Morphogenesis in the yeast cell cycle: regulation by Cdc28 and cyclins

Analysis of cell cycle regulation in the budding yeast Saccharomyces cerevisiae has shown that a central regulatory protein kinase, Cdc28, undergoes changes in activity through the cell cycle by associating with distinct groups of cyclins that accumulate at different times. The various cyclin/Cdc28 complexes control different aspects of cell cycle progression, including the commitment step known as START and mitosis. We found that altering the activity of Cdc28 had profound effects on morphogenesis during the yeast cell cycle. Our results suggest that activation of Cdc28 by G1 cyclins (Cln1, Cln2, or Cln3) in unbudded G1 cells triggers polarization of the cortical actin cytoskeleton to a specialized pre-bud site at one end of the cell, while activation of Cdc28 by mitotic cyclins (Clb1 or Clb2) in budded G2 cells causes depolarization of the cortical actin cytoskeleton and secretory apparatus. Inactivation of Cdc28 following cyclin destruction in mitosis triggers redistribution of cortical actin structures to the neck region for cytokinesis. In the case of pre-bud site assembly following START, we found that the actin rearrangement could be triggered by Cln/Cdc28 activation in the absence of de novo protein synthesis, suggesting that the kinase may directly phosphorylate substrates (such as actin-binding proteins) that regulate actin distribution in cells.

[1]  J. Cooper,et al.  Disruption of the actin cytoskeleton in yeast capping protein mutants , 1990, Nature.

[2]  E. Cabib,et al.  Synthesis of the yeast cell wall and its regulation. , 1982, Annual review of biochemistry.

[3]  P. Russell,et al.  Fission yeast p107wee1 mitotic inhibitor is a tyrosine/serine kinase , 1991, Nature.

[4]  D. Beach,et al.  The Xenopus cdc2 protein is a component of MPF, a cytoplasmic regulator of mitosis , 1988, Cell.

[5]  D. Soll,et al.  Temporal and spatial differences in cell wall expansion during bud and mycelium formation in Candida albicans. , 1985, Journal of general microbiology.

[6]  P. Nurse Universal control mechanism regulating onset of M-phase , 1990, Nature.

[7]  J. Tkacz,et al.  Wall replication in saccharomyces species: use of fluorescein-conjugated concanavalin A to reveal the site of mannan insertion. , 1972, Journal of general microbiology.

[8]  D. Soll,et al.  The involvement of cell wall expansion in the two modes of mycelium formation of Candida albicans. , 1985, Journal of general microbiology.

[9]  T. Hunt,et al.  Maturation promoting factor, cyclin and the control of M-phase. , 1989, Current opinion in cell biology.

[10]  Jonathan A. Cooper,et al.  Purification, characterization, and immunofluorescence localization of Saccharomyces cerevisiae capping protein , 1992, The Journal of cell biology.

[11]  J. Chant,et al.  Budding and cell polarity in Saccharomyces cerevisiae. , 1991, Current opinion in genetics & development.

[12]  Uttam Surana,et al.  The role of CDC28 and cyclins during mitosis in the budding yeast S. cerevisiae , 1991, Cell.

[13]  K. Beran Budding of Yeast Cells, their Scars and Ageing , 1968 .

[14]  B. Haarer,et al.  Immunofluorescence methods for yeast. , 1991, Methods in enzymology.

[15]  M. Snyder,et al.  Cell polarity and morphogenesis in Saccharomyces cerevisiae. , 1992, Trends in cell biology.

[16]  F. Cross,et al.  A potential positive feedback loop controlling CLN1 and CLN2 gene expression at the start of the yeast cell cycle , 1991, Cell.

[17]  M. Snyder,et al.  Studies concerning the temporal and genetic control of cell polarity in Saccharomyces cerevisiae , 1991, The Journal of cell biology.

[18]  P. Nurse,et al.  Gene required in G1 for commitment to cell cycle and in G2 for control of mitosis in fission yeast , 1981, Nature.

