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 Nakayama

Research output: Contribution to journalArticle

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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 - Aug 1 2003

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GA-Binding Protein Transcription Factor
Cell Cycle
Electrophoretic Mobility Shift Assay
S Phase
Genetic Promoter Regions
SKP Cullin F-Box Protein Ligases
F-Box Proteins
G2 Phase
Ubiquitination
Nuclear Proteins
Site-Directed Mutagenesis
Luciferases
Small Interfering RNA
Organism Cloning
Messenger RNA

All Science Journal Classification (ASJC) codes

  • Cancer Research
  • Oncology

Cite this

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

Cell cycle-dependent regulation of the Skp2 promoter by GA-binding protein. / Imaki, Hiroyuki; Nakayama, Keiko; Delehouzee, Sophie; Handa, Hiroshi; Kitagawa, Masatoshi; Kamura, Takumi; Nakayama, Keiichi.

In: Cancer Research, Vol. 63, No. 15, 01.08.2003, p. 4607-4613.

Research output: Contribution to journalArticle

Imaki, H, Nakayama, K, Delehouzee, S, Handa, H, Kitagawa, M, Kamura, T & Nakayama, K 2003, 'Cell cycle-dependent regulation of the Skp2 promoter by GA-binding protein', Cancer Research, vol. 63, no. 15, pp. 4607-4613.
Imaki H, Nakayama K, Delehouzee S, Handa H, Kitagawa M, Kamura T et al. Cell cycle-dependent regulation of the Skp2 promoter by GA-binding protein. Cancer Research. 2003 Aug 1;63(15):4607-4613.
Imaki, Hiroyuki ; Nakayama, Keiko ; Delehouzee, Sophie ; Handa, Hiroshi ; Kitagawa, Masatoshi ; Kamura, Takumi ; Nakayama, Keiichi. / Cell cycle-dependent regulation of the Skp2 promoter by GA-binding protein. In: Cancer Research. 2003 ; Vol. 63, No. 15. pp. 4607-4613.
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