyowo entprqruvonq v' ono- rLe(er roug dMoug dpoye Booi(eror oe puo oMomepr- rr1 en(6poor1 orrl ouvol.rr<r1 6ro;r6ppr,ton rou prBoorrrl.rotiou r1 oe dpreoo owoyrr.lvrop6 yto enrro).u- m6preveg Odoerg np6odeolg; H nopouoLd- (et pro oouvrl0torq opordrlro prero([ tcov doprltv rou tpLtotoyoUg oupnLdrou rou oprvooKutro{RNA-EF-Tu- GDP roL rou EF-G. H 6opr1 tou EF-G onoprpe(ror 11 ouvoLtr<r1 6opr1 rou EF-Tu nou npoo66veror mo Bpo- 094 KE0A,A|0 24 MeToQpoon r 1," "' ,a; t-Y -- - ^-- Ydootruon GTP rl-lll-trlr + trE I' nnD optvoorutro-tRNA H rppogurivrl qvoordiler Uv onel,eu06proon IUv0eoq nemr0rro0 0eopo0 I r,r ; * Y5ooLuon cTP EF-G/GTP -- EF-c + GDP . . To rpouorbrxo o(u 4rJ ovootetrtret rn5: . onetreu0eoulofl - 7^iq<t, V l;u";-..q,*l +j "s*e. L J'U .F H npdodeol rov nopoyovroiv EF-Tu ror EF-G evotrtroooetor ro069 ro prpootopolo 6dXovror vrlo oprvooru),0- tRNA, oXqpol(ouv nenldrroug deopouq ror peroronl(ovror. 1(ovo uno6oplg rou opuvo(dog ndvo-l oro opuvoor<ulo- IRNA. Iuvenc6g, eUtroyo ouvdyeror ro oupndpoo;-ro 6rr or ouyKeKprg5vor nopdyoweq owoyov((owor yro rnv (6ro 05oq np6o6eorlg (npo<povcig rowd orq 06oq A). To yeyov6g 6rr o Svog nopdyowog np6ner v' onel,eu- Oepor0e( nporori npoo6e0e( o dMog 6roo<potr((er 6tt ro oup.rBdwo rqg pretd<ppoorlq 0o npoyuoronoul0oriv pre rorcrrx6 rpdno. Kor oL 6[o nopdyoweg enrprrlxuvorlg e(vor 1-rovope- pe(g nporefveq nou npoo65vouv GTP, or ono(eg e(vor evepyoi 6rov 5Xouv npoo66oer GTP r<or, ow[Oeto, ove- vepyo( 6rov SXouv npoo65oer GDP. H tpr<proogopu<r1 Uopgn e(vor onopofunrrl yro qv np6o6eoq oro prBo- or,rprdrto, yeyov6q nou 6too<pol.[(eL 6rr r<d0e nopdyov- rog onorrd np6oBoorl oto prBooroprdtro pr6vo nopouo(o GTP, ro ono(o lpetd(rror vo exn).qpd;oer 14 ).enoupy(o rou. O EF-G npooddveror oro prBoocopdtLo dnou npo- dyer q peror6nrorl rr 6neno one).euOepctlveror perd q pretor<(vr;o1 rou ptBoocopor[ou. O EF-G pnope( vo npoo6e0e( mo prBooroprdrro or<6pq rr 6rov ro GTP ov- ttt<otootoOe( on6 GMP-PCP: ouvenotg q nopouofo ev6g vour<i.eort6(ou youov(vr1g e(vor ovoyr<o(o yro qv .j Boorxfg ewong
  • 186.
    Agtvooxutro-tR NA- EF-Tu-GTP H Dopl rou TprToToyo0q oupfltroKou Tou 0Utv00KU- , :RNA-EF-Tu-GTP (oprorepo) poro(er pe q doptl iou EF-G (6e- : . 0r Doprro ouvqpqpeveg neptoXdg rou EF-Tu rot rou EF-G .:vi(ovror rorrrveg rot npoolveg. To IRNA Kot I neptoxl flou -- proro(e|. orov EF-G etrovl(ovrot UtrtB. H tporoYpotp[o nopoXtrl- -:-xe ono rov Poul Nissen, University of Aarhus. np6odeorl, rrror5oo 11 u6p6luo4 tou 6ev efuot onolti- to4 onopo(rnrn yro u1 petot6nto4 (nop6Lo nou rl Ue- rm6nrorl e(vor noLU Bpo66reprl Xcop(g u6p6Luoq GTP). H u6pSluoq rou GTP e(vor ovoyro[o yro r4v one],eu06- p<oo4 rou EF-G. H ovdyxq yt' onetreuO6poon rou EF-G ovoroLti<p01- r<e on6 ng enr6p6oerq rou qouotdtroU o[5o9, ev6g ote- poer6orig ovtrBtortroU ro ono[o 'oupnt5(et' to prpooroprdrto orrlv rotdoroorl pretd q peror6ntoq (plene Err5vo 24.30). llopouo(o Eouor6txoti o[6oq, npoyporonote(ror Svog r6rLoq pretot6nto4g: o EF-G npooddverot oro ptBootrrl.t6tto, to GTP udpoLtieror Kot ro ptBoooprdtto petortve(rqt Kotd rp(o vouxtreor(6to. To gouor6tr6 o(0, oo16oo, oroOeponote[ to otipntroro prpooropor(ou-EF-G-GDP Srot drore o EF-G rot to GDP vo nopop6vouv oro ptBoooprdno ow[ v' one].eu0epdl- vowor. 'Etor, ro prBooroprdrro o6uvote[ vo npoodSoet olrwoorulo-tRNA r<or 6e prnope(, nl.5ov vo npoore0oUv 6Mo o;rwo(5o. H perot6nroq e(vot evdoyevrig t6t6r4to tou ptBooto- pcn(ou, q ono[o npouno0Sret Ueyoln petoBotrrl orn 6o- pr1 rou (Bldne Ev6trlra 24.17). Autrl 4 ev6oyev{g pruror6nrorl evepyonore(rot on6 tov EF-G oe ouvduo- oU6 pe qv u6pSLuoq rou GTP, q ono(o ouptBo(vet nptv an6'rn peror6nroq n enmolfvst t4 ;retor(vr1or1 rou pt- fuotopro{ou. I0;rqo:vo pe rov nr0ov6tepo prqXovtopr6, 4 u6p6l,uoq tou GTP npor<ol,e[ pto pretoBotrrl orq 6o;t4 rou EF-G, r'1 ono(o pre q oetpd rqg enttdooet pto oMo- Vn orn 6o;111 rou ptBootoprot[ou. Kotd rr1 petot6ntorl oupBo(ver 6poorx6g ovonpooovoroLto;tdg tou EF-G. l-lprv r4 peror6nroq, o EF-G npoo66vetot r<ord prrlroq rov 6rio prBooopurriv unopovd6tr:v. Ot neptoo6tepeg mog6g rou Ue qv 30S unopovddo 6qpuoupyotiwot trre pro neproXri nou rotrefuor neptoX4 4 (domain 4) Kot et- oXrrlpe( orq 06on A. Aurn 4 nepropl 0o pmopotioe w efvor uneU0uvn yro rnv em6nrorlrou IRNA. Mer614 p- ror6nro4, 4 nepropj 4 npooovorotrf(etor owiOgro, npoq rnv 50S uno;rovd6o. Eur<opurr;nr6 opr6Aoyo tou EF-G e(vot q npo:refv4 eEF2, q ono(o Lemoupye( rotd nop6ptoto tp6no pe Uto rpovol.ordoq e[optdrprevrl on6 qv u6p6].uoq tou GTP. H 6pdoq qg en(o4g ovoorStrtretot on6 to tpouotdtx6 o(ri. 'Evo oro0ep6 otiprn),oxo rou eEF2 ge ro GTP gno- pe( v' onopovoOe( r<or vo npoo6e0ei oe ptBootoprdtto pe enor<6trou0r1 udp6l,uoq rou GTP tou. Mto povodrrrj r6t6qro tou eEF2 e(vot 4 eun60erd rou onl 6up0epmrxrl roq(vn. H to((vq Xpqotponoe( vxo- rwol-rwo-o6evwo-6wourl,eot(6to (NAD) tog oupnopdyov- ro yro vo perorp6per pro p[(o ptBo(u].o-6r<poo<poptr<r1g o6evoo(v4g (ADPR) mov eEF2. H ou(euypr6v4 Svooq ADPR-eEF2 e(vor ovevepy6q orn pret6gpoo4. To un6- orpopo yro rnv np6odeorl e(vot 5vo oouvtl0toro optwo- [U nou nopdyerot trle tponono(r1oq rnq ron6frnlg: out6 ro oprwo[6 e(vot row6 orov eEF2 noMc0v et66v. H ADP-prBo(ul[cooq e(vot unefOuvq yto tq Oovoq- 96po 6pdor1 qg 6r<p0eprrxrlg to((vqg. H ow(6pooq e(- vot eforpenxd onorel,eoprottxrl: 6mo rt 5vo pr6pto ro[(vqg ;rnope( vo rpoTlonotrloer opr<erd p6pto eEF2 nporotrcilwog ro Odvoro tou rurrdpou. 24.14 Tpio Kro6rr6vto Tepuqri(ouv Tn Merdrppoon o To roDrrdvro UAA, UAG xor UGA reppori(ouv 14 perorppoor;. * Iro Borrnpro, ouvnOolq Xprloqonototlvtot pe oXenml ouXvo- qro UAA>UGA>UAG. M6vo 61 rprnl5reg roOop((ouv oprwo(5o. Or un6lot- neq rperq e(vor ro6rr6vto rtpportoprori (termination codons, stop codons) ro ono(o teppot((ouv q pret6- epoon. H g[or1 outtilv trov tptn].et<itv opltrd ono- deffiqre Ue Uro yevetu<r1 6ot<t1loo(o, q ono(o 66r<prve 6rio tUnou g oq ;rerorcj g pretdMo[qg : o Mto orluetorrl pretdMo(q nou pretoBdMet dvo rro6t- r6vro riore v' owrnpooone0et 6vq 6toEopenr6 oprt- vo[ri xo].e[ran Suouvletury (missense mutation). 'Evo opwo[ri owu<oOtord 6vq dI o orrlv nprote(vr1: q en(6poo4 or4 tremoupy[o rrlg npore(vqq e[optd- tor on6 t4v ew6nroq qg pretdMo[nq Kot rn gUon rqg owrrordoroonq rou oprwo[riog. . Mto onUetox4 prerdMo[q nou petoBdMet 5vo ro6t- r<6vro d4puoupydrwog 5vo on6 ro tp(o ro6tr6vto reppronopori rol.e(rot aouv1enx4 petdMaq (non- sense mutation). l-'lpor<oe( np6opo repponop5 rqq prerdgpooqq oro pretoMoypt6vo rto6u<6vto. Iro pe- roMoypr6vo r<Urropo ouw(Oetot p6vo to npri:ro pr5- poq rnq npo:teivqg. H Lemoupy[o qg npotetv4q Boorxdg 6woreq 24"'i4 Tp(o Kodrrovro Tep;roti(ouv rI MeroQpoon 69S
  • 187.
    Unope( vo Xoee((ovd).oyo, rpuorrd, Fe rrlv ew6nrorl rrlg perdMo[qg rord trrr]rog qg nporefvqg). (Iqger6ore 6u o 6pog oouv0eru<6 xro6m6vro -non- sense codon- Xpnorpoflote(ror preprr6q qop6q yro vo neprypdqer rrg rprnL5reg rep;roropro0. Iqv npoyprort- K6Tlro, o ilpoodroprop6q (oouv0elK6,, pdMov nepr- ypd<per qv en(6pooq truog prerdMoEnq o' 6vo yov6ro nopd rq o4proo(o rou rco6u<ov[ou yro 14 pretdgpoorl. O opoq xa6Lx6vLo reppartopol e(vot ro).[rcpog.) Ie xdOe yov(6ro nou 61et o).Lr1).ouXr10e(, 6vo on6 tq KodtK6vro repporrol.rori Bp(oreror oprSoog perd ro ro6u<6vro nou owrnpoorr:nefet ro C-re].rr<6 oprrvo[ri rrlg oLLqLoul(og dyptou r0nou. Or preroMd[erg lrop(g v5qpo unodrll,civouv 6n onoro6rlnore on6 ro rp(o rco- 6tx5vro rcpprotropori enopre( yro vo repgor(oet rn Ue- tdgpoorl o' 5vo yov[6to. Iuven<ilg, or oM4].ouX(eg rov tptntrerc0v UAG, UAA xor UGA e[vor ovoyro(eg xor rro- v6g vo repporioouv q prerdgpoorl e(re Bp(oxowor guorrd oro drpo tou yovrdiou e(re 6qpuoupyor3wot pt6oo prerdLloEnq prioo orr;v rrodrronorqtrrq oLLq- l.ouX[o. Ito Borcrqpror<d yov(6ro, ro UAA e(vor ro nro rorv6 xo6rr5vto reppotroprorl. To UGA Xprlorilonore(ror ne- proo6repo on6 ro UAG, nop6l go(veror vo ouprBo(- vouv neproo6repo ldOq rord r4v ovdyvroorl rou. (Evo )d0og ovdyv<ooqg ev6g rto6u<ov(ou teppotrogori, 5rov oe our6 owrororyrore( 6vo ol.rwooruLotRNA, 61er og onor6Leopo vo ouvelro0e( 4 petdgpoorl 5og 6vo dMo t<ro6u<6vro reppotroproU ri 5rrrq 6rou ro prBooroprdrto qrdoa oro 3'dr<po rou mRNA, yeyov6g nou ev66Xeror vo nporol6oet dtrl,o npoBLrjporo. l-t' outq qv nep(- oroorl, ro Borrqpro 6ro0rftouv 6vo er6rx6 RNA.) 24.15 To Kto6rr6vto T€ppoTtopo0 Avoyvtrlpi(ovTot on6 npulreTvrro0q llopdyovreq e To rto6rrovro repgortopo0 ovoyvrop((ovror ono nporreiroug nopoyovreg onetreu06ptoor1q, 6Xr on6 optvoorutrotRNA. e 0r dop6q rtov nopoyovrov one),euOepotonq Toqnq 1 ;roLd(ouv pe lg ovrlorolxeq doUeq rou oupn],orou oluvoorutrotRNA- EF-Tu xor rou EF-G. * 0r nopoyovreg onetreu0Sproor;q ro[r1g 1 onovrouv oe erDrrd rodrrovro reppolopou ror u6potr0ouv ro Deopri no),unenlD[- OUTRNA. * 0r nopoyovreg oneLeu06prrlolq ro[r1q 1 unopol8o0vror ono roug nopoyovreg onetreuS6poorlq ro[qg 2, or ono(ot e(op- r6vror ono GTP. * 0 Urlxovrogrig ervor nop6porog oro porr1pto (ro onoio dro06- rouv 6uo r0noug nopoyrlvrolv onetreu0epoolg ro[r1g 1) roL roug euxopudlreg (or ono[or 6Lo06rouv govo 6vo nopoyovro onetreuOepooqs roEriq 1 ). Irov repporropr6 rrlg prerdgpoorlg eprnl6xowor drio ord6ro. H (6ro 11 aw[6paor7 rou repparopori ouv(oro- ror oe oneLeuO6poon rnq nporeivrrqg oLuo(6og on6 ro reLeutq(o IRNA. H aw[6paor1 petd rcv rcppauop6 ouv(ororor oe oneLeu0Sptrlo4 rrov IRNA ror mRNA, xoOrig Kor oe drdo"rooq tou prBoorr:gor(ou mrg uno- ;.rovddeg rou, Kovdvo rodtx6vro repUol.ogoU 6ev owrnpooo- neUeror on6 rdnoro IRNA. Aemoupyo6v eweLrig 6ro9o- pe'nrd on5 ro un6Lornq rco6u<6vro Kor ovoyvop((owor oneuOe(og on6 nporeivrrorig nopdyoweg. (H owi6po- or1 6e Boo(eror oe ovoyvrirproq ro6rrovlou-owrxcodtxo- v(ou, ouvenc0g 6ev e(vor oogSg yror[ onorte[ror 4 oL}.qLoufo puog rprnl,Srog. Ilt0ov6v ow6 owovoxM qv e[5]l[4 rou yeverrKo0 rri6tro.) To rcoSrx6vro repUqrropro0 ovoyvrrlp((owor on6 roug nopdyoweg onetreuoSpcoonq (release factors, RF) rd[49 1. Iro E coli,60o nopdyoweg one],eu06pr* oqg rd(r1g 1 eivor erdrrof yro 6ro<poperrx6g oM4trou- 1(eg. O RFI ovoyvcop((er to UAA r<or UAG, evc0 o RF2 ro UGA ror UAA. Or nopdyoweg 6pouv orrl pgooopr- K4 06orl A xot ononofv notrunemrdu],o{RNA orn eSon P. Or RF undplouv oe nolti Xopr4L6reprl ouyr6wpoorl on6 roug nopdyoweg 6vop[r;g 11 enr;uir<uvoqq: undg louv -600 ;r6pro rd0e nopdyowo ovd rfrropo, noo6 too6rivol.ro pr'6vo pr6pro RF ovd 66xo prBooroprdtro. K6- nore nt0ov6v unrlpXe pr6vo 6vog nopdyowog ondeu- 0dpoor1g, o ono(og ovoyvdlpr(e 6Lo tq ro6rx6vro tepporrol.rori, ror opy6tepo e[e],ffiqxe oe 5uo nopd- yoweq ge er6r161410 yro ouyKer<prpr5vo rco6rr<6vro. Iroug europucbreg, undpXer pr6vo Svoq nopdyowoq onel.eu05proons rd(nq 1 nou roAe(ror eRF. H onorele- oponr5qro pe rrlv ono(o ovoyvop((ouv r' ow(mor1o xro6rx6vto-or61oug ot Borrqproro( nopdyoweg eflrlpe- d(eror on6 rtg Bdoerg orqv 3'06o11. Or nopdyoweg oneLeuO6poonq rd(rlg 1 unoBoq- OoUwor on6 roug nopdyoweg one),eu05poonq rdqnq 2, ot ono(or dev e(vor erdro( yro ouyKeKprprdvo xo6rx6- vto. Ot nopdyoweg td[qq 2 e(vor nptorefveq nou npoo- 65vouv GTP. Iro E. coli, o p6trog rou nopdyowo td(r1g 2 RF3 5yr<enor oe oneleu05pcoorl rou nop6yowo rd- [r1g 1 on6 ro prBoorrlpr&ro. O RF3 e[vor nptore'lv4 nou npoo66ver GTP, 11 ono(o oXet((eror. Ue roug nopdyov- teg enqlrlxuvorlg. l-'lop6t o yevu<5q pr4Xovropr6g rou repUorr.opori e[- vot nop6goroq orouq nporopuc0req Kor rouq europucir- req, or ol,trq).ent6pdoerg prero(ri rcov nopoy6w<ov rd[rtg 1 r<or rd(49 2 5Xouv xdnoreg 6ro<popdg. Or nopdyoweq rd(r1g 1 RF1 xor RF2 ovoyvtopf(ouv ro r<odr6vto repprouoprori Kor evepyonororiv ro prBo- oopdrro oe u6p6Luoq rou nemr6ul,o-tRNA. H 6rdono- on rou noLunemt6(ou on6 ro IRNA npoyproronoefuor pe puo ow(6poor1 ovdloyq pre q ouvq0ro;r6v4 prerogo- pd nenrt6uLoprddov, p' e(o(peo4 6rt 65rcrr1q e(vor ro vep6 ow(rou ol.uvoorulo-tRNA (BI5ne Err6vo 24.34). Ie ow6 ro oqpre(o o RFI r1 o RF2 onetreuOepdrveror on6 ro prBooroprdno ;.16oo rou nopdyowo rd(qg 2 RF3, o ono(og oXer((eror Ue rov EF-G. O RF3-GDP npoo66veror oro prBoocol"rdno npw npoyporonot40e( r1 696 KEOAAAIO 24 Merorppoorl Boorxdg evuorcg
  • 188.
    ow(6poon rou repportogoU,ev6 ro GDP owKoetord- tqr on6 GTP. Aur6 enrp6net orov RF3 vo 6tr0a o'eno- q{ Ue to ptBooto;rtr6 r6wpo trlq GTPdorlq, 6nou nporotre( rrlv ondreu0Spoorl tolv RF1/2, 6tov teppott- ore( 11 noAunemdxrl aAuo6a. O RF3 pot6(er ile rtq neptoXdq nou npo-o66vouv GTP orouq EF-Tu rot EF-G, evri ot RF1 t<ot RF2 pe qv C-teltr( neptoXd tou EF-G, to ono(o ;rq'refuot to IRNA' Aur6 uno6qtrrilvet 6tt ot nop6yoweg onetreuO5poonq ipno,uo"o,otiv tr1v (6to neproXq 4 ono(o.lpqottrlonoteG i'1,'onO touq nopdyoweg ent;r{xuvorlq' H E|(ONA 24'32 qvoseKv0et in tsooxn opXrl 6tt 6trot quto( ot nopdyov- reg SXouv t4v'(6to yevtrrl popgo}'oy(o xot rpoo66vov- .o,'*l p$ootoprdtto 6to6o1tr<d orlv (5to 06o4 (Bootxd' orrl 06o4 A ri oe ;rto neptoXq nou entrol'0rrerot eKre- vd:g pe q 06on A). d "rnoprr-rr<6q nopdyowog onetreuO6pcr:oqq to- EnS 1 eRFi e(vot 1.tto provodrrl npcotetvq q ono(o ovo- yvcop((et Kot ro tp(o t<to6tt<6vto teptrrottol'toti' H oLLnLouXfo tou 6e olet((etot Ue rouq Borcrrlproxo6q nop&yo*tq. Mnopef vo tepprotioet q letd<ppoorl in vltio Xop(q'rov nopdyowo td[4g 2 eRF2, nop6tt o eRFz'iivor'qnopofurltog aroug (upro;rUxrpeg in vivo' H 6opr( rou eRFl orotrouOe( pro ou<e(o Booxrl opXrl: 11 El- ioiil za.aa 6eftvet 6tt onoteLefrot on6 rpetg neptol6g' nou gtgo6wot rn 6opri tou IRNA' 'Evo Boorr6 lro.iBo tptdrv opwo(6cov,.GGQ, ercr(0e- rqt offlv r<opu<pri rrlg neptoXrlg 2' H ew6ntoti tou orrl 06on A owimotle( orq ouvr10r1 tonoOeo(o rou qptvo- [5og o' 6vo o1-ttvoorutro-tRNA' Iuvencirg, prnope( vo XcTlotUonoe( rq ytr'outopr(vn (a) Yto vo tonoOete[ to ,epO oe gSon xqrdl 'nLq d:me v' owtro0tmd to o1'tt- EIK0NA 24.33 0 euxoputonr6q nopdYovroq teppottoltof eRFl exer 6oUn nou ptpe(tor ro IRNA. To potipo GGO ornv oxpn rnq fle- pioXRS 2'etvot onopo[rr1to yto qv udpo]ruol qg no]runentdxnq olroiOos ono to IRNA' H rpotoYporpio nopoXtrlpri0r6e ond tov David Barford, The lnstitute of Cancer Research' vo(ri tou optwoot<u)to-tRNA orqv ow(6poor1. trlq Tlemt- dutro-rpovorpep6onq' H EIK0NA 24'34 ouyrpfuet rrlv ov- t(6pooq repUortopoti pre tq ouvri0n ovt(6poor1 geto<popdq nenrt6(ou' O tepgotropr6g petoq5pet gto uopoiuiop66q on6 to vep6, u6potrtiowoq 6tot onote- lleptopl 1 (orpo 0doq P nerntdtml oluo(60 I H-N .-, H-C,-R / H-C;R o"''{ g'''b IRNA r IRNA Inennorrn duo(60 H-N H-C.R ,lH o'" ' N' H-Ctn H ,Co 000 i*,ro ,**o I nemrdrtl atruo(60 H.N H-i-n I o'"'oH CH I cll I uuu I I eRFl r ,'a-l , m #ffnnr v :,rri;! ?,'i.::r H poptorl onopiprl0rl entrp6net oro ougn)'oxo :F-Tu{RNA, otov nopoyovro perorontorlg EF-G rot orouq nopo- , ovreq oneheuOepuronq RFl/2-RF3 vo npoobtivovrot oqv [dto pt- :coo.r1-LLrr1 neptoxrl. 0 RRF e(vot o nopoyovtog ovorurtroorlg rou :Lpootogor[ou (ptr6ne Etrovo 24 35). :1{Sl'JA :4,3{ H ;reroqopo rou nenrtdfou xot o repportopdg e[- voL nopopoteg ovrtdpooetg, ortg onoieg Uto Boorl oro r6vrpo qq nenlDu),orpovoqepoonq nupo0oreI pto ovr[Dpootl trans-eoTep0- noirlorlq peoo dtoonoolq evoq 6eopo0 N-H n 0-H onetreu0epot- vovtog N n 0 Yto vo dLoonooet ro 6eopo ge ro IRNA ?4.15 To Ko0Kovro TeppOrtOporj Avoyvtrlpt(ovrot ono [lptlreivtrou-c l-lopoyovreq 697 5ruilnxuvcI K6vtpo nenrLdutro- lp0voq0poqq Teili-rn'iiti;*r, K6wpo nenttDutro- rpqvogepdonq DUUII' or$r P ).'r$ /,,'.o'0'-'q ,
  • 189.
    Aeoporrrd ro Seopdnennd(ou-tRNA (Bl,5ne ELx6vo 24.48 yto to ncbg ),enoupye( ro rSwpo rqg nenndulo- rpovogepdorlg). MeroA.d[erg oro yov(6ro RF e].orrdlvouv rrlv ono- rel.eopotLr6rrlro rou repporro;rori, 6notg Srqnror6ve- tor on6 rqv ou(qprdv4 rr<ov6r4ro ouv5Xtor;g rrlg prerdqpoorlg n6po on6 ro ro6u<6vro repprotroprori. H unepSr<ppoorl rou RF1 4 rou RF2 ou[dver qv ono6on- x6qro rou replrouopro6 oro rodrr6vro oro ono(o enr- 6porJv. Aur6 uno6rlLriver 6rt 4 ovoyvriproq rou rodu<ov(ou on6 roug RF1 r<ot RF2 owoyrov((eror pre r' opuvoorulo-t R NA nou ovoyvop ((o uv ).ov0oo;uivo ro rrodu<6vro repporrol-rori. Or nopdyoweg oneleu05pco- onq ovoyvop((ouv opr<etd onoreleoporrrd trg ow(- ororxeq oMql,o u1(eg-or6xo u g. H owbpooq rcpprorrollori neprLotrrBdver oneLeu06- poorl rou oLoxlr;poprdvou noLunemr6(ou, oMd ro'ro- Lo(ner 6vo onoorulrrogrlvo IRNA r<qr to mRNA ouv6edeg6vo oro prBoorrlprdtro. H Ilt(0NA 24.35 6e(X3rer 6rL 4 6rdoroon rov un6i.orncr:v ouororrrcilv (IRNA, mRNA, 30S ror 50S unopovd6eg) onome( tov nopdyov- ta avaxlxiaorlg rou ptBooaparbu (ribosome recycling factor - RRF). O RRF evepye[ po(( pre rov EF-G oe Uro ow(6pooq 11 ono(o Xprlorponore( qv u6p6Luon rou GTP. Avo<popu<d pre roug dMoug nopdyoweg nou eU- nA6rowor mrlv oneLeu0Spcooq, o RRF 51er nop6proro 6oprl pe to IRNA, oMd 6e 6ro0fter too6rivol-ro pe qv 3' neproXrl np5o6eoqg rou oprvo(rlog. En(oqg onome(tot o IF-3. O RRF enrdpd m4v 50S unopovd6o, evcb o lF-S oqorpe( ro onooruLropr6vo IRNA on6 rqv 30S unogo- vd6o. M6).rg 6roXatprorofv or unotrrovd6eg, o lF-B e(o- :a t: i: ;.:. .:l:tifiiti:,:-,-, :lt i)tia? xol,ouOe( vo Xpeto(eror yro vo nopepnod(oer rrlv eno- voorivdeo4 roug. H ouyr<p(ver rrg opro).oy(eq oq npoq rn Letroupy(o Kor rlv oMq).ou1(o rov npoKopurotrrtrtv r<L europurrltrxrJtv nopoydwrov pretdqpoorl g. 24.16 To PrBootrll.lK6 RNA Aronepvd Kot Ttq Ado PrBootllUtKdS Ynopovo6eq e Ko0e rRNA rp6per notrtr6g droxpn6g neproX6g nou dlntrtirvov- ror ove[dpqro, r lxeddv 6treg or prpooroprr6g nporetveg 6pXovroL o' enorprl ge ro rRNA. e 0t neprooorepeg enoq6g pero[f rov prpootoprrdrv unopovo- Dov npoyporonorouvror UeToEu 165 ror 23S rRNA. To drio rp(ro qg pd(og rou BorcrqpLoxori prBoocopor[- ou onoreLoUwor on6 rRNA. H nro nfulpogopror<r1 trr5- 0o6o9 yro rnv ovd).uoq qg deureporoyo0g 6oprnq rcov preydLcov RNA efvor q oriyrproq rov oMq],ouXuilv ttr;v ow(ororlrov rRNA oe ouyyeve(g opyovroprorig. 'Ooeg neprol6g e(vor oqprowrrcig yro rn deweporoyrl 6oprl 6toqpo0v qv u<ov5rrlro v' oM4trenrdporiv 96- oo (euyopc0goroq rcov Bdoe<ov. 'Eror, ov 5vo (eriyog Bdoecrlv e(vor onopofrr1ro, prnope( vo oxni]orrore( or4v EIK0NA 24.35 0 nopdyovrog onetreu06poor;g (RF) reppor[(er q peroqpooq onetreu0ep6vovT0q Trlv flpo- reivxrl otruo[60. 0 nopoyovrog ovorurtroorlg rou prpo- oopor(ou (RRF) onetreu0ep6ver ro retreuro[o IRNA rot o EF-G onetreu0eptiver rov RRF nporotrdrwog dtdmo- or1 rou prpoorlporiou. q1 -#qD@ -P ? F*-sruF*"6-B< 698 KEOAAAI0 24 l/leToqpool Boorreg ewoleg #
  • 190.
    ) 1::,ii: r,--.:':1.,':: . ,:,:. tF-1 tEo*t tF-3 flporopu6reg EF-TUt, EF-Gt trE T. EF.GS [6to q4enrcj 05or1 oe r<dOe rRNA. Aur{ q pr60o6og 6Xet enrpSQer vq KorooKeuooroUv l,emoprepq prowSLo 16- oorou 165 6oo Korrou 23S rRNA. Ko0dvq on6 ro r<6pto rRNA prnope( v' ono6o0e( pe pro deuteporoyrl 6opn pre noM6g 6tor<prdg neproX6g. T6ooeprg yevu<69 neprol6g oXq;rot(owor on6 to 165 rRNA, 6nou nep(nou or puo6g Bdoerg qg oM4trou{og e(vor (euyopropr6veg (BL5ne Etr6vo 24.46).'E[t yevr6g neproXdg oglprot((owot on6 to 23S rRNA. Ot [eXopt- m6g 6(x).roveg neproX6q ouvrl0oq e(vot BpoXe[eg (<8 bp). :uXyd or drnl.ooroopr6veg neprol5g 6ev e[vot t5treeg xor neptSXouv npoe(o16q pre o(euydptoteq Bdoetq. Iuy- xp[oqro prow6].o 61ouv oXedtoore( yo to l.utolovdptord rRNA (ro ono(o e(vor BpoX0rcpo rot nept5louv Lty6re- peg neproX6g) Kqr ro euKopuorxd rurroponlool.tolrcd rRNA (to ono(o e(vor ;ror<pritepo xot 6to05touv neproo6- repeg neproX6g). To peyo).Utepo ;rqr<og rov euKopuorrt- xriv rRNA rup(cog ocpeiletor otqv on6rrqoq ol),q- )'.ouXuiv nou owrnpoocone6 ouv enrnp6ooeteg nepto16g. H xpuorotrrrn SoUn rov prBoorollodrov 6e(1vet 6r oe xdOe unopovd6o ot neprol6g tou r0ptou rRNA 6tn).d:- vowor ove(dprrlro Kr 51ouv 6toxptr6g tono0eobg. 'EXouv Fpe0e( 6rotpop5g mqv u<ov6tqto tou 165 rRNA v' owrdpd ;re X4pur<o0g nopdyoweg 6rov or 30S unopovddeg ouyrp(vowor pe rq 70S ptBootopdtto: undpXouv en(o4g StoEop5g ;reto(rj trrlv etre00epcov pt- Booropor[ov rr ere(vov ilou oupperSXouv mq prerd- ------l------..* - r ^ ! elFlA ! Anogpdooet q gdol A ! 0 elFl A ^unopoq0ei Tov elF2 vo npotoOtl_oet nlv npdoOeon tou Met- i tRNAiMet oro 40S. Entonq npooyer rn dtdotoorl Tov unopovo0ov elF2, elF3, elFsB* I ffooOoq rou evop*nptou IRNA | 0 elF2 eivot Uto GTP6on . 0 elF3 oreyerpet ro oXlgolopri rou TprloToyouq oupnldxou, qv npdodeon Tou 0To 40S, r<006q enlolq qv npdodeorl xot oopcoorl rou mRNA elFl, elF4 oripntroro, , lpoo6eon rrlq ptxprjg 0 elF5B epntrSrerot oqv e(oo6o Tou evopKnlptou tRNA t<ot elvot elF3 ; unopovobog oro mRNA ltto GTPdorl i , !--,+ i;=* i ; I eEFlo+ i npooDeon GTP i I eEFlp, eEFly I AwoMoyrl GDP ! i eEF2E I Merordnrol rou prpootogoriou i ' t -_-L-*ffi ---4 i flpoxopudrcq i Europudlreg RF1 : eRFl , Avoyvrirprorl UAA/UAG : : Rtz eRFl Avoyvoprorl UAA/UGA RF3- eRF3 Ardyepon otrtrou Ctov) RF(s) *OL lF-2 xot elFSB dXouv ouotroyio otrtrr1trouXiog -01 lF-2, EF-Tu, EF-G rot RF3 dxouv ouotroy(o otrtrr1trou1(oq '01 EF-Tu roL eEFl o dXouv ogotroy[o otrtrntroux(oq r0L EF-G xot eEF2 6Xouv ogotroy[o otrtrntrouxioq Aeuoupyrreg oUotroy(eq Tov np0Kopuolr6v rr europurrrur6v nopoyovTolv Tnq UeroQpoonq. epoon. MeroBoLdg mqv owrdpoortr6rlto tou rRNA ouprBo(vouv xord rlv np6o6eoq tou mRNA, roJV uno- ;rovd6ov r1 rou IRNA. Oprop5veg ;retoBol6g owtKoro- nrp((ouv puo dpreo4 olL4l,en(6poorl rou rRNA pe to mRNA r1 to IRNA, evdr 6LLeg npoxolotiwot Spgeoo on6 dMeg peroBoLdg orrl 6o;tq tou ptBoool-tor(ou. To r<ripro or1;re(o e(vor 6n 1 dropr6prpcoon rou ptBootol.totl- ou e(vor eudtrucrq rord t1 perdgpoorl, t6[oq qq UtKp]iq unopovd6og, enet6rl np6net vo etr5yXet qv orp(Beto tou (euyopcirporog rr.r:6txov(ou-owtrro6u<ov[ou. 'Evo lopom4ptonr6 rnq npororqyoUq 6oprrlg rou rHNA e(vor 11 nopouo[o pe0u].topr6vov roto].o(ncov. Yndplouv -10 pre0ulogddeg qro 165 rRNA (r<up(tog npoq ro 3'drpo rou prop(ou) t<ot -20 oro 23S rRNA. Ito rUrropo rov 04trootrrirv, to 18S rot 28S rRNA peto- <p5pouv 43 t<ot 7 4 pre0uLopd6eg, owtoro(Xog: ouvencbg -2Y" rav vourl.eort6(ov e(vot preoutrropr5vo (nep(nou rprnldoro noooor5 ouyrprnxd pre ro Borcrrlpto). H peydLq prBooroprr4 unopovddo en(oqg nepulXet 6vo pr6pro 5S RNA 120 B6oeov (oe 6Lo to ptBootoprdtro ercr6g tclv ;.utoXovdpror<cbv). H oMqLouX(o tou 55 RNA e(vor Lry5repo rol.d ouwrlprlpr6vrl ouyrprtrd pre rg otr- Lrlloulieg rov r<ripuov rRNA. 'Ol.o to ;t6pto tou 53 RNA ep<pov[(ouv pro 6o;14 pre (euyopropr5veg Bdoetg oe pe- y6l.r16mooq. Iro ptBoocoprdrro tou euKopuortKo0 xurroponl,6- opqroq, 5vo dMo pu<pd RNA Bp(oreror orq preydtrrl .:,,,r To Pipoorr,rptro RNAAronepvo rotrtgA0o PtpootopLxec Ynl-:,::.: 6gg l-evml Aenoupy(o
  • 191.
