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J. Biol. Chem., Vol. 280, Issue 21, 20860-20866, May 27, 2005

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Activation of the Rat Renin Promoter by HOXD10·PBX1b·PREP1, Ets-1, and the Intracellular Domain of Notch^*

Li Pan, Sean T. Glenn, Craig A. Jones, and Kenneth W. Gross

From the Department of Molecular and Cellular Biology, Roswell Park Cancer Institute, Buffalo, New York 14263-0001

Renin gene expression is subject to complex developmental and tissue-specific regulation. A comparison of the promoter sequences of the human, rat, and mouse renin genes has revealed a highly conserved sequence homologous to the DNA recognition sequence for CBF1 (CSL/RBP-J/Su(H)/LAG1/RBPSUH). Electrophoretic mobility shift assays document that As4.1 cell nuclear protein complex binding to the putative rat renin CBF1-binding site (-175 to -168 bp) contains CBF1. Transient transfection analyses in COS-7 cells further document that a CBF1-VP16 fusion protein and the intracellular domain of Notch1 robustly activate a promoter containing multiple copies of the rat renin CBF1-binding site. An Ets-binding site (-143 to -138 bp) has also been identified in the rat renin promoter by sequence comparisons and electrophoretic mobility shift assays. Transcription factor Ets-1 is capable of activating the rat renin promoter through the Ets-binding site. Mutation of the CBF-binding site significantly increases transcriptional activity of the rat renin promoter in Calu-6 and COS-7 cells but not in As4.1 cells, whereas mutation of the Ets-binding site reduces promoter activity of the rat renin gene in all three cell lines. Finally, we show that the intracellular domain of Notch1, Ets-1, and HOXD10·PBX1b·PREP1 activate the rat renin promoter cooperatively in COS-7 cells. These results strongly suggest that the renin gene is a downstream target of the Notch signaling pathway.

Received for publication, December 28, 2004 , and in revised form, March 22, 2005.

^* This work was supported by National Institute of Health grant HL48459 (to K. W. G.) and funds from the Bruce Cuvelier Family. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Dept. of Molecular and Cellular Biology, Roswell Park Cancer Institute, Elm and Carlton Sts., Buffalo, NY 14263-0001. Tel.: 716-845-4572; Fax: 716-845-8169; E-mail: gross{at}acsu.buffalo.edu.

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