[19]  D. Botstein,et al.  Requirement of yeast fimbrin for actin organization and morphogenesis in vivo , 1991, Nature.

[20]  S. Reed,et al.  Direct induction of G1-specific transcripts following reactivation of the Cdc28 kinase in the absence of de novo protein synthesis. , 1992, Genes & development.

[21]  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.

[22]  B. Carter,et al.  Genes which control cell proliferation in the yeast Saccharomyces cerevisiae , 1980, Nature.

[23]  Daniel J. Lew,et al.  A cyclin B homolog in S. cerevisiae: Chronic activation of the Cdc28 protein kinase by cyclin prevents exit from mitosis , 1991, Cell.

[24]  Kim Nasmyth,et al.  Positive feedback in the activation of Gl cyclins in yeast , 1991, Nature.

[25]  V. Farkaš,et al.  Autoradiographic Study of Mannan Incorporation into the Growing Cell Walls of Saccharomyces cerevisiae , 1974, Journal of bacteriology.

[26]  J. Pringle,et al.  Use of a screen for synthetic lethal and multicopy suppressee mutants to identify two new genes involved in morphogenesis in Saccharomyces cerevisiae , 1991, Molecular and cellular biology.

[27]  J. Pringle,et al.  CDC42 and CDC43, two additional genes involved in budding and the establishment of cell polarity in the yeast Saccharomyces cerevisiae , 1990, The Journal of cell biology.

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

[29]  Paul Russell,et al.  Negative regulation of mitosis by wee1 +, a gene encoding a protein kinase homolog , 1987, Cell.

[30]  I. Herskowitz,et al.  A yeast gene (BEM1) necessary for cell polarization whose product contains two SH3 domains , 1992, Nature.

[31]  S. Reed,et al.  Different G1 cyclins control the timing of cell cycle commitment in mother and daughter cells of the budding yeast S. cerevisiae , 1992, Cell.

[32]  S. Reed,et al.  Cyclin-B homologs in Saccharomyces cerevisiae function in S phase and in G2. , 1992, Genes & development.

[33]  S. Reed The selection of S. cerevisiae mutants defective in the start event of cell division. , 1980, Genetics.

[34]  F. Cross,et al.  DAF1, a mutant gene affecting size control, pheromone arrest, and cell cycle kinetics of Saccharomyces cerevisiae , 1988, Molecular and cellular biology.

[35]  J. Cooper,et al.  Effects of cytochalasin and phalloidin on actin , 1987, The Journal of cell biology.

[36]  J. Broach,et al.  The Molecular biology of the yeast Saccharomyces : metabolism and gene expression , 1982 .

[37]  David Botstein,et al.  Phenotypic Analysis of Temperature-sensitive Yeast Actin Mutants , 2022 .

[38]  B. Futcher,et al.  The Cln3‐Cdc28 kinase complex of S. cerevisiae is regulated by proteolysis and phosphorylation. , 1992, The EMBO journal.

[39]  Curt Wittenberg,et al.  An essential G1 function for cyclin-like proteins in yeast , 1989, Cell.

[40]  S. Reed,et al.  Isolation of three novel human cyclins by rescue of G1 cyclin (cln) function in yeast , 1991, Cell.

[41]  B. Haarer,et al.  Purification of profilin from Saccharomyces cerevisiae and analysis of profilin-deficient cells , 1990, The Journal of cell biology.

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

[43]  B. Haarer,et al.  Cellular morphogenesis in the Saccharomyces cerevisiae cell cycle: localization of the CDC3 gene product and the timing of events at the budding site , 1991, The Journal of cell biology.

[44]  Eric T. Rosenthal,et al.  Cyclin: A protein specified by maternal mRNA in sea urchin eggs that is destroyed at each cleavage division , 1983, Cell.

[45]  C. Ballou Yeast Cell Wall and Cell Surface , 1982 .