    AUtroro ntrorioro os RNA Ell(0llA24.37 H 30S unopovddo 6Xer pro Ksqqtrn nou 6roXop(e_ r0r ono ro orirpo ond 6vov ouX6vo rr en(o4g dLo06ret pro npoe[ri_ Xouoo e[6dpo. unopovd6o. flp6reuor yro ro 5.gS RNA, rou ono(ou 4 oMrlLouX(o owrororle( cro S'dr<po rou npoKopucrlrxorj 23S rRNA. O ptopr6veg prBootrrplr<dg npoteiveg npoodrivowor tqXupd oe onopovo;rdvo rRNA. 7Meg 6ev npooddvov_ tor o' eLeU0epo rRNA, oMd onorrorjv ourrl t4v rrov6_ ulro og6rou npoo6e0oUv dtr)eg nporeiveg. Aw6 uno_ bql,d;ver 6rt 4 6ropr6peoon rou rRNA r<o0op((er ov undpXouv neproX6g np6o6eorlg yro opropdveg nprorei_ veg. Ko0c0g npoo66vetor rd0e nporetvq, endyer peto_ BoLSg 6rop6pqoonq oro rRNA, or onofeg enmp6nouv r4v np6o6eor1 dL),rov nporeivdrv. Iro E. coli, oXe66v 6Leg or 30S prBoocrtpu<dg npoteiveg oAL4lenr6poriv (prolov6tr oe noxfi.oug Bo0potlg) pre ro 165 rRNA. Ot neproxeq npdodeoqq ndvtrr oflg nporetveg e;rgov((ouv eu pe(o no rril,(o Soprr<civ XqpqKrn p rortrcilv, yeyov6 g nou uno6rll,cbver 6rt or ptlxqvropo( ovoyvriproqq npo_ reivc0v-RNA prnope( vo 6rog6pouv. - _lo Z0S prBoocoprdno 6Xer oo0pr;rmpq xoroor<eur1. H Ell(0NA 24.37 nopouord(er pto qXrlpronrrl bop4 r4g iOS unotrrovdSog, 11 ono(o unoErotpefuor oe r6ooeprg nepro_ X6g: r<ego),rg, ou15vog, ocilpo ror e[d6po. H ElKOl,lA Z4.gg nopouoto(er pro nop6proro oneu<6woq qg 50S unopo- vddog, 6nou ro 6rio rUpLo XopoKrrlprourd e(vor 11 xev_ tpxrl npoe(oXn (6nou ronoOerefuor ro 55 rRNA) xor o Urq(oq (ouw(Oeror on6 noMon).d owrypogo rnq npo_ retv4g L7). H EIK0NA 24.39 deipet 6n 4 e[d6po (ptat_ form)_qg UrKpnq unoprovd6og egopp6(er prdoo or4v npoe[op1 (protuberance) rnq ;reyd].qg unopovd6og. Yndpler truo r<od,6r4ro ;rero(rj rov unopovd6cov, r1 ono(o neptdXer opr4r6veg on6 ng oqprowrdg 06oerg. H 6oUn qg 30S unopovddog oro).ouOe( t4v opyd_ voorlrou 165 rRNA pre rd0e 6optr<d Xoporrqproru<d v, owrororle( oe Uro neproX4 rou rRNA. To oripro Foor(e_ tor ndvo ornv 5'neproXrj, q e[d6po ndvo orqv Kewpr_ EIK0NA 24.39 H e[e6po rng 30S unopovodog etpopg6(er p6oo oqv evrogq qg 50S unogovodog oX4por(ovrog ro 70S ptpooor_ goTto. r{ nepropl r<ot 4 rego},4 ndvor u1v 3'neproXrl. H EIK0llA 24.40 6e(yyer 6n 11 30S unopovd6o 61er oori[;rerpn Ko_ rovoprq RNA ror npolreiV6v. 'Evo orlpowu<6 XopoKnlpr_ ortK6 e(vqr 6n 4 e[66po rrlg 30S unoprovddog, q onoio nopdXer q 6rdrpoorl Ue rnv 50S unoprovd6q, ouw(Oetor o1e66v onori.ermrrd on6 RNA. Kowd m4 6rdgoo4 Bp(or<owor ro noLrj 6[o nporeiVeg Bp(or<owor (dvo pr_ xp6 rpr;pro tqg 57 xor (ocog U6poq.rnq S12). Aw6 or;_ pofver 6u 11 orjvdeoq xor 6rdmoo4 rtov prBooropu<cilv unogovddcov np5ner vo e[oprdroL on6 ol,Aq],ent6pd_ oerq Ue ro 165 rRNA. H oM4Aen(6poorl rov unopov6_ 6ov en4ped(eror on6 puo prerdMo(4 o, 6vo Bp61o rou 165 rRNA (or4 06o11 791) nou ewon((etor orq dtdgoorl rov unopovddov. Me nerpdporo rponono(4oqq/nopep_ polnS, ono6erX04re 6l eprnLdrowot rqt dllo voux.leo_ r[dto rou 165 rRNA. Auq q noporrlp4oq unooqp((er qv 165o 6r ro prBoorrtprdno nr0ov6v e(ell10qr<e on6 6vo oropror(6ro onorel,oUgevo gdMov on6 nruR nopd npcore(v4. H 50S unoprovdSo 6Xer neproo6repo ro6trprq xcno_ vo1.r4 ouorortxcilv ouyr<pnrxd pe qv 30S unogovd6o, pe porpd poB6(o 6(r<Arovou RNA nou 6ronepvoriv 14 6opr1. To RNA qXqpot((er puo pd(o ogrXrd ouoKeuo_ opr6vov el(rov. H e[orepu<q enrgdvero onote].efror xu_ p(trlg on6 npcotetvrl, trr' e[o(peor1 ro rdvrpo rqg nemrdul.orpovogepdoqg (F).6ne Ev6r4ta 2a.1 9). Z14e_ t;l{*tiA :4.3t H 50S unopovodo eXet gro xevrptrrl npoe[oXr1 onou evron[(erot ro 55 rRNA ror droXopr(eror pdoo prog evro;u1g ono ro ploXo nou ouvrlOeror ono ovriyporpo rrlg nporeivqg 17. :;i:r':: -: .:,,.:: H 30S pLpOOtogrrl UnopOvodO e[vOr rivo prpovOu- rtreonporeivuo ooporOro. 0r nporefveg errovl(ovror urptveg, H qoroypotp[o nopoXoprl0rlre on6 rov V. Ramakrishnan. Medical Research Council (UK). 700 KE0AAAI0 24 Merorppoorl
  • 192.
    1 65 rRNA ir!,i11.1ic.+l ToorlueiueiloQriqUeToEuTovrRNAevroni(ovror oe60oneproX6groul65rRNAror pLoneptoXrl rou23SrRNA,Avonopo- yerorono: M. M. Yusupov, et al., Sclence 292 (2001):883-896 lhttp://www.sciencemag.org]. H rptrrroyporpio nopoXrrlprl0rlre ono rov Harry Noller, University of California, Santa Cruz. 66v 6Lo ro qrrlproro rou 23S rRNA oMqLenrdporiv pe nprrrtetveg, oLi.d noMdg on5 rrg npcoreiVeg efuot q4err- xd ovopydvoreg. H ou;rBoL4 ro:v unopovddrov oro 70S prBooroprdrro nepiloprBdver enorpdg ;reto(ri tou 165 rRNA (noLl,5g orrlv neproXrl qg e[66poq) ror tou 23S rRNA. Yndp- louv en(or1g oproprd,veg oMr;lenrdpdoerg gero[6 rRNA xdOe unotrrov6doq rot npo:reivcirv qq dlJtqq, roOri:q rr eLdXrmeq enoq6q ;rero[rj nporeivtirv. H EIK0NA 24.41 rouronore( ro oqpre(o enogrlg ndvro mrg 6o;169 rou rRNA. H EIK0NA 24.42 ovoryer tr1 6o;rq (qowoorefue rqv 50S unopov66o neptorpoppSvrl optmep6mpogq Kor qv 30S unopov66o neptorpo;r;rdvq 6e[t6orpoeo yti- pro on6 tov 6(ovo nou uno6etxvUetot orrlv etr6vo) no- pouord(owog r1v ew6ntoq tcov orlprefov enocp4g ndvto orrlv enrgdvero rdOe unoprovd6og. 24.17 Io PrBootopdrro ileprdxouv noA d Evepyd Kdvrpo r 0t olLnLenLdpooeLq orrq onoleq epntr6reroL to rRNA e(vor rp(orpo g6pog rrlg tretroupy(og rou ptpootrtgor[ou. * To nep0otrtrov rov 06oeorv npoodeoqg rou IRNA ro0op((erot ruplog on6 ro rRNA. To Boorr<6 XopoKrnprfitr5 tou ptBootol-iot(ou nou np6ner vo Ouprdror rove(g e(vor 6rL np6rettot yto ou- vepyoru<r1 6oUn, n ono(o e(oprdtot on6 pretoBo).69 ElK$ilA 24,42 Enoq6q pero{rj rtov prpooropLr6v unopovodtov ruproq npoypotonoto0vtot ond ro RNA (etrov[(erot UtoB). 0t enoq6g nou nepilogpovouv nptote[veg euov(ovroL r(rprveg. 0r duo unogovo6eq neptmp6qovrot gorpLo 11 pio on6 qv o]rtr4 onoro],Unrowoq Ttq ert- rpdveeg dnou npoyporonorouvror or enorpdg: onrl 6vo en[ne6o enorprlg ro0ero oqv o06vq, t1 50S unopovodo neptorp6qetot 900 optme- pdorpoQo, evtir 1 30S unopov60o nepLmp6qeror 900 6e[r60Tpoqo (r1 exdvo ug nopouotd(et oe owlorporpo npooovorotrtop6 ond to ouvri0rl). H rptrlroypogro nopoXropl0rlre qno rov Harry Noller, University of California, Santa Cruz. f4.1? To Prpoorr:porLo [ept51ouv tlotrtro Evepyo Kevrpo 701 23S rRNA Boorrdg dworcg - i
  • 193.
    oilq ox6oerq Ueroqrlrov evepyci)v Kdwpov rnq Kord rn Uerdepoon. To evepyd K6wpo 6ev e(vor ptr<pdg, 6tor<pn5g neproX5g 6nc,:g ro evepyd r6wpo rcov ev(U- 1.uov, oMd peydLeg neproXSg, trrlv ono(ov q 6opui r<or ot dpdoerg e[oprcitwor 16oo qn6 ro rRNA 6oo ror on6 rrg prBooropur6q nporeiveg. Ot xpuoroLlm6g do- ;r5g rolv [eloprmc0v unopovddov Kor rov Borcr4pro- r<dlv prBooropor(otv npoorpri,pouv pto r<ol.r1 ew6notoq yro rrl ouvolrrrl opydvcooq xor tov[(ouv ro p6].o rou rRNA. H nro np5o<porn 6oUn, oe ovdLuoq 3.5 A, rou- ronoe( ooepcbg lg ewon(oerq rov IRNA xot rrr:v Ler- toupyrxd:v neprolcitv. Mnopotipre nl6ov vo e[r1yr1oou- pe noLL6g ).etroupy(eg rov prBoorol-ror(rr:v on6 u1v dno{.rr1 qg 6opqg roug. Or prBooropur<69 Lemoupy(eg neptorpd<powor yUpco on6 rlv otrl,rllenbpoorl Ue ro IRNA. H EIK0NA 24.43 no- pouord(er ro 70S prBooroprdno Ue lq ewon(oerg rcov IRNA ong rperg 06oerg np6o6eo4g. To IRNA oug 05- oetg A ror P e[vor oXe65v nopdtrtrql.o ;rero[ri toug. Kor to rp(o IRNA eu0uypopp((owor, Ue rouq Bp5Xoug rov owxo6u<ov(cov rouq vo npoo66vowor oro mRNA ;r5oo orqv orjloxo ndvot or4v 30S unopovd6o. To un6Lorno rpritro r<dOe IRNA npooddvetor or4v 50S unopovd6o. To neprB6Mov yripo on6 rdOe IRNA nopriXetor rup(rrlg on6 rRNA. Ie r<d0e 06oq, ro rRNA r<or IRNA 5pXowor o' enorprl toq rgqproro rnq 6opnq, nou yevrrd e(vor ouwrl- pnuSvn. 'Htov ndwore ;rey6).o o(vry;ro vo rorovo{oouge n6g prnope[ vo lopSoouv 6rio oyrcl6q IRNA ro rivo 6(- nLq oro dMo r<ord qv ovdyvotoq yemovrr6v rco6txo- vicov. H r<puoroMu<r1 6opr4 eprpov((er puo r<oprn4 45o oro mRNA prero[ti tov O5oeov P ror A, 4 ono(o enmp6ner Ell(0NA 24.43 To 70S prpootoluirto onoretrefror on6 r4v 50S unogovo0o (oonpo) K00r Trlv 30S unogovddo (UoB) Ue rp(o IRNA nou ewon((ovtor entqovetoxo: r[rprvo or4 06011 A, pntre oqv 06- or1 P ror npdotvo orn eeon E. H rporoypotp(o nopoXtoprl0rlre ono rov Harry Noller, University of California, Santa Cruz. 06oq A d Iumpoqr'1 I ,. r-i r t;' .', .i /', ' 3* t;iI ,] .,:, q:i-.r- ' ':j. + EIK0NA 24.44 To tpio IRNA 6Xouv drorpopertrri npooovorotrrolrd novo oro prpootrlgolo. To mRNA nepLmp6EeroL pero[u rov 06- oetov P ror A enupenovrog 6ror oro ogrvooru),0-tRNA vo npoo- 66vovror oe 6rn)ovo rodrrrivro. H rporoyporp(o nopoXtop404xe on6 rov Harry Noller, University of California, Santa Cruz. oro IRNA vo eEdmowor, 6ntog cpo(veror orrlv qg ElK0- NA 24.44. To IRNA ong 06oerg P r<or A Bp(or<owor un6 y<ov[o 26o pero[ri rouq Ue r<6wpo t' owrro6u<6vrd roug. H orev6rep4 npooSyyrorl prero[ti rcov ore].er6v rcov IRNA ougBo(ver oro 3'dr<po, 6nou on6Xouv g5).q 5 A (rdOero oq npoq ro en(ne6o r1g oel(6og). Aur6 enr- rpSner orqv nemr6u<4 ol,uo(6o vo perorpdperor on5 ro nemrdu).o-tRNA qg 05or1g P oro oUrvooKULotRNA rqg 05o49 A. To oprwoorulotRNA evoo;rordlveror orn 05orl A pe- oo rou EF-Tu ror ro (euydpoprd rou pe ro rro6x6vro e(- vor ovoyro(o 6ore o EF-Tu vq u6polrioet to GTP r<or t' oneleu0eprrr0e( on6 ro prBoorrl;rdtro (Bl,5ne Ev6rqta 24.10). O EF-Tu opXu<6 ronoOere( to ol.rwoorul,o-tRNA orrl prrp4 unopov66o, 6nou ro owu<rrl6x6vto (euyoptr> ver Ue ro rc,r6x6vto. H peror<tvr1on tou IRNA efvor ono- pofuqr4 6ore vo ;rero<pepOe( o).6x),qpo ;rdoo orq 05oq A, 6rov to 3'dr<po rou erodpXetor oro r6wpo rrlq nerrn- 6 ul,orpovorpepdoqq or4 peyo),q unopovd6o. YndpXouv 6to<popenr<d prowSLo yro ro n6g npoyporonoefuor qu- q q 6repyoo(o. liprqovo pr' 6vo on6 oLnd, o).6r<L4po ro IRNA neprorp6geror, 6ro1 6ore o oyr<rivog qg 6o- prlg oXql:orog L, r'1 ono[o oX4pror[(erot on6 roug Bpo- X(oveg D ror TIUC, vo UeroKrve(rot prSoo mo prBo- oolpdrro, enmp6nowog oro Bpo{ovo TIUC vo (euyoprrl- oer Ue ro rRNA. li;r<pcovo p' rlvo dtri.o ;row5Lo, n roo- repx4 6oprl rou IRNA peroBdMeror Xorlorponot6wog ro Bp61o rou owu<ro6u<ov(ou oov oppr6, evr.ri ro un6l,or- no IRNA neptorpdqeror on6 pro 0rio4 6nou oropd(etor orrlv 3'nleupd rou Bp6Xou rou owu<ro6u<ov(ou oe Uro dMn 06on, 6nou oropd(eror orrlv 5'ntreupd. Merd rr; peror6ntoq, o EF-Tu u6poL6er ro GTP enrp6nowog vo ouvelrore( 11 o0vOeoq rou nemr6[ou. H perot6nrorl nepil.oprBdver preydl.eq perorrvqoetg ong 06oerq rcov IRNA pr6oo oro prBoocopdrto. To drpo rou owu<co6rxov(ou rou IRNA perorrveftor -28 A on6 rr1 l 06on E 702 KEOAAAI0 24 MeToqpool
  • 194.
    06o11 A orrl06o11 P xor orq ouv6lero perorrvefuor enr- nLriov 20 A on6 rr1 05on P mq 06or; E. E[on(og r4g ytov(og r<dOe IRNA cDq npoq ro owrrcodrx6vro, o 6yrog rou IRNA peroxrvefuot r<otd no),ri preyoArjtepeg onootd- oetg: 40 A on6 q 06oq A orr'1 05o4 P r<or 55 R on6 r1 06orl P orrl 05or'1 E. Aw6 uno64l.c0ver 6l q preror6nrorl npouno06ra 6poonr<q ovo6ropydvoon rnq 6oUriq. l-ro noMd Xp6vro enrrpotodoe 4 peror6nron Llno- po{oe vo npoyl-loronorrl0e[ p6vo nopouo(o rou nopd- yowo EF-Tu. Om6oo, ro owrBronr<6 onopoopur<(v4 (sparsomycin) (to ono(o ovoor6l,).er q 6pdo4 rnq ne- mr6uLorpovogepdoqg) nupodote( rr1 peror6nrorl. Au- 16 uno64),civer 611 4 evSpyeLo nou npootOe( 14 peror6nror; ornv fl poyponr6rlto onoO4reUeror oro ptBoorr:prdtro oe6rou olrlprorrme( o nenrrdrr6g 6e- op6g. O EF-G ouvr'1Ocog enr6pd oro ptBooro;rdno one- )reu0epri:vowog our4 r4v evdpyero ror ro0ror6wog ro rxov6 vo npoco0foer 14 peror6nto4, roorSoo rl oTropoo- guxftn4 6Xet rov [6ro p6tro. H onopoopur<(v4 ovoord].- Aer rrlv nemr6u).orpovogepdoq pr6oo np6odeorlq fio nemtduLotRNA: our6 qvoordl er qv oM4Len(6poo4 rou Ue ro oprvooKuLo- IRNA. l-I0qv6v 64ptoupye( puo 6rop6pgolo4 nou prord(er ;re t4 ouvqOro;rdv4 6rop6p- gtooq prerd 14 peror6nrorl, q ono(o pre q oerpd r4q npodyer tr1 peror[v4o4 rou nemr6utro- IRNA. To oq- gowx6 orlpreio efuor 61 4 peror6ntorl onorele( ev6oye- vr; rdr6qro rou ptBoocol.ror[ou. ljpr<provo ge to prowSLo rov uBptdu<rilv r<otoordoe- rov, 4 perot6nrorl prnope[ vo ouprBofver oe 60o srd6ro, Ue rn Uro prBoooprrrl unopovd6o vo petorrve[tor oe oxSon Ue qv dtr.q c6ore vo 6r1puoupy40e( 6vo evdrdpe- oo mddro mo ono(o undpXouv uBpr6u<rig neptolSg np5o6eoqg rou IRNA (50S E/30S P ror 50S P/30S A) (Blene Erx6vo 24.29).Iuyrp(oerg r4g 6oprlq rou prBo- orol.ror[ou pero[U trov roromdoeov nprv ror perd tr1 petot6nrorl, r<oOcig xor ouyrp(oerg qg Sropr6peoonq tou 165 rRNA gero(rj el,eU0epov 30S unopovddorv rot 70S prBoocol.ror(rov uno6rllrilvouv 6tr 6op4 SXer 5wovr1 xlqrtr6rrlto r6(cog ong neprol6q qg r<eqo).rig Kor rrlq efd6pog rng 30S unoprovd6og. Mto ev6rogdpouoo 6v- 6er(q yo to 1-row6l,o trov uBpr6u<rilv r<oroordoeov npo- 6pleror on6 ro yeyov6g 6rr noM5g Bdoerg rou rRNA nou epnfuirowor orrl o[v6eoq rov unopovddtov, Bpi- oKowor rowd oe Bdoerg nou epnl.6xowor mrlv oMrl- Aeni6poorl pre ro tRNA. Aur6 uno64lcirver 6rr ot neproXSg np6odeoqg rou IRNA Bp[orowor rowd or4 6t6rpooq pero[ti rov unopovd6otv, ev6 en(o4g unovoe( 6n oMoy6g orrlv oM4Len(6poor; rov unopovddcov 0o ynopo0oe vo ouv66owqr Ue rrl preror<(vqor1 rou IRNA. Meyd),o rUngo qg 6op{g rou prBootrlpor(ou roro- LopBdveror on6 to evepyd r6wpo rou. H oXqpolr<4 onem6vro4 r<ov ptBooropur<riv 06oetov orr'1v EIK0NA 24.45 6efiyer 6tr onoreloriv nep(nou ro 6Uo tpfuo qg 6oUnq rou prBoocol.ro{ou. 'Evo IRNA eto6pXeror orn 06oq A, prrog6,peror prSoco perot6nronq orn 06o4 P rr Snero uyxmo.trefuer ro prBooroprdtro on6 q 05or1 E. Or 06oerq A xot P ercre(vowor rord ;r4r<og Kor rov 6tio prBooro;u- xtiv unopovd6ov: ro IRNA (euyopriver ;re to mRNA E|(0NA 24.45 To prpootoporto dLo06rer notrtro evepyri r5vrpo. Mnope( v' otrtrr;trenrdpo Ue Uro Uepppovq. To mRNA npoyUoTonor- e( gro neprotporprl roOdrg DLepleror ond lg 06oerg A ror P, or onofeg pp[oxovrot un6 yov(o 4 Uro oq npoq qv otr1. H 06oq E pp(oreror n6po ond U 05oI P. H 0eo1 qg nenldutrorpovoqepo- onq (dev onerxovf(eroL) exre[vrrot droprioou rtov xopurptirv rotv OSoeorv A xor P. M6pog rr;g 06or;q 6nou npooD6vovror or EF-Tu/G ppforeror oq pdo4 rov 0tioealv A xor P, m4v 30S uno;.rovd6o, otrtrd ;rerogopd rou nenrtd[ou ouprBo(vet ornv 50S unopovddo. Or 06oetg A ror P Bp(- oKowor 6[n].o-6[n],o, 6reurol.rjvowoq rrl ;reror<(v4oq rou IRNA on6 q puo 05or1 orqv dM4. H 05oq E ewonG (eror rowd orrl 05o4 P, evdr ro r6wpo tqg nemrduLo- tpovoqepdorlq ornv 50S uno;rovddo, xovrd oro ol.rwoorulrrd drpo rov IRNA ong O5oetg A ror P (B).6- ne Ev6trya 24.18). 'OLeg or nporeiveg nou npoodrivouv GTP ot ono(eg Xpnorponororiwor orrl pretdEpooq (EF-Tu, EF-G, lF-2, RF1, RF2 ror RF3) npoo66vowor orqv 6ro 06o11 (pept- 169 <popdg rq^,e(ror r<6wpo GTPdo4g), yeyov6g nou prdMov nupodore[ qv u6p6truor1 rou GTP. Aur4 4 05- oq ewon(eror ar4 Bdorlrou prioXou trlg preydLqg uno- povd6og, 4 ono(o onotel,efuor on6 rtg npotefveg L7 xor 112. (H L7 e[vor nopoMoy4 qg L12 r<or nepdXer pro oretulopd6o oro o;rworetrrr6 dxpo). Er<r5g on6 out4 qv neproXri, ro oUpnl.oKo rnq npcoreivrlg 111 p' 5vo rpqpo 58 Bdoeorv rou 23S rRNA nopdXer plo 06oq yro rrlv np6o6eoq opropr5vov owrBrorrr6v nou enrlped- (ouv qv evepy5r4to rqg GTPdoqg. It4v npoypotrr6- tqto, r<oprd on6 our5g rrg prBoocrlpru<69 6opr5g 6e 6ro05ret evepy6r4ro GTPdoqg, coor6oo e(vor xor or 6rio ovoyxo[eg yr' our4 rr1 6pdoq. O p6].og rou prBooroprot! ou efuor vo nupo6oreirrlv udp6Luoq rou GTP on6 no- pdyoweq npoodede;rdvouq orrlv ow(mo rXq 05or1. H oplrr<4 np6o6eo4 rrr:v 30S unogovd6ov oro mRNA qnore( qv nporeffi 51, 11 ono(o 61er roXupq ouyydvero yro prov6r<trtovo vouxleivrrd o[5o. H nporei'- v4 51 6rorqpe( oe prov6r<l,ovq xotdmoorl ro mRNA nou efuor npoo6e6e;rdvq orrlv 30S unogovd6o. Auq 11 6pdoq efuot onopofrrlrr'1 d;ore ro mRNA vo prlv npoo- ).dBer pro 6rop6p<pcoo4 pe (euydptopo Bdoeo:v nou 0o rlrov or<ordl.L4Lrl yro q pretd<ppoor1. H Sl 5Xer puo e[orpetu<d enrprrlrrl 6opr4 r<or ouv66eror pe trg S18 r<or S21. Or rperg nprrrreiVeg ouvroroUv pto neptopl 11 ono[o Odon A -:-_-*- -- | r.,l:,:5J" I ?:+.'; t Ta Prpooolpono llepreXouv il0trtro Eveoyo (a,, :: 703
  • 195.
    epni.6reror m4v oplrxrinp6o6eor'1rou mRNA Kor ornv np6o6eor1 rou evoprerlptou IRNA. Aut6 rono0ete( tr1 06o11 np6o6eonq rou mRNA xowd orq oxropr| rnq Ur- rpqg unoprovddog (plene Exdvo 24.3). To 3'drpo rou rRNA, to ono(o (euyopcbver pe rq 05or1 6vop[qq rou mRNA, ewon((eror oe ourrlrqv neproXr;. Or nopdyoweg 6vop[4q npoo66vowor orqv (6ro ne- proXri rou prBoorollor[ou. O lF-3 prnope( vo 6roouvde0e( pe to 3'dxpo tou rRNA, 6ntr;g en(o4g ror pe opret6g pt- Bootopur<69 nporeiveg, prero[ti d],],rov r<or npo:reiveg nou gdMov epnL6xowor onlv np6o6eorlrou mRNA. O lF-3 0q pnoporioe vo moOeponore[-r4v np6o6eorl mRNA-30S unoprovddog: crTl ouv5lero erron((etor, ;16- l.rg ogr1Oe( q 50S unopovd6o. H evoopdtroon rou 55 RNA oe 50S unoprovd6eg nou ouvoppol,oyoriwot in vitro e(oprdror on5 rrlv xo- v6rrlro tpuiv np<rl'reiv6v -L5, L8 ror L25- vo oXqpo{- oouv rivo ororxeloperprr<6 oUpn].oxo pro(( tou. To oUgnl,oro npoo66vetor oro 23S rRNA, r<drr nou rov6vo on5 ro [eXopror6 ouoronrd rou 6e pnope( vo ilpoy- poronorrloer. To o0gnl,or<o Bp(oreror rowd ortg O5oerg P ror A. 'Evo veoouwtOriprevo nol,unem(6ro ercreiveror 6ropr5- oou rou prBoorol.ror(ou, prorprd on6 ro evepyd r6wpo, npoq rnv neproXq 6nou ro prBootoprdrro [orog ouvdmov- rot pe preprBpdveg. Mro nolunemrdxri oAuo(6o ovo60e- ror qn6 to prBooroprdrto 6roprSoou ev6g 6roriLou e[56ou nou o6r1ye( on6 q 06o4 ulg nemrdul.orpovogepdorlg orqv enrgdvero rrlg 50S unopovddog. O 6(oul,og onore- Le[ror rupfrog on6 rRNA. E[vor oprerd orev6q -e0pog 1- 2 nm- rr 6Xer UnKoq -10 nm. To veoouvrr06prevo noi.unem(6ro ovqdUetot on6 ro ptBooroprdrro -15 A pro- rprd on6 to xriwpo r4g nemr6uLorpovoEepdoqg. O 6(- ouLog prnope( vo ouyxporrloer -50 qrwo(6o ror ;rdMov neprop[(er tr1v no].unemrdxrl o].uo(6o riore vo Un pno- pe( vo 6mLo0e( nporo0 eyxorotre(rler rrlv neproXq e(6- 6ou, poLov6rr ev66Xeror vo oxnportororiv l.(yeg deweporoye(g 6opdg. 24.18 To l6S rRNA Ato6popoTi(€t Evepy6 P6Ao orn Merdrppoon c To 1 65 rRNA OrodpoporL(er evepyrl potro olg treuoupyieg rrlg 30S unopovodog. Atrtr1trentDpo opeoo pe ro mRNA, qv 50S unopovodo r0r r' ovlruldrrovlo rorv IRNA olg 0eoerg P ror A. To ptBooropduo opXLrd Oe<rlpr10r;xe toq Uro ou).).oy4 nporreivtov pe norr<(l.eg xoroLurrr5g evepy6trlreg TIou ouyKporo6wqr pSoo oLLrlLentdpdoeov 16oo trreto[rj npcoteivov 6oo ror geto(ri RNA xor npctlrei- vrrrv. Qor6oo, 11 ovor<dlurf4 prop(cov RNA pe rorou- lrrl evepy5tqto (BL6ne KegdLoro 21) oo<pc0g uno- 6qLcirver 6rr ro rRNA (oog 6ro6popror(er nro evepy6 p6lo orq Letroupy[o rou ptBooropor(ou. YndpXouv nl.5ov ev6e([erg 6tr ro rRNA o].Lq].enrdpd pre ro mRNA ( to IRNA oe xd0e orddro qg trrerd<ppoonq Kr 6tr or np<o"retveg e[vor ovoyr<o(eg oiore ro rRNA vo 6rorqpq0e( oe Urq 5oprl orrlv ono(o gnope( vq enme- Le( trg r<oroLutrr6g l,enoupy(eg rou. l1otr).59 otri.qLe- ntdpdoerg nepri,oprBdvouv erdrr<rlg neproX6g rou rRNA: o To 3' dr<po rou rRNA otr4lenrdpd oneuOe[og pre ro mRNA rord qv 6vop[r1. o Erdu<6q neprolSg rou 165 rRNA o).l,r1l,enr6po0v oneuOe(og Ue rrq nsproX6g owrro6rrov(ou trov IRNA 16oo or4 06o11 A 6oo ror m4 06o11 P. l-lopo- pto(og, ro 23S rRNA oMqLent6p6 pre ro CCA drpo tou nemrdulotRNA 16oo or4 05oq P 5oo xor orrl 06ortA. o H qI ntren(dpooq rorv unopovddov nepil.opBdver oLtrrll,enrdpdoetg peto(ri 165 r<or 23S rRNA (9).5ne Ev6rrTta 24.1 6). l-loLLdg nA4pogop(eg oXeru<d ;re ro (eloprord Fri- Uoro rnq perdtppoorlg oro Bolcrlpro npo4l0ov on6 tr1 Xpnon owrBrorxc0v ro onofo ovoor6Mouv t4 6epyo- o[o oe ouyrer<prpSvo orddto. O or6Xog ev6g owtBrorr- rof prnope( vo rouronotqOe( on6 ro oumorrr6 mo ono(o ouprBo(vouv peroMd(erg nou npood[5ouv owo- Xrt oro owrBronrd. Meprrd owtBronrd enr6porlv oe (e- Xrrlprordg prBoorotrur6g npo:reiveg, evtil noLl,d 6pouv ndvo oro rRNA, yeyov6g nou uno6qkivet 6l ro rRNA epnLdr<erot oe noM6g 11 or6pr1 r<or oe 6treg ng l,etroup- y(eg tou prBooroproilou. Or Lenoupy(eg tou rRNA rilouv 6tepeuv40e( pe 6tio 6rogoperu<59 npooeyy(oerg. Or 6o;ur<rig preLSreg 6e(;youv 5tr t6ro(repeg neproX6q rou rRNA ewon((ov- ror oe o4;rowu<59 06oerg rou prBooutl.ror[ou ror 6t or Xqgrr<59 rpononorrloetg ourcbv rrov 06oeov nopoKo- Lriouv ouyxerptpr6veg prBooalpur<69 Leno u py(eg. Enr- nL6ov, or prerd.1,d(erq rouronotoriv Bdoerg rou rRNA onopo(rqreq yro ouyKerptpr6veg prBooropl169 Ler- roupy(eg. H EIK0NA 24.46 ouvor|((er ttg O5oerg mo 165 rRNA nou rowonororiwor Ue rn ouyrerpr;.uivn npo- odyyroq. Mro 5v6er[q yro rrl orlproo(o rou 3' dr<pou rou 165 rRNA nop6leror on6 r4v euoroOqo(q rou oro 0ovorn- <p6po nopdyowo r<ollolvq E3 (colicin E3) nou nopdye- ror on6 opro;r6vo Borrrlpro. H ro)lo(vq dtoond -50 vour<,l,eor(6ro on6 ro 3'dxpo rou 165 rRNA rou E. coli. H drdonooq r<oropye( ewelcirg rrlv 5vop(4 rqg prerd- epoonq. l-1oLf.5g o4prowrrdq Lemoupy(eg onoroUv rqv neproXri nou droondror: q np6o6eon rou nop6yowo lF-3, n ovoyvtilpto4 rou mRNA r<or 4 np6odeon rou tRNA. To 3'dr<po rou 165 rRNA eprn).5r<eror dpeoo mqv ow(6poor1 6vop[qg prSoo (euyopd:prorog pre qv o].].r1- LouX(o Shine-Dalgarno on1 06oq rou prBooropror(ou yro np6o6eon oro mRNA (BL6ne Etr6vo 24.12).'Evoq 704 KEOA Al0 24 Merorppoo1 Boorxeg ewong
  • 196.
    flposroteuUevo ond ToP{RNA I (tl ltnl ti Kovr6 oqv npdodeon -rou EF-G Kewptxfl nepLolrl IIvdeoq unogovddtov I :.:lr1 opeoog p6Log tou 3' dr<pou tou 165 rRNA mq -::cepoorl ovodetKvUerqt on6 ttg t6r6qteg petol'l'oy- -::Lrv ov0erttrorv mqv KooouYKopur<(vq (kasugamy- : ^ ro ono(o 6e 6to06touv optoprdveq tpononotloetq -:- 165 rRNA. H KooouyKoUUr(vr1 ovoot6L).et tqv . .:clr1 rqq pretdgpooqg. Av0errtrd preroMdyUoro rou -- .ou ksgA eprgov((ouv ovendpreto prtoq preOuLdoqq -: r srodyet t5ooeptg ;reOutroprddeg oe 60o yetrovtt<5q :Ii,,'[veq oe Uro neproX4 xowd oro 3' drpo tou 163 'r"A, H preOul.(too1 drlprtoupye[ qv e[otpettr<d ouwq- :-*evl o).Lq).ou1(o 6-602A-m6zA, 11 ono(o Bp(or<etot -:lc oe npoKopuortKd 6oo r<ot o' suKopuolrrr<d pLrpd ':'rA. H peOuLttopr6vrl ol'r;).ou1(o egnl'6xerot mqv ., *cl rov 30S rot 50S unopovd6cov, q ono(o pe r1 :..:o rqg en(or1q oXet((etot pe tr1 6tottlpnon rou . . :crrrlpLou IRNA mo n).r1peg prBootl;rdtto. H xo- 3'gei(ov nepLoXrl oouyr<opux(vq nporoLe( oneleu 05ptoon rou f Met-tRNAr on6 ro euo(o04to (pe0ul.ttrlprSvo) ptBootoprdrto, oMd t' ovOerrtrd ptBootogdtto e(vot tt<ovd vo dtornpoUv ro evoprcr4pro tRNA. 'Orov ro ptBoo<,Dpdrto dpoorrlptonotoUwot orn Ue- rdgpoorl, ougBotvouv oL).oy6q orrl 6opti rou 165 rRNA, 6nog eo(verqt on5 rrlv npocrroo[o ouyrexptpr6- vtov Bdoeov 6vowt XnUKIiq en(Oeor1g. Ot (eloptor6g 06oetg Kqrordooowor oe Liyeg opddeg, ot ono(eg ouy- rewpc0vowot orrlv 3'etrdooovo Kor rnv r<ewpxti nepro- 1q. MoLov6tt ot evron(oetg e(vot dtdonopreg otrl ypoprpu<r1 otrrlLoufotou 165 rRNA, orrlv npoypronr<6- rnro or 06oerg ttov Bdoeov nou epnldxowot orqv (6to tretroupy(o prnope( Bp(orowot t<owd 11 ;rto orrlv dtr'q finv rprroroyn 6oUn. Optop6veq on6 rtg ol,oy6g mo 165 rRNA nupo- ll lll Bprjxoq llllll ll lll I 2. orn 06on A Kotomotrn 0dor1 492 orn e6on A TNV Ue0utrfoon oro ksgl - IuuntrnpouoTtKo Ue To mRNA - AKp(pero 3'eldooov neproxn : r ;i: MepLr6q 06oeLg rou 165 rRNA npoororeuovror ono Xrlgtro0g ovLyveur6g 6rov oL 50S unopovodeg ouvduooro0v pe rtg :,: lnoUovodeg 1 ro optvoorutrotRNA npoodeQouv oU Oeon A. Atrtreg etvot 0eoetg perotrtro(eov nou enqpeo(ouv q peroqpool. 0t 05- ,. , <oroorotrflq TERM pnope( vo enlpeo(ouv rov repporLopo oe pepLro rt notrtro roDtxovto In[lS. To UeVotro SYXptlUo ntro[oto defivouv - : eoosprq neptolSg rou rRNA. J,.: 1= To 165 rRNA Aro6po;rori(er Evepyo Potro oq Merorppool 705 Korqorotrn TERM
  • 197.
