HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 278, Issue 42, 40815-40828, October 17, 2003

This Article Full Text Full Text (PDF) All Versions of this Article: 278/42/40815    most recent M306683200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Chow, J. P. H. Articles by Poon, R. Y. C. Search for Related Content PubMed PubMed Citation Articles by Chow, J. P. H. Articles by Poon, R. Y. C. Social Bookmarking                      What's this?

Differential Contribution of Inhibitory Phosphorylation of CDC2 and CDK2 for Unperturbed Cell Cycle Control and DNA Integrity Checkpoints^*

Jeremy P. H. Chow, Wai Yi Siu, Horace T. B. Ho , Ken Hoi Tang Ma , Chui Chui Ho and Randy Y. C. Poon ¶

From the Department of Biochemistry, Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong

Inhibition of cyclin-dependent kinases (CDKs) by Thr¹⁴/Tyr¹⁵ phosphorylation is critical for normal cell cycle progression and is a converging event for several cell cycle checkpoints. In this study, we compared the relative contribution of inhibitory phosphorylation for cyclin A/B1-CDC2 and cyclin A/E-CDK2 complexes. We found that inhibitory phosphorylation plays a major role in the regulation of CDC2 but only a minor role for CDK2 during the unperturbed cell cycle of HeLa cells. The relative importance of inhibitory phosphorylation of CDC2 and CDK2 may reflect their distinct cellular functions. Despite this, expression of nonphosphorylation mutants of both CDC2 and CDK2 triggered unscheduled histone H3 phosphorylation early in the cell cycle and was cytotoxic. DNA damage by a radiomimetic drug or replication block by hydroxyurea stimulated a buildup of cyclin B1 but was accompanied by an increase of inhibitory phosphorylation of CDC2. After DNA damage and replication block, all cyclin-CDK pairs that control S phase and mitosis were to different degrees inhibited by phosphorylation. Ectopic expression of nonphosphorylated CDC2 stimulated DNA replication, histone H3 phosphorylation, and cell division even after DNA damage. Similarly, a nonphosphorylation mutant of CDK2, but not CDK4, disrupted the G₂ DNA damage checkpoint. Finally, CDC25A, CDC25B, a dominantnegative CHK1, but not CDC25C or a dominant-negative WEE1, stimulated histone H3 phosphorylation after DNA damage. These data suggest differential contributions for the various regulators of Thr¹⁴/Tyr¹⁵ phosphorylation in normal cell cycle and during the DNA damage checkpoint.

Received for publication, June 24, 2003

^* This work was supported in part by the Croucher Foundation, the Philip Morris External Research Program, and Research Grants Council Grant HKUST6129/02M (to R. Y. C. P.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Croucher Foundation Scholar.

Recipient of a grant from the Hongkong Bank Foundation Local Bursary and Scholarship Program.

^¶ Croucher Foundation Senior Fellow. To whom correspondence should be addressed: Dept. of Biochemistry, Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong. Tel.: 852-23588718; Fax: 852-23581552; E-mail: bcrandy{at}ust.hk.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				Y. W. Chan, K. F. On, W. M. Chan, W. Wong, H. O. Siu, P. M. Hau, and R. Y. C. Poon The Kinetics of p53 Activation Versus Cyclin E Accumulation Underlies the Relationship between the Spindle-assembly Checkpoint and the Postmitotic Checkpoint J. Biol. Chem., June 6, 2008; 283(23): 15716 - 15723. [Abstract] [Full Text] [PDF]

				F. Hu, Y. Gan, and O. M. Aparicio Identification of Clb2 Residues Required for Swe1 Regulation of Clb2-Cdc28 in Saccharomyces cerevisiae Genetics, June 1, 2008; 179(2): 863 - 874. [Abstract] [Full Text] [PDF]

				J. Zamborszky, C. I. Hong, and A. Csikasz Nagy Computational Analysis of Mammalian Cell Division Gated by a Circadian Clock: Quantized Cell Cycles and Cell Size Control J Biol Rhythms, December 1, 2007; 22(6): 542 - 553. [Abstract] [PDF]

				H. Hochegger, D. Dejsuphong, E. Sonoda, A. Saberi, E. Rajendra, J. Kirk, T. Hunt, and S. Takeda An essential role for Cdk1 in S phase control is revealed via chemical genetics in vertebrate cells J. Cell Biol., July 10, 2007; 178(2): 257 - 268. [Abstract] [Full Text] [PDF]

