Phosphorylation of p21 in G2/M Promotes Cyclin B-Cdc2 Kinase Activity

ABSTRACT Little is known about the posttranslational control of the cyclin-dependent protein kinase (CDK) inhibitor p21. We describe here a transient phosphorylation of p21 in the G2/M phase. G2/M-phosphorylated p21 is short-lived relative to hypophosphorylated p21. p21 becomes nuclear during S phase, prior to its phosphorylation by CDK2. S126-phosphorylated cyclin B1 binds to T57-phosphorylated p21. Cdc2 kinase activation is delayed in p21-deficient cells due to delayed association between Cdc2 and cyclin B1. Cyclin B1-Cdc2 kinase activity and G2/M progression in p21−/− cells are restored after reexpression of wild-type but not T57A mutant p21. The cyclin B1 S126A mutant exhibits reduced Cdc2 binding and has low kinase activity. Phosphorylated p21 binds to cyclin B1 when Cdc2 is phosphorylated on Y15 and associates poorly with the complex. Dephosphorylation on Y15 and phosphorylation on T161 promotes Cdc2 binding to the p21-cyclin B1 complex, which becomes activated as a kinase. Thus, hyperphosphorylated p21 activates the Cdc2 kinase in the G2/M transition.

[1]  J. Harper,et al.  The Cyclin E/Cdk2 Substrate p220NPAT Is Required for S-Phase Entry, Histone Gene Expression, and Cajal Body Maintenance in Human Somatic Cells , 2003, Molecular and Cellular Biology.

[2]  T. Burns,et al.  Silencing of the Novel p53 Target Gene Snk/Plk2 Leads to Mitotic Catastrophe in Paclitaxel (Taxol)-Exposed Cells , 2003, Molecular and Cellular Biology.

[3]  J. Harper,et al.  The Cyclin E/Cdk2 Substrate and Cajal Body Component p220NPAT Activates Histone Transcription through a Novel LisH-Like Domain , 2003, Molecular and Cellular Biology.

[4]  Robert L Sutherland,et al.  Regulation of the ubiquitin‐conjugating enzyme hHR6A by CDK‐mediated phosphorylation , 2002, The EMBO journal.

[5]  A. Zeiher,et al.  Glycogen Synthase Kinase-3 Couples AKT-dependent Signaling to the Regulation of p21Cip1 Degradation* , 2002, The Journal of Biological Chemistry.

[6]  W. El-Deiry,et al.  Enhanced Sensitivity of G1 Arrested Human Cancer Cells Suggests a Novel Therapeutic Strategy Using a Combination of Simvastatin and TRAIL , 2002, Cell cycle.

[7]  M. Winey,et al.  The Mouse Mps1p-like Kinase Regulates Centrosome Duplication , 2001, Cell.

[8]  S. van den Heuvel,et al.  S and G2 Phase Roles for Cdk2 Revealed by Inducible Expression of a Dominant-Negative Mutant in Human Cells , 2001, Molecular and Cellular Biology.

[9]  P. Jackson,et al.  Cyclin E Uses Cdc6 as a Chromatin-Associated Receptor Required for DNA Replication , 2001, The Journal of cell biology.

[10]  E. Nishida,et al.  Polo-like kinase 1 phosphorylates cyclin B1 and targets it to the nucleus during prophase , 2001, Nature.

[11]  Peter D. Adams,et al.  HIRA, the Human Homologue of Yeast Hir1p and Hir2p, Is a Novel Cyclin-cdk2 Substrate Whose Expression Blocks S-Phase Progression , 2001, Molecular and Cellular Biology.

[12]  M. Hung,et al.  Cytoplasmic localization of p21Cip1/WAF1 by Akt-induced phosphorylation in HER-2/neu-overexpressing cells , 2001, Nature Cell Biology.

[13]  W. El-Deiry.  Akt takes centre stage in cell-cycle deregulation , 2001, Nature Cell Biology.

[14]  Erich A. Nigg,et al.  Cell division: Mitotic kinases as regulators of cell division and its checkpoints , 2001, Nature Reviews Molecular Cell Biology.

[15]  R. Bernards,et al.  Distinct Initiation and Maintenance Mechanisms Cooperate to Induce G1 Cell Cycle Arrest in Response to DNA Damage , 2000, Cell.

[16]  K. Kinzler,et al.  Cooperative effects of genes controlling the G(2)/M checkpoint. , 2000, Genes & development.

[17]  K. Ball,et al.  Reversible Phosphorylation at the C-terminal Regulatory Domain of p21Waf1/Cip1 Modulates Proliferating Cell Nuclear Antigen Binding* , 2000, The Journal of Biological Chemistry.

[18]  K. Khanna,et al.  Caffeine Abolishes the Mammalian G2/M DNA Damage Checkpoint by Inhibiting Ataxia-Telangiectasia-mutated Kinase Activity* , 2000, The Journal of Biological Chemistry.

