Nucleocytoplasmic trafficking of G2/M regulators in yeast.

Nucleocytoplasmic shuttling is prevalent among many cell cycle regulators controlling the G2/M transition. Shuttling of cyclin/cyclin-dependent kinase (CDK) complexes is thought to provide access to substrates stably located in either compartment. Because cyclin/CDK shuttles between cellular compartments, an upstream regulator that is fixed in one compartment could in principle affect the entire cyclin/CDK pool. Alternatively, the regulators themselves may need to shuttle to effectively regulate their moving target. Here, we identify localization motifs in the budding yeast Swe1p (Wee1) and Mih1p (Cdc25) cell cycle regulators. Replacement of endogenous Swe1p or Mih1p with mutants impaired in nuclear import or export revealed that the nuclear pools of Swe1p and Mih1p were more effective in CDK regulation than were the cytoplasmic pools. Nevertheless, shuttling of cyclin/CDK complexes was sufficiently rapid to coordinate nuclear and cytoplasmic events even when Swe1p or Mih1p were restricted to one compartment. Additionally, we found that Swe1p nuclear export was important for its degradation. Because Swe1p degradation is regulated by cytoskeletal stress, shuttling of Swe1p between nucleus and cytoplasm serves to couple cytoplasmic stress to nuclear cyclin/CDK inhibition.

[1]  E. Bailly,et al.  Differential cellular localization among mitotic cyclins from Saccharomyces cerevisiae: a new role for the axial budding protein Bud3 in targeting Clb2 to the mother-bud neck , 2003, Journal of Cell Science.

[2]  Daniel J. Lew,et al.  A Morphogenesis Checkpoint Monitors the Actin Cytoskeleton in Yeast , 1998, The Journal of cell biology.

[3]  P. Silver,et al.  Regulated nuclear localization of stress-responsive factors: how the nuclear trafficking of protein kinases and transcription factors contributes to cell survival , 1999, Oncogene.

[4]  Josep Clotet,et al.  Phosphorylation of Hsl1 by Hog1 leads to a G2 arrest essential for cell survival at high osmolarity , 2006, The EMBO journal.

[5]  Jennifer K. Hood-DeGrenier,et al.  Cytoplasmic Clb2 is required for timely inactivation of the mitotic inhibitor Swe1 and normal bud morphogenesis in Saccharomyces cerevisiae , 2006, Current Genetics.

[6]  D. Lew,et al.  The morphogenesis checkpoint: how yeast cells watch their figures. , 2003, Current opinion in cell biology.

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

[8]  E. Bi,et al.  ZDS1 and ZDS2, genes whose products may regulate Cdc42p in Saccharomyces cerevisiae , 1996, Molecular and cellular biology.

[9]  M. Donzelli,et al.  Regulating mammalian checkpoints through Cdc25 inactivation , 2003, EMBO reports.

[10]  Hiroyuki Osada,et al.  M-phase kinases induce phospho-dependent ubiquitination of somatic Wee1 by SCFbeta-TrCP. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[11]  Chandra L. Theesfeld,et al.  Determinants of Swe1p degradation in Saccharomyces cerevisiae. , 2002, Molecular biology of the cell.

[12]  R. Sikorski,et al.  A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. , 1989, Genetics.

[13]  R. Sikorski,et al.  p62cdc23 of Saccharomyces cerevisiae: a nuclear tetratricopeptide repeat protein with two mutable domains. , 1993 .

[14]  Daniel J. Lew,et al.  Differential Susceptibility of Yeast S and M Phase CDK Complexes to Inhibitory Tyrosine Phosphorylation , 2007, Current Biology.

[15]  Ingrid Hoffmann,et al.  Cdc25b and Cdc25c Differ Markedly in Their Properties as Initiators of Mitosis , 1999, The Journal of cell biology.

[16]  M. Snyder,et al.  Nim1-related kinases coordinate cell cycle progression with the organization of the peripheral cytoskeleton in yeast. , 1999, Genes & development.

[17]  Steven P. Gygi,et al.  Cdk1-Dependent Regulation of the Mitotic Inhibitor Wee1 , 2005, Cell.

[18]  David P. Toczyski,et al.  Securin and B-cyclin/CDK are the only essential targets of the APC , 2003, Nature Cell Biology.

[19]  Catherine G. Takizawa,et al.  Control of mitosis by changes in the subcellular location of cyclin-B1-Cdk1 and Cdc25C. , 2000, Current opinion in cell biology.

[20]  M. Grunstein,et al.  A search for proteins that interact genetically with histone H3 and H4 amino termini uncovers novel regulators of the Swe1 kinase in Saccharomyces cerevisiae. , 1996, Genes & development.