				T. K. Fung, H. T. Ma, and R. Y.C. Poon Specialized Roles of the Two Mitotic Cyclins in Somatic Cells: Cyclin A as an Activator of M Phase-promoting Factor Mol. Biol. Cell, May 1, 2007; 18(5): 1861 - 1873. [Abstract] [Full Text] [PDF]

				H. T. Ma, K. F. On, Y. H. Tsang, and R. Y.C. Poon An inducible system for expression and validation of the specificity of short hairpin RNA in mammalian cells Nucleic Acids Res., February 28, 2007; 35(4): e22 - e22. [Abstract] [Full Text] [PDF]

				J. R. Biggs, L. F. Peterson, Y. Zhang, A. S. Kraft, and D.-E. Zhang AML1/RUNX1 Phosphorylation by Cyclin-Dependent Kinases Regulates the Degradation of AML1/RUNX1 by the Anaphase-Promoting Complex Mol. Cell. Biol., October 15, 2006; 26(20): 7420 - 7429. [Abstract] [Full Text] [PDF]

				A. Csikasz-Nagy, D. Battogtokh, K. C. Chen, B. Novak, and J. J. Tyson Analysis of a Generic Model of Eukaryotic Cell-Cycle Regulation Biophys. J., June 15, 2006; 90(12): 4361 - 4379. [Abstract] [Full Text] [PDF]

				Y. P. Ng, Z. H. Cheung, and N. Y. Ip STAT3 as a Downstream Mediator of Trk Signaling and Functions J. Biol. Chem., June 9, 2006; 281(23): 15636 - 15644. [Abstract] [Full Text] [PDF]

				V. Mayya, K. Rezual, L. Wu, M. B. Fong, and D. K. Han Absolute Quantification of Multisite Phosphorylation by Selective Reaction Monitoring Mass Spectrometry: Determination of Inhibitory Phosphorylation Status of Cyclin-Dependent Kinases Mol. Cell. Proteomics, June 1, 2006; 5(6): 1146 - 1157. [Abstract] [Full Text] [PDF]

				C. C. Ho, W. Y. Siu, A. Lau, W. M. Chan, T. Arooz, and R. Y.C. Poon Stalled Replication Induces p53 Accumulation through Distinct Mechanisms from DNA Damage Checkpoint Pathways Cancer Res., February 15, 2006; 66(4): 2233 - 2241. [Abstract] [Full Text] [PDF]

				A. Lindqvist, H. Kallstrom, A. Lundgren, E. Barsoum, and C. K. Rosenthal Cdc25B cooperates with Cdc25A to induce mitosis but has a unique role in activating cyclin B1-Cdk1 at the centrosome J. Cell Biol., October 10, 2005; 171(1): 35 - 45. [Abstract] [Full Text] [PDF]

				K. M. Regula, M. J. Rzeszutek, D. Baetz, C. Seneviratne, and L. A. Kirshenbaum Therapeutic opportunities for cell cycle re-entry and cardiac regeneration Cardiovasc Res, December 1, 2004; 64(3): 395 - 401. [Abstract] [Full Text] [PDF]

				I. Bartova, M. Otyepka, Z. Kriz, and J. Koca Activation and inhibition of cyclin-dependent kinase-2 by phosphorylation; a molecular dynamics study reveals the functional importance of the glycine-rich loop Protein Sci., June 1, 2004; 13(6): 1449 - 1457. [Abstract] [Full Text] [PDF]

				W. Y. Siu, A. Lau, T. Arooz, J. P.H. Chow, H. T.B. Ho, and R. Y.C. Poon Topoisomerase poisons differentially activate DNA damage checkpoints through ataxia-telangiectasia mutated-dependent and -independent mechanisms Mol. Cancer Ther., May 1, 2004; 3(5): 621 - 632. [Abstract] [Full Text] [PDF]

				C.-P. Ng, H. C. Lee, C. W. Ho, T. Arooz, W. Y. Siu, A. Lau, and R. Y. C. Poon Differential Mode of Regulation of the Checkpoint Kinases CHK1 and CHK2 by Their Regulatory Domains J. Biol. Chem., March 5, 2004; 279(10): 8808 - 8819. [Abstract] [Full Text] [PDF]