[19]  T. Pandita,et al.  Ionizing radiation activates the ATM kinase throughout the cell cycle , 2000, Oncogene.

[20]  C. Smythe,et al.  ATR is a caffeine-sensitive, DNA-activated protein kinase with a substrate specificity distinct from DNA-PK , 1999, Oncogene.

[21]  K. Kinzler,et al.  14-3-3σ is required to prevent mitotic catastrophe after DNA damage , 1999, Nature.

[22]  S. Reed,et al.  Deregulated cyclin E induces chromosome instability , 1999, Nature.

[23]  J. Roberts,et al.  Evolving Ideas about Cyclins , 1999, Cell.

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

[25]  M. Nakanishi,et al.  Cell cycle-dependent and ATM-independent expression of human Chk1 kinase , 1999, Oncogene.

[26]  James M. Roberts,et al.  CDK inhibitors: positive and negative regulators of G1-phase progression. , 1999, Genes & development.

[27]  David O. Morgan,et al.  Regulation of the APC and the exit from mitosis , 1999, Nature Cell Biology.

[28]  James M. Roberts,et al.  The p21Cip1 and p27Kip1 CDK ‘inhibitors’ are essential activators of cyclin D‐dependent kinases in murine fibroblasts , 1999, The EMBO journal.

[29]  K. Miyazono,et al.  Apoptosis inhibitory activity of cytoplasmic p21Cip1/WAF1 in monocytic differentiation , 1999, The EMBO journal.

[30]  S. Leach,et al.  p21Waf1/Cip1 Inhibition of Cyclin E/Cdk2 Activity Prevents Endoreduplication after Mitotic Spindle Disruption , 1999, Molecular and Cellular Biology.

[31]  L. Hengst,et al.  Complete inhibition of Cdk/cyclin by one molecule of p21(Cip1). , 1998, Genes & development.

[32]  K. Kinzler,et al.  Requirement for p53 and p21 to sustain G2 arrest after DNA damage. , 1998, Science.

[33]  B. Ducommun,et al.  Interaction with cyclin-dependent kinases and PCNA modulates proteasome-dependent degradation of p21 , 1998, Oncogene.

[34]  J. Sarkaria,et al.  Inhibition of phosphoinositide 3-kinase related kinases by the radiosensitizing agent wortmannin. , 1998, Cancer research.

[35]  B. Schulman,et al.  Substrate recruitment to cyclin-dependent kinase 2 by a multipurpose docking site on cyclin A. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[36]  B. Stillman,et al.  Cyclin-Dependent Kinase Inhibitor p21 Modulates the DNA Primer-Template Recognition Complex , 1998, Molecular and Cellular Biology.

[37]  E Warbrick,et al.  PCNA binding through a conserved motif. , 1998, BioEssays : news and reviews in molecular, cellular and developmental biology.

[38]  J. Erhardt,et al.  p21WAF1 induces permanent growth arrest and enhances differentiation, but does not alter apoptosis in PC12 cells , 1998, Oncogene.

[39]  S. Reed,et al.  Nuclear Accumulation of p21Cip1 at the Onset of Mitosis: a Role at the G2/M-Phase Transition , 1998, Molecular and Cellular Biology.

[40]  W. El-Deiry.  p21/p53, cellular growth control and genomic integrity. , 1998, Current topics in microbiology and immunology.

[41]  O. Bachs,et al.  The cell cycle inhibitor p21CIP is phosphorylated by cyclin A-CDK2 complexes. , 1997, Biochemical and biophysical research communications.

[42]  M. Shimizu,et al.  G1 phase accumulation induced by UCN-01 is associated with dephosphorylation of Rb and CDK2 proteins as well as induction of CDK inhibitor p21/Cip1/WAF1/Sdi1 in p53-mutated human epidermoid carcinoma A431 cells. , 1997, Cancer research.

[43]  J. LaBaer,et al.  New functional activities for the p21 family of CDK inhibitors. , 1997, Genes & development.

[44]  D O Morgan,et al.  Cyclin-dependent kinases: engines, clocks, and microprocessors. , 1997, Annual review of cell and developmental biology.

[45]  Marc W. Kirschner,et al.  How Proteolysis Drives the Cell Cycle , 1996, Science.

[46]  W. Sellers,et al.  Identification of a cyclin-cdk2 recognition motif present in substrates and p21-like cyclin-dependent kinase inhibitors , 1996, Molecular and cellular biology.

[47]  John Kuriyan,et al.  Structure of the C-Terminal Region of p21WAF1/CIP1 Complexed with Human PCNA , 1996, Cell.

[48]  P E Wright,et al.  Structural studies of p21Waf1/Cip1/Sdi1 in the free and Cdk2-bound state: conformational disorder mediates binding diversity. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[49]  Bert Vogelstein,et al.  Uncoupling of S phase and mitosis induced by anticancer agents in cells lacking p21 , 1996, Nature.

[50]  M. Dorée,et al.  p21 contains independent binding sites for cyclin and cdk2: both sites are required to inhibit cdk2 kinase activity. , 1996, Oncogene.

