Cell cycle-dependent regulation of the Skp2 promoter by GA-binding protein

Hiroyuki Imaki, Keiko Nakayama, Sophie Delehouzee, Hiroshi Handa, Masatoshi Kitagawa, Takumi Kamura, Keiichi I. Nakayama

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71 Citations (Scopus)

Abstract

Skp2 is the F-box protein component of an SCF-type ubiquitin ligase that interacts specifically with p27Kip1 and thereby promotes its ubiquitylation and degradation. The abundance of Skp2 mRNA oscillates in a cell cycle-dependent manner, being maximal in S and G2 phases. The regulation of Skp2 transcription was investigated by cloning the promoter region of the mouse gene and determination of its activity in a luciferase reporter assay. Deletion analysis identified a minimal ∼0.3-kb promoter region with marked transcriptional activity and a 105-bp essential sequence within this region. Electrophoretic mobility shift assays indicated the presence in nuclear extracts of proteins that bind to this sequence. Site-directed mutagenesis revealed that the core binding motif, CACTTCCG, which is similar to that of GA-binding protein (GABP), is essential for Skp2 transcription. "Supershift" analysis indicated that the protein-probe complexes detected by electrophoretic mobility shift assays contain GABP. Endogenous GABP bound to Skp2 promoter element in a cell cycle-dependent manner. Furthermore, overexpression of GABPβ increased Skp2 promoter activity, and suppression of GABPα or GABPβ by a small interfering RNA resulted in the reduction of Skp2 promoter activity. These data suggest that the cell cycle-dependent binding of GABP to the Skp2 promoter plays an important role in the regulation of Skp2 expression and cell cycle progression from G1 to S phase.

Original languageEnglish
Pages (from-to)4607-4613
Number of pages7
JournalCancer Research
Volume63
Issue number15
Publication statusPublished - 2003 Aug 1
Externally publishedYes

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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  • Cite this

    Imaki, H., Nakayama, K., Delehouzee, S., Handa, H., Kitagawa, M., Kamura, T., & Nakayama, K. I. (2003). Cell cycle-dependent regulation of the Skp2 promoter by GA-binding protein. Cancer Research, 63(15), 4607-4613.