JBC Transcription and Nuclear Factor Monoclonals

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Originally published In Press as doi:10.1074/jbc.M008361200 on October 31, 2000

J. Biol. Chem., Vol. 276, Issue 5, 3597-3603, February 2, 2001
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Retinoic Acid-mediated Activation of the Mouse Renin Enhancer*

Qi ShiDagger , Kenneth W. Gross§, and Curt D. SigmundDagger

From the Dagger  Departments of Internal Medicine and Physiology & Biophysics, The University of Iowa College of Medicine, Iowa City, Iowa 52242 and the § Department of Molecular and Cellular Biology, Roswell Park Cancer Institute, Buffalo, New York 14263

Previous studies demonstrate that the mouse renin gene is regulated by a complex enhancer of transcription located 2.6 kilobases upstream of the transcription start site which is under both positive and negative influence. We demonstrate herein that a positive regulatory element (Eb) is repeated 10 bp upstream (Ec), and both are required for baseline activity of the enhancer. The Eb and Ec core sequences are identical to the consensus sequence for the nuclear hormone receptor superfamily of transcription factors, and transcriptional activity of constructs containing the enhancer is increased after treatment with retinoic acid. Maximal induction requires both Eb and Ec. Expression of endogenous renin and a renin-promoter controlled transgene in As4.1 cells, and kidney renin mRNA in C57BL/6J mice was induced after retinoid treatment. Gel mobility supershift analysis revealed the binding of RARalpha and RXRalpha to oligonucleotides containing both Eb and Ec. Reverse transcriptase-polymerase chain reaction analysis revealed that As4.1 cells express both receptor isoforms, along with RARgamma , but do not express RARbeta , RXRbeta , or RXRgamma . Co-transfection of an expression vector encoding wild-type RARalpha increased enhancer activity, whereas a dominant negative mutant of RARalpha significantly attenuated retinoic acid-induced activity of the enhancer. These results demonstrate the importance of the Eb and Ec motifs in controlling baseline activity of the renin enhancer, and suggest the potential importance of retinoids in regulating renin expression.

* This work was supported by National Institutes of Health Grants HL48058, HL61446, and HL55006 (to C. D. S.) and HL48459 (to K. W. G.). DNA sequencing was performed at the University of Iowa DNA Core Facility.The costs of publication of this article were defrayed in part by the payment of page charges. The 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: Molecular Biology Interdisciplinary Program, Director, Transgenic and Gene Targeting Facility, Dept. of Internal Medicine and Physiology & Biophysics, 2191 Medical Laboratory, The University of Iowa College of Medicine, Iowa City, IA 52242. Tel.: 319-335-7604; Fax: 319-353-5350; E-mail: curt-sigmund@uiowa.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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