[46]  L. Meijer,et al.  Cyclin is a component of the sea urchin egg M‐phase specific histone H1 kinase. , 1989, The EMBO journal.

[47]  A. Barton Some aspects of cell division in saccharomyces cerevisiae. , 1950, Journal of general microbiology.

[48]  D. Lew,et al.  A proliferation of cyclins. , 1992, Trends in cell biology.

[49]  G. Fink,et al.  Methods in yeast genetics , 1979 .

[50]  A. Bretscher,et al.  Characterization of TPM1 disrupted yeast cells indicates an involvement of tropomyosin in directed vesicular transport , 1992, The Journal of cell biology.

[51]  S. Aaronson,et al.  Catalysis of guanine nucleotide exchange on the CDC42Hs protein by the dbloncogene product , 1991, Nature.

[52]  D. Beach,et al.  Population explosion in the cyclin family , 1991, Current Biology.

[53]  W. L. Fangman,et al.  Cell cycle phases in the unequal mother/daughter cell cycles of Saccharomyces cerevisiae , 1984, Molecular and cellular biology.

[54]  J. Pringle,et al.  Staining of actin with fluorochrome-conjugated phalloidin. , 1991, Methods in enzymology.

[55]  G. Tokiwa,et al.  The WHI1+ gene of Saccharomyces cerevisiae tethers cell division to cell size and is a cyclin homolog. , 1988, The EMBO journal.

[56]  J. Pringle,et al.  Roles of the CDC24 gene product in cellular morphogenesis during the Saccharomyces cerevisiae cell cycle , 1981, The Journal of cell biology.

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

[58]  J. Pringle Staining of bud scars and other cell wall chitin with calcofluor. , 1991, Methods in enzymology.

[59]  K Nasmyth,et al.  Control of the yeast cell cycle by the Cdc28 protein kinase. , 1993, Current opinion in cell biology.

[60]  S. Reed,et al.  A family of cyclin homologs that control the G1 phase in yeast. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[61]  H. Piwnica-Worms,et al.  p107wee1 is a dual-specificity kinase that phosphorylates p34cdc2 on tyrosine 15. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[62]  J. Pringle,et al.  Molecular characterization of CDC42, a Saccharomyces cerevisiae gene involved in the development of cell polarity , 1990, The Journal of cell biology.

[63]  G. C. Johnston,et al.  The Saccharomyces cerevisiae MYO2 gene encodes an essential myosin for vectorial transport of vesicles , 1991, The Journal of cell biology.

[64]  J. Pringle,et al.  Cellular morphogenesis in the Saccharomyces cerevisiae cell cycle: localization of the CDC11 gene product and the timing of events at the budding site. , 1991, Developmental genetics.

[65]  Curt Wittenberg,et al.  G1-specific cyclins of S. cerevisiae: Cell cycle periodicity, regulation by mating pheromone, and association with the p34 CDC28 protein kinase , 1990, Cell.

[66]  David Beach,et al.  cdc2 protein kinase is complexed with both cyclin A and B: Evidence for proteolytic inactivation of MPF , 1989, Cell.

[67]  J. Labbé,et al.  MPF from starfish oocytes at first meiotic metaphase is a heterodimer containing one molecule of cdc2 and one molecule of cyclin B. , 1989, The EMBO journal.

[68]  S. Reed,et al.  Control of the yeast cell cycle is associated with assembly/disassembly of the Cdc28 protein kinase complex , 1988, Cell.

[69]  Sergio Moreno,et al.  Conservation of mitotic controls in fission and budding yeasts , 1989, Cell.

[70]  L. Meijer,et al.  cdc2 is a component of the M phase-specific histone H1 kinase: Evidence for identity with MPF , 1988, Cell.

[71]  S. Reed,et al.  Mitotic role for the Cdc28 protein kinase of Saccharomyces cerevisiae. , 1990, Proceedings of the National Academy of Sciences of the United States of America.