Redistribution of the CDK inhibitor p27 between different cyclin.CDK complexes in the mouse fibroblast cell cycle and in cells arrested with lovastatin or ultraviolet irradiation

Mol Biol Cell. 1995 Sep;6(9):1197-213. doi: 10.1091/mbc.6.9.1197.

Abstract

The cyclin-dependent kinase (CDK) inhibitor p27 binds and inhibits the kinase activity of several CDKs. Here we report an analysis of the behavior and partners of p27 in Swiss 3T3 mouse fibroblasts during normal mitotic cell cycle progression, as well as in cells arrested at different stages in the cycle by growth factor deprivation, lovastatin treatment, or ultraviolet (UV) irradiation. We found that the level of p27 is elevated in cells arrested in G0 by growth factor deprivation or contact inhibition. In G0, p27 was predominantly monomeric, although some portion was associated with residual cyclin A.Cdk2. During G1, all of p27 was associated with cyclin D1.Cdk4 and was then redistributed to cyclin A.Cdk2 as cells entered S phase. The loss of the monomeric p27 pool as cyclins accumulate in G1 is consistent with the in vivo and in vitro data showing that p27 binds better to cyclin.CDK complexes than to monomeric CDKs. In growing cells, the majority of p27 was associated with cyclin D1 and the level of p27 was significantly lower than the level of cyclin D1. In cells arrested in G1 with lovastatin, cyclin D1 was degraded and p27 was redistributed to cyclin A.Cdk2. In contrast to p21 (which is a p27-related CDK inhibitor and is induced by UV irradiation), the level of p27 was reduced after UV irradiation, but because cyclin D1 was degraded more rapidly than p27, there was a transient increase in binding of p27 to cyclin A.Cdk2. These data suggest that cyclin D1.Cdk4 acts as a reservoir for p27, and p27 is redistributed from cyclin D1.Cdk4 to cyclin A.Cdk2 complexes during S phase, or when cells are arrested by growth factor deprivation, lovastatin treatment, or UV irradiation. It is likely that a similar principle of redistribution of p27 is used by the cell in other instances of cell cycle arrest.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 3T3 Cells / cytology*
  • 3T3 Cells / drug effects
  • 3T3 Cells / metabolism
  • 3T3 Cells / radiation effects
  • Animals
  • Base Sequence
  • Cell Compartmentation
  • Cell Cycle / drug effects
  • Cell Cycle / radiation effects
  • Cell Cycle Proteins*
  • Cyclin D1
  • Cyclin-Dependent Kinase Inhibitor p27
  • Cyclin-Dependent Kinases / antagonists & inhibitors*
  • Cyclins / metabolism
  • Interphase / drug effects
  • Lovastatin / pharmacology*
  • Mice
  • Microtubule-Associated Proteins / metabolism*
  • Mitosis
  • Molecular Sequence Data
  • Nocodazole / pharmacology
  • Oncogene Proteins / metabolism
  • Tumor Suppressor Proteins*
  • Ultraviolet Rays*

Substances

  • Cdkn1b protein, mouse
  • Cell Cycle Proteins
  • Cyclins
  • Microtubule-Associated Proteins
  • Oncogene Proteins
  • Tumor Suppressor Proteins
  • Cyclin D1
  • Cyclin-Dependent Kinase Inhibitor p27
  • Lovastatin
  • Cyclin-Dependent Kinases
  • Nocodazole