p27Kip1 induces an accumulation of the repressor complexes of E2F and inhibits expression of the E2F-regulated genes

Pavel Shiyanov, Steven Hayes, Ningyu Chen, Dimitri G. Pestov, Lester F. Lau, Pradip Raychaudhuri

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

p27Kip1 is an inhibitor of the cyclin-dependent kinases and it plays an inhibitory role in the progression of cell cycle through G1 phase. To investigate the mechanism of cell cycle inhibition by p27Kip1, we constructed a cell line that inducibly expresses p27Kip1 upon addition of isopropyl-1- thio-β-D-galactopyranoside in the culture medium. Isopropyl-1-thio-β-D- galactopyranoside-induced expression of p27Kip1 in these cells causes a specific reduction in the expression of the E2F-regulated genes such as cyclin E, cyclin A, and dihydrofolate reductase. The reduction in the expression of these genes correlates with the p27Kip1-induced accumulation of the repressor complexes of the E2F family of factors (E2Fs). Our previous studies indicated that p21WAF1 could disrupt the interaction between cyclin/cylcin-dependent kinase 2 (cdk2) and the E2F repressor complexes E2F- p130 and E2F-p107. We show that p27Kip1, like p21WAF1, disrupts cyclin/cdk2- containing complexes of E2F-p130 leading to the accumulation of the E2F-p130 complexes, which is found in growth-arrested cells. In transient transfection assays, expression of p27Kip1 specifically inhibits transcription of a promoter containing E2F-binding sites. Mutants of p27Kip1 harboring changes in the cyclin- and cdk2-binding motifs are deficient in inhibiting transcription from the E2F sites containing reporter gene. Moreover, these mutants of p27Kip1 are also impaired in disrupting the interaction between cyclin/cdk2 and the repressor complexes of E2Fs. Taken together, these observations suggest that p27Kip1 reduces expression of the E2F-regulated genes by generating repressor complexes of E2fs. Furthermore, the results also demonstrate that p27Kip1 inhibits expression of cyclin A and cylcin E, which are critical for progression through the G1-S phases.

Original languageEnglish (US)
Pages (from-to)1815-1827
Number of pages13
JournalMolecular biology of the cell
Volume8
Issue number9
DOIs
StatePublished - 1997

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

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