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Structure of cell nucleus,  nuclear receptors and intracellular signalling
Nucleus : stru c tur e  a nd  func tion ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Ultrathin section Freeze fracture ne nucleolus np
Nuclear DNA is packaged into  chromosomes ,[object Object],[object Object],[object Object],Mitotic chromosomes as seen in a scanning electron microscope
Chromosome:  a complex of DNA and proteins (chromatin).  Basic units are  nucleosomes Nucleosomes     protein core made of 8  histone molecules:   2 copies of  H2A,H2B, H3 a H4     DNA  Histones: Small proteins with a high proportion of positively charged amino acids – lysine and arginine. These positive charges help the histone bind tightly to the negatively charged sugar-phosphate backbone of DNA
Levels of chromatin packing 1.  naked DNA molecule -  does not occur in vivo  2. beads-on-a-string occurs teporarily during  transcription 3. nucleosomes condensed into 30 nm thread due to a binding od H1 histone 4. Organization of the  300 nm thread from loops along a central axis. Non-histon proteins are involved 5. Formation od secondary loops to form a 700 nm thread 6. Metaphasic chromosome 1400 nm thick ( role of condensin)
Lidské chromosomy Mitotic chromosomes from  human lymphocytes Chromosomes:  homologous nonhomologous Chromosome identification:  banding FISH: fluorescence in situ hybridization
The three DNA sequence elements  needed to produce a eucaryotic chromosome that can be replicated and then segregated at mitosis
Interphase nucleus:  nuclear envelope  nuclear lamina  nuclear matrix nuclear pores IF ER MT Heterochromatin highly condensed, transriptionally inactive   Euchromatin decondensed in interphase condensed during mitosis
The nuclear pore complex The nuclear envelope is perforated b y n. pores – the gates for entering or leaving molecules of RNA or proteins NP is composed of about  100 different proteins and forms a short channel with protruding protein fibrils Small molecules – nucleoti-des, ATP and others enter freely. Large molecules  require  an appropriate sorting signal  (nuclear localization signal)
Topolog y of biochemical pro c esses   in the nucleus Chromosom al domains  - each chromosome occupies a defined sector of the nuclear space „Movement“   of  chromosom es  is experimentall y proved –  tagged  centromeres  or   telomeres  change their position  to the NE Transcription: RNA polymerase I –  nucleolus   RNA polymerase II and  I II –  nucleoplasm   (small compartments  40-100 nm in diameter) A ssembly  of  ribosomal subunits:  nucleolus  DNA replication:  replication foci  along the chromosomes     1  replication focus  contains some 5 replicons, velocity of the   replication - 2kbp/min    during S phase some 10 000 replication foci are   operating in the nucleus
Topology  of interphase chromosomes  in the nucleus Each chromosome occupies a particular region of the nucleus so that the chromosomes are not entangled with each other. The chromosomes are attached to a NE or nuclear lamina via  proteins  of the inner membrane NE:  lamin B receptor  (LBR) or lamina – associated protein (LAP2). To these proteins  c hromatin  and  lamins  are bound
Nucleolus   a region where the parts of different chromosomes carrying genes for ribosomal RNA cluster together. Here the genes are transcribed and ribosomal subunits are assembled (using ribosomal proteins imported from the cytoplasm
Nuclear receptors ,[object Object],[object Object]
Nucleus and signal transduction pathway
Nuclear receptors as transcription factors
Nucleus and signal transduction pathway
Nuclear receptors ,[object Object],[object Object]
Nuclear receptors as transcription factors
Key terms  1. Overall structure and function of the human cell nucleus 2. Chromosome structure as seen in the optical microscope. 3. Cytochemical identification of single chromosomes  4. Three sequence  elements of chromosomal DNA 5. Molecular arrangement of DNA and histones in the  chromosome 6. Levels of chromosome condensation  7. Ultrastructure of the nuclear surface and nuclear pores  8. Nucleolus.  9. Topology of chromosomes in the nucleoplasm 10. Nucleus and intracelular signalling 11. Nuclear receptors  as transcription factors
 
Vyštěpující tělísko Heterochromatin PcG tělísko Cajalovo (svinuté)  tělísko   a  Gem Chromosomální teritoria PML tělísko Jaderné skvrny SAM68 tělísko Perinukleární  kompartment Jadérka  Periferní jaderná lamina RNA pol II transkripční faktor OPT doména Komplex jaderného póru JADERNÉ DOMÉNY