    llemduho-tRNA Ilemdtr1 oluoi6o I H.N I H.C.R I ,C. 00I r IRNA I V flenldrxfldtuo[6o I llemdml duo(60 I H.N I H.C.R iH 0" '11 ' H-i-nI H ,C.. 00'0It IRNA IRNA EIK0NA 24.4S 0 oXlgologog nenldxoU OeogoU npoUno0Srer o[eo-Booxrl KoTotruon, K0T0 Trlv ono[o 6v0 dropo H UeTqqdpeTor o' 6vo pooxo KoTotrorno. 24.2O Or PrBoourprKdq AopCq AAAd(ouv 'Orov luv6uooTo0v or Ynopov66€q & '0r0v oXqpor((ovror ntr(pq pLpootol.rotto, 11 regotr1 rtllv 30S unopovoDtr.rv neprorp6qerol yupo ono rov ouXrlvo. * To evepy6 revrpo qg nenrrDutrorpovotpepoolq orrlv 50S unopovodo elvot nro dpoortro oro ntrr1pr1 prpoorolrolo ouy- rpurro ge rtq pepovrr:p6veq 50S unouovooeq. l-1o).).59 Sprpreoeq evdei(etg uno6qtrtrrvouv 6tt q 6opq rtov felcoptmtrlv unopovd6rov oL).d(er notrU 6rov outdg ouv6uomoUv yro vo oXrlpor(oouv 6vo nqpeg prBooroprduo. Or 6rogop5q crlq npoq rqv euoLo0qo[o rrr.lv rRNA o' e[oyeve(g nopdyoweg e(voL 1-rto on6 rrg toXup6repeg ev6eferq (B].6ne Ev6rrlto 24.18). llto dgeoo, ouyr<p(oerg r<pumoLLrr<cbv 6oprorv uQr;- l.r1q ovdl.uonq rov [eXcoprorcbv unogovddtov pe 14 dopq logqL6reprlg ovdLuorlg o6r<).qpou tou ptBo- orrr po-r[o u uno 6r1].rilvo uv rnv Un o p [r1 oq povrtr<rrrv 6rogopd:v. Aur5g or on6Qerg enrBeBorcb0qrov cm6 1.rro xpumoMrxrl 6opu1 rou prBooropot(ou rov E. coli oe en(ne6o ov6Luoqg 3.5 A, q ono(q enrnAsov rqu- 'ronore( 6Uo 6ro<popelr<69 dropropgcioerq rou prBo- o<r: gor(ou nou (o<.r:g owrnpoo0Jne6ouv dto<popefl r<d orddro orq prerdqpoo'r1. O xpUoroMoq nepr6Xet 6rio prBoorr:prdno ovd govd- 6o, ro xo06vo pre 6rorpopenx{ 6ropr6p<ptooq. Or 6ro<po- p6g ogefi.owor oe oMoydg orq 6tdto[r1 rov neproloiv p6oo oe rdOe unogovd6o. Irrlv nro o41.rowu<ri otroyri, q regotrr] rnq UrKpriq unoprovd6og neprorp6geror rotd 6" yfprrr on6 rqv neproXri"rou ou16vo npoq rn 0dor1 E. En[or1g, Uro neprorporp{ 6" r<ord qv ow(Oerr1 xote6- 0uvoq noporrlpefuot orrg 6o;159 6ogdg rrov ptBoorol.ro- ilrov rou Thermus thermophilus nou npoo66vowor oro mRNA Kqt 6ro06rouv IRNA 16oo orrl 06oq P 5oo r<ot orrl 05oq A, Aur6 unodqlc0ver 6n q regoAq pnope( vo neprorpoqe( ouvoAxd r<ord 'l2o ovd),oyo pe ro orddro rqq gerdrppoonq. H neprorpogrlrqq xego).r1q oxotrou- Oei qv nope(o rov IRNA 6rop6oou rou prBoorotrlo{ou, eye(powog rqv nt0ov6r4to vo puOpr((er q perox(v4oq tov mRNA ror IRNA. Or oll.oy6g r4g 6ropr6pqoonq nou ouprBo(vouv 6rov ouvduomotiv ot unoprovd6eq e(vor notr0 nro 5164- treg orqv 30S nopd ornv 50S unoprovddo. Or oMoy6g ;rdMov oXet((owor Ue rov 6leyXo rqg 06oqg Kor rnq perox(vqoqg rou mRNA. H oqpowrx6repq oMoyrl orqv 50S unogovd6o o<popd oro r<6wpo rqg nemrdul,orpov- oEepdoqq. Ot 50S unopovd6eg e[vor -1000 <pop6q ].t- y6repo onoreLeoporu<5q crrlv rordLuoq qq oUv0eoqq nemr6u<ri:v 6eopuiv ouyr<prxd pe to nLtjprl pgooopd- rro: o tr6yog (oog e[vor 1.uo oMoyrlqg 6opr1g nou ro- noOere( to un6orpopro nro onotetreoportrd oro evepy6 r6wpo rou nLrlpouq ptBoorr:1.ror(ou. 24.21 llepiAnQn 'Evq rodx6vro rou mRNA ovoyvrrlp((etor on6 6vo o1-uvooxulo-tRNA, ro ono(o 6rq06tsr 6vo owu<ro6rr6- vro ouprnlqpcoporrx6 rou rol6u<ov(ou r<ot peroq5per ro oprwolrl nou owrororXe( oro xodrx6vro. 'Evo er6r- r<6 evopr,crjpro IRNA (fMet-tRNAt orouq npor<opuri:- req Kor Met-tRNAi orouq europuciteg) ovoyvrop[(er ro rodrK6vro AUG, ro ono(o XprlorUonorefuor orrqv 6vopEn 6trcov rrov r<o6 r rono rqrrxcilv oLLrlLo uXrrriv. Itoug nporopucirteq Xpnorgonore(ror en[or1g ro GUG. M6vo rq Ko6rK6vrq repgoloUo0 (Xrop(q v6r1- ;ro) UAA, UAG ror UGA 6ev ovoyvopi(owor on6 opr- voorui.o-tRNA. To prBooroprdlo oneleuOepcivowor on6 q pretd- gpoon xor ero6plowor oe Uro 6e[opevrl e].eU0epov pr- Booropor(crlv, ro ono(o Bp(or<owor oe rooppon(o ge [epopror6g pur<p5g xor geyd].eg unopovd6eg. Ot trru<p6g unopovd6eg npoo65vowor oro mRNA r<ot mq ouv6- Xeto evc0vowor trre peydLeg unopovd6eq nopdyowog 6vo nl,rlpeq ptBooroprdrto nou ovoLopBdver q prerd- 708 KEOA Al0 24 Merdqpoorl &Bdon Boorxdg euvoreg
  • 198.
    qpoon. H ovoyvc0proqUroq npoKoputonrqg ogeqp(oq nepil.opBdver rrlv np6odeoq puog oLLrlLouX[og mo 3'dxpo rou rRNA oro pror(Bo Shine-Dalagarno nou Bp[- oKeror npw on6 ro ro6u<6vro AUG (n GUG) oro mRNA. H ovoyvtirptoq ev6g europuolxo6 mRNA nepil.qrBdver np6o6eoq mqv 5' rol.Unrpo: mr1 ouv61ero, 11 purprl unopovd6o perovooreOer orq 06o4 6vop[qg ovo(rpdrv- tog xo6u<6vro AUG. M6Lrg ovoyvrrlp(oet 6vo rordMqLo xtodrx6vto AUG (ouv4Ooq, ro npcirro nou ouvowd), ou- 6ud(eror ;re puo preydLrl unoprovddo. 'Evo prBoorrlprdno pnope[ vo pretorpSper rour6lpovo touLdXrorov 6rio o1-rwooxulo-tRNA: n e6on P rotoLopr- Bdveror on6 6vq noLunemr6u).otRNA, ro ono[o pero- rp5per rqv nol,unenrr6rrrl oLuo(6o nou 5Xer 4611 ouweOe(, evcil q 06o11 A 1pr1or;ronoe(ror yro qv e(oodo ev6g o;uvooxutro{RNA, nou preroqdper to en6pevo o1.u- vo[ri ro ono(o 0o npomeOe( orrlv o].uo(6o. To Bor,rrl- ptord prBooopdrro en(oqg 6ro06touv puo 06o4 E, 6ropr6oou qg ono(og 6rdpXeror onoorutruoprSvo IRNA, nporori oneLeuOepco0e( perd rn Xprlorpono(qor1 rou orrl perdgpoorl. H notrunemrdu<{ o},uoi6o orq 05or1 P petog6petot npoq ro ol.rlooxulo-tRNA rrlg 06oqg A, 6rlproupyc0wog 5vo onoorul,uog5vo IRNA orq 06oq P xt dvo nemr6u).otRNA orq 06o4 A. Merd rq oUv0eoq nemr6xoti deopoil, ro ppooroprd- ro peroron((erot rord 6vo rodx6vro n6vo oro mRNA, perorwcilwoq ro onooKulru:;r5vo IRNA pSoo orq 06o11 E r<or to nemr6ulo-tRNA on6 q 05or1 A orrl 06oq P. H petot6ntorl roroL0eror on6 rov nopdyowo enrpn'1xuv- oqg EF-G ror, 6nog noMd dMo orddro rrlg )reroup- y(og tou ptBoorrrprot(ou, onone( u6p6Luoq GTP. Kqrd rr1 peror6nroq, ro prBoorrlprdno 66p1etor on6 puo uBpt- 6x4 xordmoon ornv ono(o q 50S unoprovd6o peroxt- ve(rot tog npog rnv 30S unoprovd6o. H perdqpoorl efuor 6onovqpq 6upyoo[o. ATP pq- orponore(ror yro nopoXrl ev5pyerog oe noMd m66tq, preto[6 dltrrov ro rp6prrr:1.ro rou IRNA pe ro o1.rwo(ri rou r<ot ro l,eilNypo rou mRNA. Ercrq-rdror 6tr 5trrg ror 90% 6l<ov rrov gop(olv ATP nou ouw(0ewot o' 5vo Bomrlpro nou ou(dver ypnyopo r<orovotrc0vowor orq ouvopprol.6- ynon rov o1.rwo[5rr:v oe nporetvegl Ie rd0e srddro rqg ;rer6<ppoorlg onomo0wqt ent- np6o0eror nopdyoweg. Ko0op[(owot on6 rqv xur].trr1 o0v6eoq ror 6rdoroori roug on5 to ptBootrlprdtto. Or nopdyoweg 6vop[r1g epnMrowor orqv dvop[r1 dvop[q orouq npoKclpurireq. O lF-3 onotte(tot yto rrlv np6o6e- orl rov 30S unopovd6<ov mo mRNA rt en(o4g efuot uneriOuvog yro rn 6rofipr1or; qg 30S uno;rovd6og o' eLe60epq Uopgn. O lF-z onone(ror yro rrlv np6o6eoq qq fMet-tRNAt or4v 30S unoprovddo ru eivot une0Ouvog yro rov onoxtreropr6 6trtrrov o1-uvoorul.o-tRNA on6 qv ow[6poor1 6vop[qq. To GTP udpoLtietot, og6rou to evoprcr{pto IRNA npoo6e0e( qro oU;rn}.oro Svop(qg. Or nopdyoweg 5vop[49 np6ner v' one].euOepo0otiv drore q geyor4 unopovddo vo prnope( vo ouvduoote( pre ro o6prnLor<o Svop(r1q. Iroug europutineg 4 Svop[r1 nepil.oprBdver preyo].rj- repo opt0pr6 nopoy6wrov. Oprop6vot epntr6xowor orrlv op1Kn np6o6eor1 rqg aOS unoprovdEoq orqv xofurintpo rou 5'6rpou tou mRNA, oqpe[o oro ono[o ro arupr<rr1- pto IRNA efuor npoo6e6eprSvo pe dM4 o;166o nqpay6rF tov. Merd on6 ourl qv oplrrrl np6o6eorl, n Utxp{ unopovd6o oopc0ver ro mRNA 5og 6rou ovoyvcopfoer to o<oo16 r<ro6rx6vto AUG. 'Enerto, ot nopdyoweg 6vop[qg onetreuOeprilvowor rr 4 605 unogovd6o ouv- 6ud(eror pe ro o0pnl,oxo. Or npoxopurrlnro( nopdyoweg enrtrr{r<uvoqg epn},6- Kowor. orqv enrprqruvoq. O EF-Tu npoodSver opuvooru- ).otRNA oro 70S prBooroprdrro. GTP u6pol0eror 6rov oneLeuOepc0veror o EF-Tu, evrir o EF-Ts ononefuor yto v' ovoyevrrqOe( q evepy6g Uopgri rou EF-Tu. O EF-G onomefuor orq peror6nroq. Iro ppoocoprdrro npoo66- veror efue o EF-Tu e[te o EF-G, yeyov6g nou 6roorpo],G (er 6n rdOe Bqgo np6ner vo o),oxLqpoOe( nporoU (errv{oer ro en6pevo. O repporroll6g ouprBo[ver oe onoro64nore on6 ro tp(o er6rd ro6x6vro UAA, UAG ror UGA. Or nopdyov- reg oneLeuO6poonq rd(rlq 1, nou ovqyvop(ouv er6u<d to rrrl6rr6vro repprortopro0, evepyonoro0v ro prBootogd- no oe u6p6Luon rou nemr6ul,o-tRNA. 'Evog nop6yov- rog RF rd[49 2 qnore(ror yro rnv one],euO5poon rou nopdyowo RF rd[49 1 on6 ro prBootoprduo. Or npoo- 6e6ep5vor pe GTP nopdyoweg lF-2, EF-Tu, EF-G ror RF3 6Xouv nop6poro 6op(: or 6tio rel,ewo(ot ;.u1.roriwor q 6ogrj RNA-np<oreivcbv rurv 6Uo np6rrov 6rov e(vor npoo6e6eg5vor oro IRNA. 'OLor rouq npoo66vowor orrlv (6to ptBooogrr( 06o1, tr1 06oq np6o6eonq rou nopdyowo G. To prBoocoprdrro efvor otr:poil6ro prBovoux],eonpro- reivdrv, oro ono(o ro preyolUrepo prSpog qg pd(og no- pdXetor on6 ro rRNA. To oXqporo 6Lcov tcov prBo- orrlpo{tov yevu<d e(vor nop6trroro, oor6oo to Borcrrlpro- xd p$ooco;rdtro (70S) dXouv neprypoge( Lemopreprirq. H pxpri unopovd6o (30S) 6Xer oupneopSvo ofnUo, U' 6vo "oriuo", nou nepdXa nepkou ro 60o rp(o rqq pd- (oq rot 6roXrr:p((eror on6 rqv "xego[rl" p6otrr ptoq oXt- oUnS. H geydLq unopovddo (50S) e(vor neptoo6tepo orporprrrj pre 6vo npoe[61owo ";r(o1o" mo de[td t<ot Uto (KewptKli npoe[oX(". Ot ewon(oetg 6ov tcov npol- reivrilv orrl ptr<pq provdSo e(vot yvoor6g rotd npoo5yyt- on. Kd0e uno;rovd6o neptdXet 5vo pr6vo x0pto rRNA, 165 r<gr 23S oroug nporopucbteg, 18S xot 28S oroug europurireg. Yndplouv en[or1g etrdooovo rRNA pe ne- proo6repo o[roorlpre(<oto to 55 rRNA orrl peydtrrl uno- provdSo. Kor ro 6rio xrjpro rRNA e;rrpovf(ouv ertero- ;i6vo (euydpcotrro Bdoeov, rup(cog un6 propgt'1 FpoXd- cov, orelr6g (euyoptog6vtov drnl,drv pr[oXutv Ue Uov6- xLrovoug Bp6Xoug. Iuwqpqpr6vo Xopocnptortxd oto rRNA towonoroUwqr ouyxp(vowog ttg ollqtrouX(eg pe rrg deureporoye(q 6ogdg nou pnope( vo oledto- oroUv yro ro rRNA noxfi.tov opyovtoprcbv. To 163 rRNA 5Xer r6ooeprg 6ror<ptr5q neproX6q, evd: ro 23S rRNA 6[r. To europurottrd rRNA 6ro06rouv entnp6o0eteg ne- pro16g. H rpuoroMu<n 6oUri 6e[Xyet 6tt q 30S unopovd6o 24:i l]eprtrnUn 709
  • 199.
    61et ooriprperprl r<orovopuiRNA r<or nporeivc0v. To RNA ouyrewpriveror orrl 6rdgoo4 Ue rnv 50S unoprovddo. H 50S unogovddo 6ro06,rer gto entEdvero npulteiVrlq, pe gor<pd poBS(o 6k).ovou RNA ro ono(o 6too{(ouv q 6o911. H oriv6eoq 30S pe 50S nepfiotrrBdver oqpre(o enogqg prero(ri rrov 165 rRNA r<or 23S rRNA. Aopru<5q oL).oy6q ouprBo(vouv Kor orrq 6Uo unopovddeg 6rov ouvdud(owor. ylo vo oXrlpor[oouv rivo n].r1peg prBooro- prdrro. Kd0e unoprovddo 6ro0drer nolLd evepyd rdwpo, ro ono(o ouyrewpdrvowor oill UrroepoqrrKq nepro- pi rou prBoocol.ror(ou 5nou ouw(0ewor or nporeiveg. To no).unenr(6ro eyr<oroLe[nouv ro prBoocoprdno 6ro- pr6oou qg neproXqg e[66ou, q ono(o ;rnope( v' ol,l.4- Lenrdpdoer pe Uro pepBpdvq. To rripro evepyd r6wpo e[vor or O5oerg P r<ot A, 11 05o4 E, or neproX6g np5o6e- orlq rov EF-Tu ror EF-G, q nenrr6utrorpovo<pepdorl r<or 4 06oq np6o6eoqg rou mRNA. H 6rog6prpoo4 tou prBootrrl-ror(ou gnope[ v' o],].d(et oe drdrpopo ord- 6ro r4g gerdrppoorlg: 51ouv ewonrore( dro<poprig oq npoq rnv npooneloorp6rqto ouyrerprg5vov nepro- 1d:v rtov r0puov rRNA. To IRNA orrg 06oerg A r<or P efvoL nopdMq],o peto- [0 roug. Or Bp61ot rrrlv owrrco6rxov(rov npoo6Svowor mo mRNA oe Uro qU^,oKo qq 30S unoprovddog. To un5trorno rpnpo rdOe IRNA npoo66verot orqv 50S unopovd6o. Mro evotrtroyr1 6rop6prpoo4g tou IRNA p5- oo orrl 05or1 A efuor onopofuqr4 c0ore ro ol.rwooruAu<6 dr<po rou vo Bpe0e( 6(ntro mo drpo rou nenrrdul,o- IRNA or4 05or1 P. H 05or1 qg nemr6ulorpovorpepd- onq nou ouv66er rrg O5oerg np6o6eor'19 P r<or A onorcLefuor on6 23S rRNA, ro ono(o eprpov((er xorotru- tu<r1 evepy5rqro nenn6ulorpovoqepdonq, pol,ov6rr yto vo 6r1proupyr10e( 4 ooorri 6opri prdMov onotro0v- ror Kor. npa:reiveg. O evepy6g p6log rtr:v rRNA mr1 pretd<ppoorl uno- 6errvUetot on6 peroMd[erg nou enqped(ouv rq Ler- roupy(o rou prBooro;ror(ou, on6 olLql,enr6pdoerq Ue to mRNA r<or ro IRNA, 6noq en(oqg ror on6 qv ovdy- xq drorlprlonq Eexoprfitirv oMq).ent6pdoeo:v (euyo- prilprorog Bdoecov pre ro IRNA xor ro mRNA. H 3'rehr<4 neprolq rou rRNA (euyopdrver pre ro mRNA crrrlv Svop- En. Or eororeprr6g neproX6g npoyporonororiv r6to(re- peg eno<pdg ;re ro IRNA 16oo orq 06oq P 6oo ror rq 06oq A. To prBooo;ur6 RNA e(vor or6Xog opropdvrr:v owtBronrcbv r1 dMrov nopoy6wrov nou ovoor6trl.ouv rrl prerd<ppooq. BrBAroypagio 24.4 H 'Evop[r1 0T0 BoKrrlpro Xpero(etor 30S Ynogovodeg Kr EnrKouprKouq nopoyovTeq .'1:,;j1;r 1,1 1 I 11;: I IVlaitra, U. (1982). Initiation factors in protein biosynthesis. ,4nnu. Rev. Biochem. 51, 869-900. Noller, H, F. (2007). Structure of the bacterial ribosome and some implications for translational regulation. ln "Translational Contr: in Biology and IVledicine." (Mathews, M. 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    24,20 0t Prpootopx6gAoUeq AIId(ouv'Orov luvDuoororjv or YnogovdDeg AvL:gcpdq Schuwilrh, B. S., Borovinskaya, M. A., Hau, C. W., Zhang, W., Vila- Sanjurjo, A., Holton, J. M., and Cate, J. H. (2005) Structu':. :' the bacterial ribosome at 3.5 E resolution. Science 310 82;- 834. Btptrroyporpio 713
  • 203.
    Avotun60qre 0nd J.Struct. Bioi., vol.157, K. Y. Sanbonmatsu and C.-S, Tung, High performance computing in biology , pp.470-480. Copy- right 2007, rordniv obeIoq ono Elsevier Ihttp://www.sciencedirect.com/scrence/journall1047B477). H rporoypogro nopoXcoprl0lre 0n6 rcv Kevin Y. Sanbonmatsu, Los Alamos fJational Laboratory. Tou ovTrKodrKov(ou Kor TIq rpiTnq Boorlg rou KodrKov(- ou nportinrer 0rt0 U X0tr0p6repq enurlplorl rou (eu- y0p6U0T0q ono T0 vouK)e0T[6r0 rou rRNA oTri prpoooprKn 0eon A. ffi fo IRNA l-lopoyovror pe Ene(epyooio Meyotrurepov llpri- 6poUtov Mop[rov * 'Evo rirprUo IRNA dlproupye[ror pe ercqepyoolo svdq npo6popou lLopiou. a To S'oxpo Dlptoupye(ror peoo droonooqg ono rr;v ev- dovourtredorl RNAdol P. e To 3'oxpo 6lproupyeiror geoo notrtrontrov ev6ovou- rtreotrunxdrv rt e(tovourLeotrurtrdrv droonooeov, ng ono[eg oro],ouOei q npoo0qrrl rou rorvou rep;lorxof rprvourtreond[ou CCA" ffi fo IRNA l-lepr6Xer Tpononoulpeveq Booerg * To IRNA neprdXouv novo on6 90 rpononotqlreveq po- 0etq. s H rpononolr]on ouvn0oq ouvlorotor oe dpeol otrtroyr1 rotv nporoyevtirv pooeov rou IRNA, oorooo undpXouv oprop6veg e[op6oetg rinou pro poorl otpotpe(ror xr ov- rlro0rororor on6 Uro otrtrn. O I-eveTrK6q I(66lroq Iohn Perona *iHg ttooY(')Yn ffiff9 Iuyyevfl Ko0rxovLo Avlnpoooneuouv Xqgrrog l-loptlpoto A;uvo[60 * EEnvro pto on6 rtg e(rlvro rdooeprg rprntr6reg rtodrro- norouv e(xoor ollvo[eo. e Tp[o rtodrrdvro dev ovrrnpootoneuouv oprvol6o ror nporoLouv repportop6. e 0 yevelroq 166rro9 unoyooe, oe nptrtrl.to orodro rqg e(elt[1q ror elvor orroupevxog. a To neprooorepo ollvo(60 ovlnpooone0ovror onri 6uo 11 neprooorepo rtodtr6vto. * To notrtrontro xrrrdtxovro yro evo oluvo[u ouvqOorg eivor ouyyevrl pero(u rouq. * XrluLrtlq nop6poro olitvo[eo ouXvo rlXouv ouyyevrl ro- 6ixrivto, etroXLoronordrvrog eror rtg enrnr6oetg Tov Ue- rotrtroIeov. ffi H Avoyvoprol KroDrrovrou-Avlxtodrrov(ou llepiloggover Aoro0eLo * l-lotrtrontro xrodx6vro rou ovTrrrpooone0ouv ro [6ro o1u- vo[t1 ouXv6repo drorpepouv oq 06or; rqg rprqq pooqq. e Aoro0ero oro (euyopcolro pero{f qg npdrr4g pooqq 714 KEOA Al0 25 0 l-everuoq Ktil0rxog ,@, ,! lr I I t'tt l, i, i-. ?a' .i F- '-- r-- . *'t::>- * tF4 &- t/ - @t,
  • 204.
    ffig* e 0r TponoroulueveqPooerq npoo6[60uv ouEnudvn oT0- 0eprlqro oro tRNA ror pu0p[(ouv t1v ovoyvtitptorl rouq ono nporelveq rot otrtro RNA rqq peroqpoorurlg ouoreuflg. 0L Tpononoulpeveg Booerg Eqpeo(ouv ro Zeuyoptolto AvlrolD lrov(o u - Krr:d rrov(o u e Tpononorrloetg oro ovtrrtrldrrrivLo enrqpeo(ouv ro npo- TUno Tou ooro0orlg (euyoptirporog rot ouvendtg e[vol olpovnr6g yro rov xo0optogo rrlg etdtrdrrpog tou tR- NA. YnopXouv InopodrxSg Tpononou;oerg rou Otroupevtrou K6drro e Ie oproutlvo erdn 6xouv ouppei otrtroytlg oto yevenrd rdl6rro. * Aur6q ot otrtroydq e[vor ouvl0eoTepeq oT0 UrToXov- dpLoro yovtduirl-roro, rinou pnope( vo rorooreuoote( 6vo rputroyevertro devrpo yro rLg otrtroy6g. * Iro nuplvuo yovrduirporo, ot otrtroy6g ouvrl0tog enq- ped(ouv p6vo to xoDrrovro reppolopori. Neo Aprvo[60 Mnope[ vo Evotrlgoro0ouv oe 0propdvo Ktrl6xrjvro Tepgortopou * H evootuoroorl qg oetrlvoruoreivrlg oe oplop6vo ror- drxdvro UGA onoue( rr; Dpool evoq oouvl0rotou IRNA oe ouvduoogo Ue notrtr6q nptorelveg. e To oouvrl0roro ogrvo[u nupotruolvr1 ;lnope[ vo evoouo- rol0el oe oprop6vo rtrldrrrjvro UAG. c lro 0treQoptdoQtipo nporo(oo Euplotes crassus, T0 roduovro UGA ro0opt(er rrioo rr1 oetrlvoruoreiV4 tloo rot rrlv xuoretVl. ffi fo IRNA Zeuyopdrvouv Er),exlrd pe Aprvo[60 Meoto Aur- voorutrotRNA Iuv0eroodrv I 0r ourvooKutro{RNA ouvOerooeg e[vor 1uo orroy6veto ev(0Ur,rv, ro ono(u npood6vouv oluvo[eo oro tRNA nu- pdyovrog ol.uvoorutrotRNA oe pto ovridpool 611o pq- pdrtov nou Xprlorgonore[ ev6pyeto onrj ro ATP. * Kd0e outvoorutrotRNA ouvOeroorl oluvoorutruiver dtro ro IRNA oe pro roodernr1 opo6o nou ovlnpoooneUer 6vo ouyrerpq6vo oluvoIu. * H ovoyvdrproq rou IRNA ond rtg tRNA ouv0erdoeg po- ol(eror oe pLo ouyrerpqr6vq ogo6o vourtreortdltov, qv (oud60 tourdqrog, rou IRNA, ro ono[o ouXvo ouy- revrptirvovror oro ot6treXog rou onod6rrrl ror ro ppo- Xo rou ovlrtodxov(ou. ffi 0'AgrvooxutrotRNA luv0erooeg Kororoooovror oe Arjo 0rroy6vereg I 0r ourvooKutro- IRNA ouvOetooeg Dtorp(vovrot ortq ot- roy6veteg ro[qq I rot ll pe pdoq rdrofrepo Uor(00 otrtrn- trouXuirv xot rdtolrepeg 6opxeg neproX6g. ffi O, Iuv0erooeg Xprlorgonoro0v q Ar6p0ooq Aortplov yto vo Be),rtiloouv rnv Arp(BeLo * H etdrrouro rou (euyoptirlrorog pero[f oprvo[6ov xot IRNA puOpi(eror ;.r5oor ovrtdpooetov 6rop0oorlg 6orr- p[ov or ono[eg udpotruouv troOoq oxrlU0logeveg opr- voorutro-odevutrx6g evdroerg ror oluvoorutro-tRNA. ffi fo Koroorotrlro IRNA l-iepreXouv Metotrtroy;r6vo Avrtro- drrovLo, ro 0no[o Aropo(ouv N6o Kodtrovro e'Evo xoroorotrlrd IRNA runrro qeper pLo perotrtr0[r1 oto ovrtraldxovto nou otrtro(ei t0 ro6txov,o J:i : ,:,": onovro. * Otov ro vto ovlrtodtrovto ovrtorotXel oe rol6t<o;r, reppottoltou, evotopordtverot 6vo optvo[0 rot q no],tr- nenl6rrrl otruoido enerre[vetot n6po ond ro xtr.tdtxcl- vro TepUoTtopou. Aurd o61yei oe oouvOertr1 K0T0o-I0- trrl oe Uro 06o11 oouv0ertrnq Uerotrtro[lq 1 oe ovoyvtrt- o1 nepo ono 6vo rovovtrd rotdtrovto repportopou. * Auouv0elu'l rotoorotrrl ouppoivet dtov ro IRNA ovo- yvtop(er dvo drorpopenrrj rtodx6vto on6 ro ouvrl0to;t6- vo, onore 5vo olLtvo[tj ovrtro0tororot ono ronoto dtrtr0. ffi*,} Ynopxouv Aouv0erxol Kotomotrelg yto Kd0e Ko:Dtrovto Teppolopou. s Ko0e runog oouv0ertrou xodtrov(ou roroor6trtrerot on6 6vo tRNA pe perotrtroygevo ovrtrotdtrdvto. * Oprourivo onovto roroorotrlrd IRNA nept6Xouv perotr- tro[erg oe dtrtro rpt1poro tou ltoplou. ffi O, Koroorole(g Mnopei v' Avroyovi(ovror qv Kovovtrq Avdyvororl rou Ktildrro * To roroorotrtrro IRNA ovroyovl(ovror ro IRNA oypt- ou runou nou nept6Xouv ro (6to ovrtrtrldtr6vto yto rr;v ovoyvtrlol rou ovr[ororxou rordrrov(ou (tl 1(l]6txo- vitov). e H onoretreoUonrrl roroorotrfl eivor en$tropr1q, enetdrl odlye[ oe ovoyvtrloq n6po ono r0 rovovrro rodtxrivto reppolo;rori. * To rtrldtr6vro UGA epqovr(er droppon xor dtopo(eror eoqotrp6vo ond ro Trp-tRNA pe ouXvoqro 1 -3%. ffi fo Prpooorgorto Ennpeo(eL rrlv Arpipero qg Merdqpooqg a H 6o[rl rou 1 65 rRNA ong 06oerq P ror A rou ptpooo- por(ou eqpeo(er rqv oxp[pero qq petoqpootlg. ffi: Merordmorl Tou ll),oto(ou Avoyvoorlg Mnopeivo luppei oe 0trrofipeg AtrtrntrouXieq * To ntro(oto ovoyvtrlorlg enrlpeo(eroL ono rrlv otrtr1trou- Xio rou mRNA xor ro prpootolrLr6 nepLpotrtrov. a 0t otrtoOnpeq otrtrntrouxieq enlrp6nouv o' 6vo IRNA vo peroroni(erot roro pro Boon og6rou (euyoprioer pe ro ovlrodrrdvrd rou, petopotrtrovrog 6rot ro ntro(oto ovo- YV(00nq. er H UerdQpool optop6vtov yovtdltov eloprorot ono rrlv roxnx1 npoypopUoTropdvrl perordnroq rou ntrotolou ovoyvtoorlg ffi afio luppovro Enovoxodrrono(norlq: Merorppoortrfl l-lo- poropqr; ror o MlXovrolrdq rou tmRNA yto qv Anetreu- 06ptrro4 rtov Ko0qtrtop6vrov PLpooropot[ov e H noporouqlrl ouvfororor oqv xovtirr[o rou ptpooto- ;ioriou vo reppori(er q peroqpootl, v' onetreuOepdtve- ror ono ro mRNA rot vo (ovopX((et rI Ueroqpootl nep(nou 50 vourtreor6ro ro0o6rro. * To pL0ootouorto nou e(vol xo0r1trotg6vo novo oro mR- NA pero rr; peprrrl ouv0eorl prog nporreiVlg pnopei v' onetreu0epo0o0v ono rr1 6poor1 rou tmRNA, evoq po- vodxori RNA pe Xoporqprorxo rooo rou tRNA 6oo rot rou mRNA. ffi nepfi1qrl 715
  • 205.
    25.1 ErooyoVri H ol,i.4).ouX(oev6g ro6rronor4rrroU r<),c0vou DNA nou droBd(eror oe rore60uvoq on6 Shpog 3'qnore- l,e(ror on6 rprni.6reg vou xl.eorr6(rov (rrrl6x6vto) owi- ororxo pe rnv oMrlLoul(o o;uvo{6rov ptog nporetvrlg nou 6toBd(eror on6 ro o1-rlorelrx6 npog ro xopBo(u- tetrrr6 drpo. H oMq),oriX4orl rou DNA Kor rov npo- teiVciv enrpdner qv dpreoq oriyrproq rov vouxl,eo- rt6xc6v Kor rov opuvo[rr6v oM4].ouXrc0v. Yndplouv e[qwo r6ooepo rco6rx6vro (xoOrivo on6 rq rSooepo nr0ov6 vourtreor(dro prnope[ vo r<oro],6Ber pro on6 rg tpetg 06oetg tou rodu<ovfou, 6qpuoupyc0wog 43 = 64 ntOovdg oMrl),ou1(eg rpwoux,treorr6(ov). Ko06vo on6 ourd ro xo6u<6vro 6Xer er6u<5 v6qpro orq perdEpooq: el{wo 5vo r<ro6rxdvro owrnpoooneUouv ol.uvofdo, evci ro un6l,orno rp(o enr<p6pouv rov repporropr6 r4g perdgpoor;9. To v6qgo ev6g xur6xoviou nou owrnpooone0er 5vo o1-uvo(r3 r<o0op((eror on6 ro ow(mot1o IRNA, evdr to v64pro rcov rcodxovfrov repgonoproU r<oOop((eror dpeoo on6 nporeivu<oriq nopdyoweg. H onorpumoypdrprloq rou yeverrKoU xril6u<o opXt- xd 66er[e 6r or yeverrr5g ilrr1po<pop(eg ono0qr<eUowor und poprpfl vouxLeorr6rxrilv rprnlerriv, om6oo 6ev onoxdLurle noro opuvo[ri xo0op((eror on6 rd0e 11116r- r6vro. [lprv on6 r4v 5].euor1 qg oMq],otixnonq, ro v6r1- Uo rov r<rodrrov(ov npoodtopr(6rov pe Bdorl 6Uo r0noug preLercilv in vitro.'Evo o0m4;ro nou Boo((eror on1 perdqpooq ouvOenxriv noluvou xLeortd(cov eroriX- 0qre ro 1961, 6rov o Nirenberg 66er[e 6n to no],uoupt- 6uLu<6 o[6 [no].u(U)l roreuOrlver rrl ouvoppo].6yr1or1 qg <porvuloLov(v4g oe noluEorvul,oLov(v4. Aur6 ro onor6l,eopo oqpo(ver 6rr ro UUU npSner vo e[vor ro rco6rx6vro yro rrl eorvul.ol,ov(vq. I' rivo 6e0repo, Uero- yev6orepo o0mr1po, 6vo rptvouxLeor(6ro Xprlorpoilor- q0qre yro v' onoprprlOe[ 6vq rcodu<6vro, rdvowog ro ow[mor1o ol.uvoor<u],o-tRNA vo npoo6e0e( oro ptBo- tlpc,tq pdorl UCAG oropdrro. Towonorc0wog ro opuvo(ti rou ol.rloorulo. IRNA prnope( vo BpeOe( ro v6q;ro rou r<ro6rrov(ou. Mo(i, ot 6rio re;yrr6q npoo6r6proov ro v6rlgo 6trcrlv rrov xro6r- rov(rov nou owlnpooconeUouv oprwol5o. E[rlwo 6vo qn6 ro e[rlwo r6ooepo rrrlSrr6vro owt- npooconeUouv o1.uvo(5o. To un6Lorno rpfo enrg6pow rov rspporroU6 rnq ;rerdgpooqg. H on66oo4 oprwo[6- ov oro r<rrl6x6vro 6ev efvor rulo(o, oMd unoyopeUerur on6 q46oerg ong ono(eg q rpfuq Bdorl 51er pu<p6tepr1 en[6poor1 mo v64pro rou r<ro6rxov(ou. Enrn].5ov, ]3p1. r<drg nop6poro oprrvo[6o ouXvd owrnpoooneUowor on6 ouyyevrl ro6u<6vro. 2 5.2 Iuyyevfi Kto6rr6vro AvTtnpoooJnEUouv Xnprrtiq [op6Uoro Aprvo[6o * E[nvro p(o on6 rLq e{qvro r6ooeprg rptntr6reg rtoDrronolouv e[rooL o;-uvo(eo. * Tp[o xroDrrovro dev ovrrnpooulneuouv olrtvo[60 Kor ttpoKo- trouv reppoloprl. * 0 yevelrdq 166rro9 ((r0y00eD oe npdrrpo orodro r4g e[etrr- [1q xor ervor orxoupevtrog. * To neprooorepo oprvo[eo ovrtnpooone0ovrot ono 6uo n ne- ptoo6repo rtrldrrovro, * To notr),ontro xrodtrdvro yLo tlvo o;uvo[0 ouvrl0oq e[vor ouy- yevl gero[u roug. $ XnUrKdq nopo;.roro oluvo[60 ouXvo eXouv ouyyevrl rodtxovro, e),oXroronouirvrog 6ror ng entnrtloerg rov perotrtroIeov. O rdl6u<og ouvotf((eror orr;v IIH*tiA 15."i. Yndplouv ne- proo6repo rto6tr6vro on6 opuvo[5o: dpo, oXe56v 6].o t' oprvo[6o owrnpooone0owor on6 drjo q neproo6te- po r<rodtx6vro. Movodrx6g e[orp6oerq e(vor 11 preOerov(vq Kor n rpurrogdul. To rcodrr6vro ge ro (6ro v6qpro r<o- Lorjwor ouvtirvupo (synonymous). O yeverrx6g r<or6L- Koq ornv npoy;,rotrx6tqto 6roBd(eror ndvo oro mRNA r<r StoL ouvq0og nepLypdgeror oe o16o4 pe lg rSooe- prg BdoeLg nou undpXouv oro RNA: U, C, A r<oL G. To r<co6Lr6vro nou owrnpooolneUouv to (6to n XnUr- r<org nop6proro opuvo(6o ouvrlOotq 61ouv nop6proro otr- l.rllouX(o. IuXvd, n pdon orrlv rp(rr1 05or1 evdg r<ro6tr<ov(ou 6ev e(vor orlpowu<r1, ener6q to rriooepo rr,,.t- 6tr6vLo nou drorpdpouv pr6vo orqv rp(tr1 Bdoq owrnpo- ooneUouv to (6ro opuvo[ri. H ;reulp6vq erdrxdqro orqv relewo(o 06oq e[vot yvotmrl otg exgr3trroq rpir4g B6- oqg (third-base degeneracy). Fro vo perorppoore[ 6vo ro6u<6vro rou mRNA, r] npcD- eiK&l{A :5.1 Otreg or rprntr6req-rtodrrovto 6Xouv vdqpo: e[rlvro gio ov- rtnpoooneuouv o;-uvo[eo Kor Tperq nporotrouv i1i1 (ST0P) FO b.6 =a P= UUU l_Phe ucu I UAU -LTyr UGU -LCys UUC -] F UCC I Ser UAC -] Y UGC -] C UUA LLeu UcA f S UAA l_(rnD UGA SToP uuc _r L ucc _.1 UAG I "'"' UGG lll CUU I CCU I CAU _LH|S CGU I 696 | Leu ccc I Pro cAC I H cGC I Rrs CUA f L ccR f ecnnleln CGA [n CUG -] CcG I CAG -] O CGG -I AUU l ,- ACU I AAU I Asn AGU I Ser AUC l": ACC Im,nncf ru AGCl-s tut I g, ffit l-' titl'il' t3t lT GUU I GCU I GAU I ASP GGU I GUc I va Gcc I Aa GAc f D ccc I GIy cuA I v GCA l- A GAA lGtu GGA f G GUG -] GCG -] GAG -] E GGG -] 716 KEOMA|0 25 0 l-everxdg Ktildrrog Boorxeq ewoleg
  • 206.
