Dual regulation of the yeast CDC28-p40 protein kinase complex: Cell cycle, pheromone, and nutrient limitation effects

A 40 kd polypeptide that coprecipitates with the CDC28 gene product in immune complexes is specifically phosphorylated by the CDC28 protein kinase. Using this reaction, we detect activity only in extracts from dividing G1 phase cells. Exit from G1 by entry into S phase or the preconjugatory state induced by mating pheromone correlates with loss of p40 phosphorylation activity. Inactive extracts from cdc28 mutants complement extracts from cells arrested in S or M phase, suggesting that non-G1 cells are deficient in an exchangeable activating factor. Stationary and pheromone-treated cultures are rich in this exchangeable factor, but possess an inactive kinase that is not activated by complementation. cAMP-deficient mutants resemble stationary cells.

[1]  D. Botstein,et al.  Isolation and Characterization of Mutations in the β-Tubulin Gene of SACCHAROMYCES CEREVISIAE , 1985 .

[2]  L. Hartwell,et al.  Genetic Control of the Cell Division Cycle in Yeast: V. Genetic Analysis of cdc Mutants. , 1973, Genetics.

[3]  Components of microtubular structures in Saccharomyces cerevisiae. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[4]  E. Bücking-Throm,et al.  Purification and partial characterization of -factor, a mating-type specific inhibitor of cell reproduction from Saccharomyces cerevisiae. , 1973, European journal of biochemistry.

[5]  C S McLaughlin,et al.  Rate of macromolecular synthesis through the cell cycle of the yeast Saccharomyces cerevisiae. , 1978, Proceedings of the National Academy of Sciences of the United States of America.

[6]  S. Reed,et al.  Primary structure homology between the product of yeast cell division control gene CDC28 and vertebrate oncogenes , 1984, Nature.

[7]  S. Reed,et al.  Protein kinase activity associated with the product of the yeast cell division cycle gene CDC28. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[8]  M. Slater Effect of Reversible Inhibition of Deoxyribonucleic Acid Synthesis on the Yeast Cell Cycle , 1973, Journal of bacteriology.

[9]  P. Nurse,et al.  The cell cycle control gene cdc2 + of fission yeast encodes a protein kinase potentially regulated by phosphorylation , 1986, Cell.

[10]  L. Hartwell Sequential function of gene products relative to DNA synthesis in the yeast cell cycle. , 1976, Journal of molecular biology.

[11]  L. Hartwell,et al.  Sequential gene function in the initiation of Saccharomyces cerevisiae DNA synthesis. , 1974, Journal of molecular biology.

[12]  M. Osley,et al.  Cell-cycle regulation of yeast histone mRNA , 1981, Cell.

[13]  L. Hartwell Saccharomyces cerevisiae cell cycle. , 1974, Bacteriological reviews.

[14]  P. Nurse,et al.  Functionally homologous cell cycle control genes in budding and fission yeast , 1982, Nature.

[15]  Richard A. Lerner,et al.  Immunogenic structure of the influenza virus hemagglutinin , 1982, Cell.

[16]  J. D. Levi Mating Reaction in Yeast , 1956, Nature.

[17]  J. Kyhse-Andersen Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. , 1984, Journal of biochemical and biophysical methods.

[18]  S. Reed,et al.  Subcellular localization of a protein kinase required for cell cycle initiation in Saccharomyces cerevisiae: evidence for an association between the CDC28 gene product and the insoluble cytoplasmic matrix , 1987, The Journal of cell biology.

[19]  I. Herskowitz,et al.  Control of yeast cell type by the mating type locus. I. Identification and control of expression of the a-specific gene BAR1. , 1981, Journal of molecular biology.

[20]  M. Wigler,et al.  The S. cerevisiae CDC25 gene product regulates the RAS/adenylate cyclase pathway , 1987, Cell.

[21]  L. Hartwell,et al.  Genetic control of the cell division cycle in yeast. , 1974, Science.

[22]  K. Matsumoto,et al.  Control of cell division in Saccharomyces cerevisiae mutants defective in adenylate cyclase and cAMP-dependent protein kinase. , 1983, Experimental cell research.