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. 2005 Aug 19;94(5):431–436. doi: 10.1111/j.1349-7006.2003.tb01460.x

hCDC4b, a regulator of cyclin E, as a direct transcriptional target of p53

Takashi Kimura 1,2, Mitsukazu Gotoh 2, Yusuke Nakamura 1, Hirofumi Arakawa 1,4,✉
PMCID: PMC11160198  PMID: 12824889

Abstract

To identify p53‐target genes we have been using a cDNA‐microar‐ray system to assess gene expression in a p53‐mutated glioblas‐toma cell line (U373MG) after adenovirus‐mediated transfer of wild‐type p53 into the p53‐deficient cells. In the work reported here, expression of hCDC4b, which encodes one of the four sub‐units of the SCF (ubiquitin ligase) complex responsible for degradation of cyclin E, was dramatically up‐regulated by infection with Ad‐p53. An electrophoretic mobility‐shift assay and a chro‐matin immunoprecipitation assay indicated that a potential p53‐binding site (p53BS) present in exon 1b of the hCDC4 gene was able to bind to p53, and a reporter assay confirmed that this p53BS had p53‐dependent transcriptional activity. Expression of endogenous hCDC4b, but not the alternative transcript of this gene, hCDC4a, was induced in a p53‐dependent manner in response to genotoxic stresses caused by UV irradiation and adria‐mycin treatment, suggesting that each transcript has a different functional role. These results suggest that hCDC4b is a previously unrecognized transcriptional target of the p53 protein, and that by negatively regulating cyclin E through induction of hCDC4b, p53 might stop cell‐cycle progression at GO‐G1. This would represent a novel mechanism for p53‐dependent control of the cell cycle, in addition to the well‐known p21WAF1 machinery. (Cancer Sci 2003; 94: 431–436)

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