    rn 96on rounpSnet vo (euyopdroet ge rqv npc0rr; Bdorl rou owtrrodrxov(ou tou ow[orotXou ol.twooru].o-tRNA. Aw6 ro (euydpropro ouprBo(ver pr5oo oro p$oocoprdtro, 6nou 11 oM4Len(6pooq ;reto[ri oupdrqpoptottt<tilv tpt- vouxleon6[tov oroOeponotefuot pSoto e[openrd ourrrl- prlpr6vov voux.l.eott6(tov tou 165 rRNA orrl 05or1 A. H ouorqpn enrnpncnl rou ouvolu<o0 o;q4porog tcov (euyo- polpr6vov Bdoeov on6 to rRNA enmp5net vo npqYporo- noq0e( 51t pr6vo ouprBcnrx6 (euydpopro peto[0 A-U t<ot G-C ong 6rio npcirreg 06oetq tou ro6tt<ov[ou qI d Kt en[- np6o0ero (euyoprilproro or4v rptrrl Bdor1, 6nou ot eTIo- <p6g rou rRNA e(vot Lty6tepo ouprntrqpoprorxdq. Iuve- nri.rq, 5vo ol.rwooruLo-tRNA prnope( v' ovoyvrop((et ndvol on6 5vo r<todx6vto pt5oto rov enmp6o0erov, Ul'l Kovovt- r<drv (euydrv nou enrp6nowot on;v rpkq 06o11. Enmtr6ov, or olrltrenr6pdoetg tou (euyopdr;rotog (oroq enrlped(ov- ror on6 rrl pero-Ueroypogrrrl tponono(qoq rou IRNA, rd(oq ;16oo r1 og6otr:g 6inLo oro owxtodu<6vto. H rdoq Xqpxtirg nop6ptouov oprvo[5tov v' owmpooo- ne0owor on6 ouyyevn rto6tr6vto el.oXtoronote[ lq ent- mcboerg rcov gerotrtrd[eov. Au(dvet rrlv nt0ov6rr[o Uto pr6vo ru1o(o oUoyt Bdoqg vo prlv npoxotr6oet owxotd- orooq oprwo[6og r1 vo o6qyrjoet oe qwKordqroorl nou nepil.oprBdver optwo(5o nop6protou loporcqpo. l-to nopd- 6erypo, 4 predMo[r1 CUC npog CUG 6ev 6Xet xdnoro en[- nroorl enet6rl rorro d[o ro6tx5vto qwmpoooneUouv rq leur(vr1. H perdllo[q CUU npog AUU 5Xa og onor6ls- o1.ro owu<otdoroon rnq treur<(vr1q on6 tooLeurtvq: ourd ro 6rio oprvo[5o e[vor u6p5tpoBo rot nt0ov6v 6todpopro{- (ouv nop6proro p6Lo ornv rtodxonototlprevrl nponetvq. H Ell(0NA 25.2 nopouotd(et puo ypqqtxli nopdoroorl rou opr0proU ttov rordrrov(ov nou owtnpoooneUouv r<dOe oprwo[0 oe oX6o1 pre q oulv6tnro pe tqv ono(o ro ouyKeKprp6vo opuvo(ri Xprlogonote(tot ortg nporei- veg (oro E. coli). l-evu<d, ro ouvq0Sorepo ogtvo[6o ov- nnpooconeUowor on6 6Uo rl neptoo6tepo ro6tt<6vtq. Aw6 uno6qtr6vet 6l ouv6B4 rdnoto Betrttorono(qoq Leu . rou yeverrKoU r<ci6u<o ovorpoptr6 Ue rn Xprlotgototqtrl rrov o1.rwof6tov. To tp(o xcr:6tr6vto (UAA, UAG xot UGA) nou 6an or rnpoorone6ouv o1.rwo(5o XpnotUonototiwot et6xd yto rov repuqnopr6 qg pret6gpooqg. 'Evo on6 our6 to xor6r x6vro rep gorro;.rof (stop codons) orl poto6orc[ to r6Log rdOe ovotXro6 ntroto(ou ovdyvooqq. Iuyr<p[oerg otr],r1].ouXtc0v DNA pe trg ow(motleg notrunemrdx6g o).Lql,ouX(eg onorol,0mouv 5r rq Kto- 6tr6vto 6Xouv ro (6to v5rlpro t6oo qro porrtlpto 5oo xot orouq euKqpurilreg. Iuven6g, mRNA ev6g e(6oug ouvrl- 0og prnope( vo Ueroepome[ ocoor6 in vitro rl in vivo on6 ro UsrogpoortK6 pr4Xovtog6 ev6g 6Mou e(6oug. 'Etor, To ro6rr6vro nou Xpnotponototiwot oro mRNA ev6g efdoug 6Xouv to (6ro v6rlpro yto ro prBootopduo xor to IRNA otrrov er66v. H orroupevtr6rrlro (p' el,dlrmeg e(otp6oetg) tou xci6rro e(vor 5vder[r1 6rt r<o0tep604xe notr6 nprilrpro ro- rd rqv eESLtEn. 'lorog o r<cir6xoq (ex[vqoe ge pro npcot6- yovrl prop<prj, orrlv ono(o Lty6repo r<o6x6vto owtnpo- odlneuqv ouyxpttu<d l,iyo o1.two[6o, (oolg or<6ptr1 rot pt' 5vo ro6rx6vro yro onorodqnore pr6trog ;rtog opr66oq opuvo(6cov. ArprB6orepo vorlproto t<to6trov[tov Kt eTIt- np6o0ero oprwo(6o eroq104r<ov opy6tepo. Mto nt0ov6- r4ro e(vor 6r opXu<6 Xpnoqonotoriwov g6vo 6tio on6 nq rperq Bdoetq r60e ro6u<ov(ou: 11 6tdxproq orrlv tp(- q 06o11 (orog e(eLiX04re 6rPrpo. H e[6h[q tou t<til6xo prnopoUoe vo 6Xet (noY(ioet,, o' 5vo oqpre(o rord ro ono(o rc oUorrlpro eile r<oroore( 16- oo notr0nLoro tiore onotodrlnore ;retoBotrrl oro v6qpro rcov r<rodu<ov(rrlv 0o dtordpoooe rtg undpXouoeg npotei'- veq ;r6ou: prq ono6errcilv ogrvo(u<riv unorcnoordoeov. H oxoupevx6rqro rou rtil6u<o uno6rltrdlvet 6n ow6 np6net vo ouv6Brl oe 16oo npr6tpro orddto riore 6trot ot dprBtot opyovrol.ro( npo6pXowor on6 rqv (6to 6e(opevq opX6yo- v<ov rurrdpov 6nou ouvSBq ro .mdyoUo, tou rc06u<o. Or e[orp6oetq rou otKoupevtxoti yevettxoti rril6tro e(vot ondvreg. Ot oMoySg oro v6rlpro tou np<rltopltroU yovr6uill.rorog ev6g e6oug ouvqOcog orpopotiv oro Ko- 6rx6vto repportopoti. l-ro nopdderygo, o' 5vo My- coplasma, to UGA xtrl6tronote( tpunto<pdvn: oe opropr6vo e(6r1 rov BLeqopt6or6v Tetrahymena Kot Paramecium, ro UAA r<or UAG rro6tronotoriv yLoutqt[- vq. Ium4potu<6q oMoydg tou x66tro 6Xouv ouprBe( p6vo oro prolovdptor6 DNA (BL6ne Ev6trya 25.7). 25.3 H Avoyvtirpron Ko6troviou- Avrrrtrr6rroviou ileprAoUB6ver Aord0eto * l-lotrtrontro rordtrovro ttou ovTrnpooone0ouv ro Ldto ol.ttvo[u ouXvorepo 6rorp6pouv oq 05or; rqq tpiqq poo1q. * Aoro0eto oro (euyoprrrlro pero[0 qq npriqg poolg rou ovrt- ro6trov[ou ror qg rp(rr1g Boonq rou rodtrovlou nporunret ono rr1 Xotropdrepq enu1prlorl rou (euYoptitporog ono ro vourtreor(dro rou rRNA oq ptpootogurl 06oq A. 10 F .= P o =b d d .a P o b o E B Glu . ^ Lvs. o ' lle. ^ Met. g;s. a Cvs. Trp . . AIa . Gly Ser . . Val . Thr . Pro Arg .Aso . Gld 'Asn . 4 Phe. Tyr . a '.rr:;, ri, :, n.o*r0,.il'';o,;H.iJ.,, qs ouxvoqros Xprlolq oprvoE6tov ortg nptorelvsq Us Tov optOpo rtov rotdtxovltrlv, nou ro0opf(ouv ro oprvo(u. YnopXet pto e[o(peo1 Yto T0 optvo- [eo, nou roOopf(ovror on6 duo rodu<ovto, ro ono(o urpiorovrol oe eupu tpoouo ouxvorr[ov. 25.S H Avoyvtirprorl Ko:dLxovlou-Avrxtodrrov(ou I-lepilqUBdvetAm60eto 711 Boorreg evvorcg
  • 207.
    UUU UUC UUA, UUG CUU cuc CUA CUG AUU AUC AUA AUG GUU GUC GUA GUG UCU ucc UCA UCG ccu ccc ccA CCG ACU ACC ACA ACG GCU GCC GCA GCG ?iirt_( UAU UAC UAA UAG CAU CAC cAA. CAG AAU AAC *AA AAG GAU GAC GAA GAG 1 r:i:;:r:. lii;:;:::- i;,:1..t i:i UGU UGC UGA UGG CGU CGC CGA CGG AGU AGC AGA. AGG GGU GGC GGA GGG U,C,A,G U,C,A e I 0r nouprveq droqripouv A n G G _l or0 Trq flupruro[veq U n C ffi Movu0rxrl 0L rpireg pooerg eXouv Tnv r),oxroTn rnppo1 0T0 v0np0 rov KtldrK0v(01v. To roulo unodorKvuouv opodeg rodrro- v[ov, evrog Tov 0n0[0]v I sKQutrron Tnq Tpiqq poorlq e[ooQotri(eL r1Tt ro vrlrlpo 0o nopoperver ro Oro. 718 KEOAAAI0 25 0 levertroq K66rK0q H ).eroupyio rou IRNA orrl petdgpoorl ernLrlpcirvetor pr6Lrg ovoyvopfoer ro r<ro6rx6vro orr'1 prBooroplr<r; 05- oq A. H ol).ql,en(6poo4 preto(ri owtKodrKov(ou ror rro6tr<ov(o u npoyporoTro te(ror USoo (euyopril poro g tcov Bdoeov, oor6oo unoxoUer oe rov6veg or ono(ot eneKrefuouv ro (euydpropro n6po qn6 ro ouv4Oro;uivo (efyq G-C ror A-U. O (6ro9 o yeverr6g r<d:bu<og ono6(6et oprogdvo or1- gowu<d ororle(o yro rn 6repyoo[o qg ovoyvripro4g rcu6rrov[ov. To np6runo r4g exrpUtrronq rnq rpftr1g Bd- onq nopouord(erot ornv EIK0NA 25.3, 4 ono(o defiyer 6rr oe 6Leg oXe66v trg neprmciroerg e(re 4 rpfurl Bdorl e(vor odtdqoprl e(re y(veror 6rdr<pro4 p6vo prero[6 nouprvriv ror nuprl.u6rvcbv. YndpXouv orcr6 orxoy6vereg xco6rrov(<ov, ong ono(- eq Kor rq r6ooepo ro6x6vro nou popd(owor rtg 6rJo npcbreg Bdoerg 6Xouv ro (dro v6r1;ro, pe onor6leopro r1 rpfu4 Bdorl vo pr4 6todpoproil(er oXe66v r<ov6vo p6l,o orov ro0opropr6 tou opwo(6o9. Ie emd (etlyrl ro6xo- v(rov to v6r1po e(vor [6ro oveloprrpotg qq nuprprd(vqg nou Bp(or<eror firlv rpfuq 05oq. Ie n5we (e0yq, to opr- vo[U nou xrodrronoe(ror 6e peroBdMeror 6noto rr ov efuor 4 noup(vr1 orqv tpfuq 06o4. Yndplouv ;r6vo rperg neprmcrioerg rord rg ono(eg ono6(6eror 1.rovo6rr6 v6rlpro ond rrlv nopouo(o Uroq ouy- rerpqlSvrlg Fdonq or4v rpfuq 06o'q: AUG (pe0erov(v4), UGG (tpumo<pdvr) r<or UGA (tep;rotrop6g). 'Eror, or C ror U nord, 6ev 51ouv ;rovo6u<6 v6r1po oqv rpfuq 0dor.1, ev6 q A norS 6ev r<oOop((er 6vo provodu<6 optvo(ri. To owxcodrr6vro e[vor oupn],4pco1.rorrr6 rou ro6rro- v(ou: ouvendlg, 11 npcirq gdoq ornv oMrl),ouXfo rou ov- rtxro6rov(ou, nou ypdEeror ouprBoru<d pe r<oref0uvor1 lprInq Apr0prig rto0wovltov on6 5'npog 3', e[vor ereftr1 nou (euyopd:ver pre rrlv rp[rr1 F6on ornv otrq),ouX(o rou r<ro6xov(ou, 4 ono(o ypdee- ror r<ord rqv 6ro oriprBoorl. Enoprdvcog, o ouvduoopr6g Kcodu<6vro 5'ACG3' Awrco6u<5vro 3'UGC5' ouvqOcog ypdgeror og rcodu<6vro ACG/owtro6rr6vro CGU, 6nou q oM4).ouX(o rou owu<co6rKov(ou droBd- (eror owf0eto ci:ore vo e(vor oupn].r1pcoprorx4 pre-ro ro6u<6vro. l-to v' onoEeuX0e( oriyXuo4, 0o 6roqp4ooupre rr1 ouvr104 oriprBoor;, origqrrlvo Ue rnv ono(o 6).eg or otr-4- ).ouX(eg ypdrpowot oe roreU0uvoq 5'-3', evtir 0o uno- 6etxv6oupe rrg olLql,ouX(eg rcov owrro6rr<ov(ov pe ovdmpoqo B5).og 6ore vo unevOup((oupre q oxdcnl rouq Ue to ro6rx6vro. 'Etor, ro nopondvco (eUyog ro6r- xov(ou/owu<to6rrov(ou 0o ypdgeror ACG r<or CGU<-, owtmorxrog. KdOe r<codu<6vro onore( dpoye 5vo r6tofrepo IRNA p'tivo ouprnL4proporrr6 owxto6u<6vro 11 pqncog ptnope[ 5vo pr6vo IRNA v' owrororlef Kor. oro 6rjo pr6],r1 ev6g (etiyoug ro6u<ov(rrlv (ror' en6rroon, Kor mo oUvoAo rcov pel.cilv, n.1. 4, ;uog orroydverog r<trr6xov[rov); Iu1vd, 6vo IRNA prnope[ v' ovoyvrrlp(oer ndvro on6 6vo ro6rr6vro. 'O).o ro rrodx6vto nou ovoyvop((er rivo ouyrerprpuivo IRNA npdner vo e[vqr rour6oqgo ortg 6Uo nptireg 06oerg. Aw(Oero, n Bdon orrlv npc0rq 06o11 rou owtrodrrov[ou rou IRNA prnope[ vo (euyopciver p' evoMorcnr6g Bdoerg or4v ow(morXq rp(rr1 05oq rou r<ro6rxov(ou. Ie outrl rr1 05or1 ro (euydpr.upro Bdoeov 6ev neprop(eror oro ouvr104 (efy4 G-C ror A-U. Or rov6veg nou drSnouv to np6runo ovoyvriprorlg ouvoqu((owor orqv un<5oeon rnq oordoelog (wobble hypothesis), 11 ono(o 64Lrivet 6rr ro (euydpropro perofri ro6rrov(ou ror owrrcodu<ov(ou orrg 6Uo nprireg O5oerg rou xcodu<ov(ou or<oLouOe( ndwo roug ouvqOerg rovo- veg, oLl,d orqv rp[tr1 05oq ouprBofvouv oouvrlOroreg (oordee€q,. Aord0eto nporfmet ener6rl n 6oUn rnq pF Booopx4q 05o49 A, 6nou ouprBo(ver to (euydpoLlo Ko- 6trov(ou-owrral6rrov(ou, enrpdnet ou[r1;r5vr1 eue].r[[o orrlv npdrq Bdorlrou owu<o6xov(ou. To ouXy5repo prl oupBorrxd (e6yog nou Bp(or<eror oe ourrlq 05o4 eivor ro C-U (E|K0NA 25.4). l-ro nopd6er4ro, ro owrxro6u<6vto UUG oro tRNAGrn ovoyvropl(a ror ro 6r]o r<ot6rx6vto rrlg yl,outopr(v4q CAA ror CAG ror ro qwrKodrr6vro GUG oro tRNAHi" ovoyvcop((er ror ro 66o rco6tr6vro rnq ron- 6rraq CAU rqr CAC. iMo pr4 ouprBorrd (eUyr1 nou e(vor ovemd orqv rp[rr1 06o4 trov r<co6rov(rov nepil.qrBdvow rpononoLqpdveg O5oerg (B).5ne Ev6qta 25.6). Autrl q u<ov6rlro rrlg rp(rr1g 06oqg rov xo6u<ov(rov v' ov5Xowot (eriy4 G-U drlproupye( 5vo pror(Bo (euyo- pciprorog Bdoecov mo ono(o 4 A 6e prnope( nl.6ov vo 51er povo6u<6 v6qpro oro r<ro6u<6vro (enerdrl q U nou rrlv ovoyvop(et npSner eniorlg v' ovoyvrrtpioer qv G). llopopo(cog, 11 C 6ev 51er nl,5ov provo6x6 v64pro (ener- 6n n G nou rrlv ovoyvrop((er npdner v' ovoyvrop(oer en[- orlq Kor rnv U). H EIK0NA 25.5 ouvorf(er ro np6runo rqg ovoyvdr prorl g. Iuvenrri g, l.rovod rd rod rx6vro ovoyvop(- t_ I l0lrn ll00n J ob,o,popn' an 3 14 10 2
  • 208.
    H a:.1:,.:.::.:: HC ,C .C lll Zuvn9(e(ty Bdotov oe dleg Trq Odoslq To IRNA ouvn9oq ouw[Oewor alq npd6popeq ofrrf 6eg pre fip6oeero uAu<6 oro dvo 4 xor ro 5fo titlpo -roug. Irqv EIKONA 25.6 nopouord(owor or enmp6o0ereg otr4i.ouXieg nou oqoporiwor on6 ouvduoopotig w- dovo u r<.tr eol.uru<cilv r<or ef ovo u t<.),eoLurtrciv 6 p6oeov. To rp(o vourLeo{6ro oro 3' drcpo, nou oXe66v ndwq e(vor q rptntrSro CCA, prepu<5q qop6q 6ev rto6tronot- ofwot oro yovr6(copro. Ie ow5g ttg neptmcioetq, npo- or(Oewor oro ntro(oro rqq ene!,epyoo(oq tou IRNA. To S'dr<po rou IRNA drlpuoupyefuor p5otrl dtdonooqg 11 ono(o r<orol(erot qn6 ro ptBovourtreonpcoteivtr6 6v- (upro ptBovouxhedo4 P. Aw6 to dv(upro ovoyvop(et r1 ogoprxrl 6oprq q4(trrorog L rou IRNA xot u6poLUer erdt- xd ro groogodreorepu<6 6eqr6 nou onel,euOepcivet to cbprpro S'drpo rou ;.top[ou, xototro(nowoq Uto 5'-Qo- oqopx4 opr66o. Zto E. coli, q RNAdo4 P onotelefuot on6 6vq RNA 377 voux..eott6(cov Kot Uto nptotetvrl 17.5 kD, evri ro evepy6 xSwpo qg ouw(Oetot on6 RNA. /n vitro, ro RNA prnope( p6vo tou vo r<otolfer tr1v ow(- 6pooq ene(epyoo(og tou IRNA. (Aur6 onoreLe( 5vo no- pddery;ro p$oev(l1tou; BLSne Keqdl,oto 23.) H l,enoupy[o qg nporreivrr<r1 g unopovddog ouv(motot mr1 mo0epono(4or1 prog dropr6pqoonq tou evepyori To npddpogo tRNA neprdXet erurpdoeereq 5'roL 3'q),trnloux(eq Ev6ovoudedo1 To 3'oxpo tfrotrrDi(eror -N. H. "0 ll -r tc -6 -N til 'H_N.ffi I H * [ouov[wl :CH ? tdKXopo cH" ouooKrtrn nc{h+20.. IdKXoPo 0 H.N,H ,Mrou"''n"rsft,*:! . IoKXopo t;;,.:li,;: i_t 1,1*:: lr !i:,.J1,,:,.t:t ri.C.Jt: .iit.j :::..,,:i ,1aai, 7.1.:;. tilr::1.r.C':,::ii H un 'rC 'n ..0 {. "., ,'Y Y /t-fr-r-r...0 roKxopo r...n_n,.d.._r n *,[.-r-i -,'u),n t" ).0Kx0p0 - 'r ..,, r. .l r H ooroOeto oro (euyoprrrlro rov pooeolv enttp6ner ro oXlpolopo (euytirv G-U UeroE[ Uq tpiqg pooqq rou xodtro- v(ou rot qg nptlqq poolq rou ovlrtobrrovlou. (owor ;rdvo 6rov orrlv rp(-r1 06oq Bpior<erot G rq U. 0o16oo, p6vo ro UGG rot AUG onoteLoUv nopo6ery- Uoro Uroq rdrorog provodu<r1 g ovoyvrrrprorl g. ffi ITo 5', 3'dKpo DroofldTor 25.4 lo IRNA llop6yowor MeyoA0repov np66poUov U' EneEepyooio Mopitrlv + 'Evo rirprUo IRNA dlproupye[ror p' ene[epyooio evog npddpo- l.rou lrop[ou. * To 5'orpo 0qptoupyeirot ptlotrl dtoonooqg ono rrlv ev0ovou- rtreooq RNAoort P. " To 3'orpo drlgtoupyeuot geoo notrtrontr6v evdovourtreotrul- r6v rt e(orvourtreotrulrtiv Droonooeorv, ttg ono(eg orotrou- Oet 1 npooOlq rou rorvo0 repprolro0 rprvourtreortd(ou CCA. **;1 *tii';':t ?iy;rs *i* E+fl ,-*i_q) rtl *vc'lv,*f i(eiti, c ""i,t,;ri *rix*5iit*,:icx 6{1! t3":l i;C! r0! (iraid;ic! To s'dKpo Droondror Ev6ovourtreoorl (RNdon P) * ffi 5',3', tlpoor(0erot CCA 3''Ev(uuo(o) fl npoo0rlrrlq 5'I rou 3'drpou EIK0NA 25.5 To (euydporlro xtrl6lrov(ou-ovlrodrrovtou eprntr6- ret qv ooro0eto orrlv rptq 06o4. ,. . To 3' orpo rou IRNA dqproupye[ror prioo ovldpo- oetov erro;.ri1g (ev6ovourtreotrurrrdv) ror ilotrrdiouoToq (e(orvou- rtreotrurtr6v), nou orotrouOouvror ono qv npooOflr1 CCA ro 5' orpo dqproupye(ror peotr.r luog orprgoug evdovour<),eotrulrqg Dro- 0n00rlq. U C A G An G G Uovo U Uovo cnu ?i.;i To tRNA I-lopoyovror p' Ene[epyoo(o lVleyotrurepov tlpoDpol.u,.rv ["lo0l,-:', 719 Boorxfg eworeg
  • 209.
    Kdwpou rou RNAn ono(o e[vot ouprnLrlpoporrr{ rou np66poprou IRNA. Oo e(eroore( nepor6pto anlv Ev6- uyto 23.8. To 6v(upro nou ene[epyd(owor ro 3'drpo 61ouv ne- prypoge( roLrirepo aro E. coli,5nou puo ev6ovoudr,e6- or1 nupo6ore[ qv owbpooq 6roonrilwog ro np66popro pr6pto r<o0o6u<d r<r 6nero noMSg e(ovouxi.edoeq Uoh- 6[(ouv ro dxpo oe roteU0uvoq 3'-5'. H ene[epyoo(o rou 3'dr<pou rou IRNA orouq euKopurireg en(oqg neprl,op- Bdver noMd 6v(u;ro. H npooOrirrlrou 3'-CCA r<orol0e- tor on5 ro 5v(upro IRNA vouxi.eon6ul,orpovorpepdorl, nou l,emoupye( o4 RNA noLupepdo'q ove[dprqrq on6 erproye(o. Me dMo L6yro, ro 6v(u;ro npoo0Srer er6rd C, C ror A cnnv oMqLouX(o 1op(g vo (euyopriver qv r<utoo[vr1 Kot rrlv o6ev(vr1 pre rrg ouprn].qpoporu<69 Bd- oerg youov(vqq Kqr oupoKiLqg ndvo o' 5vo erpoye(o. Aw( yt' ou"r6, 4 6ro 4 6opri rou ev(6prou enopre( yro vq qXq godoet 6ro6oXx6g oupntr4 prrrporx6q O5oerg np5o- 6eor1q yo rtg C, C ror A. Ko0c0g npoor(0ewor rcl vou- r.treor(6ro, ro oOpntror<o ev(Ugou- IRNA peroBdMet tr1 dtopr6pgroo4 rou rilore vo xoroore( oupnArlpcol.ronr6 pe xdOe 6ro6oXtx6 vour<Leor(6to. To rp(o voux.Leor6tq npomiOewor on6 r4v IRNA vourLeorr6uLorpovogepdoq, 6rov 6ev rodrronororjv- ror orn yovr6torci oM4Loul(o rou IRNA. E(vor evdrotp6- pov 6tr ro 5v(upro 6ro6popor((er en(o4q Boorr6 p6Lo orqv enrdr6pOoorl roreorpoprpr6vcov 3'dxprov rou IRNA oe opyovroporig, 6ntog to E. coli, nou xolduronorcAv to CCA. Ie owo0g rouq opyovropro0g, ovoyvropi(owot rp(o 6roqoperrrd IRNA unoorpc0proro: lrop(g CCA, Ue 31C i ue 3'-CC. Or IRNA vourcl.eonduLorpovogepdoeg 6rorp(vowor oe 50o roqyop[eg nou 61ouv oqprowrrrj opror6qro o1.rwo[6rov pr6vo ortg neproldg rov evepy<ilv rdwprov. To 6v(upro rnq Kornyop(og I Bp(or<owor or' opXo[o, evdr ro Borrqproxd rr europuorrrd 6v(upro ouvonotetroUv q defrepq roqyopfo. Ie opropr5vo Bor,cripro, n npo- o0qrq r4g CCA xorql0eror on6 6Uo orevd oXetr(6pe- vo 5v(upro roqyop(og ll: ro 6vo on6 ourd ro 6v(upro npooO5rer -CC, evdr ro dMo npoo06rer qv 3'-re).rxrj A. 25.5 To IRNA tleprdxer Tpononolnpdveq Bdoerq * To tRNA nept6Xouv ndvo ond 90 rpononoulp6veg pdoerg. r H rpononoulon ouvn0toq ouv(ororor oe ogeorl otrtroyr| rtov nporoyevtirv Bdoeov rou IRNA, oordoo unopXouv oprogeveg e[orpeoerg onou pto pdon oQoLpe(rot u ovnxoOrordror on6 pro 0trtr1. r 0t rpononornp6veq pooerg npood0ouv ou[nUdvn oroOeporrl- T0 oT0 IRNA rot pu0p(ouv qv ovoyvtitpro1 roug on6 npoTsl- veg ror otrtro RNA rqq peroqpoorxrlg ouoreurlg. To perogoprr6 RNA e(vor 1.rovo6u<6 ;rero{ti rov vou- 720 KEOAMI0 25 0 levelrdg Kdrdrrog rLeivrrrilv o(6ov oq npoq rrlv nepreKnr<6qrd rou oe rpononorrlp6veg Bdoerg. Mro tpononor4;r6vr1 Bdo4 e(- vor onoroo6rlnote dorcrritrrog noup(vqg 11 nuprpu6ivqg em6g rov ouvri0cov A, G, C r<or U, on6 rrg ono(eg ouw(0ewor 6l.o rq RNA. 'Oeg or dtrLeg Bdoerg nop6- yowor on6 peropetoypoqu<4 rpononoirloq puog on6 trg rdooeprg Bdoerg, orp5rou our4 evoo:1.rorro0e( oqv notruprBovouxi.eotr6rrrl qluo(6q. To odxlopo rnq pr- B6(119 oprop6vcov vour<.l.eol6(tov rou IRNA en(oqg pre- 0uAr6veror oro 2'-udpo(rjro o6qyri:vrog 6ror oe 2'-O-Ue0u(coon. Evcir 6),eg or xorqyop(eg rou RNA eprgov((ouv rd- noto Bo0pr6 rponono(r1or1g, to eUpog rrov l4prrciv pe- toBoLriv orrg Bdoerg e[vor notrrj preyo].Jrepo oro IRNA Ot tpononorqoetg rupro(vowor on6 on).rl pre0utr(rr:oq 5cog yevtrri ovo66;rr1or1 rqg Bdorlq. Tpononorrloerg oupBo(vouv oe 6Lo ro ttrrrlproro rou tRNA. l-lorril,ouv no).rj rog npoq rrlv 5r<roor1 rrlg ouwqp4oriq roug prero- [ri rcov 6rdgopr,:v IRNA, 6n<og en(oqg Kor oq npoq rrlv ew6nroq roug pr5oo oro ;r6pro. Tpononot4oerg er6txdg yro ouyKeKprpr6vo IRNA q prrpdg unoo;rddeg 1RNA yevr- xd efuor ondveg. Yndplouv en(oqg opro;r6vo np6runo rpononorrloeov erdrrd rou e(6oug. IuvoLtrd, ro IRNA neprdXer ndvco on6 70 6rorpopertroUg rpononorqpSvrov Bdoecrlv-r6noug. Kd0e IRNA rpononoe(ror rotd p6oo 6po oe noooor6 15-20y. rcov Bdoecivrou. To tpononorrlpi6vo vourLeoo(dro ouw(Oewot on6 erdu<d 6v(u;ro rponono(r'1onq rou IRNA. To ou0ewrrd voux.treoo(6ro rdOe Orioqg prnope( vo xoOoprore( eke pre oriyxpro4 r4g oMrll.ouX[og rou IRNA ge r;v oMrli.ou- X(o rou ow(ororXou yovt6(ou r1 ().ry6repo onoreleopo- lxd) pre onop6vcoorl np66popr<ov prop(r.ov Xop(g rdnoro r1 6),eq rrq rpononouioerg. Or oAlqtroul(eg r<ov np6- 6popur:v 1.rop(rov deiXvouv 6tt 6ro<popelxdg rpononoL- 4oerg erodyovror oe 6ro<poperrxd otd6ro xord r1v rr:p(provorl tou IRNA. To no).).d 5v(upro tponono(r1onq rou IRNA norx(- l,ouv noLU oq npoq r1v erdrr6r4to. Meprrdg qop6q 5vo tiv(upo nporoLe( Urq ouyKeKprp6vrl rpononofqor1 oe Uto pt5vo 05oq. 'AMore, 6vo 6v(upro prnope( vo rpo- nonorrloer Bdoerg oe oprerSg 6rogoperrrd,g 06oerg-ord- Xouq. Opro;r6vo 5v(upo ovo).oprBdvouv povdg owtdpdoerg pre [eX<r:prord IRNA, evcb dtr].o 6Xouv 6ro- qopo unoorpoiproto. Kdnoreg rpononotqoetg npouno- 06rouv 6ro6o1u<q 6pdoq noMcov ev(0grov. Aenrop6pereq yro rn 6opurri Bdoq r1g rponono(- nonq rou IRNA on6 5v(upo p6llg dpXroov v' ovo6riov- tor. 'Evo ewunoloror6 nopd6etypro e[vor o prqXovropr6q p6oa; tou ono(ou 11 opXoroo(vr1 (archaeosine), puo rpo- nonorqg6vq G, erodyerot oro Bp61o D ouyr<exptpdvutv IRNA lov oplo[orv. I-ro vo npooeyy[oer q Bdoq nou np6xetror vcl rporonorq0e(, q ono[o rqvovrrd Bp(oxeror ewocptootrr6vrl pr5oo qrov rprroroyrl nuprlvo tou IRNA. to 5v(upo frans-yLuxo(ul.doq tRNA-youovrv4g (tRNA guanine transglycosylase) euodriver pro 6poonrc1 eno- vodtdro[r; rou IRNA, dlpuoupycitwog 6ror gro evoMo- mrrrl rptrorqyi dotrlq yvrrlmrl cog 6o94 tr. Enovo6ro- td(erg rqg 6op4g tou IRNA mo nLq(oto enoy6prevqq npooopproyr;g (induced fit) 51ouv en[or1g noporrlprlOet Boorxeg evuorcg
  • 210.
    Kor oe dMo6v(u;ro rponono(qoqg ror e(vor r<ow6 Xo- porrrl pronr6 orrlv ovoyvd:ptoq. Or rpononot4p5veq Bdoerg npoo6(6ouv ou(r1p5v4 oro0ep6qro oro IRNA ror puO;r((ouv qv ovoyvcilptorj roug on6 npolreiveg ror dMo RNA oro prllovroprd qg petdgpooqg. l-ro nopd6erypo, oe oprerdg neptnrdr- oetg, 61ouv oorpcig npoodroprore( p6i.or yro nq rpoflo- nor4pr6veg Bdoerg mqv ovoyv6proq on6 rrq oprvoo- xuAo-tRNA ouvOerdoeg (Bldne Ev6tryeq 25.9 6o9 25.11).Ie noM6g neprm6oerg, roo16oo, o BroLoyrx6g p6),og tqq rponono(r1onq rou IRNA nopog5ver dyvo:- sroq. H EIK0NA 25.7 nopouord(er opropdveg on6 tq nro xow6q rpononor4;rdveg Bdoerg. Or tpononor1oag rrrlv nuptptdrvcbv (C r<or U) yevrrd e(vor fuy6repo nol,rjn).o- xeg on6 erefueg t<ov noupwcbv (A r<ot G). H ouv406orepq rponono(qon rnq oupr6(v4g Kot rrlq rutoo[vr1g elvor q pre0u)r(rrloq, q ono(o ;rnope( vo ouprBe( oe notrl,5g 6ro<poperu<69 O5oerg rou 6olctu).[ou. H peOu- tr(roor1 or4 06oq 5 r4g oupor<fi.r1q 6qpuoupye( q prBo- 0upu6(vq 0. H Fdon qg 0upu6(vr1g e(vor row5or;prq pre rr1 0upr6ivq oro DNA, oor6oo oro IRNA e(vor npoo6e- 6ep6vr1 oe prB6(rlnopd oe 6eo(uprB6(q. Awri 4 0upu6(- vq undplet qXe66v oe 6l.o ro IRNA arq 05o4 54 tou pp6Xou TqrC. H rfeudooupr6(vq efvor 1.uo ewunootoxr; rponono(r1oq ouprd(vqg nou nporUnrer pe drdonooq rou yLur<o(nrxoU 6eoproU, rrlv onotc c<3:.:-:a -:::. propr6vr; neprorpogrl rou oneLeu0rpc0Us,,,o- 51.-:-, : _ xr enov6voo4 rou dvOpor<o C5 pe rov ovOpci<: l' -: ptF6(rlS.'Etot, q ql,reudooupr6(vr1 6lo0ereL evov I'r-. r.,. r<o(ttu<d 6eopr6. I1e66v 6).o ro IRNA neprdXouv q,er5.- oupr6(vr1 mr1 06oq 55 rou Bp61ou fqJC. H 05o1 56 en(or1g e(vor e[orperrr<d ouwrlprlprdvrl oq xuroo[vqt 1 o).).4Lou1(o TQC mrg 06oetg 54-56 nopSXet rr1 Boorl yto rrlv ovolloro0eo(o ouroU rou rpr4potoq rou tRNA. H drudpoouprd(vq (D) nou 6rlpLoupye(tor p5oco r<o- peoproti tou 6Lnl.oU 5eo;roU nou ouv66er ro C5 xoL C6 qg oupor<fi.r1q e(vot oxe66v ou<oupevrrq oro Bp61o D rotv IRNA. H ogo(peoq rou 6Ln).ori deoproU qg D xoto- orpSqer rnv opopolr<6qto ror ro en(ne6o oXqpro rou 6or<ruL(ou qg oupor<il.qq, ono6(6owoq ULo oouvq0L- orrl 6opq, q ono(o tpononore( e).orpptrtg to o"1r1po tou o<porpu<oU nuprlvo rou IRNA. To vour<Leoo[6ro tvoo(vq (l) Bp(or<eror rovovu<d oro x0rropo og ev6rdpeoo npoi6v r1g Broouv0etu<r1g o6ori rotv nOuprvtr-rv. Om6oo, 6ev evOotl.iotrbve-rOr dpeoo mO RNA. Aw(0ero, 11 6nop[r1 rou efoprdror ond rnv rpo- nono(4o4 qg A. H evocogdtooq qq I orr; 06o11 rou 5'- ov-rrrot6tr<ov(ou ouvero<p6pet no).ri oro ooro06g (euydpopro Bdoe<lv oqv rp(q 05or; tou r<codtxov(ou rou mRNA (B,l.dne Evorrlro 25.6). Ot rpononorqoetq rov A r<oL G oupd nopdyouv rv- Tponrniririuive_t $ciueit 00 H H Prpo0ugtdivrl g1 Atudpooupr0irtr (D),l,.rOoorp16rvn (q.r) 4-0ttooupt0[vrl 0uotohoyrdg pdoeLg 0 ,lt Hl{cffi0H o'ffi' 0uprdivrl NH, t' ,'@; KurtD[vr1 NH, I HN -ctc-Nrt A 11 )cH ,HN'"lt1-u-N' . -'-Y3l ( ntH-ciiocf . -lFl Cli.iir1C:. H^ o :l ;;'"";"r I Lh; I g5 -c-6-N., i*-'tr- H / ,ll )bH ;r.-, . J )H 2HN'u1r],-u-N/ l/x1,'"' -11'zHN' Iouovoofu1 7-ge0utroyouovoo[v1 Krouoorvrl(0) touoioo(vr1 (Y) Kor or reooeprg pooerg rou tRNA pnope( vo rpononorl0ouv. 2S.5 To IRNA lleprdXer TpononoqgSveg Booerg 721 N--:il, xloC-6-t',t. nd=*..8-N)bn UeOutrodevootvn(m6R; trt6
  • 211.