[21]  M. Longtine,et al.  Regulation of septin organization and function in yeast. , 2003, Trends in cell biology.

[22]  J. Pines,et al.  Four-dimensional control of the cell cycle , 1999, Nature Cell Biology.

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

[24]  J. Pringle,et al.  The septin cortex at the yeast mother-bud neck. , 2001, Current opinion in microbiology.

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

[26]  K Nasmyth,et al.  Destruction of the CDC28/CLB mitotic kinase is not required for the metaphase to anaphase transition in budding yeast. , 1993, The EMBO journal.

[27]  L. Hartwell.  Genetic control of the cell division cycle in yeast. IV. Genes controlling bud emergence and cytokinesis. , 1971, Experimental cell research.

[28]  Chandra L. Theesfeld,et al.  Septin-Dependent Assembly of a Cell Cycle-Regulatory Module in Saccharomyces cerevisiae , 2000, Molecular and Cellular Biology.

[29]  E. Nishida,et al.  Myt1 protein kinase is essential for Golgi and ER assembly during mitotic exit , 2008, The Journal of cell biology.

[30]  S. Reed,et al.  Morphogenesis in the yeast cell cycle: regulation by Cdc28 and cyclins , 1993, The Journal of cell biology.

[31]  W. Dunphy,et al.  The decision to enter mitosis. , 1994, Trends in cell biology.

[32]  P. Silver,et al.  The Saccharomyces cerevisiae cyclin Clb2p is targeted to multiple subcellular locations by cis- and trans-acting determinants. , 2001, Journal of cell science.

[33]  A. Lindqvist,et al.  Characterisation of Cdc25B localisation and nuclear export during the cell cycle and in response to stress , 2004, Journal of Cell Science.

[34]  N. Offner,et al.  Compartmentalization of the functions and regulation of the mitotic cyclin Clb2 in S. cerevisiae , 2007, Journal of Cell Science.

[35]  B. Futcher,et al.  Relationship between the function and the location of G1 cyclins in S. cerevisiae. , 2001, Journal of cell science.

[36]  M. Kirschner,et al.  Properties of Saccharomyces cerevisiae wee1 and its differential regulation of p34CDC28 in response to G1 and G2 cyclins. , 1993, The EMBO journal.

[37]  Jung-Eun Park,et al.  Concerted mechanism of Swe1/Wee1 regulation by multiple kinases in budding yeast , 2005, The EMBO journal.

[38]  Douglas R. Kellogg,et al.  Regulation of Mih1/Cdc25 by protein phosphatase 2A and casein kinase 1 , 2008, The Journal of cell biology.

[39]  Chandra L. Theesfeld,et al.  A monitor for bud emergence in the yeast morphogenesis checkpoint. , 2003, Molecular biology of the cell.

[40]  D. Lew,et al.  Phosphorylation-Independent Inhibition of Cdc28p by the Tyrosine Kinase Swe1p in the Morphogenesis Checkpoint , 1999, Molecular and Cellular Biology.

[41]  D. Lew,et al.  Control of Swe1p degradation by the morphogenesis checkpoint , 1998, The EMBO journal.

[42]  J. Ferrell,et al.  A positive-feedback-based bistable ‘memory module’ that governs a cell fate decision , 2003, Nature.

[43]  J. Thorner,et al.  Hsl7 Localizes to a Septin Ring and Serves as an Adapter in a Regulatory Pathway That Relieves Tyrosine Phosphorylation of Cdc28 Protein Kinase inSaccharomyces cerevisiae , 1999, Molecular and Cellular Biology.

[44]  P. O’Farrell,et al.  Triggering the all-or-nothing switch into mitosis. , 2001, Trends in cell biology.

[45]  O. Ozier-Kalogeropoulos,et al.  A simple and efficient method for direct gene deletion in Saccharomyces cerevisiae. , 1993, Nucleic acids research.

[46]  D. Lew,et al.  Cdc28 tyrosine phosphorylation and the morphogenesis checkpoint in budding yeast. , 1996, Molecular biology of the cell.

[47]  Jung-Eun Park,et al.  Coupling morphogenesis to mitotic entry , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[48]  U. Surana,et al.  Cdc20 is essential for the cyclosome-mediated proteolysis of both Pds1 and Clb2 during M phase in budding yeast , 1998, Current Biology.

[49]  Chandra L. Theesfeld,et al.  The Morphogenesis Checkpoint in Saccharomyces cerevisiae: Cell Cycle Control of Swe1p Degradation by Hsl1p and Hsl7p , 1999, Molecular and Cellular Biology.