Buněčný cyklus - principy regulace buněčného růstu a buněčného dělění

1. Buněčný cyklus - principy regulace buněčného růstu a buněčného dělění

2. Mitóza Dr. B. Duronio, The University of North Carolina at Chapel Hill

3. Průběh mitózy v buněčné kultuře fibroblastů Dr. B. Duronio, The University of North Carolina at Chapel Hill

4. Buněčný cyklus Dr. B. Duronio, The University of North Carolina at Chapel Hill

5. Kinázy závislé na cyklinech kontrolují buněčný cyklus Dr. B. Duronio, The University of North Carolina at Chapel Hill

6. Kontrola vstupu do mitózy - výsledek využití různých buněčných modelů

7. Experimental Systems Important for Cell Cycle Studies Saccharomyces cerevisiae Arbacia punctulata Schizosaccharomyces pombe Xenopus laevis Dr. B. Duronio, The University of North Carolina at Chapel Hill

8. Protein Level cyclin A cyclin B Time M M M Cyclin was Discovered in Sea Urchin Embryos can stimulate to lay lots of eggs Dr. B. Duronio, The University of North Carolina at Chapel Hill

9. Frog life cycle OOCYTE GROWS WITHOUT DIVIDING (MONTHS) FERTILIZED EGG DIVIDES WITHOUT GROWING (HOURS) FERTILIZATION 1 mm sperm tadpole feeds, grows and becomes an adult frog

10. The Maturation of Frog Eggs An Assay for Maturation Promoting Factor (MPF) Yoshio Masui, 1971

11. MPF Activity Peaks Before Each Cell Division Moreover, MPF has kinase activity

12. Dr. B. Duronio, The University of North Carolina at Chapel Hill

13. Dr. B. Duronio, The University of North Carolina at Chapel Hill Purification of MPF: The Birth of Cyclin Dependent Kinases This is cdc2+!! (Cdc28 in S. cerevisiae) This is cyclin!! Which = cdc13+ in S. pombe

14. Fission yeast: Schizosaccharomyces pombe Dr. B. Duronio, The University of North Carolina at Chapel Hill

15. Budding Yeast Saccharomyces cerevisiae Dr. B. Duronio, The University of North Carolina at Chapel Hill

16. Cdc Mutants Arrest at the Same Cell Cycle Phase Permissive (low) temperature Restrictive (high) temperature Dr. B. Duronio, The University of North Carolina at Chapel Hill

17. Cdc Genes Encode Proteins Needed for the G2-M Transition: Studies in S. pombe cdc2+ encodes a kinase Moreover = cdc28 in S. cerevisiae! Dr. B. Duronio, The University of North Carolina at Chapel Hill

18. Phosphorylation of CDK Targets Changes Their Activity Now performs a cell cycle function Dr. B. Duronio, The University of North Carolina at Chapel Hill

19. Jak jsou CDK regulovány? • prostřednictvím syntézy a odbourávání cyklinů • fosforylací • pomocí CDK inhibitory proteins (CKIs)

21. Cyclin Destruction is Controlled by Ubiquitination Dr. B. Duronio, The University of North Carolina at Chapel Hill

22. Představují cykliny jediný způsob regulace CDK?

23. Exprese cyklinů v jednotlivých fázích BC Dr. B. Duronio, The University of North Carolina at Chapel Hill

24. CDK jsou regulovány fosforylací

25. Conformational Changes Associated with CDK Phosphorylation Free CDK CDK + Cyclin T161 phosphorylation The T-loop blocks substrate access Binding of cyclin moves the T-loop Phosporylation moves the T-loop more

26. Cyclin Dependent Kinase Inhibitors (CKIs)

27. Dr. B. Duronio, The University of North Carolina at Chapel Hill active inactive CDK CDK + p21 p21 Cyclin Cyclin p27Kip1 CDK2 Cyclin A The p21 Family of CDK inhibitors (p21CIP1/WAF1, p27KIP1, p57KIP2) Cyclin A CDK2 Jeffrey et al. (1995) Nature 376:313 Russo et al. (1996) Nature 382:325

28. Dr. B. Duronio, The University of North Carolina at Chapel Hill active inactive CDK4/6 CDK4/6 + + Cyclin D INK4 Cyclin D INK4 Russo et al. (1998) Nature 395:237 Brotherton et al. (1998) Nature 395:244 The INK4 Family of CDK inhibitors (p16INK4a, p15INK4b, p18INK4c, p19INK4d)

29. CKIs Regulate the G1-S Transition (p16) (p21, p27) Dr. B. Duronio, The University of North Carolina at Chapel Hill

30. Nature Reviews Cancer 2001, 1:222

31. Dr. B. Duronio, The University of North Carolina at Chapel Hill

32. pRB Binds to the E2F Transcription Factor Dr. B. Duronio, The University of North Carolina at Chapel Hill

33. G1 Cyclin CDKs Phosphorylate pRB Dr. B. Duronio, The University of North Carolina at Chapel Hill

34. Proliferation Signals p16 Regulates pRB Phosphorylation Dr. B. Duronio, The University of North Carolina at Chapel Hill

35. Nature Reviews Cancer 2001, 1:222

36. Kontrola buněčného cyklu úzce souvisí s: • kontrolou buněčného růstu; • přítomností růstových faktorů a dalších růstových stimulů a živin; • působením ostatních buněk populace a mezibuněčné hmoty.

37. Growth with No Cell Division Cell Division + Growth = Proliferation! Cell Division No Growth The Difference Between Growth and Cell Division

38. Growth with No Cell Division: A Differentiated Neuron Dr. B. Duronio, The University of North Carolina at Chapel Hill

39. Cell Division with No Growth: Early Development 1 mm sperm Dr. B. Duronio, The University of North Carolina at Chapel Hill

40. Nature Reviews Cancer 2001, 1:222

41. Growth Factors Induce Cell Cycle Progression Growth Factors act at the Restriction Point Dr. B. Duronio, The University of North Carolina at Chapel Hill

42. Nature Reviews Cancer 2001, 1:222

43. Dr. B. Duronio, The University of North Carolina at Chapel Hill pm = post mitotic ps = pre-synthetic Zetterberg and Larsson, PNAS 82:5365 (1985)

44. Growth Factors Induce Gene Expression Dr. B. Duronio, The University of North Carolina at Chapel Hill

45. Growth Factors Induce Cyclin D1 Expression Sherr and McCormick, Cancer Cell, Vol 2, 103-112 (2002)

46. Mitogen Induced Cell Cycle Progression in Cell Culture Dr. B. Duronio, The University of North Carolina at Chapel Hill

48. Cell Cycle Checkpoints

49. Cell Cycle Checkpoints Improve Cell Viability Weinert and Hartwell, Science 246:629 (1989)

50. How do Cell Cycle Checkpoints Work? Zhou and Elledge Nature408, 433 - 439 (2000)