    re6q v6eq 6op6q(Ex6vo 25.7). [-to nopddetygo, 6[o notr0nhoreg oerp5g vourtreott6(tov Boo((owqt ornv rponono(r1on rnq G. Ot Bdoetg Q, 6n<oq 11 r<touoo(vt1 (queuosine), nept6louv 6vov np6o0eto donrUAto Tlewe- vutr(ou nou npoor(Oeror p6oco ev6g deopoti NH orq pe- 0utropd6o qg 7-;re0uLoyouovoo(vr1q. O 6ortUltoq newevutr(ou gnope[ vo ;retog6pet notrileg dMeg o;16- 6eg. Ot Bdoetg Y, 6nog 11 ouoioo(vq (wyosine), nep6- louv 6vov np6o0ero dol.c0Lro nou ouwtlretot Ue rov (6ro ro 6qhir0fuo nouplvqq. Aut6g o np6o0eroq dorcrri- Ltog ;rerorp6per Uto puo proxptd ovOpoxtxti otruo(6q, orqv ono(o gnope( vo npooreOorlv 6togopettr6g, t<otd nep(molor1, opd6eq. 25.6 Or lpononornp6v€q Bdoelq Ennped(ouv To Z€uydpopo AvTtKoOtKoviou-Ko6lroviou r Tpononouloerq oTo ovrtrodxtivto ennped(ouv ro npdruno rou ooro0o0g (euyopriUotoq rot ouvendlg e(vot or1povru6g yro rov xo0optopd rnq etdxdrnrog tou IRNA. Tpononorfoerq rou IRNA p6oo t<ot 6(nlo oro owtKo- 6u<6vro enrlpe6(ouv rqv tt<ov6tqtd tou vo (euyopdrvet pe ro rrrl6u<6vlo rou mRNA. Ot neptoo6repeq rpono- notrloerg owo0 rou e(6oug ewon((owor orrg 06oetg 34 t<ot 37 tou Bp6Xou tou qwtKo6txov(ou t<ot yevtxd LetroupyoUv neprop(owoq ro e0pog r4g 6ro06otpqg Krvrlonq oro owu<to6x6vto. Me rq oepd tou, outS 6t- euroLUver r4v oyt<upo6eo(o tou IRNA orq 05o4 A tou prBooopror(ou. Aut6g ot rpononot(oetg enqped(ouv ro (euydpcopo rov xodtrov(ov rt 6tot enrlpe6(ouv dpreoo nc0g 0o ep;tqveUoet to xtittopo to v6r1po tou IRNA, Ot rpoTronotnU6veg Bdoerg enmpdnouv enrnp6- oOeto np6runo (euyoprilprorog erc6g on5 ere(vo nou Boo((owor oro ouvqOtogdvo xot to ooro069 (euyd- popo rov A; C, U xot G. H tvoo(vr1 e(vot t6to(tepo orlprowrri 6tov Bp(oretot orrlv np6tr1 06oq rou owmo6xov(ou (vouxl.eotbto 34 orqv oMqlouX(o), enet6ri e(vot tt<ovtl vo (euyopdroet pe onorod{nore on6 ng pdo'etq U, C rqt A (ElK0l'lA 25.8). O p6Log qq woo(vqg ovo6exv6etot ornv qfloKodxono(- rlon rov r<co6u<ov(tov t4g tooLeur(vqq. E6ri ro AUA t<to- 6rxonote[ r4v too].eur(vq, evr6 to AUG tq trre0etov(vq. l-to vo 6roBdoet rrlv A ornv tprn 06on tou rto6troviou, to IRNA 0o Xpero(6tov 1.uo U orqv np6rn 06oq rou ov- rrxrodxov(ou - coor6oo owrl4 U orrlv ooroOf 06o11 0o (euydptrrve ovoyroortrd Kot Ue rqv G. 'Etot, onoto6rl- note IRNA Ue Uto 5' U oro owtt<to6x6vt6 tou 0' ovo- yvripr(e t5oo to AUG 6oo Kot ro AUA. Aur6 to np6p)qpo enil.[etot pe tr1 o0v0eor1 ev5q IRNA tootreu- rfvqg nou 6ro06rer A34, qv ono[q qKo].ou0e( q rpono- no(r1oq qq A34 oe 134 on6 to 6v(upro 6eoprwdo4 rqg o6evoo(vqg rou IRNA (IRNA adenosine deaminase). 722 XEOA,A|0 25 0 [eveudq Kdtdtxog --.2@1i , :qKxqpo u'nr6ffi* lvos(vn ,ryIdrXopo I6Kxopo Adevivrl IoKXopo flqili,i& I5.& H rvoolvrlpnope(vo (euyoptitoetge rqv U m, , - rrlv A. Irrl ouvdXeto, q 134 prnope( v' ovoyvop(oet Kot rq rpic rco6rr6vto rqg LooLeur(vqg: AUU, AUG t<ot AUA. Ttg neproo6repeg <popdg, 11 U mrlv npcbq 06o11rc- owtxro6u<ov(ou en(oqg petotpdnerot oe Uto rpononc.- 11p6vr; pop<p4 pre 6torpoperu<59 r6t6qteq (euyop6uc- rog. To nopdyo.lyo qg U nou neptdXouv m4 06oq 2 puo 0eurrl optddo ow( rou o[uy6vou eprgov((ouv ou[r- ;r5vr1 ex).ercrtx6t4to mo (euydp<opo Ue rnv A oe o["- Kpron Ue rqv G {;:{lll&. j:I !l Avrrr<to6tr6vLo ga 'Evaq 6esU6q Dav enupxei :oKXopo --@: ,. IdxloPo S 0 I H-5- I H :orxqpo H 2-OerooupoKiln (euyqptivet Udvo Ue A CH" t' HCffi"O.. ,w-,.:x* Adev(vn 'TIlj.1,,'.-* HWH ru**ooo Ell(ONA 25.9 H rpononoinon orrl 2-Oetooupt6ivrl neptop((et ro (euydptopo g6vo otlv A, dt6rt gdvo 5voq deopdq H pnope( vo oXrlpolore[ge qv G, Boorx6g evvoreg lvoorq
  • 212.
    ouprdrvo-5-o(uo[rx6 o[ri r<orouyyeve(g rpononor4oerg orqv npdrq 05o4 51ouv r1v o[roor1;re(crrrrl rdr6rrlro vo enmp6nouv oro povo6rrd IRNA vo 6roBd(er rp(o 11 or<6- Uq Kor ro rdooepo ouvrilvul.ro ro6x6vro NNA, NNC, N- NU xor NNG. Aw5g roOc0g r<or dl}eg tdr6qreq (euyoprirprotog 6e(Xvouv 6rr undpXouv notrl.o[ rp6nor yro rqv Koro- or<euq ptog o;166o9 IRNA u<ovri:v v' ovoyvrop((ouv ro 61 t<codtt<6vto nou owrnpoocone[ouv opuvo[6o. Aev ent- rpore( rdnoro rdro(repo np6runo oe xdnorov opyovr- o;16, nop6tt q onouo(o proq ouyKeKprprivrlg o6o0 rpononofioqg prnope( vo epno6(oer rn Xpion oprqr6- vrov nporUnov ovoyvripror1q. 'Eror, pro ouyrexprpr6vrl orroySvero rcodu<ov(rov droBd(eror on6 IRNA pe Srogo- pertrd owxro6 rrdvro oe 6 ro<popetu<o ri g opyovLopro rj g. IuXvd ro IRNA dro06touv enrrol,umdpeveg rxov6- rqteg ovdyvoonq ouyKexprl.uivcov ro6tr<ov(otv, 6ror riore rivq ouyxerprprdvo xr.o6rr6vro 6ropd(eror on6 6[o 11 neptoo6tepo IRNA. Ie ow5q lg neprmc0oerg (ocrrg undpXouv drorpopdg oq npog rlv onoreleoporrr6rq'ro rov evol,l,omu<rilv owrdpdoeolv ovoyv6proq g (yevrrd, ro rodtx6vro nou XpnorponoroUwqr ouXvd, ouv{Ocog droBd(owor nro onoreLeoprotxd.) Or npoBl,6gerg tou ooro0orig (euyopriporog efvor noL0 ouprBotdg ;re ro nerpol.rotrrd 6e6oprdvo yto 6Lo oXe66v ro IRNA. Oqr6oo, undplouv e[otpSoerg, ong ono(eg ro xro6rr6vto nou ovoyvop((owor on6 6vo IRNA dtoE6pouv on6 ere(vo nou npoBLdnowor on5 rouq Kq- vSveg qg omdOerog. Aur6 [oog npor0nrer on6 rqv en(6poor1 yemovu<cilv Bdoeov Kor/ri rnq 6rog6pgoo4g tou Bp6Xou rou owrxrodrrov(ou orrl ouvo).rx4 rprroro- Wt 6opn rou IRNA. l-'lepottSpo ev6e[[erg yro rn onUo- o(o rqg neprBdl},ouoog 6oprrlg nop6Xowot on6 rrlv onop6vooq onopodxri:v preroMoyprfuov orcl ono(o 11 olr.l.oyri ptog Bdoqg oe xdnorq dMrl neproXrl tou prop[- ou peroBdLLet ulv u<ov6rrlro rou owrrro6rr<ov(ou v' ovoyvro p (er rcr:6 rr6vto. GCU 2 5.7 Yndpxouv Inopo6rr6q lpononorlroerq Tou OrKouu€vtKorj Ku)6rro c Ie optou6vo ei6q 6Xouv ougper otr)royrig oro yevenrd rtirdr- KO, c Aur6q or drtroy6q etvor ouvrl06orepeq or0 pnoXovdprord yo- vtduirporo, 6nou pnopei v0 KoTooKruoore[ 6vo rpu)oyeveux6 d6vrpo yro ilg otr)roy6g. e Iro nupnvuo yovrduirgoro, ot otrtroy6q ouvn0oq enrlpeo(ouv privo ro rodx6vro repponopou. H ou<oupevrr6r4ro rou yoverrorl r<cbdu<q e(vor o(ro- orlge(oq, coor6oo undpXouv oproprdveg e[orp6oerq. Aur6g ouvrl0rrlg en4pe6(ouv ro xrodx5vro nou eprnl,6- Kowor mrlv Svop[q ( rov tepgorropr6. Or o]}.oy6g oro xripro yovr6rilproro (Borctrlprord q nuprlvu<d) ou- voQ(owor or4v EIK0NA 25.10. IXe66v 6Leg or otrl,oy6g oro nuprlvrKd yovt6uil;roro nou enrtp6nouv o' 5vo r<o:6rr6vto v' owrTrpooroneUer 6vo ogtvo[ri enqped(ouv to rro6x6vto reppotrogo[: o Irov nporopurirq Mycoplasma capricolum, ro UGA 6ev Xp4orponorekor orov repporr.opr6, oMd ro6xo- noef r4v rpumoEdvq (Trp). Irrlv npoyporr5r4ro e(vor ro r<up(opXo ro:6u<6vro qg Trp, evcb ro UGG Xpnorponorefuor ondvrq. Yndplouv 611o ei64 TrptR- NA, ro ono(o neprdxouv ro owu<rodrx6vto UCAe (6roBd(er ro UGA xot UGG) ror CCAe (6roBd(er pr6- vo ro UGG). o OproUSvo Blegopr6orp6po (provor<rjrropo npot6- (roo) 6roBd(ouv ro UAA r<or UAG rog ytrouro;r(vr1 ov- r( yro orlproro repUorropori. To Tetrahymena thermophila, 6vo Ble<popr6og6po, nep61a rp[o ei- 6n tRNAot". 'Evo tRNAGrn pe owx<o6rr6vro UUG qvo- UUB 'n, ,IU: "."INil,, 'iy33 ,,, UUt ,,, 'iUt "''" NUtt sT0P-n',,,o iUBt sT0P-Trp'cvsser cuu lccu lcAU p;q HICGU ffi .'' 'issl ".'isii::: :issl ^,' 919 !:1rryt i999 l:19 i:" itl tlgi: AUU I ncu i RRu ^^^ ^, i AGU ^-- AUa 1e r I ncc -u" , I AAC utn * I AGC ue' AUA tte-ry6x1g IRCR ""'lAAA,,,^ ,, lRcR Rrg-N6NE nud ii,r"rt '''"'"M i AaC I nno tv' K ! roc nt[ WCS W 3i3 *' D GAA ^n^ Glu E uAu GUU GUC ,,, GUA JiI: GUG GCC v GCA ^'o ^ : trUU GGU GGC GGA GGG Glv 1 i , r:, -'i.: 0t peroBotr6q rou yeveruou r66txo oro porqproro fl euxopuolro nuprlvrro yovrdrrirporo ouv10otg onodr6ouv ogtvo[50 ,. <oDtrovto Il(nS I geropotrtrouv evo rodtrovto 6rot 6ore vo Unv ro0op(er ntr6ov ronoro olrrvo[u. Mto Ueropotrn oro vorlgo ono evo ,- ,,0[u oe ronoro otrtro elvor oouvl0Loq. 25"? YnopXouv Inopodrreg TpononoLrloerg rou 0rxoupevrxou Krrrdrx: 723 Boorxeq eworeq G
  • 213.
    yvop((et ro ouvnenKodK6vto CM r<ot CAG yto 11 yl.outopr(vr1, 6vo 6erlrepo e(6og pre qwtro6tr6vto UUA ovoyvtopi(et t6oo to UAA 6oo t<ot to UAG (oripqrovo Ue rnv un60eoq tqq oord0etog), ev<ir to rpfuo e(6og pe owrrodtt<6vro CUA ovoyv<op((et p6- vo to UAG. O neptoptol.t6q qq et6r6rrltog rou no' pdyowo oneleu06ptoorlg eRF, cborc v' ovoyvop((et pr6vo to ro6x6vto teppotropo0 UGA, en(oqq Xpet- 6(eror c0ore vo Un ouUPe( np6topog repprottopr6q oro veonpoo6topt(6prevo KtoSK6vtq yLourog(vr1g, o Ie rdnoto dIIo BLeqopr6og6po (Euplotes octacari- natus), to t<todu<6vto tepprottol-toti UGA enovonpoo- drop[(etot oe ruorefvq. M6vo ro UAA lprlotgonot- e[tot tog xo6tr6vto tepporropoti, evdl 6ev undpXet UAG. H oIIoVn oro v6rlpro tou UGA prnope[ vo ont- reuX0e( pSoto tponono(r1oqg tou owtt<to6tt<ov(ou rou tRNAcv' pe 134, 6rot coare vo gnope( vo 6toBd(et ro UGA pro(( ge to ouvriOq rro6tr6vto UGU t<ot UGC. To UGA 6Xet 6ml'6 v6rlpro oro Euplotes cras- sus (BL6ne Ev6trya 25.8). o I'5vo (u;roprir<r1to (Candida), to CUG enovonpoo- drop(etot oe oep(vq ow( qq Leuxffiq. Aur6 e(vot 6vo ondvto nop6Setypro enovonpoo6roptopori on6 5vo rodx6vto pe v6qPro o' 5vo d.tro. l-evrxd, yro v' onorcnloet xcodtronorrlurrl Aenoupy(o 6vo t<to6tr6vto repportoUo0 np6net vo oupBotiv 60o r0- nor otr),oycirv: 6vo IRNA np6net vo petoMol0e(cirore v' ovoyvtop(et ro rrodtx6vto rot o nopdyowog onetreu06- poonq rd[4q I np6ner vo ;reroMol0ef 6ore vo Urlv rep- 1.roil(et oe ow6 ro xu:6tr6vto. O dMog ouvriOlg t0nog otrtroycirv e[vor 11 onciLeto tou IRNA nou owtorotXe( o' 6vo t<to6tx6vto, 6tot dlore to rodu<6vto vo Urlv r<oOop[- (et nl,6ov xdnoto opwo(tl. 'Otreq ow5q ot oMoy6g e(vot onopo6u<5q, yeyov6q nou o4go(vet 6rt pr6Mov ouv5B4oov ove(dpqto oe et- 6u<6q e[e].rrcrtr5g oep6g. Iuyxewpdlvowot oro t<trl6u<6- vro repportotrtoU (orog enerdri oe our6g ttg 06oetg 6ev undpXet owtxotdmoorl ev6g oprtvo(dog on6 xdnoto dLlo. M6).tg xo0teprirOrlre o yevettr6g r66trog, oe npcilq-ro ord6ro rrlg e[6].t(qg, onotod{note yevtrrl peto- Bo).4 oro v6qtrro ev6g rodtrov(ou 0o nporol'o[oe owt- rordmoo4 oe 6leg ttg npore(veg nou neptSxouv ro ouyxexptgSvo o;rwo(0. l-lt0ov6v 4 petoBoLq 0o rpov enrBloBrlg yto optop5veg on6 out5g ttg npotelveg, on6te 0o eniley6tov opvrlnrd. Ot onor.tr'(vouoeq Xpn- oerq rov rodtxov(tov teppottopo0 (oog owtnpooto- ne0ouv rn "oUIInqJn" touq pe oron6 t4v rovovtrrl rrodtxonoirloq. Av optopr6vo xto6tr6vto teppottopo0 Xpnotponototiwov ondvto, 11 entorpdteuo{ toug yto xio6rronofqor1, g6oto o]'Loycirv sro IRNA nou enttp6- nouv rov enovonpoo6toptop6, 0o ntov nro nt0ovt1. E[otp6oetg orov otKoupevtr6 yevettr6 rtil6tro en(- oqq ouprBoivouv oro prttoX6v6pto noMrirv et6riv. H El- K0NA 25.11 nopouotd(er prto cputroydveto (e[e].trtrr<6 66vdpo) tcov oMoydrv. H txov6tqro KorooKeunq ptoq guLoydvetog unodqtrcivet 6tt unrlp(e 6vog ott<ougevt- 169 xri6xog nou pretoBtrrl0rlre oe 6td<popo orlpe(o r<o- 0txoupevudg rtiDtroq EIK0NA 25.1"1 0r Ueropotr6q rou yeveuo0 xtit0txo oro ptrol6v- Dpro pnopouv vo ewontototjv oq rputroy6veto. 0 etrdxtmoq opt$ U6q ove[oprntov petopo)rtirv npo5pXerot onri qv undOeorl 6n u Ueropol6q AUA=Met roL AAA=Asn npoyporonototjvrot 1 roOe- pio ove[opqto 6uo Qop6q rot 6tt 11 npdrtpq geropo)rn AUA=Met owtorpdqrlxe mo eXwddePPo. rd qv e(6LrEn tov l.tttoXov6p(trlv. H npotp6teprl oMe V{ ntov n Xpnoqoflohon tou UGA yto u1v rcodmonoi- non rnq rpumorpdv4g, 4 ono(o e(vot t<ow{ oe 6tro ro pr4 <punxd groX6vdpto. Optopr6veg prroXov6ptordg oMoydg rdvouv tov ni6t- Ko qnlo0orepo pe rrlv owxotdoroorl 60o rurl6xov[trw nou e(Xov dtorpoperr6 v6rlpro on6 6vo (e0yog nou dletro (6ro v51pro. Ze0yr1 ro6xov(tov nou owqretoln(oqrov rqd ow6v rov tp6no nepr.ogBdvouv to UGG rq UGA (rot ro d6o yto rnv rpunroqdv4 ow( to 5vo yto rqv tpumog6vrl ror ro dMo yto repUortoU5), 6no4 entonq rot to AUG xa AUA (r<or to 60o yto rr1 geOetovwn ow[ to 5vo Yto rn UE- Oerovfvq rot to dtrtro Ytq rnv toolsur(vq). l-rot( ot oMoy6g tou t<ti6u<q prnopo0oe vo e(etu1- 0otlv eur<ol.6tepo oro prmoX6vdpto oe o0yr<ptor1 Pe rov nuprlvo; To prrol6vdpto ouv06ta l,iyeq npoltetves (-10) xt 6tot to np6BLqtrro rqg onopp00prtoqg ).6yto otrLoytilv mo v6qpo e(vot notr0 l'ry6tepo ooBop6. [lt0ov6v ro rpononoqpr5vo xto6u<6vto 6e lp4otponoqOrlrov eme- vri:g oe 06oetq 6nou ot owtxo-roordoetg oprwo(6rov 0o rpov entBtroBe(q. Etigrprovo Ue rnv un60eo4 qg oordOetoq, 6voq eA6- Xrorog opt0trr6g 31 IRNA (erc6g tou evoprcr{ptou) eivo onopo(rqrog dlore v' ovoYvoJptqro[v rot to 61 r<ro6xo- vro (touldlto,ov dUo IRNA onqroUwot yto rd0e oro- y6veto teoodptlv rodtrov(rov xt 5vo IRNA e(vot ovoyt<o(o ovd (erjyog rcodrov(ou r1 [e1optor6 ro6rG vro). To onodonx6 truolovdptor6 yovtd(trlpro t<ov Oqtro- ottxtirv, toor6oo, rtodxonote( trrcvo 22 IRNA. To IRNA, nou ro6txonotoUwqt oro nuprlvK6 yovt6(olpto, 6ev et- odyowot mo puoX6v6pto rov 0rlLoortr<rilv: ouvenc0g, efdyetot to oupndpoopro 6tt npSnet vo undpla rdnotq rponono(qor1 oroug t<ov6veq rnq oordOetoq yto vo ei- vot 6uvorlq pretd<ppooq oro gttoXovdptor6 p$ootopr6- tto. E(vot ev6totpSpov to yeyov6g 6tt oro prroX6vdpto Uro pn tpononot4pt6vrl oupr6(v4 or4v npriltrl 06sn tou qwrxo6trov(ou e(vqt trov{ vo (euyopdrvet Kot Ue rtq r6ooeptg Bdoetg orqv tpfur1 06oq tou t<to6xov[ou. Mto Inov0utroro 'EwoUo Motrorro EXrvddepgo Nrlgortil6erg 11troru6trptv0eg Murrlteg ZUUeq 0uro AUA 724 KE0A Al0 25 0 levettxdq K66Lr0g
  • 214.
    rdroro Url rpononotnpdvnoupt6[vr1 undpXet oro IRNA nou qwmpooonefouv Kot rtq orcril tetpol.teLe(g oxo- ySvereg ro6trov[tov: Pro, Thr, Ala, Ser, Leu, Val, Gly t<ot Arg. Aur6 elorrcbva ro ouvohr6 opt0pt6 ttov IRNA nou onomotwot oro pmoX6vdpto t<otd ormdr. H petorponq t<lv AGA rot AGG oe rol6x6vto tepgortoporj oro ptro- l6vdpro tov 0r1troonr<drv (B).6ne Ex6vo 25.11) ovorpe( rqv ovdyrq yro 5vo entnp6o0ero IRNA, on6te o ouvolt- 169 onomoUprevoq opr0pr6g IRNA e(vot orptBri:q 22. H gerotponrl tou AUA oe preOetovfuq getrivet nepom6p<o rrlv ovdyxq yto rponono(noq rnq rvoo(vr1g orq 05oq 34 rou tRNAIr" (BL5ne Ev6r4ta 25.6). Or 6rogopertro( rov6veg oordOetog yro 14 petdrppo- on oro ptroX6v6pto Kot rov nup(vo nr0ovdrg nporU- nrouv on6 6rogop5q orrl Lentopreprl doprrl to:v ow(morlrov prBoorrlpor(cov nou Uerogpd(ouv to 60o yovtduill.roro. Ito xurroponLool.totrd p$ootoprdno, ot rpononor{oerq orrlv U34 Xprlorpronototiwot rirore vo eneKro0e( q trov6rrlto onor<o6trono(qoqg optop5vtr:v IRNA (BI5ne Ev6trpa 25.6). Aw(Oero, oro 1.ttto1ov6pto- r<d prBooro;rdrro, ot rpononoqoetg orqv U34 Xpnoqo- nororivror yro vo neptop(oouv to (euYdpopo oro xco6u<6vro nou neptdXouv A ( G orqv tp(tr1 05oq, o0pt- govo pe toug ouvqOetg rov6veg qg oord0etog. Tpono- nouloerg orrlv U34 6wcog undpXouv oe Uroxovdptord IRNA nou owrnpooonefouv oprwo[5o yto 6rpre].e[g oprd- 6eg rco6u<ov(cov rilore v' onogeuX0e( to rorodtdBoopto nou 0o ouv6Bowe oe ow(Oerr1nep(mcoor1. 25.8 Ndo Aurvot6o Mnopel vo Evotoyorul0oriv oe Opropdvo Kol6rr6vro Tepprorroporj + H evotouorool rrlg oetrrlvoruotefvr1q oe optop6vo roldtrovto UGA onoue[ rn 6poon evog oouvrl0torou IRNA oe ouvduool.to pe notrtreq nporetveg. " To oouvrl0roro oprvo[0 nupotruolv1 pnopel vo evootportl0el oe opropdvo ro:dtrovto UAG. * Ito gtreQoptdotpopo npo:ro(oo Euplotes crassus ro rtrl6tr6vto UGA ro0opi(et tooo q oetrqvoruorelvl 6oo rot qv ruorelv1. Yndplouv 60o yvocrrd nopo6erypoto oro ono(o 6vo r<ordrr6vto teptrrotrol-toU Xpnotponolefuot yto vo xoOo- ploer dvo oouvrlOtoro oprtvolri 6togopertr6 on6 ro 20 xlooorxd optvo[6o. M6vo ouyxer<ptptdvo t<tl6tr6vto reptrrorrotrroU enovepUnveUowot ror' out6v tov tp6- no. Aw6 6e(yyer 6l ro v6r1;io rrlg rprnL5tog enqped- (eror on6 qv tout6rqro dMtrlv Bdoecov rou mRNA. 'Evo r6roro 6116 v6r1po ev6g ouyrerpt;t5vou xtodtro- v(ou np6ner vo 6Lor<pr0e( on6 tov ove(dptqro on6 ro ou pup po(6 ptevo nlrl pr1 enovonpoo6toptopt6 tr.ov t<tl6t- rov(ov oe optop6vouq opyovtogoUg 11 oro pttox6v- 6pto, 6nog neprypdqetot orrlv Evdr4ru 25.7. H oelrlvoruoteivrl, orqv ono[o to Oe[o rqg Kustd- vqg 61er owrKorooro0e( on6 oetrrivto, evoo4rcnthrcrot oe ouyKeKptg6vo xtodu<6vtq UGA oe yovi6to nou xto6t- ronororiv oeLrlvonpcoretveg rot ortq rpetq enxpdreeg rnq (oflq. Iuvr'10oq out6g ot npo:retveq rorohtlouv ofer6oovoytr:yx5q owtdpdoetg. To t<otd.tr otno oetrrlvo- ruoretvqg runtrd ewon((etot mo evepy6 r6wpo, 6nou Sreurotrfvet dgeoo rn )nUtKIi ow(6poor1. l-ro nopd6ety- 1.ro, ro xtr:6tr6vto UGA r<ooop((et tq oetrqvoruoretvq oe rp(o yov(6to rou E. coli nou rco6tronotoUv toodv(upto qg deu6poyovdoqg rou UUpUnYrtroU: to evooporo- pr6vo oe).r1vro ouvdmet dpreoo 6vo rorol.uttr<6 r6v ptol'u- B6ow(ou mo evepy6 rdwpo. Ot opyovtopro( nou prnoporiv vo xco6tronotrjoouv rrl oelrlvoruore"wq 6to05rouv 5vo oouvrlOtoro IRNA, yvco- o16 og IRNAS"", to ono(o 6Xet prlxog n6vo on6 90 vou- rl.eor(6ro rot nept6Xet ot5LeXog ono66rtq rot or6treXog T noxfi.ou pqroug. Aw( yto emd (eUyr1 Bdoe- ov oro or6treXog tou qnodSKrq rot n5we oro or6e1oq T (pto 6opd 7/5), to Bor,crlptoK6 tRNASu" 6to06tet pto 6oprl 8/5, evc0 ro tRNASec ttov oplo(tov Kot rov euKo- pucorrilv nr0ov6v 6ro05tu prto 6ogrl 9/5. Autd to IRNA neprdXouv en[o4g 5vo owtro6tt<6vto S'-UCA, nou rouq entrp6na vo StoBd(ouv to UGA, Ie 6Loug rouq opYovt- opro0q, ro tRNASec npdrto ol.twoorutrtc0verot Ue oepfu4 on6 rq oepul,otRNA ouvOetdorl (SerRS) nopdyowog oepuLotRNAsec. ftq Borctripto, ro 6v(upro ouvOdoq qg oel4voruorefv4q pretotp6net to Ser-tRNAS"" oneuOe(og oe oetrrlvor<uoreivulo (Sec)tRNAS"c Xpnotponotc0wog o:g 66q oe).4v(ou to oetrqvogtootpopx6. Ito opXo(o Kor rouq europucilreg, to Ser-tRNASec op1x6 Qoogo- puLuliveror on6 rrlv rtvdorl PSTK nopdyowqq Qooqo- oepul.o(Sep)-tRNAsu". I' 6vo 6eritepo Bqtrlo, ro Sep-tRN4s"" perorp6neror oe Sec{RNAsec on6 ro 6v(u- po SepSecS. H e(otpenrr'1 et6u<6qto qg PSTK efuot o[roo4;re(rorq: to 6v(upro e(vot xov6 vo <poorpopufuril- ver onoretreopottrd to Ser-tRNAsec, evd) onoxle(et to rovovtr6 Ser-tRNAS"'. H orot6Ltrr1l4 qroo<popuL(rooq rou Ser-tRNAser on6 qv PSTK 0o prnopotjoe vo npoKo- Moer evool.rdrroorl qg oeLqvoruorelv4g oe rco6u<5vto oepivnq. H enil.oyri rov rrodu<ov(tov UGA nou np6remot vo epprlvewoUv org oetrrlvoruore(vn roOop((etot on6 qv tonrr4 6eureporoyrl 6oprr] tou mRNA. 'Evog Bp6Xoq goupr<6tog r<o0o6u<d tou rodtxov(ou UGA, nou xole(- rot motye[o SEC/S, onomefuot vto rrlv evoropdtrrlorl t4g oeLrlvor<uoreiVrlq Kqt rov onorLetop6 qg np6o6eorlg nopoy6wcov onetreu06pooqg. Eto Borcr{pto, ro fiol.- 1e(o SECIS Bp(oretot oxptBcirg 6(nlo oro rto6tr6vto U- GA, evrir oro oplo[o Kot rouq europucilteq orrlv 3'-oper6gpqorq neproXri tou mRNA. Iro E coli, o e[et- 6rreupr6vog nopdyowog enttrrrptuvorlq SelB, oMrltrent- 6pd onortrermrxd pe ro SectRNASec 6q1 pe rov6vo dLl.o o1-uvoorutrtco gdvo IRNA, ou pneprl.oprBovo;t5vou rou np66po;rou Ser-tRNAS"'. O SelB enloqq npoo66ve- rot dpeoo mo ototle(o SECIS. To onot6leoUo rnq dpdonq rou SelB e(vot 6tt pr6vo to xto6tx6vto UGA nou en(oqq nept6Xouv 6vo r<otdtrtr4l,o tonoOerqpdvo SECIS 25.8 N6o Aprvo[60 Mnope(vo Evoroltotto0otjv oe 0ptop6vo KodrrovroTeppolopou 725 Boorxeq eworeq
  • 215.
    npoo6{ivovror nopoyoytKd oroSectRNASec oq prBo- orrrpxq 05o4 A {EiK0NA 25.12}. To oplo(o Kor or euKo- pucbreq 6ro06rouv 5vo og6Loyo rou SelB, toor6oo yro vo enmpdQouv oro prBooro;rdno vo erooydyer q oetrq- voxuorefvr1 onomo[v enrn].6ov rrlv npoteivrl SBP2. 'Evo dMo nopdderypro evoropdrtoorlg ev6g er6u<ori ol.uvof6og e(vor q ronoO5rqoq qg nupoLuolvqg oe ouyrerpr;r5vo rodrr6vro UAG rou y6vouq Methano- sarcina rcov opXo[rov, 6ncog en(oqg Kqr oe ].ryo Borcrr1- pro. Iro Methanosarclna, nupol,uo[v4 undpXer oro evepy6 r6wpo rcov geOulorpovogepoocirv pre0uLopr(- w1g, 6nou nof(er oqprowtr6 p6],o orrl plpur<r1 ow(6po- or1. H evotol.rdrcoorlr4g nupol,uo(vqg npo0no06rer pro e[erdrreup6vr1 o;uvooru].otRNA ouv0erdo4, rrlv nu- poLuoul,otRNA ouv0erdo4 (PylRS), q ono(o oprvoo- r<uLrc0ver 5vo efer6u<eup6vo tRNAPyl pe nupoLuo(v4. To tRNAPvr nepdler ro owrxro6rr6vro S'-CUA, fiou ro Kq0ror6 xov6 vo 6roBd(er to UAG. 'Onorg Bp60qr<e yro ro tRNASec, ro tRNAPvr en[oqg eprgov[(er oouvq0tmo 6oprrd XopqKrnprorrrd nou 6ev undplouv oe dl'l.o IRNA: yro nopdderypro, 6e 6ro06rer ro moOep5 vou- x.l.eor(6ro U8, evc0 o Bp61og D ror o peroBAqr6g Bp6- Xog efuor oouvqOtoro Bpole(q. O prr;lovropr6q Ue rov ono(o ouyrer<pr1.r6vo rodrr6vro UAG droBd(owor oq nupoLuofurl 6ev SXer 6rol,eurov0e(, enet6rl 6ev rot6- orq 6uvor6 vo rouronorqOe( oogrilg 6vo ororXe[o beu- reporoyorlg SoUnq oe 6Lo rq mRNA ro ono(o evoo- porcbvouv ro opuvo{ri. Enrn}.5ov, 6ev 61er ovoyvopr- me( rdnorog er6rx6g nopdyovrog enrprp<uvoqg trre or6Xo ro Pyl-tRNAeu. i1o),6 np6ogorq qvoKo^.rig0qr<e 6n ro rodrx6vro UGA ro0op((er rrlv evoopdrolor; e(re qg ruore(vqg ei re rrlq oei.qvorume'lvqq mo B].egopt6og6po npcor6- (ao Euplotes crassus. Arrn Xprion rou UGA Bp60rp<e 6l ouprBofver or6;rq ror pr5oo oro (6ro yov(6ro, evril q enti.oyrl rou oprwo[rlog nou 0o evocol.rorcoOe( e[optdror on6 rr1 5op{ rqg 3'-oprerdcppoorqg nepropig rou mR- NA. To UGA npoodtop(er qv r<uorefv4 yevu<d cro Eu- p/ofes ror 6e Aenoupye( crlg xrodrr6vro teppoloproti. Iuvenc0g, ow6 uno64Lriver 6rr er6x4 rog npog 14 06o11 6rrq Xprloq prnope[ vo ouprBe( pr6oo oro n].o[oro ev6g rodxov(ou nou 6ev Xcnorpoflorekor erddMrrlg yro rep- prorropr6 oro ouyKeKprp6vo opyovrop6. npood6ver mRNA f lX&N,q :;"1t 0 SelB e(vor evog nopoyovrog enrprlruvorlg, nou npoo66ver etdrrd oro tRNASec 6vo rodrr6vlo UGA, ro onolo oxo- trou0e(ror ono pro 6opr1 oretreXouq-ppoXou oro mRNA. 726 KEOAAAI0 25 0 levelrog K66u<og 25.9 To IRNA Zeuyopdrvouv ErAerrrrd pe AprvoE6q Mdoro AprvooruAo-tRNA Iuv0erootirv * 0r optvoor<utrotRNA ouvOerooeg e(voL 1uo orroy6veto ev--- Uov, r0 ono[o npoo0rlvouv olrrvo[rlo oro IRNA nopoyovi:, o;.uvooru),otRNA oe pro ovr6poo1 6uo pqporov nou Xprl- gonore( evepyero ono ro ATP. * Ko0e ogrvoorutro{RNA ouvOerool optvoorutruiver otro ro :r. NA oe pto loo6exrlrq opoOo nou ovlnpooorneuer dvo oul.,e. xprpdvo oluvo[u u H ovoyvtirprorl rou IRNA ono lg IRNA ouv0erooeg Boor(e:: 0e Uro ouyKeKptpevl ogoDo vourtreol0[rrlv, Trlv (opo00 TC-- rorqrog,, rou tRNA, ro ono(o ouXvo ouyrevrp6vovroL : - ot6treXog rou onod6rrrl ror ro pp6Xo rou ovrlrodxov(ou To optvo[6o etod,plowot orn Uerogpoorrxq o6o ;,: otrl rnq dpoorlg rov oUrvooKUIo-tRNA ouv0erooc, or ono(eg nop6louv to onopoLrrlro onorodrr<onorr..- 16 Bqpo nou perorpriner rrq nlrlporpop(eq rov vo-- r).eivrr<rov o[5crrv mqv no].unenrLdrr<r1 o].q].o u Xi : Otreg or ouvOerdoeg 6pouv pr6oor ev6g Bnxovrofrc- 6[o Bqpdrov, nou nopouord(mor ofllv f i{*iiir r: :.] o To otuvoErj npoto owLdpd pe to ATP on6re oXqu:- t((eroL 5vo ollrvoorul.o-o6evuLrr<d ev6rdpeoo ncc- i6v, evur one).eu Oepoveror n u poqoo<prr-lprr<d. M d,c c : r1g evdpyerog nou oneleu0epurveror orrlv udpoi.-- oq rou ATP noyr6eferot un6 Uopgn 6eopoli ;rrx.:- ovudp(q ut{lnlnq ev6pyetog orrlv o6evu).u<r1 oproE: o Irn ouvdxero, e(re | 2'-OH e(re 11 3'-OH o;ro6o r.c- ewon((owor oro 3'-A76 vour<.treor[6ro rou IRNA c.- rL6pd pe to xopBovu).tr<6 dropro dvOpor<o rou *.- r<roU ovudp(rr1, nopdyowoq oprvooKu).o-tRNA *: row6Xpov4 one).eu06,prooq AMP. OL r5ooeptg IRNA ouv0etdoeg nou e(voL er6rxrig 1',: rq y).outopr(v4, to yLoutol.ur<6, trlv opyrv(v4 rot rq Luc.- v4 onoLroriv rlv nopouo(o IRNA yro vq ouvOdoouv r: ol.rLvoorulo-odevuLtr<6 ev6Ldpeoo. Ze ourd ro dv(uprc 4 IRNA ouvOerdorl oomd 0eope(tor rog 6vo prBovo"- xLeonpoteivrr<6 ootl-rot[dto (RNP), oro ono(o }.error..rc- y(o rou RNA e(vor vo Boq0d tr1v npore'tvr1 vo 6torr1pa. 1-uo r<oroi.urtxd enopr<q 6roprdpgooq. Iro deUrepo Bn- po rrlq oprvoor<uLirr;orlq, ro rUnpo rou o;-rLvo(5og ro- o1-rrvooru).o-o6evu).tr<oU gero<p6peror oto RNA ro" RNP (6't).o6n, oro IRNA). Kd0e IRNA ouvOerdorl e(vor ex).ercrtrr1 pr6vo yLo dvc opttvo(ri pero[ti 6].otv rrrtv oprLvol5orv rnq KUTroprKf c 5e[opevr1g. En(or1q 6tor<p(veL 6].o ro tRNA rou Kurrc- pou. Iuvrl0rog, r<dOe opLvo(0 owrnpoooJneUeror onc neproo6tepo rou ev6g tRNA. I-loLLon).d tRNA (ocr-rc lperd(owot yro v' onowotiv oe ouvrovul.ro r<ro6rrdvLc evri prepu<dq eopdq 6rocpopelr<d IRNA (euyoprbvouv ;.-rs to (6Lo xo6Lx6vto. lloL).qn^.d tRNA nou owrnpoot.rJ- Boorxeg eworeg
  • 216.
