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PMID: 1396589 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Cell cycle regulation of CDK2 activity by phosphorylation of Thr160 and Tyr15.

The EMBO journal ·Vol. 11 ·No. 11 ·1992-11-00 ·Pages 3995-4005

Gu Y, Rosenblatt J, Morgan DO

Abstract

We have examined the role of phosphorylation in the regulation of human cyclin-dependent kinase-2 (CDK2), a protein closely related to the cell cycle regulatory kinase CDC2. We find that CDK2 from HeLa cells contains three major tryptic phosphopeptides. Analysis of site-directed mutant proteins, expressed by transient transfection of COS cells, demonstrates that the two major phosphorylation sites are Tyr15 (Y15) and Thr160 (T160). Additional phosphorylation probably occurs on Thr14 (T14). Replacement of T160 with alanine abolishes the kinase activity of CDK2, indicating that phosphorylation at this site (as in CDC2) is required for kinase activity. Mutation of Y15 and T14 stimulates kinase activity, demonstrating that phosphorylation at these sites (as in CDC2) is inhibitory. Similarly, CDK2 is activated in vitro by dephosphorylation of Y15 and T14 by the phosphatase CDC25. Analysis of HeLa cells synchronized at various cell cycle stages indicates that CDK2 phosphorylation on T160 increases during S phase and G2, when CDK2 is most active. Phosphorylation on the inhibitory sites T14 and Y15 is also maximal during S phase and G2. Thus, the activity of a subpopulation of CDK2 molecules is inhibited at a time in the cell cycle when overall CDK2 activity is increased.

Related Genes
CDK2
MeSH Terms
Amino Acid Sequence CDC2 Protein Kinase/metabolism CDC2-CDC28 Kinases Cell Cycle/physiology Cell Cycle Proteins Cyclin-Dependent Kinase 2 Cyclin-Dependent Kinases Cyclins/metabolism Fungal Proteins/metabolism HeLa Cells Homeostasis Humans Kinetics Molecular Sequence Data Mutagenesis, Site-Directed Peptide Mapping Phosphates/metabolism Phosphopeptides/isolation & purification Phosphorus Radioisotopes Phosphorylation Protein Kinases/genetics,metabolism Protein Serine-Threonine Kinases Sequence Homology, Amino Acid Threonine Tyrosine ras-GRF1
Chemicals
Cell Cycle Proteins Cyclins Fungal Proteins Phosphates Phosphopeptides Phosphorus Radioisotopes ras-GRF1 Threonine Tyrosine Protein Kinases Protein Serine-Threonine Kinases CDC2 Protein Kinase CDC2-CDC28 Kinases CDK2 protein, human Cyclin-Dependent Kinase 2 Cyclin-Dependent Kinases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Gu Y
Department of Physiology, University of California, San Francisco 94143-0444.
Rosenblatt J
Morgan D O
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1992-11-00
Pages
3995-4005
Language
English
Region
England
NLM ID
8208664
PMCID
PMC556910
Subset
IM
Grants
NCI NIH HHS · R01 CA52481 · United States
External Links
PubMed source PMC Full Text View journal details
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