2. A briefe history of the cell cycle 18xx Interphase-Mitosis 1951 Discovery of a distinct S-phase 1970 Cell fusion experiments 1971 Discovery of MPF 197x Yeast cdc mutants 1983 Discovery of the first cyclins 1988 Cyclin B+cdc2 = MPF 199x Ubiquitin and the cell cycle 1999 Sister chromatid separation

3. The most important role of cell cycle control is to ensure accurate chromosome segregation M G1 S G2

4. General principles and methods G1 - To cycle or not to cycle S- Replicating the genome M-Segregating the genome Cytokinesis - Making two cells

5. ONCE and ONLY ONCE

6. Rao and Johnson 1970

7. Fluorescent Activated Cell Scan/Sorter (FACS)

8. A typical FACS analysis

9. Methods for cell synchronisation

10. Model organisms I - Budding yeast (S.cerevisiae) L. Hartwell

11. Model organisms II - Fission yeast (S.pombe) P. Nurse

12. Identification of cell cycle ts mutants

14. Generation of temperature sensitive budding yeast mutants

15. Analysis of ts cell cycle mutants G1 S M M M M

16. Maturation promoting factor

18. Oscillations of MPF

23. Cyclins and cyclin dependent kinases (cdks) - in Budding yeast Cdk1 (cdc28) (S.p. cdc2) Cln1,2,3 G1 Clb5,6 S Clb3,4 G2 Clb1,2 M

24. Cyclins and cyclin dependent kinases (cdks) - in multi-cellular organisms Cdk1 (S.p. cdc2) Cyclin D1,2,3G1Cyclin E Cyclin A S Cdk4,6 Cdk2 Cyclin A Cyclin B1,2,3 M

25. Cdc2/28 (=cdk1) is regulated by: 1. Binding of a cyclin 2. Inhibitory phosphorylation 3. Activating phosophorylation 4. Binding of CKI 5. Cellular localization

28. G1 - To cycle or not to cycle S- Replicating the genome M-Segregating the genome Cytokinesis - Making two cells

31. P Cyc D cdk4 pRB E2F P P P P Cyc E cdk2

33. G1 - To cycle or not to cycle S- Replicating the genome M-Segregating the genome Cytokinesis - Making two cells

34. The human genome is encoded by 3,000,000,000 bp (3x10 9 ) which must all be replicated within a few hours with the highest fidelity. The loss of a bp is irreversible and a single mistake could potentially lead to the death of the organism.

35. Analogy One page contains about 3000 letters. Imagine that you have to copy one million pages in a few hours and have to bind them into 23 volumes. You can use as many typist as you like but you are allowed no mistakes -no missing letter, no letter twice, and no substitutions.

38. G1 - To cycle or not to cycle S- Replicating the genome M-Segregating the genome Cytokinesis - Making two cells

41. The story of the two blind men Two blind men go into a shop and buy each five different pairs of socks. Unfortunately all ten pairs were packed in the same bag. How can the two men separate the socks so that each of them will get five pairs ?

42. The three factors that enable sister chromatid separation The glue - cohesins The pulling force - the mitotic spindle The timing mechanism - spindle checkpoint

43. The three factors that enable sister chromatid separation The glue - cohesins The pulling force - the mitotic spindle The timing mechanism - spindle checkpoint

45. Sister chromatid separation from prometaphase to anaphase Securin Separase Cohesin metaphase Securin Separase Cohesin prometaphase APC Check point Separase APC Securin Separase anaphase Check point

46. Non-degradable securin does not prevent cytokinesis but interferes with complete chromatide separation histon-GFPHoechst HeLa securin KAA-DM securin KAA-DM NIH3T3

47. The three factors that enable sister chromatid separation The glue - cohesins The pulling force - the mitotic spindle The timing mechanism - spindle checkpoint

52. Anaphase Promoting Complex/ Cyclosome

53. APC Checkpoint cdk1 Cyclin B Fzy Checkpoint Fzr Polo Prophase

54. APC cdk1 Cyclin B Fzy Checkpoint Fzr Polo Metaphase

55. APC cdk1 Cyclin B Fzr Fzy Fzr Fzy Anaphase Polo cdc14 Metaphase

56. Yes (chrom. movement) Metaphase Chrom. alignment on metaphase plate Xkid ?TelophaseMitosisE2 of the APC/CUbcH10 No Telophase - G1 MetaphaseAPC/C activatorFzy Yes (cyclin A+B degr.) PrometaphaseG2, prophaseKinaseNek2A NoG1SReplicationCdc6 ? Telophase - G1 MitosisMitotic kinasePlk1 Yes (cytokinesis) MetaphaseProphase- metaphase Mitotic kinaseCyclin B1 Yes (chrom. separation) MetaphaseS-metaphaseChromatid cohesion maintenance Securin Yes (metaphase) PrometaphaseS, G2, prophase Kinase (+cdk1/2)Cyclin A2 Degrad. essential Degradation initiated ActiveRoleSubstrate