Fission yeast is a model organism for study of the molecular biology of cell cycle control. A gene of particular signifcance in controlling the rate of progression through both the G1 and G2 phases of the cell cycle (cdc2+) has previously been identified genetically. DNA sequences analysis and site-specific mutagenesis of the gene strongly suggest that it encodes a protein kinase. This proposal is concerned with three aspects of the molecular biology of cdc2+. (1) Since all known protein kinases are themselves phosphorylated and since genetic evidence suggests that the activity of the cdc2+ product is modulated during the cell cycle, the modification of the cdc2+ product by phosphorylation will be investigated. The possibility that the modification of the cdc2+ product might vary during the cell cycle will be particularly investigated. (2) The probable protein kinase activity of the cdc2+ product from yeast and after over-expression in E. coli will be investigated. (3) Targets of cdc2+ protein kinase phosphorylation will be sought. This will involve direct biochemical experiments, study of phosphoprotein changes during the cell cycle, analysed by 2-D gel electrophoresis and also classical genetics and analysis of cdc2(ts) suppressors. This proposal concentrates almost entirely on the cdc2+ gene product in the belief that it is of particular significance in the control of the cell cycle and that understanding its biochemical activity would open many new areas of cell cycle research.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM034607-03
Application #
3285917
Study Section
Genetics Study Section (GEN)
Project Start
1986-09-05
Project End
1989-08-31
Budget Start
1988-09-01
Budget End
1989-08-31
Support Year
3
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Cold Spring Harbor Laboratory
Department
Type
DUNS #
065968786
City
Cold Spring Harbor
State
NY
Country
United States
Zip Code
11724
Lundgren, K; Allan, S; Urushiyama, S et al. (1996) A connection between pre-mRNA splicing and the cell cycle in fission yeast: cdc28+ is allelic with prp8+ and encodes an RNA-dependent ATPase/helicase. Mol Biol Cell 7:1083-94
Connolly, T; Beach, D (1994) Interaction between the Cig1 and Cig2 B-type cyclins in the fission yeast cell cycle. Mol Cell Biol 14:768-76
Hofmann, J F; Beach, D (1994) cdt1 is an essential target of the Cdc10/Sct1 transcription factor: requirement for DNA replication and inhibition of mitosis. EMBO J 13:425-34
Samejima, I; Matsumoto, T; Nakaseko, Y et al. (1993) Identification of seven new cut genes involved in Schizosaccharomyces pombe mitosis. J Cell Sci 105 ( Pt 1):135-43
Molz, L; Beach, D (1993) Characterization of the fission yeast mcs2 cyclin and its associated protein kinase activity. EMBO J 12:1723-32
Matsumoto, T; Beach, D (1993) Interaction of the pim1/spi1 mitotic checkpoint with a protein phosphatase. Mol Biol Cell 4:337-45
Matsumoto, T; Beach, D (1991) The spil GTPase interacts with RCCl in cell cycle dependency. Cold Spring Harb Symp Quant Biol 56:385-98
Matsumoto, T; Beach, D (1991) Premature initiation of mitosis in yeast lacking RCC1 or an interacting GTPase. Cell 66:347-60
Lundgren, K; Walworth, N; Booher, R et al. (1991) mik1 and wee1 cooperate in the inhibitory tyrosine phosphorylation of cdc2. Cell 64:1111-22
Ducommun, B; Draetta, G; Young, P et al. (1990) Fission yeast cdc25 is a cell-cycle regulated protein. Biochem Biophys Res Commun 167:301-9

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