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L04 nucleus

  • 1. Structure of cell nucleus, nuclear receptors and intracellular signalling
  • 2.
  • 3.
  • 4. Chromosome: a complex of DNA and proteins (chromatin). Basic units are nucleosomes Nucleosomes  protein core made of 8 histone molecules: 2 copies of H2A,H2B, H3 a H4  DNA Histones: Small proteins with a high proportion of positively charged amino acids – lysine and arginine. These positive charges help the histone bind tightly to the negatively charged sugar-phosphate backbone of DNA
  • 5. Levels of chromatin packing 1. naked DNA molecule - does not occur in vivo 2. beads-on-a-string occurs teporarily during transcription 3. nucleosomes condensed into 30 nm thread due to a binding od H1 histone 4. Organization of the 300 nm thread from loops along a central axis. Non-histon proteins are involved 5. Formation od secondary loops to form a 700 nm thread 6. Metaphasic chromosome 1400 nm thick ( role of condensin)
  • 6. Lidské chromosomy Mitotic chromosomes from human lymphocytes Chromosomes: homologous nonhomologous Chromosome identification: banding FISH: fluorescence in situ hybridization
  • 7. The three DNA sequence elements needed to produce a eucaryotic chromosome that can be replicated and then segregated at mitosis
  • 8. Interphase nucleus: nuclear envelope nuclear lamina nuclear matrix nuclear pores IF ER MT Heterochromatin highly condensed, transriptionally inactive Euchromatin decondensed in interphase condensed during mitosis
  • 9. The nuclear pore complex The nuclear envelope is perforated b y n. pores – the gates for entering or leaving molecules of RNA or proteins NP is composed of about 100 different proteins and forms a short channel with protruding protein fibrils Small molecules – nucleoti-des, ATP and others enter freely. Large molecules require an appropriate sorting signal (nuclear localization signal)
  • 10. Topolog y of biochemical pro c esses in the nucleus Chromosom al domains - each chromosome occupies a defined sector of the nuclear space „Movement“ of chromosom es is experimentall y proved – tagged centromeres or telomeres change their position to the NE Transcription: RNA polymerase I – nucleolus RNA polymerase II and I II – nucleoplasm (small compartments 40-100 nm in diameter) A ssembly of ribosomal subunits: nucleolus DNA replication: replication foci along the chromosomes 1 replication focus contains some 5 replicons, velocity of the replication - 2kbp/min during S phase some 10 000 replication foci are operating in the nucleus
  • 11. Topology of interphase chromosomes in the nucleus Each chromosome occupies a particular region of the nucleus so that the chromosomes are not entangled with each other. The chromosomes are attached to a NE or nuclear lamina via proteins of the inner membrane NE: lamin B receptor (LBR) or lamina – associated protein (LAP2). To these proteins c hromatin and lamins are bound
  • 12. Nucleolus a region where the parts of different chromosomes carrying genes for ribosomal RNA cluster together. Here the genes are transcribed and ribosomal subunits are assembled (using ribosomal proteins imported from the cytoplasm
  • 13.
  • 14. Nucleus and signal transduction pathway
  • 15. Nuclear receptors as transcription factors
  • 16. Nucleus and signal transduction pathway
  • 17.
  • 18. Nuclear receptors as transcription factors
  • 19. Key terms 1. Overall structure and function of the human cell nucleus 2. Chromosome structure as seen in the optical microscope. 3. Cytochemical identification of single chromosomes 4. Three sequence elements of chromosomal DNA 5. Molecular arrangement of DNA and histones in the chromosome 6. Levels of chromosome condensation 7. Ultrastructure of the nuclear surface and nuclear pores 8. Nucleolus. 9. Topology of chromosomes in the nucleoplasm 10. Nucleus and intracelular signalling 11. Nuclear receptors as transcription factors
  • 20.  
  • 21. Vyštěpující tělísko Heterochromatin PcG tělísko Cajalovo (svinuté) tělísko a Gem Chromosomální teritoria PML tělísko Jaderné skvrny SAM68 tělísko Perinukleární kompartment Jadérka Periferní jaderná lamina RNA pol II transkripční faktor OPT doména Komplex jaderného póru JADERNÉ DOMÉNY