[51]  Anindya Dutta,et al.  A 39 amino acid fragment of the cell cycle regulator p21 is sufficient to bind PCNA and partially inhibit DNA replication in vivo. , 1996, Nucleic acids research.

[52]  J. Maller,et al.  In vivo regulation of the early embryonic cell cycle in Xenopus. , 1996, Developmental biology.

[53]  G. Hannon,et al.  Subcellular distribution of p21 and PCNA in normal and repair-deficient cells following DNA damage , 1996, Current Biology.

[54]  T. Guadagno,et al.  Cdk2 Kinase Is Required for Entry into Mitosis as a Positive Regulator of Cdc2–Cyclin B Kinase Activity , 1996, Cell.

[55]  James Brugarolas,et al.  Radiation-induced cell cycle arrest compromised by p21 deficiency , 1995, Nature.

[56]  Stephen J. Elledge,et al.  Mice Lacking p21 CIP1/WAF1 undergo normal development, but are defective in G1 checkpoint control , 1995, Cell.

[57]  J. Massagué,et al.  Cell-cycle inhibition by independent CDK and PCNA binding domains in p21Cip1 , 1995, Nature.

[58]  K. Kinzler,et al.  p53-dependent and independent expression of p21 during cell growth, differentiation, and DNA damage. , 1995, Genes & development.

[59]  S. Elledge,et al.  Inhibition of cyclin-dependent kinases by p21. , 1995, Molecular biology of the cell.

[60]  M. Kirschner,et al.  Separate domains of p21 involved in the inhibition of Cdk kinase and PCNA , 1995, Nature.

[61]  David O. Morgan,et al.  Principles of CDK regulation , 1995, Nature.

[62]  D. Lane,et al.  A small peptide inhibitor of DNA replication defines the site of interaction between the cyclin-dependent kinase inhibitor p21WAF1 and proliferating cell nuclear antigen , 1995, Current Biology.

[63]  S. Elledge,et al.  p53-independent expression of p21Cip1 in muscle and other terminally differentiating cells , 1995, Science.

[64]  J. Maller,et al.  Phosphorylation and activation of the Xenopus Cdc25 phosphatase in the absence of Cdc2 and Cdk2 kinase activity. , 1995, Molecular biology of the cell.

[65]  C. Sherr.  G1 phase progression: Cycling on cue , 1994, Cell.

[66]  G. Hannon,et al.  Differential effects by the p21 CDK inhibitor on PCNA-dependent DNA replication and repair , 1994, Nature.

[67]  E. Karsenti,et al.  Activation of the phosphatase activity of human cdc25A by a cdk2‐cyclin E dependent phosphorylation at the G1/S transition. , 1994, The EMBO journal.

[68]  S. Elledge,et al.  Cdk-interacting protein 1 directly binds with proliferating cell nuclear antigen and inhibits DNA replication catalyzed by the DNA polymerase delta holoenzyme. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[69]  Y. Xiong,et al.  Cell cycle expression and p53 regulation of the cyclin-dependent kinase inhibitor p21. , 1994, Oncogene.

[70]  G. Hannon,et al.  p21-containing cyclin kinases exist in both active and inactive states. , 1994, Genes & development.

[71]  G. Hannon,et al.  The p21 inhibitor of cyclin-dependent kinases controls DNA replication by interaction with PCNA , 1994, Nature.

[72]  G. Hannon,et al.  p21 is a component of active cell cycle kinases. , 1994, Cold Spring Harbor symposia on quantitative biology.

[73]  David Beach,et al.  p21 is a universal inhibitor of cyclin kinases , 1993, Nature.

[74]  J. Trent,et al.  WAF1, a potential mediator of p53 tumor suppression , 1993, Cell.

[75]  S. Elledge,et al.  The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases , 1993, Cell.

[76]  D. Beach,et al.  Proliferating cell nuclear antigen and p21 are components of multiple cell cycle kinase complexes. , 1993, Molecular biology of the cell.

[77]  Charles J. Sherr,et al.  Mammalian G1 cyclins , 1993, Cell.

[78]  K. Kohn,et al.  G2 delay induced by nitrogen mustard in human cells affects cyclin A/cdk2 and cyclin B1/cdc2-kinase complexes differently. , 1993, The Journal of biological chemistry.

[79]  Hui Zhang,et al.  D type cyclins associate with multiple protein kinases and the DNA replication and repair factor PCNA , 1992, Cell.

[80]  D. Morgan,et al.  Human cyclin-dependent kinase 2 is activated during the S and G2 phases of the cell cycle and associates with cyclin A. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[81]  A. Murray,et al.  Cyclin is degraded by the ubiquitin pathway , 1991, Nature.

[82]  P. Roach,et al.  Formation of protein kinase recognition sites by covalent modification of the substrate. Molecular mechanism for the synergistic action of casein kinase II and glycogen synthase kinase 3. , 1987, The Journal of biological chemistry.