DNA Replication Cell Division Chromosome Segregation

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DNA Replication Cell Division Chromosome Segregation

G1 division xerox DNA replication cell machine collating S machine DNA chromosome segregation M G2 Alternation of DNA replication and chromosome segragation DNA replication + chromosome segregation The logic of the cell cycle: cell fusion experiments (Rao & Johnson) S phase cells contain a factor that can trigger DNA replication in G1 cells: S phase Promoting Factor The logic of the cell cycle: cell fusion experiments (Rao & Johnson) block of re-replication The Cell Division Cycle division 2 mass growth 1 2 DNA nucleus 1 Balanced growth and division TC = cycle time TD= mass doubling time 4 3 TC > TD mass 2 TC = TD Cytoplasmic 1 TC < TD 0 0 1 2 3 4 time Chromosome cycle Growth cycle - DNA replication - cytoplasmic growth - mitosis - cell division (precise replication (approx. doubling and segregation) and halving) Dissociation of growth and chromosome cycle Oogenesis Embryogenesis Saccharomyces cerevisiae Lee Hartwell Isolation of temperature sensitive cdcts mutants Phenotype of cdc mutants asynchronous culture cdc mutant with problem of mitotic exit Cdc28 is responsible for the first genetically controlled event cdc28 Cloning of cdcts genes Cdc28 is a protein-kinase Schizosaccharomyces pombe S G1 M G2 Sir Paul Nurse Murdoch Mitchison cdc2ts cdc25ts wee1ts wildtype cdc13ts cdc2ts ts wee1ts cdc25 wildtype ts wee2ts = cdc2D cdc13 cdc2 is required at two points during the cell cycle cdc2 cdc28S.c. and cdc2S.p. are functional homologs Do human cells have the same gene? Nature, Vol, 327, No. 6117, pp. 31-35, May 7, 1987 Melanie Lee Cell, Vol. 45: 145-153, April 11, 1986 Cell, Vol. 49: 559-567, May 22, 1987 inject cytoplasm Yoshio Masui, Ph.D., FRS Professor Emeritus of Zoology University of Toronto Ontario, Canada Yoshio Masui, Ph.D., FRS Professor Emeritus of Zoology University of Toronto Ontario, Canada Periodic proteins (cyclins) during early embryonic development Tim Hunt Western IP Jim Maller MPF is a cyclin-dependent protein-kinase Cdk1 CycB Cyclin and MPF levels during early mitotic cycles Questions: What causes the fast disappearance of cyclins? What is the reason of the delay between cyclin and MPF? Cycling cell free extract Anaphase Promoting Complex or Cyclosome chromosome segregation C d c 2 APC 0 Kim Nasmyth Anaphase Promoting Complex or Cyclosome chromosome segregation C d c 2 APC 0 Cdk1 Cdk1 CycB CycB degradation + degraded CycB The G2/M transition is regulated by post-translational modification Wee1 P Cdk1 Cdk1 CycB CycB Cdc25 inactive active (preMPF) (MPF) Wee1 is a tyrosine-kinase Cdc25 is a tyrosine-phopshatase Start of DNA replication is also controlled by proteolysis degraded CKI G1 Cdk P Cdk Cdk CKI CKI CKI cyclin cyclin cyclin inactive Cdk phosphorylated multiubiquitaned Cdk/cyclin CKI CKI cyclin complex active Cdk/cyclin complex Regulation of CDK activity Cyclin Cdk P Cdk Cdk Cyclin Cyclin Cdk CKI Cyclin CKI.

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