    'Ev(ugo neproXn ogrvo[6og lleproXli tRNA [ieoLo vn AipH ouv0eroorl nePLdXet 3 neptoxdg npdodeonq fl "-H-C-NH" Y -u- l O-U.U -o-p=o DU-T=U + AEevocivti To opwo[t rot ro ATP oplgori(ouv opwooruLo-AMP To tRNA npoo06veror To tRNA tDoprriverut Ue ouwo[ri n H-C-NH2 c"t+B:.'0-H r G,f 0 S Aoevootvrl R I H-C-NH I QDtro 'u lCS { EIK0NA 25.13 MLo optvoorutrotRNA ouv0etdorl entqop{(et to IRNA pe 6vo oluvo(ti. neriouv ro (6to optvoqf KoloUwot loodeKrlKd IRNA (isoaccepting IRNA). 'OLo rq IRNA eprgov((ouv u1v rovovtrrl tpttotoyri 6oUn qfriUotog L (Bldne Keqdtroto 24, MetdEpao4), To IRNA 6rnldrvetot t<otd t6toto rp6no c0ote to cr6LeXoq- ono65r,cr1q Kor ro or6^exoq T vo q(4Uor(ouv 1"tto ouvo- (ovrrui orriLrl, evdt to ar6lexoq D rot to or6leXoq tou qwKo6txov(ou vo qXq;roil(ouv pro((tov xdOeto BpoX(o- vo oX4prorog L. O Bp61oq tou qwtrodtxov(ou Kot ro drpo-onob6rcqq CCA tono0etoUwot oe ow(0eto dxpo rou 1-rop(ou rot onSlouv nep[nou 40 A. H otpotptrcl ne- ptoXrl rou oppoti rou IRNA, q ono[o ouv6Set rrq 60o r<d- Oereg orr1l,eg, onorctre(tot on6 to 9p6Xo D, to Bp6Xo T, ro ;retoBtrqt6 Bpo{ovo rt 6vo dtoorrltrrorr6 tptlpto 60o voux.treort6(rov preto[ti rou oreMXoug-ono6drctq Kot rou oretr61oug D. To neptoo6tepo IRNA nepdXouv ;rxp5q geroBLrlrsg nepto16g, nou onote^,o0wot on6 5vo Bp6- Xo 4-5 vouxLeortd(ov, evc0 eldXtoreq too6emtr6g opt6- 6eg e;rqov((ouv 6vo ;reyol0tepo pretoBtr4t5 BpoX(ovo, nou oupnepiloprBdvet 6vo (euyoptogSvo (6(xl'tovo) o16- LeXog, ro ono(o npoe(51et on6 6vo otpotptx6 nupfvo. To xorv6 oxnpo L t<ov IRNA e(vot 0eprel,tdr6eg yto rqv oLl.ql,en(6pooq 6l,tov ttov IRNA Ue rouq nopdyoweg enrprlruvorlg rot to ptBootoptdtto. Iro nLo(oto owr;g rqg r<otvqg 6opnq oXipcroq l- nou evtq4detot on6 rrlv nopouo(o ouvrrlprlpdvtrlv tpr toroycbv oMrlLent6pdoeov pt6oo crro ogotpx6 nuprl- vo, or oLtrrllouX(eq rou IRNA onoxtr[vouv mtg neptoo6tspeg O5oetg Kqt rov teoodptov Bp6ltov tou proplou. Auri 11 notxt),6rr1to drlptoupye[ ].em6g dtogo- p6g orn yov(o peto[ti trov 6Uo BpoXt6vov tou L t<ot, to or11.rowtr6tepo, odqye( oe nopoNoy5g orqv nope(o rou notruvouxtreor6txoU oxel,erorj xotd pr4xoq rou po- p(ou. Aufi orprBdrg q 6opxrl notril.6tqto e(vot 11 Bdorl yro q Srdxptorlrov ;rop(<ov on6 ng IRNA ouv0etdoeq. Ot IRNA ouv0etdoeg 6torp(vouv to IRNA p6oo 6tio yevrxcOv prrlXovto;uilv: dpeo1 @irect) t<ot 61tpeor7 avd- W@o4 @direct readout). Etrlv dpeoq ovdyvtoo4, to 6v- (upro ovoyvtop((er dpreoo Lettoupytt<5q oprddeq et6tr6g yr.o ouyKeKprp6veg Bdoerq-n.X. 6vo entgovetor6 opttvo- [rj puog IRNA ouvOfidoqg pnope( vo 5eX0e[ 6vo 6eopr6 u6poy6vou on6 rrlv e[orux].tr<ri oprwopddo rqg youov(- vnq Gnv N2 qg G), pto oprd6o qg eLdooovog otitrot<og nou 6ev undpXet ortq dLLeg tpetg Bdoerg, Aw(0ero, orrlv 6;rpeorl ovdyvoorl, to 6v(upro npoo66vetot d;reoo oe pq etdu<d rprlporo tou IRNA, ro ooKxopoogooqopt- 16 oreLer6 rot gr1 et6xd tprr]goto ttov vourtreortdxrbv Bdoeu:v. l-to nopd6etypto, oll,rlLouX(eq oro pretoBLrlr6 Bpo(ovo r<ot ro Bpol(ovo D ev6g IRNA prnope( vo 6qgt- oupyrjoouv 1.uo t6to(tepq entrpdveto, ouprnlrlpoprotrr{ pr6vo ge qv ow(orotXrl IRNA ouv0etdorl rot rop[o d],- lq. Kot' our6v tov tp6no, voux.treoil6to to ono(o on6- louv on6 q 6tdcpooq ev(UproutRNA 6rlprtoupyorjv ;tto 6o;ui drdqoonq nou ge r4 oepd r4g npooS6vetot dge- oo. T6oo n dueon 6oo rot q 5prpreo4 ovdyvtoo4 ouvti- 0tog trettoupyotiv cno ntro(oto oprotBo(og enoy6pevqg npooopproyrlg: ot oMoy6q dropr6prptoorlq t6oo oro tR- NA 6oo Kot ro 6v(upro ouptBo(vouv prerd qv opXtrrl np6odeorl oXq;rot((owoq 6vo nopoytoyx6 xototrurtx6 ofgntroro. Kot oroug 66o pqlovtopotig ou;yd en(or1g oullper6Xouv pr6pto vepo6 orrlv entrpdveto enoqfg ge- ro[ti tou IRNA ror rou ev(Ugou. l-to nopd6ety;ro, 5tov q ouvOerdorl tou yl,outol.ttvul'o-tRNA (GlnRS) npoo66- ver ro tRNAGrn, 60o neptol6q tou ev(Uprou neptorp6rpov- ror rl pto oe oX5or1 Ue rnv dlLr1. Tout6Xpovo, to 3'- pov6x).rovo drpo xot o Bp6Xoq tou owtxtodtrov(ou tou IRNA uE[orowot onpowtr6g oMoy6g 6topr6prpoorlg oe oriyxpro'q pe ttg unoOerx5q 6opr6q rouq orn Un ouv6e- 6epr5vr1 r<otdoroorl. Ie noLtr5q neptntc0oetg, ot xo0optor6g tou IRNA nou e(vot ovoyxo(ot yto er6trcrl ovoyvdrptoq ewon(owot oro drpo rou pop(ou, 6r1Lo6n crro or0reXog-ono65rt4 Kor ro Pp5Xo tou owtrodtrov(ou. Yndplouv oor6oo nopo6efuproro qro ono(o to vourtreot6to rou rptroro- yoUg nuprivo nop6louv to orlpoto tout6t4rog. 'Evo dMo xow6 vourtreot(dto toutono(r1or1g e(vot n *pdon 6rdr<ptor19" ("discriminator-base") orrl 05or1 73 tou tR- NA, 11 ono[o ewon((erot oprSoog 5'qq 3'-re].tt<{q qI n- l,ouX(og CCA. E(vil ev6tocp5pov 6tt q otrLrlLouX(o owt- rro6u<ov(ou rou IRNA 6ev e(vot ovoyxoortxd onopofu4- rn yro rnv ovoyvcirpto4 et6trriv IRNA ouvOetoociv. l-evt- r<d, 11 oprdSo tour6ultog rou IRNA e(vot tdtofuep4 yto rdOe IRNA ouvOerdoq. 25.9 To IRNA Zeuyoptirvouv Er),exnrd pe ApLvo[60 M6oro AgwoorutrotRNA luv0eroodtv 727
  • 217.
    Or roOopror5g rout6qtognorrilouv toq npoq rn oqproo(o rouq Kor preprx5g gop6q ouwqpoUwor r<ord qv e[5].tlr1. H ouwrlprlorl rov ororxe(ov rout6rqtog ovo6erKv0erqr on5-r4v rxovSrpo noMc0v IRNA ouv0e- roorilv v' otruvooruLrcilvouv IRNA nou npo6pXowor on6 drogoperxoriq opyovroUorig. Yno06oerg oXenxd Ue rrlv opd6o rcov ororle(tov row6ul-rog rou IRNA nou e(vot onopo[r4to yro rnv ex].er,crr4 ovoyvdrprorl on6 puo tR- NA ouvOetdoq npo6pXowot on6 preldreg rpumdMov IRNA ouv0eroo6v ;le nep(O).ooq orcr(vtov X, r<).oorrc] ye- verLxrl ovdl.uon Kor rn vfuo trreroMo[ty5veor1. H oprorxrl on66er(f 6n gro ogd6o tow6r4rog rou IRNA e(vot ro- Ld xo0oproprSv4 ),opBdveror qn6 nerpdproro preropr6- oxeuorlq, oro ono(o 11 nr0ovo).oyoti;revr1 oprddo vou- rl,eol6(ov evocopor6veror o' 5vo IRNA on6 pro 6ro9o- penr4 roodercnrrl oprd6o. I-ro nopd6erypo, q owrrord- orooq 15 vouxLeon6[ov oro or6].e1og-ono66rcrq ror to 9p6Xo rou owrxrodu<ov(ou rou tRNAAsp on6 r' ow(- ororxo voux.eor[6rq rou tRNAGln en6rper+re orq ouv0e- tdorl rou yl,ouroprvu).otRNA (GlnRS) v' optvooKu- Luioet ro rpononorqpSvo tRNAAsp pre yl,ouro;r[v4 pre onoreleogorrx6rrlro rr erLerrx6rrlro ouyr<p(orpeq Ue rtg ow(ororleq ornv rovovu<r1 ow(6poo( qg. l-1oLl,6g IRNA ouv0erdoeg opuvoor<uLuilvouv erdu<d 1.uo "1-uvui),xq" ("minihelix") rou IRNA, q ono(q qnore- Lefuqr on6 ro BpoX(ovo-ono65rcrq xor ro Bpol[ovo TrfC rou pop(ou. Ie opropr5veq neprmriroerg, rog undorpcol.ro pnope( vo l,eroupyrioer Uro purpo6lrro rou IRNA, nou onoretrefuor ;r6vo on6 6vo orrileXog-onod5rcrq xlerqr6 oro dnro drpo rou on6 5vo oro0ep6 retpontr6 Fp6Xo. T6oo or pwdlrreg 6oo ror or pxporiLxeg oprwoor<uLui- vowor fuy6repo onorel,eoportxd qn6 ro or5poro IRNA. Autd rq nerpdproro 51ouv rdnoro orltrroo(o, rosr6oo, yto rnv e[e],trcnxr1 ovdmu[r1 tolv ougn].6r<rov r<ov IRNA ouv0erood:v. I' 5vo np6r;ro e(efur<rx6 ord6ro, ro IRNA (oog onote}orjwov pr6vo on6 ro BpoX(ovo-ono65r:rr1 tou oriyXpovou prop(ou. 25.'l O Or AprvooruAo-tRNA luv0erdoeq KorordooovTor oE Ario Orroy6vereq * 0r oprvoorutro{RNA ouvOerooeq dtorp(vovTor oTrq orroy6ver- eq To[nq I ror ll pe goorl rdroirepo porLBo otrtr1trouXulv ror tdr- olrepeg 6opLreg neproX6g. l-iopd rrlv rorvrl enoupy[o roug, or ouvOerdoeg e(vor opr<etd erepoyevqg oprddo ev(riprcov. Atoxp(vowor oe 6r-io orxoy6vereg. Or IRNA ouv1eraoeq Tar1q / e(vor r<upftog provoprepe(g r<r epr<pov((ouv 6oprr<d nop6proreg neproXSg (nruX6g) Rossmann (Rossmann fold do- mains) cro oUrvoreLrr<6 dxpo 11 xowd oe our6, ot ono[eg oupprer5Xouv mr1 6ropr6pgoon rou evepyori r<5,wpou. H nruXrl Rossmann onoteLe(ror on6 5vo B- 728 KEOAAAI0 25 0 levettroq Kdrdtxog muX<ot6 g0Mo 5-6 nopdMrlLcov muXc0oeov Ue otr vernx5g SLrreg. Aurq q neproX4 e(vor opr6Loyn rnq neptoXrlg rou evepyo0 r<6wpou rrov deudpoyovoorirv ror unefOuvn yro rnv np6odeor; rou ATP, rou oprvo- [6o9 r<ot rou 3'dxpou rou IRNA. 'O].eg or ouv0erdoqg TdEnS I nepr6louv puo neproXrl "np6o6eonq rou ono- 66rcrq", q ono(o evo<rt1lor6veror srnv (6ro 05oq rqq rnuxnq Rossmann oe Uro rotvri 06o4, 6nou en(oqg npooS5veror ro l.rov6xLrovo drpo-onod6rcrqq rou tR- NA. Ie peprrd 6v(upro, ourl 11 06oq en(or1g nepdXer 6vo evepy6 xSwpo ene[epyoo(og (BI6ne Ev6r4ta 25.11). Or xopBo(urelrxSg neprolrlg rotv ouvOerooriv tdlqg I npoo66vowqr ornv eoo:reprx( yrov(o rou tR- NA oXqgorog L xor oro Bpo(ovct rou owrKo6rxov(ou xor q Aenoupy(o roug e(vor vo drorp(vouv ro IRNA. AUo rotvd BpoX6o ;ror(Bo o).Lr1).ou1rc6v qg nrulqg Rossmann egnl6rowor ornv np6o6eoq rou ATP. Ercr6q on6 rdnoro nepropropr6vq oproi,oy[o prero[6 ].i- yrrlv ev([;rov, 6ev undplouv orlprowrx6g opror6qreg orrl 6op( 4 qv oLlql,oul(o rov ev(rigtov rdfirlg l, err6g qg nruxnq Rossmann. Ot fRNA ouv9xdoeq Tdlryq il e(vor en(or1g erepoye- ve(q. H tetoproroyrls 6opn roug yevxd e(vor 6rprepqg, roor6oo prepu<69 qopdq oXrl;.ro{(ouv oprorerpogepq 4 ozBz ereporetpouepn. 'Onog ro 5v(upo td(r1g l, ot tR- NA ouvOerdoeg rd[qg ll 6ro06rouv puo 6o;rxd ouwn- pnUSvn neproXrl evepyori r6wpou - ornv npor<erpdvrl nep(nrororl np6retror yro pro prrrcrl o/B neproXrl nou 6rog6per on6 r1v muXr'1 Rossmann. To evepyd xSwpo rov IRNA ouvOeroocilv rd[rlg ll ewoni(owor npoq ro t<opBo[ure]lx6 dr<po rov no],unemr6(cov. Ie ourrl r1v oroy6vero e(vor ouwrlpqpr6vo rp(o BpoXSo ;ror(Bo ol,- l,4Louluilv orqv neptoXri rou evepyoti r6wpou: ro 5vo on6 outd Letroupye( mov no)r,upepropr6, evdl r' dMo 6Uo 51ouv r<orolunr6 p6).o. YndpXouv 23 <pu).oyevmrrd 6rorptr5g orroydvereq IRNA ouvOmoorilv. 'Ewexq xorordooowor orqv td[q l, ev6 or un6lotneg 12 e(vor dv(upo rdEnq ll (ElK0]lA 25.14).'EXer ev6rog5pov 6r undpXouv 6rio 6togopetu<o( rfnor ev(rjpov LysRS nou ov{rouv oe 6rogoperrxdg rd- (erg. l-1 p6orporo en(or1 g ovor<o).ri g0r1 rov 6 [ o orroydver- eq Url Kovovrrtiv IRNA ouvOetoorilv nou neprop((owor oe oprol-uivo e(6r1. Aurd ro 6v(upro e(vor 11 nupol.uoul,o- IRNA ouv0erdoq (PylRS) rd[r1g ll (neprypdgetor ornv Ev6r4ra 25.8) xor n eooeooepuLotRNA ouv0erdorl (SepRS)td[49 ll. H SepRS neprop((etor oro pre0ovoy6- vo (puo un6ro[r1 rov oplo(rov) ror ro orevd ouyyevx5 Archaeoglobus fulgidus. llpoo6Sver gologooep(vr1 (Sep) oroug ono66meg rou tRNACys nopdyowog 5vo peprrtirq oxuLuop6vo Sep-tRN4cv". 'OLor or opyovtopro( nou 6ro05rouv SepRS en(or1g 6to05rouv to SepCysS, 6vo ove(dprqro rrlq gtrrogopK4q nupr6o[d],qg ouvo66 5v(u;ro ro ono(o perorpdner ro SeptRNAcv" oe Cys-tR- NACys. O 66rqg Oe(ou nou Xpnorponorefror on6 ro SepCysS in vivo e(vor dyvotmog. E(vor o[roorlpe(trrro 6rr opropr6vo pe0ovoy6vo 6ro0drouv qv o66 6Uo B4- ptdrov rolv SepRS/SepCysS rot, nop6M4Lo, ro rovovr- r<6 5v(upro CysRS. l-1op6n undpXouv 23 rpuloyeverrxd 6rorpudg orro- Boolxeg ewocg
  • 218.
    Apwooxu),0- tRNA ouvOerdotg TdqnI Td[n ll EIK0NA 25.14 AroXoptopog rov IRNA ouv0er0o6v oe d0o Td- [erg, nou otoo5rouv onor]retmtxd opotpo(eg oU66eq UoTiBov otr- )rnlouxtdrv rot 6optrd evepytiv neptoXtitv. Irlgetdtvetot 11 reroprotoynq 6oprl rou ev(upou. tlol)ron)to( 6e(xreg unodet- xvoouv ofl 4 retoptoroyrlq doun dtotp6pet oe nox0rouq opYuvt- oUouq. H retoprotoyrlg 6o911 qq PyIRS 6ev 6Xet xo0optore[ oorptirg. ydvereq IRNA ouvOetoooiv, ot neptoo6rcpor opyovtol.to( dto06rouv g6vo 18 6v(upro. H GnIRS Kot rl ooilopoYtvu- tro-tRNA ouv0etdorl (AsnRS) tunrd onouotd(ouv on6 ro penepr6pto rov opYovto;rc0v. l-to vo ouv06oouv Gln- lRfrlfiotn Kqr AsntRNAA"n, ouro( ot opYovtouo( dto06touv [eX<oprmd 6v(upro trr' evepy6rrlto y],ouroputro{RNA ouvOerdoqg (GluRS) Kot oonopruLo-tRNA ouvOetdoqg (AspRS), to ono(o 6ev epgov((ouv xov6rlto dtdrptorlq (nondiscriminating-ND). H GIuRSND ouv06tet 16oo Glu- tRNAGru 5oo rot pepxdrg oxutrttopt6vo Glu-tRNAGrn, evr6 4 AspRSND ouv06ret 16oo AsptRNAA"p 600 rot peptrcig oxuluol.uivo Asp-tRNAA"n. Itrl ouv61eto, to peptxcOg orulrcopdvo IRNA ;retotp6nowot oe GlntRNAGrn 6q1 Asn-tRNAA'n pre q opdorlq Utoq oprt6orpovotpepdoqg nou e[optdtot on6 to IRNA (amidotransferase-AdT). Ot AdTs e(vot o[tooqpre(<oro noLupreprl 6v(upro nou 6ro05- rouv rperq 6ror<pm6g evepy6qteg (ElK0l'lA 25.15)' flprirro nopdyouv opr;rorv(o o' 6vo evepy6 x6wpo pr6oco onog(- 6ooqq ev6g 66q o(tirou, 6noq 11 yLoutopr(vq 4 r1 oonopoy(vq.'Enstto, 11 oprprcov(o pretog6petot 6to;t6oo u ev6g evdoptoptorori 6to[].ou rot ovo6tletot on6 pto deUrcpq 05or1, 6nou npoo66vetot oro 3'-drpo tou trd- 0og or<uLuo1l6vou IRNA. Iro de0repo evepy6 x5wpo, pro rrvdorl y-gtrrotpopuLttilvet rqv r<opBo[u]'oprd6o rrlg ni.eupu<r1q oLuofdog rou ogwo[6oq oro Glu-tRNAGh n ro AsptRNAA'n. T6Log, q oprpov(o ercron(et ro Qooqo- prr6, g4qpror((owoq Gln{RNAGrn ri AsntRNAA'n. Yndp- louv [eXcoptordg oroydveeq AdT nou enevepYoUv rot sro 6Uo Ld0og orultorp6vo IRNA n neptop((owot pt6vo sro oxrlUorropt6 Gl n-tRNAGr". Or ouv0erdoeq rd(nq I r<ot ll 6tor<p[vowot lemoupyt- xd on6 oprerSg on6q.retg. [lptirtov, ro 5v(upo tdfiqq I ol.rlooxullc0vouv ro IRNA or4 2'-OH e6on qg A76, evcir ro 5v(upro rd[qg ll yevtrd opttvoot<utrtcilvouv to IRNA arnv 3'-OH. H 05on qg opXtxrlg ogwooruL(trloqq q(eil- tBNAGrn >-Gru.tRNAcrnKGlu + ATP Gln + ATP tRNAAsn Asn-tRNAAsn y-Asp-tRNAAsnN Asp + ATP Asn+ ATP Asp EIK0NA 25.15 Mryovrogo(yto rn ouvOeon GlntRNAGln xqt Asn- tRNAAsn. Ie xdOe nepinroorl, 11 dvto nope[o unodexvUet qv odti ev6q Bnporoq nou rotqluerot on6 q oupponxn IRNA ouv0etdol. 0t xor6repeg o0o( Dtlo Bnpdttov onovroUv moug neptoodrepoug opyovroporjg. Anoretrouwot ono pto pr1 dteurptvtortxrl IRNA ouv- Oerdor1, nou orotrou0e(rot onri rq 6poor1 ev6g tRNA-e[opr6pevou ev(rjpou opt6orpovorpepoons (AdT). (eror ge tqv roteUOuvo4 np6odeonq rou IRNA ndvo oro 6v(u;ro. Ot ouv0etdoeg t6[r1g I npoo6Svouv IRNA orrlv nl,eupd rqq eldooovog otiLor<oq tou oretr61oug- qno66Krn rot onoro6v to pov6xl,tovo 3'-rel,x6 drpo vo 61et 6opuj <poupr<6tog yto xotdMqtrrl owtnopd0eoq pe to opwo(ri ror to ATP cro evepy6 r<6wpo (EIK0NA 25.16). Aw(0ero, ot ouvOetdoeq td[qg ll npoo66vowot orrlv nleupd t4g pre((ovog orilorog tou oreMXouq-ono66tc4 rou IRNA rqt 6ev onomoUv vo oxnportare( goupr6to peto[ri tou 3'-dxpou Kot rou evepyoti t<6wpou. Yndplet en(onq Uto Unxovtorxrl 6rogopd nou np6ogoto ovoKo- , 0r xpuorotrtrLrdg 6op6q unodetrvuouv ort ot opl- voorutrotRNA ouv0erooeq ro(nq I rot ll npoodtlvovrot orlg ovr[- 0ereq oQetq rorv tRNA unoorptlportov T0uq. To tRNA onerrovl(erot epuOpo rot 1 npotelvrl ruovrl. H tporoypotpto nopo- Xopn0lre ono tov Dino Moras, lnstitute of Genetics and Mo ecular and Cellular Biology (IGBMC). Asn Asp Ser His Lys Thr Pro Phe Ala Glv Sep Pvl Gln (o) Glu (o) Arg (o) Lys (o) Val (o) lle (o) Leu (o) N4et (0,02) Cys (0, o2) Tyr (02) Trp (02) (oz) (q2) (oz) (oz) (oz) (oz) (oz) (0, oz0z) (qz, o+) (oz, oz9z) (0, ) (?) Gln + ATP Asn + ATP J*.1& 0L A;rrvoorutrotRNA luv0erdoeq Kororoooovrotoe Auo OLxoYsveLec 729
  • 219.
    ).0q0r1re: 11 roXUqroow(6pooqq tcov ouvOetoocilv rd- (qq I neprop((*ot on6 rrlv onetreuO6pcoon optvoorufuco- pSvou npoi5wog IRNA, ev6 or ouv0etdoeg tdEnq ll neprop((owor on6 npoql6repo Xrlpur<6 Bqgoro Kq/n eu- orr6g ovo6rord(erq oro evepyd xdwpo. 25.1 I Or Euv0erdo€q Xpnorponoro0v Tn Ar6p0oon Aorrpitov yro vo BeArrdoouv rnv ArpiBero r H er6xriuru rou (euyoptirlroroq uero[0 ogLvo[6ov roL IRNA pu0gi(eroL p6orrl ovndpooeov 6r6p0too1q 6orq[ov ot ono(eg u6potrtiouv trd0oq oxnUottop6veg optvooxutro-odevu)txdg ev6oeLg roL oluvooxutrotRNA. Or oprwoor<utrotRNA ouv0etdoeg np6net vo 6toxp[vouv 6vo er6u<6 opuvo[ri on6 rrlv rurroprr( 6e(opevrl tov oprwof6tov Kot rov ouvorpdrv prop(tov, 6nog en(,cr1g rot to ouyyevrl IRNA oe prcl ouyKeKprprdq toodererxrl opd- 6o (runtrd 5vo 6ro9 rp(o) on6 to otlvotro tcr:v IRNA. H Srdrproq rou IRNA prnope[ vo enmeul0e( ge Bdoq ng 6rogop6q ong 6op5g oXrlprotog L (BMne Ev6tqra 25.9). Aur6 ou;rBo(ver t6oo r<otd to opXx6 Bf go np6o6eoqg 6oo xqr mo en(ne6o rrlg enoy6gevnq npoooppoyiq. To prl ouyyevrl IRNA nou npo6pXowor on6 dtrtreg too- 6erctx6g oprd6eg 6e 6ro06touv rqv ntuipq oprd6o vou- r.l.eol6(rov rout6rrltog. 'Eror, 6ev efuor oe 05or1 vo enovodtord[ouv rrl 6opr( roug c0ore vq utoOetjoouv puo 6rop6prpoon ornv ono(o to owt6poortr6 telu<6 drpo CCA eu0uypoprpr[(etor r<otdLtrr1tro Ue rnv xopBo- [u].opd6o tou ogwof6oq Kot ro o-gooeoptK6 tou ATP. Autq 11 on6ppr{.rr1rov Un ouyyevd:v IRNA o' dvo ord6to qg ow[6pooqg nprv on6 rr1 orJv0eoq tou pepmrig ot<u- Luog6vou IRNA prepu<69 ovog5petor og rtv4nx{ 6l6p- 0roo4 6orrp(ov (kinetic proofreading). H o6uvo;t(o rtov Un ou1ryevc0v IRNA v' oxolou0rloouv to Xqprxd Brl- Uoro rnq oploorutr[oo4g npo6plerot qn6 to yeyov6g 6rr ro IRNA dr(ororot on6 to Sv(upo notrU toX[tepo on' 6,rr pnope( v' owt6pdoet (EIK0NA 25.17). Aw(Oero, or IRNA ouvOerdoeg o6uvotoUv vo dror<p(- vouv oproptdvo 6opxd nop6ptoto oprtvo(6o rotd r1v no- pe(o rrlq o[vOeoqg rov oUtvooKu].otRNA nou oup- Bo(ver oe 6rio Brlpoto. E(vor r6tofuepo d0orotro Yto ro 6v(upro vo 6tot<p(vouv 60o opuvo[6o ro onofo 6tog6- pouv pr6vo oq npoq to prrjr<og tou ovOpot<tt<of ordr.e- to0 (6ql,o6ri rord pro opd6o CHr) ti optvo(6o (6tou prey60ouq nou droqdpouv ;t6vo oe Uto 06oq. Fto nopd- 6ery;ro, o 06l,oro9 np5odeo4g optvo(5tov rqg tooleu- xutrotRNA ouv0er6orlg (lleRS) 6e prnope( vo 6tot<p[vet rqv rooLeurfrn1 on6 rr1 Bofuvn opretd rol.d c6me vo no- pepno6(oer 14 o0v0eor1 orlprowxrlg noo6rrltog ValtR- NArr". llopopo(cog, q Bo].utrotRNA ouvOetdorl ffalRS) ouv06rer Thr-tRNAVar oe orlpowx6 Po0U6. H lleRS, 11 VaIRS r<ot toutrdXrorov emd entnp6o0e- reg IRNA ouvOerdoeg (or erdu<69 yro q treurfur1, rrl Ue- 730 KEOMAI0 25 0 [everxdq Kti:Otxoq To ouyyevdq tRNA nupo0oreI otr],oyn rnl dtoutjprpoon Autvoorutr[toon Xop[q qtrtroyn orn droprlprpoorl oumo(ver opyd Aprvooxutr[oor1 oulrBoiver yprlyopo EIK0NA 25.17 H oprvoorutr(roorl Tov ouyyevtirv tRNAs on6 q ouvOetdoq pooi(eror ev g6peL oe peyotrUtepl ouyy6veto yto outd ro ei6q, oe ouvduoo;rd pe ooOevrl ouyydvero yLo p1 ouyyevrl e(6q. Enrntr6ov, ro gr1 ouyyevrl tRNAs oOuvototiv vo unooro[v ntrtlpoq ng induced-fit 0rogoprptorr6g peropotr6q, nou onououvrot Yto T0 6rfrpo rorotrunxd prlporo. Oetov(vr1, qv oLov[r4, qv npo].(vt'1, q gowu],o],ov(vr1, q Opeov[vr1 r<ot q truo(vr1) efvot txov6g vo 6top0cbvouv r' ol.rwooruLo-o6evuLtxd Kot oUtvooKUl,o-tRNA ro ono[o oX4goil(owor mo evepyd rdwpo toug ;16oro entnp6- oOerov evepyorrltov nou efue u6potr0ouv ro ol.ltvooKu- ).o-AMP onod[dowog el,erjOepo oprwo[ti xor AMP efue udpotrtiouv to preptrcbg orul.urrpdvo IRNA ono6(6owog eLeU0epo oprwo(rj ror onoorutruop6vo IRNA. H u6p6tru- orl rou otrrwoot<ul,o-AMP ovo<pdperot oS rpo-peragopt- xr1 6t6p0ao4 (pretransfer editing), evdr q u6p6l'uol1 rou ol.rwoorulo-tRNA tog usra-peraEoptxfl 6Ldp0uo4 @ost- transfer editing) (EIK0NA 25.18). Ir4v nep(mooq'rnq npo- pretogopu<r|g 6t6p0too4q, nt0ov6v 6vo pdpog qn6 ro ).dOog olqpotroprdvo ol.twoorulo-AMP 6(orotot on6 to evepy5 r<6wpo n 6nemo u6potrrietot gq ev(uprtrd oe 6rd- Lupo (o oprwoor<uLrx6g eorepx6g 6eop6g e(vot qXenxd omo0rlg). Aut6g o rUnog ow(6poor1q ene[epyoo(og ;rnope[ en(oqq vo 0ecopr1Oe( oq Uopeli rrvrlnxrlg 6t6p- Orooqg 6orrp(cov. Aw(0eto, 16oo 11 npo-pretogoptxtl udp6uor1 rou Ld0og ol.uvoorulo-o6evutrtro0 6tov npoodeOe[ oro 6v(upro, 6oo t<ot 4 roro].u6prevr1 on6 6v- (upro preto-perogogxri 6r6p0oor1 e(vot yvoor6q oq Xq- prx{ 6rrip0too4 doxrltitov (chemical proofreading). ouvbderqr opyd Boorxeg eworeg
  • 220.
    To x6wpo 6rdp0oorlge(vor prxporepo onc iJ sJ,:3...: IuveeTtK6 r6vrpo .,::,' Ktlvrp,o 0Lo:;-:-: AMP ndnNH' tcooH To q[rvooKUIo-tRNA udpol0erot ouv0erdoeg e(vor duvotri v0 rpoYpoTonote(Tot oTo ord0Lo nptv on6 qv opwoorutr[too1 (npoperoqopxn ene[epYoo(o), oro ono(o uDpotrueror n Un ouyyevnq optLvooxul,txq oDevu)xr1 opoDo. Evotr- trornro 1 enrnpdo0eto, evd6XetoL vo )ropet Xtitpo I udptjtruon tou trov0oop6vo oXtlportop6vou olttvoorutrotRNA pero q oUv0eoq rou (peropetoqoptrn ene{epYooto). Motrov6r or owtdpdoetq npo-Ueroqopu<r1q 6t6p0oorlq prepx6g qop6q ouprBo(vouv onouo(q tou IRNA (6nLo6n npw on6 qv np6o6eoq tou IRNA), 11 nopouo[o tou tR- NA yevrrd Betrruilvet nolri rqv onotel.eoportx6rqto tqg u6p5luoqg. To noto 6r6p0toor1 0o enu<pouloet, TIpo-Ue- rorpoprx4 6vowt trleto-prerocpoptxrlg, notrfi.et ovdloyo pre q ouv0et6oq. H ow(6pooq dt6p0cooqg Boo((etot orov rl.ootr6 pllXovropr5 6rn),o0 qOpoti (double sieve), 6nog nopou- ord(eror yto rrlv lleRS mrlv El(0NA 25.19, xotd tov ono(o rog B6o4 yro q dtdrptoq XpnotUonote(tot to pr6- yeOog rou optvo[6o9. H lleRS 6to06ret 6uo evepyd r5v- rpo: ouv0ert16 (r1 r<5vrpo evepyono[r1oqg), nou ewon((eror arrl ouvq0rl mupl Rossmann td(r1g l, rot x6wpo 6t6p0toor1g (n, qltu6q, u6potrutu<6), nou ewo- nf(etot orqv neptoXrl np6o6eo4g tou ono65rcrrl (B^.6ne Evoqta 25.10). H rpuoroMtrn 6oUn rqg lleRS 6efiyet 6n ro ouv0*x6 r6wpo e(vot notrri trrtr<p6 yto vo enrpS- qrer rnv e(oo6o qg Leur(vqg (r1 n).euptr{ o}.uo(6o qg Leur<(vqg 6ror<Lo6((etot oe dtoEopettr{ 06o4 oe orly- Kpron Ue rqv tootreux(vq). Itrlv npoyl.tonr6tnro, oTIo- rle(erot q evepyono(4or1 6Ltov ttov otrtwo(6cov nou eivot preyoLrirepo on6 qv tootreurfv4, ener6rl 6e gnopotiv vo ero5tr0ouv oro ouv0ettr6 r5wpo. 0o16oo, oprqrdvo pu- rp5repo o;rrvo[6o nou dtotrlpofv enoprrl trov5ulto , NH, RCH tcooH H Leu e(vor nolU peydtrrl yto vo ltrlpdoet mo ouveertKd Kdwpo @-rcu"'" H lle xopd mo ouvOerxo x6vrPo, dxt d[oq rot oro xdwpo Dtop0toonq @-w 9@H Val nepvd ond to ouv0ettrtj r6wpo mo rdvrpo 6trjp0toonq EIK0NA 25.13 H ouv0er6o1 rou tootreuxutro-tRNA 6Xet D6o evepydg neptoX6q. ApLvo[60 UeYdrUrepo qq lle Dev pnopotlv vo evepyonoulOouv, dt6l 6ev eqontovrot orrl ouvOettrrl neptoyt'1. Aprvo[60 ptxp6repo qg lle onoporp6vovrot, xoOtitg pnopotlv vo ero5trOouv onlv neptoXrl ene[epyooiog. np6o6eoqg -n.X. n Boltvn- prnopotiv vo etoSL0ouv oro ouv0errx6 r<5wpo Kot vo npoo6e0o6v mo IRNA. To ouv0ertx6 xdwpo Lettoupye( rog npcitog q0p6g. To r5wpo 6r6p0tr:o4g e(vot ptrp6repo on6 to ouv0ettx6 rqr 6e prnope( vo tpttro[evrioet rn ouYYevq tootreurivrl, comSoo npoo65vet q Bol(vq. 'Etot, ro ValtRNAlle u6potrtieror oro r6wpo 6t6p0ooqg, nou trettoupye[ og Serirepog n0U6q, evdr to lletRNAlle 6ev u6pol'Ueror. To prowSl.ou rou dmLorj rlOprod e(vot 6vo e0xol.o rot yevrrd orpp5g prowSl,o rnq Lero-Uero<poprr{g 6t6p0to- onq. :rnv lleRS, 6nog entoqq xot oe dtreg IRNA ouvOe- rdoeg 6t6p0rrlonq rd(nq I rot ll, to 66o r<6wpo (ouv0erx6 r<or 6r6p0roo4g) on6Xouv opxetd, 10-40 A. l-lpoxeqr6vou vo Trpoypcnonoq0e( 4 Uero-UeroQoprrq udp6Luoq (5t6p- Orr:on), ro drpo-onod6rcrqg rou preptr<cirq oxuLtrop6vou ol.rwoorutro-tRNA petoton((etor xotd pnKoq'rnq entqd- veroq rou ev(0Uou (EIK0NA 25.20) on6 to ouv0err6 xSwpo oro rSwpo 6t6p0coor1g. Aurn n pretot6ruoq nepr.I.oprBdvet puo peroBotrrl orq 6topr6p<poorl rou dr<pou-ono65ffi'l rou IRNA. Itrg IRNA ouv0erdoeg td(4q l, to dt<po-onodSrrqg uroOere( 6rop6prp<ooq goupr<5tog 6tov npooS6verot oro ouv0ertr6 rSwpo (Blene Ev6tryo 25.10) xt er<tetopr5vq 6oprl 6tov npoo66verot oro r6wpo 6t6p0oor1g. Meror6nrorl ev6q LdOoq opuvo(6og ndvro mrlv ent- <pdvero rqg IRNA ouvOetdorlg e(vot ntOovrl oro Unxovt- o;16 qg Utroperqeoptxrlg ene[epyoo(og enet6q to oprwo[ri e(vor ol.rorono].u<d npoode6epdvo oro IRNA. Av- r[Oero, n npoUeroeopxrl 6t6p0rrloq ouprBo(vet npototi oXrlproume( o;totonotx6g 6eo;r6g preto[ri o;rtvo[6og r<or IRNA. [-r' out6 ro ].6yo, e(vot on(Oovo vo npoYporo- noefuor npoperoqopxrj 6t6p0ooq pr6oo orqv neptopl 6r6p0oo1g rou ev(6prou, enetbq 6ev undpXet Unxqvt- o;169 pe tov ono(o vo nopegno6((etor 4 6tdorooq tou :5."i : 0r luv0erooeg Xprlorponorouv U Arop0toon AorLp[t.rv yto vo Be],rLooouv Ti'lv AxoiSsir 731 npdodeon tRNA ADevutr(toon Ydpotr[etor ro o;uvooxutro-odevutrtx6 06pTrorl tRNA Itomd oUrvoEu +
  • 221.
