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Volume 272, Number 27, Issue of July 4, 1997 pp. 16845-16851
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

A Novel Proximal Element Mediates the Regulation of Mouse Ren-1C Promoter by Retinoblastoma Protein in Cultured Cells

(Received for publication, January 28, 1997, and in revised form, April 15, 1997)

Kouichi Tamura , Satoshi Umemura , Nobuo Nyui , Satoshi Yamaguchi , Tomoaki Ishigami , Kiyoshi Hibi , Machiko Yabana , Minoru Kihara , Akiyoshi Fukamizu ^¶ , Kazuo Murakami ^¶ and Masao Ishii

From  Second Department of Internal Medicine, Yokohama City University School of Medicine, Yokohama 236, Japan and the ^¶ Institute of Applied Biochemistry, University of Tsukuba, Ibaraki 305, Japan

The protein product of the retinoblastoma susceptibility gene, RB, is a nuclear phosphoprotein that modulates transcription of genes involved in growth control via interactions with transcription factors. Renin is a rate-limiting enzyme of the renin-angiotensin system that regulates blood pressure and water-electrolyte balance. Renin gene expression is regulated in a tissue-specific and developmentally linked manner. Similarly, the expression of RB is controlled in a differentiation-linked manner. Thus, to investigate whether RB is involved in the regulation of renin gene expression, we examined the effects of RB on transcriptional activity of the mouse renin (Ren-1C) promoter. The Ren-1C promoter contains two transcriptionally important elements; the RU-1 (224 to 138) and RP-2 (75 to 47) elements. RB activated the Ren-1C promoter in human embryonic kidney cells. The promoter element responsible for RB-mediated transcriptional regulation was the RP-2 element. The results of DNA-protein binding experiments showed that RB increased nuclear binding activity to the RP-2 element, and site-directed mutation which disrupted binding of nuclear factors to the RP-2 element markedly reduced RB-mediated activation of Ren-1C promoter in human embryonic kidney cells. These results indicate that the RP-2 element plays an important role in RB-mediated transcriptional regulation of Ren-1C promoter activity in human embryonic kidney cells, thereby suggesting an interesting mechanism by which RB may modulate the renin-angiotensin system.

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