Cyclin E-CDK2 is a regulator of p27Kip1.
CDK inhibitors are thought to prevent cell proliferation by negatively regulating cyclin-CDK complexes. We propose that the opposite is also true, that cyclin-CDK complexes in mammmalian cells can promote cell cycle progression by directly down-regulating CDK inhibitors. We show that expression of cyclin E-CDK2 in murine fibroblasts causes phosphorylation of ... the CDK inhibitor p27Kip1 on T187, and that cyclin E-CDK2 can directly phosphorylate p27 T187 in vitro. We further show that cyclin E-CDK2-dependent phosphorylation of p27 results in elimination of p27 from the cell, allowing cells to transit from G1 to S phase. Moreover, mutation of T187 in p27 to alanine creates a p27 protein that causes a G1 block resistant to cyclin E and whose level of expression is not modulated by cyclin E. A kinetic analysis of the interaction between p27 and cyclin E-CDK2 explains how p27 can be regulated by the same enzyme it targets for inhibition. We show that p27 interacts with cyclin E-CDK2 in at least two distinct ways: one resulting in p27 phosphorylation and release, the other in tight binding and cyclin E-CDK2 inhibition. The binding of ATP to the CDK governs which state predominates. At low ATP (< 50 microM) p27 is primarily a CDK inhibitor, but at ATP concentrations approaching physiological levels (> 1 mM) p27 is more likely to be a substrate. Thus, we have identified p27 as a biologically relevant cyclin E-CDK2 substrate, demonstrated the physiological consequences of p27 phosphorylation, and developed a kinetic model to explain how p27 can be both an inhibitor and a substrate of cyclin E-CDK2.
Mesh Terms:
3T3 Cells, Adenosine Triphosphate, Animals, CDC2-CDC28 Kinases, Cell Cycle, Cell Cycle Proteins, Cyclin-Dependent Kinase 2, Cyclin-Dependent Kinase Inhibitor p27, Cyclin-Dependent Kinases, Cyclins, Down-Regulation, G1 Phase, Histones, Mice, Microtubule-Associated Proteins, Models, Biological, Mutation, Phosphoproteins, Phosphorylation, Protein-Serine-Threonine Kinases, S Phase, Threonine, Tumor Suppressor Proteins
3T3 Cells, Adenosine Triphosphate, Animals, CDC2-CDC28 Kinases, Cell Cycle, Cell Cycle Proteins, Cyclin-Dependent Kinase 2, Cyclin-Dependent Kinase Inhibitor p27, Cyclin-Dependent Kinases, Cyclins, Down-Regulation, G1 Phase, Histones, Mice, Microtubule-Associated Proteins, Models, Biological, Mutation, Phosphoproteins, Phosphorylation, Protein-Serine-Threonine Kinases, S Phase, Threonine, Tumor Suppressor Proteins
Genes Dev.
Date: Jun. 01, 1997
PubMed ID: 9192873
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