    EIK0NA 25.20 HouyxpumoNLx4 0opr1 qg treuruhotRNA ouvOe- rdqg (LeuRS) npoodedey6vq mo tRNAk, oq dropdprpoorlqg pe- roperoqopx4g ene[epyoo(og (oprorepd) ror q dLopdprpoon Tnq olrLvooru)(oor1g (deErd). H neproXrl ene[epyooiog Ue BooU ruovo XpOUo reprorp6qeror orlgovrLrd ;rero[rj rov 600 dropoprpdroetov, evtir ro drpo-ono66mqg Tou IRNA perorp6nemr pero[u prog eme- rop6vlg etrxoerdoug 6rog6prptoo4q @pmepo) Kor Uroq drogoprpto- o4g d(xqv rpoupr6rog (6eELd). Avoruntir04re rordnv odefog ond J.J. Perona, el al., Biochemistry 46(2007):10419-10432. Copyright 2007, American Chemical Society. 0t rptoroyporpieg nopoXrrlprl0q- rov ond John J. Perona, University of California, Santa Barbara. pr1 ouyyevorig ol.uvoorul,o-o6evul,u<orj r<oOcbq nopeUe- ror on6 ro ouv0errx6 xdwpo mo r6wpo 6r6p0oor1g. Aw(Oero, 6rov dropeooLoBe( rdnoro 6v(u;.ro, q ow6po- on rnq rrpoperoeopu<r1g 6r6p0ooqg pdMov ouprBo(ver pSoo oro 5pro rou ouv0enro6 evepyoO r<6wpou. 25.12 To KorooroArrrd tRNA [leprdxouv MeroAAoyUCvo Avrrro6rr6vro, To Onoio AroBd(ouv N6o Ktrl6tr6vro * 'Evo rorqorotrnro IRNA runuo rp6per gro perotrtro[r1 0T0 ovl- rodrrdvro nou otrtro(er ro rodrxovro oro ono(o onovro. * '0rov ro vdo ovlroDtxovro owlororXel oe rtodrrovto Tspuol- opo0, evotogortilveror 6vo o;uvo[u ror 11 notrunenrLdLq otru- o[do enerreiveror ndpo ono ro roldxdvro reppologori. Auro odqyeL oe oouvOelrl roroorotrq oe pro 06o11 oouv0elxrlg gerotrtro[1q r1 oe ovoyvtool n6po ono evo rovovtxo rodrro- vro reppoTtopou. a Auouv0enrrl roroorotrl ougpo(ver 6rov ro IRNA ovoyvtirpi(er rlvo Drorpoperrxrj rodrr6vro ono ro ouvrlOroprlvo, onore 6vo opLvo[0 ovnro0rororol ono ronoro otrtr0. H onop6vo-ro4 peroMoypr6vorv IRNA unrlp[e 6vo on6 ro roXup6repo epyo).e(o yro rrlv ovd).uor1 r1g rr<ov6- rrlroq rou IRNA v' owrororXe( oro xcr:6rr6vrd rou oro mRNA Kor rov r<o0oprotrr6 rov enL6pdoe<rlv nou 6Xouv drogopetLrd rprrlproro tou IRNA orqv ovoyvroprorl Ko- drr<ov(o u-owrxodLr<ov(ou. To peroMoyprdvo IRNA onopovcovowor 1dpr1 mqv rr<ov6rqtd roug vo [enepvoUv lg enrmo.loerq rov Ue- toMd{eotv oe yov(6ro nou ro.l6rronoro[v no].unem(6to. Iq yevrxq yevertxq opoLoy(o, ;uo pret6Mo(rl nou pno- pe( vo unepvu<qoer lg enLmcooerg ;uoq dM4g prerdMo- lqg ro).ekor xoroorotrrrx{ (suppressor). Ie ouorr;;roto IRNA r<otooroLng, 11 nporoyevrig ;re- 732 KE0MA|0 25 0 leveruog Ktirdrroq rdtrl,o(r1 oMd(er 6vo rrrl6u<6vro oro mRNA, on6re ro no- Lunemrdx6 npoi6v dev e(vor nl*iov Leroupyu<6. H 6eu- repoyevf g roromoLtrx{ per6},}.o[r1 ;reroBd].].er ro owu<to6x6vro evSg IRNA, riore v' ovoyvrop((et ro pretoA- Loyprdvo ro:6x6vro ow( rou ou0ewxo0 rodu<ov(ou-ore Xou (i e[(oou r<o].d). To opuvo[ri flou evooUorc0veror onoroOrord rq Leroupy(o tou nolunemt6(ou. Or xcno- oroLe(g ovorp6powor <rlg oouvOerrxoi (nonsense suF pressors) rl 6uouv0enxof (missense suppressors) ovdloyo Ue rn qUon rnq op4Knq prerdMo[qg. Mro oouvOerrrrl gerdMo[r1 petorp6net 5vo xco6xe vro nou roOop(er 6vo opuvo[ri o' 6vo on6 ro rp(o xor6t- r6vro tepportopoti. I' 6vo rrirropo dyptou rUnou, pro r5roro oouv0erxrl gerdMo[4 ovoyvrop((eror p6vo on6 dvo nopdyowo oneLeu05ptr:oqq, o ono[og reppori(u tr1v npolreivoorivOeorl. Oar6oo, q 6erireprl xoromo.f,n- xli gerdMo[n oro owrKo6rx6vro rou IRNA, 64puoupye( 6vo ol.uvooxuLotRNA u<ov6 v' ovoyvrrlp((et ro ro:6rxo vro repporroproU. Evorotrrorrivowog 6vo o1.rlo[0, q xo- toorolt] on6 rr1 derirepq prerd).Lo[r1 enrrp6ner rr1 ouvSXroq t4g perdgpooqg nSpo ond 14 06oq qg oouvOerrrfg perdLl,o[4q. Awrl 11 v6o 6uvot6qrq rou UeroepofirroU oumrlprorog entrpSner rrl orjv0eorl ev6g nL4poug nolunemt6(ou, 6nog go(veror mnv EF ,(0NA 25.21. Av ro o;uvo([ nou evoroporri:veror pr6oro xo- romoLrig 6rogriper on6 ro ou0ewrrS optvo[r3 orrl ouyrer<ptpr6vq 05or1 rou nol,unemrS[ou dyprou r0nou, r'1 6poorqprdrrlro rou noi,unemr6(ou prnope( v' oM6[a. Or 6uouv0eru<59 perotrLd[erg oMd(ouv 5vq xodrKG vro nou ovrr.flpoooneUer rdnoro oprwo(ri oe rro6rx6uo nou owr.ilpoocrlneUer 6vo dtri.o oprwo(ri ro ono(o o6w+ re( vo Leroupyrloer orn 06on rou ouOewu<orl. C-unxcr, onoto6{note owmordoroorl qrwo[riog ouvrord pro 6r-r ouv0enxf perdMo(r1, oo16oo, oqv npd[4 ovrXyeferor p6vo ov otri.d(et rr; 6poorqpt6rrpo rou nolunemrdiou.) H perdLl.o[q rotoordMeror qn5 rrlv evotoprdrooq eire rou oplxori oprwo[6og efue xdnotou dMou optwo(6og nou onoro0rord rn treroupy(o rou nolunemr6(ou. H Ell(0NA 25.22 6eflyet 6tr q 6uouv0enrry xcrrooroNi pnope( vo enmeul0e( 6nog ror q oouvOerrruj, dqAodli petotrl.dooowoq ro owrKo6x6vto ev6g IRNA nou 1.te- togdpu 5vo ono6err6 ogrvo[ri 6ors v' owrorotle[ oro peroMoypr5vo xtodx6vro. Iuvendrg, 4 6uouv0ertxn xe tooroltj ouvisrotor oe otri.oyrl tou prlv[proroq rou Kq> 6txov(ou on6 6vo o;rwo[0 o' 6vo 6trtro. 25.1 3 Yndpxouv Aouv0errroi KorooroAelq yro KdO€ Ko6rr6vro Tepponopo0 * Ko0e runoq oouvOertrou rodxov(ou rorooretr),eror otto evo tRNA pe perotrtroypevo ovrtrodrr6vro. * 0proU6vo onovro roroorotrtro IRNA nepteXouv perotrtro(erc oe otrtro rprlporo rou pop(ou. Or oouvOerrxo( xorooroLe(g 6roxp(vowqr oe rperq Ko- Boorxcg evvorcg Boorxfg eworcq
  • 222.
    Ayproq nlnoq: roxodxd,vto UUG 6tq0d(erfl qnd rc Leu-tRNA AUG HH8 UM Aouv0errrd [er0uoypo: ro rodtrovto UAG repUo((et _-:- - Ai,picq ruiroq: ro rodtrovto GGA dtopo(eroL ono ro Gri-:i',: ,-.r* ILeu w UAG Arjn 880 i:^i. ,rd IGlv 13 nopoyoutoq W onetreu0epurons I ^radIArg Korumolttxn uetdllo{n: otrtrd(et ro ovrLxtodtxovto tou Tyr-tRNA AUC UAA Ell(0NA 25.21 0t petotrtrd{erg Xop(q vonUo Uropotlv v0 K0T0- orolOorjv ond 5vo IRNA Ue Uer0,tr0Yp6vo ovrtroldtrdvto, ro oro[o erooyer 6vo outvoEr] oto Uer0II0YU6vo xtootrovto II0pdYov- roq uro nporetvl oh6rhnpou p4rouq, onou ro opxtxti xordtroLno Leu owrroromoOqre on6 Tyr. rqyopieg, Uro yto r<d0e t6no x<o6tt<ov(ou repportopo6. H EIK0NA 25.23 neprypd<pet ttg rdt5rlteg optopSvtov on6 ro u g rotrrite po Xopoicn p toU5vo u g r<otooroLe(9. Or rotoorotre(q rou rlLer,cpou (amber suppressors) e(vor or onl.orjorepot. Iro E. col/, toutrdltorov 6[t IRNA 6Xouv ;retotrLoX0e( c0me v' ovoyvtr:p((ouv rco6tr6vto UAG. 'OLq tq xoroorqlru<6 IRNA rlLercpou nepdlouv ro owtrto6tr6vto CUAe, TIou ot rdOe nep(moon npo- 5pXeror on6 tov 6ypto ttino tr-r5oo oMoy4g prtog pr6vo Fdonq. H pretdMo[r1 gnope[ vo ewon((etot oe onot- odrlnote on6 ttg tpetg Bdoetq rou owtK<o6tt<ov(ou, 6ncog go[vetot oro supD, supE rot supF. Kd0e Kqro- orotrnr6 IRNA ovoyvtop((et pr6vo to rcodtx6vto UAG ov- r( yro ro npoqyoUprevo xto6tr6vto ({ ro6tr6vto)' T' oprwo(6o nou evoopor6vowot e(vot oep(w1, yLoutopri- vq 4 rupoo(vn, 6nlo6n tout6orlpo pe r' oprtvo(6o nou petotp6powot on6 t' ow(orotlo IRNA dyptou nlnou. Or rorooroleig qq 6Xpog (ochre suppressors) en(- oqq nponimouv on6 pretoMd[etq oro owtt<to6x6vto. Ifto yvooro( efvot ot supO rot supG, ot ono(ot evoogo- r6vouv rupoo(vr1 rl Luo[vr1 ow(orotXo npoq ro xto6tr6- vrq rnq 6Xpoq (UAA) xot tou r1lertpou (UAG). To Knrooioi.fi: ro AGA 0to9d(etot ond ro perotrtroyu6vo Gly{RNA EIK0NA 25.22 H dtoouv0ertxn Uerdhho[n npoygotonote[tot, orov ro ovnrodxdvto rou IRNA perdrtrdooerot 6ro 6ore vo owt- ororXel oe ]roOoq xodtr6vto. yeyov6g out6 e(vot ouprBot6 Ue rnv np6p^e$n qg un6- Oeonq tr1q ocrd0etog 6tt to UAA 6e prnope( v' ovoYvo- prore( pSvo rou. 'Evog r<otootol6og UGA SXer pro onp6oprevrl t6r6t1- to. flpo5plerot on6 to tRNATrp, oIId n ;rovo6rni pre- [eveuxdg rtinog IRNA Ayptog tunoq Koromol6og supD (su1) sr.ipE (su2) supF (su3) supC (su4) Ko6trdvro/ Avrtrodrxdvto/ owtxrodtrdvto xrodtxdvro Ser UCG Ser CUA UAG UAG AUC AGA UAA UCU A UUA UAG A UUA UAG A UCA UGG Lys Trp c UAU Tyr c UAU Tyr Gln Tyr Tyr CAG Gln CUA UAG CUA UAG A GsupG (suS) Lys supU (su7) Trp ElK0l,lA 25.23 To rorqorotrffio tRNAs xtop(q v6nuq 6nuLoup- youvror on6 perotrtro[etg oto ovrtrodxdvto. Auouv0erLxrl: ro AGA dtopo(etoL ono ro Arg{RNA 25.13 YndpXouv Aouv0euro[ Kotoototre(g yto Kd0e Kodtr6vto Tepuottouou 733 AUG
  • 223.
    tdMo[n rou e[vor11 owu<otdoroo4 A ow( G mq 06o4 24. Autrl rl oLloyrj owu<oOrord 5vo (erJyoq G-U mo m5Lelog D pr' 6vo (eUyog A-U, ou[dvowog dror t4 oro0ep6qto qg 5Lu<og. H oMql,ouXfo rou owu<<odr- rov(ou nopop5ver (6ro 6ncog Kcu. orov dypro rrJno, C- CAe. Iuvencbg, q perdMo[q oro or5],e1og D npdnet pe xdnoro tp6no vo preroBdMer q 6ropr6peool.l rou Bp61ou rou qwKodtxov(ou, ennpSnowog mo CCAe vo (euyoprilver pe ro UGA p' 6vo oouvq0toro ooroOdg (euydpopo qg C pre rrlv A. To r<oroorotrru<6 IRNA e[o- r<oLou0e( v' ovoyvrrlp(er ro oUvq0eg ro6x6vro UGG. Mro nopegqeprlg rordoroon nopqrrlpe[ror or4v ne- p(mtoo4 ev6g ouyrexprpr6vou euKopuoJlKorj IRNA. To rlnop rou Bodtori nepr6ler tivo tRNASer pe ro owtrtodr- r6vro ,CCA+. Or rov6veg ootdOerog npoBl,6nouv 6rr out6 ro IRNA 0o 6npene v' owrororxe( oro rco6u<6vto rnq rpumoedvqg UGG, oMd orrlv npoyponr6rrlro ov- rtororle( oro xor6rr6vro repponoUori UGA. Iuvendlg, oe ourj rlv rordoroo4 ro UGA prnope( vo roroor6Me- ror guou<d. H yevu<q orlpoo(o ourc0v rov noporrlpqoecov 6,yxer- ror orrlv enroqpovo4 6n 11 ovoyvrilproq xo:6xov(ou-ov- rrro6rxov(ou e(re mo IRNA dyprou rUnou e[te mo peroMoyprdvo IRNA 6e pnope( vo npoBLeq0e( nl4prrrg on6 ng oXeuxdg oMqlouX(eg ro:v rprnLerriv, evri oe pepx6g neprmtrloerg prnope( vo enqped(eror on6 dMo XopoKrn proru<d tou prop(ou. 25.14 Or KorooroAeiq Mnopel v' Avroytovi(ovTol Tnv Kovovlri Avdyvroon Tou Kd,6lKo * To roroorotrrtxo IRNA ovroyulvl(ovror ro IRNA oypLou runou nou nepteXouv ro [6ro ovurotdrrrlvro yto qv ovoyvoorl rou ovrlorotXou rcodLrovlou (r1 rodrrovLrrlv). e H onoretreopolrrl roroorotrl ervor enrptroBqg, enerdrl odrlyer oe ovoyvooq nepo ond ro xovovtro xto6rr6vro TeppoTropou. * To rodrrovlo UGA egqovi(er dLoppol ror 0ropo(eror eoqotr- g6vo ono ro TrptRNA pe ouXvrirqro l -3% YndpXer puo evdrocp5pouoo 6rogopd preto[ti qq ouv4- Ooug ovoyvcrtptorlg ev6g xrodLxov(ou on6 ro ow(oror- Xo oUrvooKULotRNA Kor rrlq r<ordoroorlg orrlv ono(o 1-uo perdMo(q enrpdner o' 6vo xoroorotrrrx6 IRNA v, ovoyv<op(oer 5vo v6o rot6rx6vro, Iro xrirropo dyptou r6nou, p6vo 6vo v6rlpro prnope( v' ono6o0e( o' dvo 6e- dop5vo rco6rr<6vLo, ro ono(o owrnpooconeUer efue 6vo ouyrexprprSvo opuvo[ti e(re 5vo o1pro rep;ronopro0. I, 6vo r[rropo nou <pdper Uro Koroorortxr1 prerdMo[r;, ro peroMoypr6vo rro6u<6vto pnope( eke vo 661etor ov- royovrop6 on6 ro roroorotrru<6 IRNA e(te vo 6topd- (eror pe ro oUvq0eg v6qprd rou. 734 KEOAAAI0 25 0 l-evelroq Kcirdtxog 'Evo oouv0etx6 rotosrotrtu<6 IRNA np5ner v' clwo- yr.uvrme( roug nopdyoweg oneLeu06pooqg or onofor ovoyvtop(ouv to xrodrx6vro (-o) repprorroproU. 'Evo 6u- ouv0errr6 roroorotrrrr6 IRNA npdner v' owoyrovrore( to IRNA Trou owrororloriv oro v5o r<ro6tr6vr6 rou. Ie rdOe nep(mcooq, o Bo0pr6g rou owqyovro;rori en4pe6- (er qv onoretreoporrr6rqto rrlq KqrofioLrlg: ouvenrig, 4 ono-releoportx6r4ro ev6g ouyrerprprdvou Korooro- L6o e[oprdror 61r pr6vo on6 rr; ouyySvero prero[( rou owrxco6rr<ov(ou rou Kor rou ro6rrov(ou-m61ou, oMd ror on6 rr1 ouyrSwpoorl rou oro rUnopo, 6ntrlg en(- onq Kor on6-ng nopop5rpoug or ono[eg 66nouv rrq ov- royrovrmrxSg owrdpdoerg repprotroprof 4 evootpdrtrl- onq. H onoreleoporu<6r1to Ue rnv ono(o droBd(etor 6vq r<o6x6vto enrlped(erot on6 rqv ew6nrorl rou. Iuvendlg q 6rcrooq rlg oouv0ettx4g r<otooroLrlg on6 6vo 6e6o- pr6vo IRNA norril.er nolrj ovdLoyq Ue ro ntro(oro rou r<o6u<ov(ou. Or enr6pdoerq rov yerovxriv Bdoeov rou m R NA orrlv ovoyvcil pro4 rardtrov(ou -owrrro6 rr<oviou 6ev efuor xo.trd rorqvoqrSg, orr6oo ro nLo(oro prnope( v' o).}.d(er rq ouyy6rqro pe rnv ono(q 5vo rco6rx6vro StoBd(eror on6 6,vo ouyr<erpq6vo IRNA rord ;to rd!r1 peyS0ouq xor nl.6ov. 'Evog oouv0etrr6g rotoorol6og onotrrovcbveror Xd- pq ornv xovdr4rd rou v' onowd oe 5vo preroMoyprdvo oouv0erix6 r<trr6rr6vro. H (6ro oMrlLoul(o, oo16oo, ou- vtord 6vq on6 ro rovovrrd orlporo tepportollof rou rurrdpou. Oetrrp4ru<d, ro peroLl.oy;r6vo IRNA nou Ko- toor6Mer rqv oouv0etxrl gerdtrio(r1 npSner vo ;rnopef vo r<otoor{i[l.er rov KovovtK6 repproropr6 oro 16}ro9 rd- 0e yovr6(ou ilou Xpnorponoe( ro ouyrerprprdvo r<o6rxo- vto. H EIK0NA 25.24 6efiver 6n ourrj r1 nopoploo4 rou repponoporl roroi,4yer orq oUv0eorl ev6g prorprjrepou no).unenrr6(ou p' enrnp6o0eq xopBo(wel,rr4 oLl.r1- Merdqpool dyprou rrjnou AijG 0 nopoyovroq onetreu06ptoorlg reppor[(er rr1 ouv0eorl oro xrodrxovro repgolopori Kotomo)r1 ntrdrrpou Li;l::, To roroorotrlxo tRNA 6LoBo(er ro rrrl6rxovlo UAG ror 11 nporTeivrl enerre[veror doq ro enopevo xrodrxovro repportogou ,::i i-'r;;' -ii: ,, ,t Q1 qgyy6t.rro( rorooroLe(q enronq dtopo(ouv ne- po ono rovovuo rotDLovro repponopou, ouv0erovrog nporrelveQ. ot onoleg e(voL porpurepeg ono erelveg rou oyptou runou. UAG UAA @ I UAA @llopriyovroq onetreu06prrlorlg UAG AI.JC "$Tvr ITyr loolrcg eworcg
  • 224.
    AouX(o. To noLunent(6to0o rotoffi[et orqv en6pevrl -p*i5.o teppottopori nou 0o Bpe0e( orq qdorl tou nAoto(ou ovdyvooqg' H ruX6v erctetop6vq r<otomotrri rou repUortogoti prnope( v' onoBe( entB)'oBr1q Yto ro xfrropo', nopdyo*og prorp0tepo nolunem6to pe oI- IoyUdvn LettouPY(o. 'b, *o.o*oLe(g ril'er<rpou ouvrlOcog e(vot oXettro onoteheopottro(, yevtrd xotd 10- 50% ovdl'oyo Ue ro oJ*nuo. Awri 11 onotetreogottx6trlto e(vot nt0ovrl enetOrl' ro r<rl6tt<6vto tou riLer<rpou Xpnotponotofwot oXenxd sn6vto Yto repporLoU6 qq petdqpoo4q oro E' "bli. R*(et-o, t<otoorotre(q cbXpoq onopovtirvowot 60' sKo,o. E(vot ndwo no}'ti Lty6tepo onorcLeogottro(' pe 6poorrlpt6trlro x6ro tou 10%. 'Otrq tq rUrropo Ue Ko- .o*oLtiq drXpoq 6Xouv pdtrl'ov cprcoXrl ovdmu(q' ye- yov6g nou uno6rll'tirver 6tt 11 rotootolrl t6oo tou UAA 3oo oo, tou UAG e(vot rotootpemtr<r1 yto to E' coli' ru' 0ov6v enet6rlto rto6u<6vto 6Xpqq UAA lpqot;ronotefuot oqy6tepo oq ro Kovovtr6 orlpro teppottotrlo0' T6Loq' ro UOR e(vor to Lty6tepo onotel'eopotu<6 on6 to rco6t- x6vto teptrtottopo0: oe rUrropo dyptou t6nou, StoBd(e- rot eogol.prdvo qn6 to TrptRNA pre ou;y6trlto 6o9 t<ot 1-3%. ir/oorqUtq, ro UGA Xprloqonote(tot ouly6tepo or,6 tqu tptntr6to rll'errpou UAG yto tov repgonog6 rcov BorcrqPtor<dlv Yovt6(tov. 'Evo duouv0eur6 rotoorotrtrr<6 IRNA nou ovttppo- ne( 5vo preroMoypr6vo rrodr6vto oe pto optop6vrl 06o11 pnopef uo ,,ooydyt, plo ovent0Uptqrrl petdMolrl o' 6vo 6)tlo yov(dro.'Evoq rotoototr6og 6top0drvet gto getdl'- Ao(r1 owu<oOtorciwog 6vo oprwo[rj pr'dvo dMo orn Ue- -oi.LoVUgrn 05or1. Ee dLLeq 06oetg, toor6oo, 11 (6to owrotdmoorl owtxo0ror6 to oprtvo[r3 dyptou ttinou U' e* v6o ogwo[6. H pretoBo].rl Unopgi v' ovoore0ret rqv xovovtrq'treroupy(o tou notrunemtS(ou'- Aw6 06tet ro x0rropo oe 6il.q1-rpo: np6net vo rotooteilet 6vo opt- olrevo petof.loyp6vo t<todx6vto 1op(g 6gto9 v' oMd[et nohti to t<ovovtx6 v6qprd rou ot 6Meq 05oetq' Euve- nOS, n onouo(o roXupcirv 6uouv0ertrc0v roroorotrStov l,t" pti vo e(r1yr1Oei on6 rg rorootpemx6q entd pdoetq nou'0o nporotrorioe Uro YevKeuUdvq r<ot onotetreopott- r{ owtrordorooq oPwo(6tov. Mro pretdLtr'o(q nou 6rlptoupye( dvo- KffiomqtrttK6 IRNA gnope( uo 5Xe, 60o ouv5neteg' l-lprinov, enttpdnet sro IRNA v' ovoyvcop((et 6vo v6o xto6tr6vto' Aerjtepov' lreptx6q Qop6q nopeprno6(er to IRNA v' ovoyvop(oet ro'ro6ir6vto orq ono(o npoqyo0gevrog onowo[oe' E(- ,o, oqpo*,*6 6tr 6tr'or ot onotetreoponxo( xotooroLe(g nie'..pou npo6pXowot on6 petdMo[n eY6S owtyp6- qou p,oq nitou&qorooq opr66oq tnrul :-e ow6g ttg t"p,*Oot,S, -o ^jr.opo nepdlet noMg IRNA xovd v' onownoouu oro rodu<6vro nou onorel'e( tov ou0ewtr6 m6Xo tou IRNA dyptou t[nou' 'Etot, 11 pretdtrtro[r1 6ev rotopye( rqv ovoyvdrptorl rov ndrottitv xto6tt<ov(trlv' to onofo L(onolou0o0v vo xo}'Umowot enoprdrq on6 to tnftrn.ns opd6og. Itrlv oouvdOtotq xotdorootl xotd rn, onoto g6vo 5vo IRNA onowd o' 5vo ouyxer<ptpt6vo xto6tt<6vto, r<dOe pretdl'tro[r1 nou nopeprno6((et rnv on6rrrrloq e(vot 0 ovorlq6 Poq. H rotomotrn ouvn0oq o[ro]'oye(tot oro ntro(oto 1.uog petdMo[r1g nou oMd(et qv ovdyvooq evoq Ktltdt *ouiou. Oo16oo, oe oprop5veg xotoordoeq, 6vo Krru6t- r6vto reppottoprori 6toB6(etot tr:g o1'tloff, nopot Fe Xopu'1l,6tepr1 oqy6tqto. To npd:to yvtr:or6 nop66etypo ovor<oLrl<p0r1Ke oro yov(6to Yto rnv nporotvq rou nepF Blqgotog tou RNA qdyou QB. O oppottopr6g Loqro- VOrw ocol-tott6(ov QB npouno06tet roromolrl tou xo6trov(ou rep;ronopro0 oro drpo owoU tou yovt6(ou Ue XoUnLn orlr6tt'1-o, tilore vq nopoX0e( ;rto prtxpri no- boinio'uopiov nptotetvnq ue uto r<opBofuretrrxq en5- "roon buotoorrrd, out6 to xto6tr6vto tepgottoprori eprpov((et Stoppod, O l6yog e(vot 6tt rc TrptRNA ovo- yvop((et to t<co6x6vto pre Xo;r1trr1 ouly6trlto' ' Au&yurorl n6po on6 to t<co6tr6vto teppottoproU ou;rBoiver Kot orouq euxopuciteg, -6nou Xprlotpronoe(- roiouyy6tepo qn6 RNA to0q. Aw6 (ocog nepil'o;rBdvet oo.o*tM tov UAG/UAA on6 to Tyr-tRNA, GlntRNA 11 leu-tRNn ri rotomotrrl tou UGA on6 to Trp{RNA q ArgtRNA. H 5r<roor1 rqq preptr<r1q rotomolf,q unoYo- pt-ftaol, on6 to *nlo(oro' tou xtrl6xov(ou' 25.15 Io PlBootol..ldrto Ennped(et rnv ArpiBero rnq M€TdQPoonq o H do[rl tou 165 rRNA ottg 06oetg P xot A rou ptpootopor[ou ennped(et Uv orp(Feto tnq UetdQpo0nq' H ouyy6trpo L60oug xord rrlv evoopdtooq rov opt- vofStov oe nolunem(dto np6net vo 6torrlpefuot XoUn- In, ornv xtr(pror<o tou ev5g )rdOouq ovd 10'000 ouwoeeir, c6ote vo 6too<potrtore( 6rt ot lemoupytr6q r6r6rqteg tcov rto6tronototipevtov nolunemt6(tov 6ev oUAEoru r<otd tp6no entploBri yo to rrirropo' Ad0r1 Ut"pti vo ou;rBo6v oe rp(o yevt16 or66tq qq prer6- epqonq (F)rdne Er6vo 24.8 mnv Ev6tqta 24'3): -'. n ,ibd-,on ev6q IRNA pre to r<otdM4o opwo(0 e(- vot oorpciq xp(oq-rr1' Aut6 e(vqt Spyo qq optvooKu- r,o-tRliA ouvOerdoqq nou XopoKrrlp((etor on6 ouXv6ulto L60oug 1:105 6coq 1 :107 o;ttvooruLr6- oetg (BL6ne Ev6qteq 25.9 5trrg 25'11)' . n UeioQopd tou otoototi o1'twoorutrro:prSvou IRNA o-ri, p,B6oiru6tto, 5pyo torv nopoy6wcov 6vop(r1q f t",U,inruonq, nop6let 6vo prqXovto;r6.nou enou[6- vet'rq ouvo)ttrr1 enil'errtr6rqto' Entnl'6ov, ot ouYKe- np,giuo, nopdyovteg Boq0orlv otq dtepyoo(o oyrupo6eofoq rou ogtvoorul'otRNA o"rtq ptBootollt- x5q 06oetq P rot A. . Kpiorun en(oqq e(vot q et6x6t1to qq ovoyvrirptorlq rto6u<ov(o u -owtt<<l6tt<ov(o u. 11 op6tt o r oro0ep6 g np6o6eoqq norr[trouv ovdtroyo pre to (e0yog t<to6t- xov(ou-owrxrodu<ov(ou, 11 ev6oyev4q et6x6tqto no u oXeil(etot pe ro crxnUortopr6 gtoq ouyyevorlg dvowt gr1 ouyyevotq drLrl)rouliog tpttilv (euycirv B6oe<rlv 25.15 To PtBootoudtto Ennped(et qv Arp(peto uq MetdQpoons 735
  • 225.
    (nep(nou 101 5<og102) e(vot nol,ri 1o;r4Lri drore vo e(oo<poL((et ouyy6qro trdOoug < 10-s. An5 xop6 erle 6toruno0e[ r; un60eoq dl o Borrrl- pror6g nopdyowoq enrprqr<uvoqg EF-Tu npoo6dveror oro RNA ove[dpqro on6 qv oM4],ou{o, r<o069 npd_ ner vq Uerogdper oro prBooopdtro 6Lo t, ol.rwooruLo- IRNA (err6q rou evopxrrlprou IRNA). O EF-Tu ovoyvrop((er r5oo ro opwo[u<6 rUnpo rou 6eopo0 opr_ voor<utro{RNA 6oo ror ro o6pro rou IRNA, 6nou rup[<og npoodSveror fio ooKxopogoogoprxd oreler6 tou metr51oug-ono65rrq Kor rou meLdXoug T. MeLdreg orrg ono(eg perp4Ory<e 11 ouyy6vero np6o6eoqg rou EF-Tu oe otom6 xor Ld0og ogrvoorul.uo;r6vo IRNA 66er[ov 6tr 11 roX6g rqg np6o6eo4g oro opuvo[0 e(vor owrorp6<prrlg ovdLoy4 Ue rrlv rq40 np6o6eoqg oro oc0_ Uo rou IBNA. Me dMo L6yro, oo0evrirg npoo6e6eg6vo otruvo{6o eoreponororiwor oroord pre ogtXrd npoo6e- 6ep6vo otil.roro IRNA, evc0 orprXrd npoo6e6e;r6vo opr- vo[5o eoreponoroUwq oroqrd pre ooOevrilg npoo6e6e- prdvo oril.toro IRNA. Iuven6g, rq ooord oxul,uo;-uivo ollvoo<ulo-tRNA npoo66vouv EF-Tu pre opr<erd nop6_ 1.roro ouyydvero. H ex).eru<6tr1ro mr1 ouvoLxrl perd_ qpoon e[oorpo].i(eror ener64 11 ]d0og or<ul,(oor; ev6g oo0evrig npoodedeprdvou ogrvo[6og o, 6vo oo0evc6g npoo6e6ep6vo oc0pro IRNA nopdyer 6vo pr4 ouyyev6q ol.uvoor<ulo-tRNA, ro ono(o oMrll,enrdpd noLrj ovoro_ rel,eopo-nrd Ue rov EF-Tu. En(onq, dvo )td0og orullco- pr6vo ol.uvoor<ul,o-tRNA nou npoo66veror nto orptXrd orov EF-Tu pnope( v' onorLerore(, enerd4 e(vot 6uoxo_ l,6repo v' oneLeu0ep6oer ro oplo[x6 rord].orno rord qv oyxupo6eofo rou mo p$oocrlprdrro. 'E1er Bpe0e( 6n geroMd[elq orov EF-Tu ;rnope( vo roroorerl,ouv Ld0r1 preror6nronq rou nLoro(ou ovdyvcrl- oqg (BL6ne Ev6trlro 25.16). Aw6 unodqLdlver 6rr o EF- Tu 6e <p6per onLcilg ro oUrvooKULo{RNA orq 0dor1 A, oLid en(or1g epnldreror orrlv ronoOdrrlon rou eroepX6- Uevou oprvoor<ul,otRNA oe oX6or1 pe ro nemr6uLo-tR_ NA orrl 05or1 P. l-lopopro(og, preroLld[erq orov nopdyowo 6vop[qg elF2 r4g (ripqg ennp6nouv rqv 6vop[q qg gerdgpoorlq o' 6vq Kodrx6vro dvop(r1g, nou peroMdooeror qn6 AUG oe UUG. Aur6 uno6ql,ri- vet 6rr o elF2 Boq0d firlv oyKupodeo(o rou tRNA,r,r"t or4 06o11 P. H 6r6p0oor1 dorrp(rov (proofreading) ndvco oro pt- Booco;rduo, d:me vo evtoXu0e( n eyyevnq 1o;r4Lr1 er6r- r6rqto nou enrruyxdvetor ;r6vo on6 ro (euydpopo Bdoeov gero(ri xto6xov(ou-owrr<rr6u<ov(ou, npounoOri- rer enrnp6o0ereg o).L4l,enr6pdoeLg nou nop6lowor qn5 to ronrr6 neptBdMov qg 30S unopovddog, Kord rr1 tremoupy[o rou coq Srop0orjg 6orrp(trlv, ro prBoorrr- prdno evroX6er rrlv 4nro eyyevn enil,ercnr6rpo rou (eu- yop<ill.rorog rrrlv rprvouxl,eorr6(cov r<ord 1000 gopdq (EtK0NA 25.25). H enil.oy4 rou ogrvooKuLotRNA on6 ro prBooro;rd_ no ouprBo[ver oe notrtrd orddrq r<o16 pujrog r4g o6orj p5oro rqg ono(og ro rprroroy6g oUprn).or<o EF- Tu:GTP:o;uvoorulo-tRNA nou oxrlpor((eror prerd rqv ol.rwooxul(oo4 pretorpdper ro oprvooKuLotRNA orn pr- 736 |GOA Al0 25 0 l-everrdg Kdr6rrog Kd00 IRNA gnope( vo ero6tr0er oq 06o11 A To oooTo IRNA otr),r1trenr6po pe ro rRNA EIK0NA 25.25 Ko0e oprvooxutrotRNA pnope( vo rono0eqOei oq 06o11 A (p6otrt rou EF-Tu), otrtro p6vo ere(vo nou (euyopdrveL ge ro ovlrodrxrlvto, pnope( vo drlproupyrloer oro0eponoLrlnx6g enog6g pe ro rRNA. Xoprg oureg ng enoq6g, ro ogrvoorulo-tR- NA 0roX6eror 6[o onri rn 06on A. Booropu<{ 05oq A. llpc[rov, oX4pror((eror dvo prd].].ov ooro0Sg oplu<6 orltrrnlor<o np5o6eo4g pre ro prBooro- prdrro. Irq ouv6Xero, undpXet 6vo Bqpo ovoyvc0prorlg rou xrodrrov(ou, xord ro ono(o to op1tr6 o0pnLoro enovo6rqtdooeror enrp6nowoq ro (euydpopo rro6r- rov(ou-owxco6rxov[ou orq 06oq A. Ou;r40e(re dtr 4 6r- nLovrl 06oq P <prLo[eve( ro nenrt6utro{RNA (BA6ne KegdLoro 24, MeruEpoory). T6oo ro opXtx6 Bqpro np6o6eoqg 6oo ro enorSLou0o B4po ovoyv6ptoqg rou ro6rrov[ou e(vor owrarpemd, To ].d0og (euyop<o- ;r6vo ol.rwoor<uLotRNA prnops( v' onoBlq0o[v oe ourd rq 0166ro prSool ev5g ouv6uoollorj ou[qtrr6v4g rolrirrl- toq 6rdmooqg ro/r1 preropr6vrlg roXriqrog oriv6eoqg tov roro(euyoptoprSvtov outrrnl,6rov. Merd qv ovoyvcbprorl xo6txov(ou-owtrro6u<ov(ou, pro np6o0et4 peroBol,4 6ropr6prp<oor1g nupo6ore[ rrlv udp6truoq rou GTP. Arol,ou0e( q onel,eu06poon go- ogopxori on6 tov EF-Tu nou npoo66ver GDp. Aurl 11 oneleu0d,proorl nupo6ore( puo 6M4 ercero;r6w1 oMo- y4 6rotrr6pgoonq Ue rqv ono(o ro oUprn).oro EF- 'Evo trdOog tRNA dtol6erot errog
  • 226.
