2. PI3,4,5P3: Translocatie van Protein Kinase B (PKB) en PDK1 (kinase) PDK1: Activatie van Protein Kinase B (PKB)

3. cellcycle p27kip1 p130 Cyclin D P RTK catalase MnSOD stress PI3K PKB FOXO P Gadd45a Bim FasL P celldeath 14-3-3 FOXO P FOXO PKB fosforyleert FoxO transcriptie factoren: Gevolg: Translocatie uit de kern Inactivatie P JNK ubiquitin -catenin College Burgering

4. Translocatie van GFP-FoxO uit de kern na insuline stimulering (PI-3K activatie)

5. Many effects of PKB/Akt: Hub

6. Signalling networks centered around hubs

7. PI3,4,5P3: Doet meer, o.a. Translocatie van BTK (kinase) en PLC (lipase) BTK activeert PLC

8. Conservering van princiepes: MAP kinase “cascades Ras

9. The Ras superfamily (>170 members) Ras family (Ras, Rap1, R-ras, Ral) - Cell proliferation, differentiation, migration Rho family (Rho, Rac, CDC42) - Cytoskeleton, adhesion, polarity Rab family - Vesicular transport Arf family - Vesicle formation/fusion Ran - Nuclear import and export Regulate the formation of protein complexes at the right time and place

10. Rhes Dex-Ras kB-Ras1 kB-Ras2 NOEY2 RIG GEM REM2 Rad REM RERG RIT RIN Rap1A Rap1B Ras2A Rap2C Rap2B K-Ras4A K-ras4B H-ras N-Ras TC21 R-ras M-Ras RalA RalB Rheb The Ras family >28 different members

11. Rheb/mTor/S6 kinase pathway controls cell growth Insulin PI-3K PIP3 PKB Wt S6k-/- Tsc1/Tsc2 1763 Rheb GAP Tsc2 (tuberin) mTor Rapamycin Regulatie door remming van GAP (Tsc2) (i.p.v. activatie door GEF) S6K 4E-BP1 S6

12. EGF receptor signalering Figure 5.1 The Biology of Cancer (© Garland Science 2007)

13. De TGFb-route (1) TGFb receptoren zijn transmembraan serine-threonine kinases Actieve receptor fosforyleert receptor-regulated Smad (R-Smad) (Smad 2 of 3)

14. De TGFb-route (2) Actieve receptor fosforyleert receptor-regulated Smad (R-Smad) Binding van Co-Smad (Smad 4) en translocatie van complex naar de nucleus Binding aan DNA bindings eiwit en activatie van genexpressie

15. Wnt signalling pathway

16. Hedgehog signalling pathway functions in development and differentiation Figure 6.29c The Biology of Cancer (© Garland Science 2007)

17. Notch signalling pathway functions in development and differentiation Transcription factor Figure 6.29b The Biology of Cancer (© Garland Science 2007)

18. Ephrin-Eph Receptor (een RTK!) signalling pathway functions in repulsion during cell migration Note: bidirectional signalling!!! Figure 15-62a Molecular Biology of the Cell (© Garland Science 2008)

19. Ephrin-Eph Receptor signalling pathway functions in repulsion during cell migration GEF for Rho GTPases Switch

20. Ubiquitin: 76 aa protein: Post-translational modification of proteins Also Sumo and NEDD: ubiquitin like molecules that modify proteins

21. Sequential ubiquitination of proteins: E1, E2, (E3) ubiquitin ligases

22. Different modifications/ different functions

23. Different modifications/ different functions Protein degradation endocytosis

24. Signaal transductie: Externe signalen integreren tot een biologische response Control units: eiwit complexen die opgebouwd worden onder invloed van signalen: Post-translationele modificaties, Scaffold en adaptor eiwitten, Eiwit domeinen Moleculaire schakelaars: Kinasen-phosphatasen, G-eiwitten Positieve en negatieve feedbacks Second messengers; o.a. cAMP, Ca2+, PIP3, DAG, IP3 Beperks aantal receptor systemen met veel variatie: GPCR, RTKs Aantal meer specifieke receptor systemen: TGFb, Wnt, Hedgehog, Notch,… Ubiquitine systeem: Afbraak, endocytose, localisatie