    Tu:GDP 6r(morot on6ro oripnLor<o opuvoor<ul,o- tR- NA:ptBooogot(ou. M6vo prerd 14 dtdorooq rou EF-Tu outrrBo(vouv ot opronxSg ovo6rord[erg nou oXet((owor Ue rqv oyrupodeo(o rrlq qUrvooKu).u<1g pi(og orrl 06o4 P qg 50S unopovddog, r<o0rr1g ror q enox6l,ou04 owG dpooq peroqopdg rou nemt6(ou. Ercr6g on6 enrLoyrl mo nprirpro orddro np6o6eo4g, onoBoLrj rou LdOoq (euyopro;r6vou ogrvooKULo-tRNA prnopef enfoqg vo npoyporonorqOe( pretd ro B{po qg udp6},uorlq rou GTP. E66 onopoln ouprBo(ver enerdri q ro16111ro tr1g oprorxrlg oMoyrlg 6rop6pgrooqg e(vor noLrj Bpode(o orqv nep(mu:or1 ev6q LdOog or<utrr<oprdvou ougnL5rou. Iuvenc0g, q ouvo).x4 er6x6rrlro enou(dveror enerd4 ro IRNA npdner vo 6r5l0er on6 60o B4proro enrLoyqg npo- toU oXrlportore( o nemr6rr6g 6eqr6g. H orp(Bero rou (euyopdrporoq ro6trov(ou-owu<ro- 6xov(ou m4 06o11A drotqpe(ror pe mevq nopor<o),o0- 0r1oq rov orepeoxrlUrrrriv ror 4l.errpoororrrriv t6tottirrrlv tou rprvourcl.eorr6(ou. Tperq ouwrlprlpr6veg Bdoerq rou 165 prBoor,tprroU RNA (A1492, 41493 ror G530) oMq),enrdporiv orevd pre rrlv el,dooovo oULoxo rqg 6lrroq ro6u<ov(ou-owrrodu<ov(ou oro 6rio nprilro (e[yq Bdoeov ror e(vor u<ov5g v' owr].ogBdvowor pe orp[Bero rrlv nopouo(o r<ovovrr<drv (euyri:v Watson- Crick oe out6g ng 06oetg. Iqv rp(rr1 (ooroOri) 05orl gnope[ vo rpr].o[evr10orlv oprogSvo Url KovovrKd (eUyq, enet6q ro prBoocopux6 RNA 6ev nopor<ol.ou0e( ro (eu- y6popro 16oo mevd. Te).Lrd, o r<0prog L6yog nou ono- BdMeror ro L60og or<ul.urtprdvo IRNA nprv r1 prerd ro Fnpo rnq udp6l.uorlg rou GTP e(vor 6n 6ev owono- xp[veror n].{pog mtg onopo(rrlreg npo6toypoqdg ro- rd t4 l,enrogepr] enr0ec6pqorl on6 ro prBooropdno ornv 6,Lu<o r<o6u<ov[ou-owu<ro6rxov(ou { xot oe dMeq 06oerg. llp6orporo, ovoxol,ri<p0r1re Svog enrnp6o0etog pr4- lovro;r6g nou ouprBdMer qrrlv er6r6r4ro qg prerdrppo- orlq: ro prBoorrrprdno efvor rrov5 vo 6e[dyer 6].ey1o nor6rqrog trrerd rr1 oJv0eoq rou nemrdrxori 6eqroU. Ie otrr6 ro pr11ovtopr6, 11 o0v0eoq ev6g nemr6rro0 6e- opoU nou npoxUmet on6 6vo KoKororprooUSvo ol.rwoo- xulo- IRNA orq 05or1 A,odrlye( oe yevu<6repq onrilero a6x6rrpog orrl 06oq A. Me q oerpd rou, ow6 o6qye( oe npcbtprq InEn rnq prerdgpooqq. To prBootopdtro ovoyvrr:p((er 1.d04 gerd rq oUv0eoq nemrdxd:v 6eopuiv enrOe<opcirwog rqv orpgri ouprnLq- porporrx6trlto qg 6Lrxog xro6rr<ov(ou-owrr<o6rrov(ou orq eSon P. Iuv6nero rqq ).dOog evoopdtoo4g e[vot 11 oulq pdvq rrov6rrlro tov nopoy6wrov oneleu Odprooq g vo npoodrivowor orn 05oq A riore vo npoxoLoriv np6- opo reppouopr6, or5pr1 rr 6rov 6ev undpXer xdnoro xrodu<6vro repprouol.roti. Enrnldov, ou[dver 11 ouXy6q- ro Ad0oq ro6u<ono(4oqg or4 drnLovrl 05oq A. H eno- x6Aou0r; 6rd6oor1 rtov ).o0rilv tefurd o64ye( oe np6opo rlepgmropr6. To r5orog ulg ;rerdgpoorlg, 6nog unoLoy((eror on6 rov oprOg5 6eoprcbv uUnlnq ev6pyerog nou np5- ntt vo u6poi.u0orlv, oogdrg ou[dver on6 trg 6tepyo- oteg 6t6p0o:o4g 6orrp(ov. To np6o0ero evepyeror6 x6mog e[oprdror on6 to orddto mo ono[o onoBd].- etot ro ).dOog or<ulurtp6vo IRNA To xorcq .3, l: i- t((eror Ue rrlv onoBoLrl nptv on6 rrlv udpo,r.r;- ::, GTP orpopd ;r6vo oqv nopoyroyq tro0oq oxuALoo;rc,3, IRNA ond qv IRNA ouv0erdo4. Om6oo, ov ro GTP u6polu0e( nprv on6 q drdcrrooq rou KoKoroLprooge- vou oprvooKuotRNA, ro evepyerqK6 x6moq 0o efvoL preyolUrepo. Aorpoltrrg, to prSyroro r6orog ouvdderoL pe rov np6orpo repprortopr6 qg prerdgpoorlq nou ono- 6(6et prr; Lenoupyrr<6 npoi6v xord tov SLeyXo noLdq- rog perd rn perogopd tou nentr6(ou. Irr;v npoKer- pr6vq nep(moror;, np6ner vo r<oroBLrl0e( oAdr<L4po ro evepyeror<6 ripr4pro nou oXet((eror Ue rrl oUv0eorl rou nolunenrrd(ou drllg ro oqpre[o qg npdotprlg one].eu- 05poo4g. 25.1 6 Merqr6nron Tou nAoroiou Avdyvoonq Mnopei vo IuUBri oe ONo0np6q AAAnAouxieq * To ntroloro ovoyvoorlg enripeo(erot ond qv otrtr1trouXro rou mRNA rol ro pgooogrr6 nepgotrtrov. * 0t otrto0t'jp6q otrtrqtrouXieg enup6nouv o' evo IRNA v0 UrTo- ron[(eror roro pro pool oqorou (euyopooet pe ro ovrtrotdr- x6vro rou, peropotrtrovrog erot'ro ntro(oro ovoyvooqg. * H Uerorppool opropdvrrlv yovLdlrov e[oproror on6 rrlv rorlrrl npoyp0pUoTr0Uevq geroronrol rou n),oro[ou ovoyvtoolg. To ou;rBdwo enovoxo6rxonoi4o4g (recoding) ou- vrlOtog nepr).opBdvouv oL).oy5g oro v6rlpro ev6g ;r6vo xto6u<ov(ou. IXenxd nopoderyproro e(vor -ro gorv6pe- vo rnq IRNA xorooroXrlq @vorrlta 25.12) Kqr n oUor- onolu<4 tponono(4o4 ev6g opuvooxuAo-tRNA (Evrir4- ra 25.8). Oor6oo, rperg dMor rUnot enovoro:6rxono(- qorlg enr<p5pouv nro oLroupevtrdg preroBoLdq oro no- poy6pevo noLunentr6rr6 npoi6v. l-lp6rerroL yro rrl geror6ntoq rou n).oro[ou ovdyv<oor1g (frameshifting) (e[erd(eror oe out4 rqv Ev6r4ro), r4v nopdKopqn (bypassing) xor rr1 Xpqoq 6rjo mRNA yro rrl o0v0eorl ev6g nounemr6(ou (r<or or 6Uo e[erd(owor orrlv Evd- qra 25.17). H ;leror6nroq rou nLoro(ou ovdyvooqg oXer((eror pe erdu<d tRNA oe 6[o nepLordoerg: . OproUSvor preto).).oyprdvor IRNA r<otomoLe[g ovo- yvrop[(ouv 5vo "r<ro6rr6vroD reoodpov Bdoeorv ow( rtov ouvriOrov rprcbv Bdoeotv. o OpLou6veq ,,o).to0rlp6S" oMrlLoul(eg entrp6nouv o' dvo IRNA vo pietortvlOe( ndvo mo mRNA orrl 05or1 A r<ord pLo Bdoq e(re oe 5'e(re oe 3' xotefOuvor1. MeroL).d(erg peror6ntorlg tou n).oLo(ou ovdyvo-l- or;g nporUnrouv on6 ).dOog ovdyvurorl rou r<to6trov[- ou rou mRNA. Avr( vo 6roBd(er pro tprn).5ro, ro ptBoooprdrto 6toBd(er e(re 6vo woudro e(re svo Kou- 3i.iC Meroromorl Tou lltroro[ou Avoyvoorlg Mnopei vo Iupper oe 0trr00lp6q AtrtrqtrouXreg 7A7 Boorxeg 6woreg
  • 227.
    oprdro vouKtreorrd(ov. Ierd0e nep(moto4, q ouv51t- on rnq ovdyvrr:or1q rprnLer6v perd to ouyrerpt;l5vo ouprBdv ono6i6er 6vo notrunent(6to er<r6g droto[ou. Mro peror6nron rou noro(ou ovdyvoorlg pnope( vo xoromotre( on6 5vo IRNA rxqv6 vo 6roBd(er 5vo ro6t- xov(o 6rio 11 reoodprov B6oecov. Ir4v nepintoorl rov r<codu<ov(olv reoodpov Bdoeov, to IRNA 6to05tet 6vo exrerog5vo 0p6Xo owrrodrr<ov(ou, nou onotetre(rqt on6 or,mdl voux.treor(6ro ow( tov rovovtr6v entd. l-to nopdderygo, 1.uo G e(vor 6uvor6 vo etoXoprioet oe Uto oerpd opxerc0v ouvel6prevov Bdoecov G. O Korooro- ).6o9 qq prerot5ntorlg rou nloro(ou ovdyvtooqg e(vot 6vo IRNAGly ro ono(o nepr6let pro np6o0etrl Fdon mo Bp61o rou owrrordrrov(ou xor gerorp6net ro ov- rtr<odrr6vro on5 rq ouvqO4 rprnl,dto CCCe oto r<ouopr6ro CCCC+-. To r<oroorotrrrx6 IRNA ovoyvo- p((er 5vo "Ko6rK6vro" reoodpov Bdoeov. Opropr6vot roroorotre[g perot6nroqg rou n],oro[ou ovdyvoorlg ovoyvcop[(ouv ndvo on6 5vo (KodtK6vtoD reoodpruv Bdoeolv. l-to nopd6etypto, 6vog Bor<tqpto- 169 roromol6oq tRNALv'owrororle[ oe AAAA n AAAU ow( rou ouvq0oug rodu<ov(ou MA. 'Evoq dtr).og r<o- roorol5og droBd(et onotoS4nore "Ko6tK6vto" teood- ptov Bdoecov pe ACC otrq rperq nprirreg 06oetg: 4 en6pevrl Bdo4 e[vor o6rdEoprl. Ie owdg ttg neptntri- oerq, or evotrtror<trr5q Bdoerg nou e(vot onodertdq orrlv r6topr4 0do4 rou 1loxpfrepou (Ko6rKov(ou" 6e oler[(owor Ue rouq ouvrlOetq rov6veg rrlg oordOetog. To xqroorotrrrx6 IRNA gdMov ovoyvcop((et 6vo ro6t- r6vro 3 Bdoeov, oMd yro xdnoto dl).o L6yo -nt0ov6- repo l.6yrrl orepeoxnlJtrrlg nope;rn66ror1q (steric hindrance)- 11 drnhovrl Bdoq ono<ppdooetot. Aur6 npor<oLe( nop6l,etr.l.rr1 prog Bdorlg npototi to en6pevo IRNA Bpet 5vo rodrx6vro. Koroordoerq orrg ono(eg q pretot6ntorl tou nloto(ou ovdyvrrlorlq e(vor <p u orotroyrr6 ou ;rBdv nopor; po tiwot oe tpdyoug xot dLtroug rorig. TSroro oupBdwo (otog enrl- pe6(ouv q ouv6Xtorl r1 rov reppottqr6 qg pretdqpo- onq Kor orpeilowor orrg eyyeve(g r6r6qreg tou mRNA. Itoug petpoio[q, 4 perdgpoorlrou npciltou yovt6(- ou reppror((erqt qn6 dvo oouv0ettr6 rtodtr6vto ornv (6ro <pdoq ge ro nl,o(oro ovdyvoorlg. To derircpo yovi 6ro Bp(or<etor oe 6rorpope'rx6 ntro(oto ovdyvtr:oqg rot (oe opropr6voug ro0q) Uero<ppd(eror on6 gto perot6nt- oq nloro(ou ovdyvtoo4g nou entrp6net pto evotrtroyrl oro 6e[repo nAofoto ovdyvcooqq rt 6ro nopordpmet ro rcr6rx6vto repgortopo0 (FMne tlK0NA 26.28xt enionq lr1v Ev6t4ta 17.12). H onoreLeoporr6tlto qg preror5- nronq rou nl.qro(ou ovdyvcoorlg e(vot XogqLrl, tuntrd -5"/o. H Xopnfui onoteleotrrotx6t4to 51er oqproo(o yto t4 BroLoy[o rou rori, xoOc0g n o0[non rqg onoretreqto- ttx6tlrog prnope( v' onoBe( entBLoBrlq. H EIK0HA 25.27 nopouord(et qv nop6proto rordoroo4 rou orotle(ou Ty rnq (UUnS, orrlv ono(o to t<to6tr6vto teptrrottol.toti rou tya npdner vo TropoKopq0e( on6 pto petor6nto4 rou nloro(ou ovdyvcooqq <iore vo 6toBoore[ to enor6- Lou0o yov(6ro fyb. T5roreg roroordoerg unoypogp(ouv rr1 orlpowtxrl Meroronrorl rou ntrotofou ovoyvoorlg -1 o{o perpoid HIV n/ NNNNUUUUU IJAG GNNNNNNNN Teleuro[o xodrxovro nou 6Lopd(etor mo opXtxo ntroloto ovoyvcrloqq Ilptino xrodrrovro nou 6r090(sr0r oro v6o ntroloro qvoyvoolq Avdyvtoorl Xcoprg peroronrol rou nloto[ou ovoyvooqg NNNNUUUUU UAG GN N N NNNN N Avdyvoon gerd rr1 gerorontoq rou ntroto(ou ovtiyvtooqg NNNNUUUUU GNNNNNNNN 'Evo IRNA, nou otrroOolver roro ;.tlo Bool oro (eu- yopto;.ro pre 6vo rordtrovro, npoxo),e[ ;lerordnrol rou n],oLoio- ovoyvoorlg 11 onolo rorooretrtrer Tn InEl H onoretreoportroq,: e(vot-5%. oppl 6rL ro onovro (oMd npoBL6Qtpo) ouptBdwo "Lov- Ooop5vr1q ovdyvtooqg" e(vot 5vo ovoyt<o[o Bqpo rqc EvotrloxrtroI tp6nor [etdqpqonq ono6i6ouv Tya 11 Tya-Tyb llporeivn Tya 5fufofofufufo) EvopIn An(n tltl - I tvterordnrol rou ] 'Evop[r1 nlqro(ou ovdyvtoolg AnEn Xroprg ArgtRNA, ro To Gly- IRNA lloporoUQ1 ougpotvet orov to ptpootr.tporto pero- ruve(ror roro plrog rou mRNA, 6ro1 6ore ro nenrtdutrotRNA qc Oeor1g P onetreuOeptlverol ono ro (euyoptol.to pe ro raldtxovtc rou xr 6netto (euyoptlver ue ronoro oLtro rodlrovto oe pteyotr0re- p1 onooroorl. Xrpopxrl rpoTelvn Tya-Tyb 738 KE0AM|0 25 0 levertrdg KtitdLxog llotrunenrldro -Leu
  • 228.
    quoro}.oyrKriq Uerdepoonq. Aw6role(tor npoypqppo- nop6vrl peror6ntoq rou ntroloiou ov6yvtooqq (pro- grammed frameshifting). IupBo(ver oe ouyKeKprpSvsg neprol6g pre ouXv6qro 100-1000 eop6q peyoL0teprl on6 rq ouly6rqro Ld0oug oe Un npoypo;rporoprdveg neproX5g (-3 x 10-s ovd r<todrr6vro). Aur6g o rinog peror6nrorlg tou nLoro(ou ovdyvto- orlg epgov((er dtio rowd Xoporcrqptonxd: o Mto "oAro0lpn" otrtrntrouX(o ennp6ner o' rivo o1-u- voorui,otRNA vo (euyopciloer pe ro rto6rr6vr6 rou Kr Snetrq vo UeroKrvrlee[ +1 rl -1 Bdo4 rime vo (euyopdloer Ue Urq enu<olunr6prevq rptnldto, en(- oqq txovrl vo (euyopdloer Ue ro qwtro6rK6vr6 rou. o To prpootrrprdrto xoOuorepe( orrlv nepLoXrl rnq pe- ror6nrorlg rou nLoro(ou ovdyvoo4g oqqvowog Xp6vo oro ogrvoor<ulo-tRNA yro v' ovo6rotd(er ro (euydpogd rou. H om[o qg r<o0uor6pr1oqq ;rnope[ vo e(vor 6vo 6rnLov6 rol6rx6vro nou onotre( 6vo ondvro ol.uvooruLo-tRNA, 5vo ro6rx5vro reppon- opoU nou ovoyvrop((erot opyd on6 rov ow(ororlo nopdyowo oneLeuO6pooqg rl 6vo 6optr6 epn66ro sro mRNA (n.1. 5vog "rpeu6or6pnoq,, Uro r6ro(repr1 dropr6p<pcoorl rou RNA) ilou nopoKol,fer ro prBooto- prdrto. H o).(o0r1o4 ouv(qrqror oe preror<(vqo4 efue npog 14 pro efue npog rrlv dMrl r<ore60uvor1: peror6ntorl -1 npo- rco.l,efuot 6rov ro IRNA perorrve[rot npog ro nioo, ev6 peror5nrorl + 1 6rqv ro IRNA getorrve[ror npoq ro pnpog. Ie rd0e nep(nroor1, to onor5Leo;.ro e(vor vo ercreOe[ gro tprn].6ro ercr6g gdorlg m4 0rio4 A yro to en6pevo o;.rwoor<ulo-tRNA. H gerot6nroq tou nloro(ou ovdyvoo4g npoyUqronore(ror nprv on6 r4 orivOeorl ttov nemt6xc0v 6eopr<irv. Iro ouq06orepo rUno, nou nupo- dorefuor on6 pro o).ro0qpri oMrlLouX(o oe ouv6uoop5 pe [ro ro0o6u<r1 goupr<fto oro mRNA, or neprBdMou- oeg otrtr4).ou1ieg enqped(ouv rrlv onoreleoporu<6r4rd rnq. H petor6nrorl rou nl.oro[ou ovdyv<ooqg orrlv Eu<5vo 25.27 6etpter rrl oupnepupopd gtog runLrrlg oLroO4prlg oMq).ouX(og. H oLLrll,oul(o enrd vourl.eorrd(atv CU- UAGGC ouvq0cog ovoyvop((erqr on6 ro Leu-tRNA oro CUU, nou oxolou0efuor on6 ro ArgtRNA oro AGG. To ArgtRNA e(vot ondvro, r<or 6rov 4 onovr6rqrd rou npo- xoi.e[ r<oOuordpnon, ro Leu-tRNA otrro0o(vet on6 ro ro- 6u<5vro CUU npog trlv enu<ol,um6;revq rprn).6ro UUA. Aw5 nporoLe[ puo petor6nrorl rou nl.qto(ou ovdyvrrl- oqg, ener6rl q en6;revq rprnl,5to orqv (6ro <pdor1 ge ro v6o (euydpopq (GGC) 6roBd(stor on6 ro GlytRNA. H oL(o04or1 ouvrl0cog flpoyporonorefuor orq 06oq P (6rov ro Leu-tRNA ornv npoyUorx6r4to SXer yfver nemrdu- )otRNA nou peroq6per. rrl veoouwe0erprdvrl oluo(6o). H peror6nror; rou nl,oto(ou ovdyvroo4g o' 6vq 106r- r6vro repponopori nporol,e( en6rcroorl r4g nporeivqg. H Bdorl ornv 3' nl,eupd rou r<o6rxov(ou repgottollori enrlped(er q oXenrq ouXv6rrlro rou repUorrotrroU ror rrlg peror6nrorlq Kt 6tot enqped(er r4v onoteLeoport- r6tqro tou o(prorog repprorqrorJ. 25.1 7 AAAo lupBovro EnovoKo6lKonolnonq, MerocppooTtKn nopdKop$n Kot o Mnxovtopoq Tou tmRNA yto Tnv AneAeu0dpoon Tov KoOnAtrrpdvov P rBoooUoTiov a H n0porouUl ouv[ororor oqv uovdrrlro rou prpoorrllror[ou vo repgor[(eL rq perorppoorl, v' one)eu0epdrveror ond ro mRNA ror vo [ovopXi(er q perdqpooq nep(nou 50 vour]reor[6ro ro- 0odLro. e To prBootouono nou efvor roOqtrolg6vo novo mo mRNA pero q peprxrl ouvOeoq pLog nptoreiVrlq pnopei v' onetreu0epo- 0o0v ono rn Dpdon Tou tmRNA, ev6g povodrrof RNA ge Xo- poxrrlprorxo 16oo rou IRNA ooo xoL rou mRNA. H nopdxo;rQ4 (bypassing) neprlo;rBdver Uro perorf- vnorl rou prBooropro{ou nou oltr6(er ro ro6rr6vro ro ono(o e(vor (euyopro;rdvo pe ro nemtdutro-tRNA orrl 05or1 P. H oli.r1l,ou1(o prero[ri rov 6rio rodrrovfrov nopol.e(netor ror 6ev owrnpootoneieror oro no).une- nrt6rr6 npoi6v. 'Onrog nopouord(eror orrlv EIK0NA 25.28, out6 ennp5ner vo ouvexr.orel r'1 perdgpooq n6- po on6 onoro6rlnore ro:6x6vro repgortoprori ruXo(ver vo Bp(or<etor finv nopeprBoM6prevq nepro14. To nro ewunooror6 nopd6erypro nopdroprprlg e(vor ro yov(- 6ro 60 rou rpdyou T4, 6nou to prBooroprdrro perorrve(- tor 50 vour<,eor[6ro r<ord UriKoq rou mRNA. To r),er6( yro ro orjmqpro nopdroprqqg e(vor 6tt undplouv novopordtuno (r1 ouvcbvupro) r<o6rr6vro oe rd0e dxpo qg oLLqLoul(og nou nopoi.e(nerot. Aurd nqpoKopt|n 60 vourtreottd(tov oro yov6ro 60 rou rpdyou T4 Tetreurq[o xo0rxdvro mo oplud ntro[oro ovdyvoorlq [lptirrl xoldrrovLo oro veo ntroroto ovoyvrooqg Avdyvtooq Xorp(q gerordnrorl rou ntrqtofou ovoyvroolg GAUGGA UGAC......,.....AUUGGAUUA Avdyvoorl gero q perordnrorl rou ntroroiou ovdyvrrlorlg GAUGGA:.)".;;.tt; . . ,t.:',-::-::.r,, UUA Mro peroronrorl nLorolou ovoyvrrlolg +1 onoue[- r0r yro Trlv erqpoori rou yovrdlou tyb rou ororXe(ou Ty rotv (upo- pur1rov. H peroronLorl npoyl-roronore(Tor 0e Uro otrtr1trouX[o enro pooeov, orrlv ono[o 6uo rtoDrr<ovto Leu orotrouOouvror ono 6vo onovro xotdrrovro Arg. GAUGGA UGAC...,........AU ' .: : , '[116 Iuppowo Enovorodtxonoqolg: tVleroqpomul tloporo;lfr1 ror MqXovrolrog tmRNA yto Anetreu0epool KoOqtrot;revov PLBoor*rur i-. 739 Buorxeg eworeg
  • 229.
    UeprK6q qop5q ovqq5poworoq ileproxSq "onoye(- ooqq" Kor (npooye(o:or1g". l-lprv on6 rnv nopdKouqJn, ro prBootol.rdrro tono0ere('roL pr' 6vo nenrt6utrotRNA (euyoptop6vo ge ro rcodx6vro onoye(rrlor1q or4 05oq P, evd: 4 ddero 05o4 A ovop6ver rlv e(oo6o ev6g o;rwoo- rui.o{RNA. H EIK0NA 25.29 6e(;yer 6n ro prBoocopdrro olro0q(ver rotd pr4rog rou mRNA oe owf r4v rord- orooq Srog 6tou ro nemrdutro-tRNA prnop6oer vo (eu- yopriloer pe ro ro6rxdvro ornv neproXrl npooye(rooqg. H otrl.qLouX(o rou mRNA nupo6ote( rqv nopdr<op- Un. To or11lowu<d XopoKrnprorxd efuor to 6rlo rcodrr6- vto GGA yro rnv onoye(cooq Kor rnv npooye(coor'1, ro nopepBoll6pevo 6rdm4Uq, Uro doprl p(oXou-0ql,tdg nou nepil.o1-rBdver ro r<<o6rrdvro onoye(oor1g ru 5vo rtrl- 6x6vro repprortopro6 6fuLq oro rto6x6vro onoye(ooqg. To md6ro r4g onoye(ooqg npouno0Ster ono65' opeuon tou nemrdutrotRNA on6 ro xrrl6rr6vro pe to ono(o e(vor (euyopo;r5vo. Ar<oLou0e( puo peror<[vqorl rou mRNA nou ro egno6f(er vo [ovo(euyopr6oer. 'Ener- to, ro ptBoocoprdrro oopciver ro mRNA, 6o9 6rou ro ne- nrrdulo-tRNA (euyopciroer Ue ro rro6rr6vro otrlv ow(6poor1 npoo'ye(<oo4g.'Eneno, q nporeivoorJv0eorl [ovopX((er, pr6llg 5vo o1-uvoor<u].o-tRNA eroS].0er orn 06oq A rord to ouvr10r1 rp5no. 'Onog q peror6ntoq rou nloro(ou ovdyvtoorlg, 6ror ror 4 owfdpoon rnq nopdroprQqg Boo(eror oe pto noU- otl rou prBooroprorfou. H nr0ov6rrlro 6rdoroorlq rou ne- mrdu).o{RNA on6 ro rrodx6vr6 rou onl 06o4 P ou(dver on6 ro0umSpnon ornv efoo6o rou oprvooKutro{RNA orn 06on A. H or6p4o4 ev6g oprwo[6og ;rnopei vo nu- fficnucAc........i [lpooye[roor1 To lterTrOutro-tRNA Eovo- (euyopovet ire v5o xtodtxovto 1:'r :ii:,; Ir' .111 Iq geooDo i1g noporo;it11g, 6vo prBootoporto pe roreAlpp6vq Oeon P Unoper vo repporloer q Ueroqpool. Otrr- oOo[ver xoro pqrog rou mRNA oe ;:ro neproXti, onou ro nenri6u- tro{RNA (euyoprlver pe evo rodrrovro 0TI 0dort P 'Ene uo q Uerotppoorl ouvey[(erot. 740 KE0AAAI0 25 0 l-evelro_c Kridrroq po6orrl oet nopdr<opr{.rq oro Borcrrl pror<d yov(6Lo elottfoc rrlg r<o0um6prlolg nou ouprBo(vet 6tov 6ev undpleL 6ro0Sorpro oprLvoor<ul.otRNA yto vo eto6tr0er oq 06o11 A Iro yov(6to 60 tou <pdyou T4, Svog p6).oq qg 6opnc rou mRNA e(voL vo geuilvst rrlv onorel.eotrlorrxdrqro rou repporroprori, npoxolrrlwog Sror qv r<o0uor6pqor nou onotrefuor yto rnv ow(6poo4 onoyefooqg. H 6rdorooq rov ro0qlu:trrdvrov prBootopror(rov mo Borcrrlpro r<or oprop6vo prLtoX6vSpro ennuyldveroi prdcc- ev6g provodLr<o0 uBpt6[ou mRNA{RNA nou xotrefuoL tm- RNA rol nepLdlet 66o LeLroupyu<69 neproXdg. H pro ne- pro1q onoprrpe(tor 6vo rprrlpro rou tRNAAu, ovri r OeUrepq rordrNonoLe( 6vo ppolJ no).unem(61o. To tmR- NA opXrr<d oprvoor<ul.rcoveror ond tqv oLovuAo-tRNA ouvOerdoq (AlaRS). Ir4 ouv6XeLo npoo6dver rov nopo- yowo EF-Tu ru 5ner-ro og rproroydg orjprnl.oxo npoo- dSveror orq 05oq A rolv r<o0r1).rlr;l6vov prBoorl-l;.tot(tov Idvo oro prBooroprdlo ouprBo(veL prerogopd nemL6[or- nou ouvddeL qv olov(vq pie ro ropBo(ureLrxd dr<po rt'1c r<o0r1Lcop6vq g, veoouweOerpr5vrlq npolrelvq g : row6lpo - vo, ro mRNA nou Bp(or<eroL ndvo oro prBoorr.ll-rotto ov- lxoOrordror ond q 6e[repr1 nepLoXrl rou tmRNA. Im ouvdXero, to tmRNA tretroupye( cog erpoye(o yLo tq orjv- 0eor1 66r<o enrnpdoOerov o1-rtvoldorv, preto ro ono[c undpXet 6vo r<rodu<6vro rsppolopori yro rov repUorroUC rqg perdgpoonq Kor qv oneLeuO6pcoorl rrlg npir.lreir,lq l-i np6o0eq r<opBo(ureLu<r1 oh}.q),ouX(o 6po rog ot11-rc yro enordlou0rl ovoyvioprorl ond npr.,:redoeg, or ono(ee ono6oproriv t4v r<o).oBropr5vq npu.lrelvq. 'EroL, ro tmRNA Lenoupye( og 6vog prqXovropr6g el.6y1ou noL6trpog yrc ttlv ovorrix..rr.lo4 r<o0q).rilptdvov ptBootopor(orv Kot rrlv onopdr<puvoq xo).oBropdvrov nporeivov, ot ono(eg er- 6oMrog 0o ouootrlpe[owov. 25.1 I llepiAn(rn H oM4LouX(o rou mRNA, nou dLoBd(eror orrg rprn),e- rsq Ue rote(Ouvoq 5'->3' olsr((eror pSoo rou yevert- r<otj xrrr6rxo Ue rnv oL).r1LouX(o rc'rv oprrvo(5ov rov no).unemt6[tov, nou dtoBd(eror ond ro N- npog to C- rel.rx6 dr<po. And ng e[r1wo r6ooeprg rprn].6req, e[qv- ro Ur.o xco6rronoroiv oprLvo[5o Kor rperq nopexouv or;;roro reppoloprori. To ouvc6vupro r<ttl6rr6vro, nou owrnpooorne[ouv ro i6to opuvo[5o, 6Xouv ouyyeve[g oMrll.ouX(eg, ouyyd pre puo preroBotr4 orriv tpfur; Boon rou xro6rr<ov(ou. Aur4 q er<cprjl.roq rqq tpkqg pdolq oe ouv6uoopd U' 6vo np6runo orjp<povo pe ro onoio 1qpu<6q nop6proro opuvo[do re(vouv vo x<r.lStronototjv- ror on6 ouyyev4 rordtx6vro, eloXtoronoLe( tLg enrdpo- osrq rov pteroltrd[eo.lv. O yevettr6g r<trl6rr<og e[vot orxoupevrx6g r<or npdneL vo xo0tep60r1re no).ri nportl,ro rotd rqv e[6].1[r1. Or oMoy6g oro nupnvrKd yovrdurr- poro e(vot ondvteg, oMd oproprdveg oMoy6g npoypo- ronoLrl04r<ov r<otd rlv e(6Lt(q rov l.utoXovdp(orv. I-loMonLd IRNA prnope( v' onowo[v oro i6ro r<or6L- r<dvLo. H otr-rodo rorv IRNA nou onowoJv mo noLr(i.o xroSu<6vto yro r<dOe opuvo[d e(vor loporrqproru<r1 yto Anoyelo.lon To ptBoocrrgorLo peroxrveltol roro prlrog rou mRNA
  • 230.
    Kdee opyovroU6. Hovoyv6pton KodtKov(ou-owtKo6t- rov(ou egntr5r<et oor60eto orqv npdrtrl 05oq tou owt- rodrr<ov[ou (tpir4 06or1 rou ro6tr<ov[ou), 11 ono(o ennpSner os optopdvo IRNA v' ovoyvcop((ouv noMo- nLd r<odr6vto. Otro to IRNA nepdXouv tpononorrlprS- veg Bdoerq, ot ono(eg etodyovtot on6 6v(upo nou ovoyvrop((ouv Bdoerg-m6xouq orn 6opr4 tou IRNA. To (euydpolpo ro6trov(ou-owtrot6trov(ou enqped(etot on6 rpononoqoetg rou (6tou tou owtt<todxov(ou 6nog en(oqg xot on6 ro (ougQpo(5pevo", 6rlLo6n rtq Yero- vrx6g Bdoetq, r6(u:q orrlv 3'nLeupd tou owtt<trl6trov[ou. A[ronotdlwog t4v oor60eto ro:6xov(ou-owro6u<ovfou, to 1.troX6v6pro rov onov6utror6v opt<otiwot oe 22 lR' NA yro rrlv ovoyvdlptoq 6trov ttov rcto6tt<ov(tov oe oX6or1 pre ro ouvr]0q eL6Xtoro opt0pr6 tov 31 IRNA: oe our6 ouprBdMouv ot oll.oy6g oto l.tttoXov6ptor6 rrirdxo. Kd0e oprwo(ti ovoyvcopf(erot qn6 pto ouyrerpt;r6vr1 o1-uvoor<ul.o-tRNA ouvOetdoq, q onoio ovoyvcop[(et en(- oqg 6Lo ro IRNA nou rto6tronoto6v ro ouyrerptpttivo optvo(ri. Optopr5veq ogwoorutro-tRNA ouvOetdoeq 6to- O5touv pto trettoupy(o pretor6ntorlg tou nl'oto(ou qvd- yvoonq, 11 ono(o ent0ecope( ro oUtvooKULo{RNA t<ot u6potrUet ro ).dOog ouvde6etrrSvo ol.twooruLo-tRNA. Or oprwooxul.o{RNA ouvOetdoeg notx(ouv no}.0, oor6oo xototdooowot oe 6Uo yevx6g ogddeg nou eU- qov((ouv oprotBo(o onorlet6pevo got(Bo oM4),ouXrcilv rot np<oreivrdrv 6o;rcbv ottg rototruttxdg neptoX6g toug. Ot 66o otrrd6eg tcr:v ouv0eroodlv 6torp[vowot en[or1g on6 r4v qp1Kn neproXq rnq optvooKuL(toorlg orrlv 3'-te- hxn ptp6(n tou IRNA, t4v roreU0uvon rnq np6odeoqq rrlg Sllrog-onod6rcr4 tou IRNA rot to puOptoneptoptort- r<6 Brjpo rnq optvooKul(ooqg. Mto roOopto;.t5vq opd6o voux.treold(tov tou IRNA, nou xotrefuot o;rd5o tout6rq- roq, ovoyvop[(eror enil.erctrd on6 t1 ouv0erdorl, 4 ono(o lp4otpronoe( 5vo ouvduoop6 dpeoov ru Sppeocov pulXovtoprdrv ovdyvcoorlg. Ie nol'L5g neptnt6oetg, 11 opd6o rout6qtog tonoOetefuot oro dr<po tou owtrcodt- rov(ou Kot rou 3'-ono65rrn tou 1-top(ou. MeroMd[etg Unope( vo ennpS{.rouv o' 6vq IRNA vo 6toBd(er dto<popertrd ro6u<6vto: r'1 ouvrlO6orepn Uop- <pr1 rdrouov pretoMd[ecov ouprBo(vet mo (6to ro owtKo- 6tr6vro. H ueropotrn tou owtrcodtt<ov(ou prnope( vo enmpdrper o' 5vo IRNA vo rotoorc(tret prto petdMo(r1 o' 5vo yov(6to nou r<odtronote[ npote(w1. 'Evo IRNA nou ovoyvop((er 5vo rrr:dtr6vto repporl.opoU nop5Xer 5vov oouv0err6 rotoorol,5o, evdl 5vo IRNA nou ;retoBdMet ro oprwo[0 nou owtorotxe( o' 5vo rco6tx6vto e[vot Svog 6uouv0enr6g r<otooro5o9. Ot r<otooroe(g trrrv rco6t- rov(cov UAG e(vot onoteleo;.totx6tepot on6 touq ow(- ororxouq rov t<to6trov[ov UAA: out6 e(qyefuot on6 to yeyov6g 6tt to UAA e(vot to nto 6tode6opt5vo rro6tx6vto reppottopo0. H onotetreopottr5tqro 6trtov ttov t<oto- orol5tov, oo16oo, Boo[(etor oro (oupqpo(6pevo" rou r<dOe [eXo:ptorof r<o6xov[ou-or6Xou. Meroton(oetg tou ntroto[ou ovdyv<oor1g +1 ri -1 nt- 0ov6v npoxotroUwot on6 olloO4pdg oMrltrouX[eq oro mRNA nou enttp5nouv o' 5vo nemt6ulo-tRNA vo ol't- oOrloet qn6 to rco6tr6vt6 tou t<otd p(o Fdon efue oe 5' efue oe 3' rote0Ouvor1. OptoprSveg flpoYpopporrotrr6veg petoron(oerg tou nLoto(ou ovdyvotolq ::- ':::: -: tor on6 qv oMql.ouX[o tou mRNA onoiro- ':]. '.: 6x<ppooq puoLxtov yovt6[cov. Ilopor<o;rqq o-*p :.' . drov 6vo pLBooto;rdtto mopord rri pretdgpool Kot []e- -rorwe(ror xotd prlrog rou mRNA pe ro nemtdu).o- tRNA orl1 06ori P 6<r-r-c 6rou owd (euyopooeL p' 6vo rotoMq- Lo opuvo(ti, ondre f,ovopXi(et q pretd<ppool' H Xplort rou tmRNA nopSlet 6vo prrlXovto;r6 etr5y1ou not6tqrog yro rrlv ovot<tixlool r<o01trtl;r6vov ptBooropiot(rov xot r1v onoproxpuvoq ovent00prltov r<o}oBtlp6vov npote'[- vLr<.ov npo[dwirlv. BrBAroyporpio 25.1 ELouytoytl "Epeuvo Nirenberg, M W. and Leder, P. (1964). The effect of trinucleotides upon the binding of sRNA to ribosomes. Science 145, 1399- 1407. Nirenberg, M. W. and Matthaei, H. J. (1961). The dependence of cell-free protein synthesis in E coli upon naturally occurring or synthetic polyribonucleotides. Proc. Natl. Acad. Sci. USA 47, 1 588-1 602. 25.3 H Avoyvtoptol Ktrtdtxovlou-AvTtKoDtKov[0u l-lepilogPovet Aoro0eto 'Epeuvo Crick, F. H. C" (1966). Codon-anticodon pairing: the wobble hy- pothesis. J. Mol. Biol. 19, 548-555. 25.4 T0 tRNA IlopoYovrot pe EfleEepYooio Meyotr(repov IlpoDpdptov Mop(ov Avoor6qoq Hopper, A. K. and Phizicky, E. M. (2003). IRNA transfers to the limelight. Genes Dev. 17, 162-180. 25.5 To tRNA ilept6Xet Tpononotlpeveq B00etq Avoouinrlorl Hopper, A. K. and Phizicky, E. M. (2003). IRNA transfers to the limelight. Genes Dev. 17, 162-180. 25.6 0r Tpononoulpeveq B6oetq Enrlpeo(ouv T0 ZeuYopoU0 AvrtroDtxov(ou-Ktodtxov(ou Avoordnnon Agris, P. F. (200S). Bringing 0rder to translation: the contributions of transfer RNA anticodondomain modifications. EMB0 Repofts 9, 629-635. 25.7 Ynopxouv Inopodu6g Tpononotfloetg rou 0troupevtrou Ktl0tro iti:rlilri i: :_rl: 0sawa, S. et al. (1992). Recent evidence for evoiution of the ge- netic code. Microbiol. Fev. 56, 229-264. BrBtrLoypotpio 7 41
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