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Originally published In Press as doi:10.1074/jbc.M206215200 on October 10, 2002

J. Biol. Chem., Vol. 277, Issue 51, 49275-49281, December 20, 2002
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Transcriptional Regulation of Human Rev-erbalpha Gene Expression by the Orphan Nuclear Receptor Retinoic Acid-related Orphan Receptor alpha *

Eric RaspèDagger , Gisèle MautinoDagger §, Caroline DuvalDagger , Coralie FontaineDagger , Hélène Duez, Olivier BarbierDagger , Didier Monte||, Jamila FruchartDagger **, Jean-Charles FruchartDagger **, and Bart StaelsDagger **Dagger Dagger

From the Dagger  UR 545 INSERM, Institut Pasteur de Lille, 1 Rue Calmette, 59019 Lille, § Groupe Merck, Centre de Recherche et Développement, 115 Ave. Lacassagne, 69003 Lyon, ** Faculté de Pharmacie, Université de Lille II, 59006 Lille, and || UMR 8117, Institut de Biologie de Lille, Institut Pasteur de Lille, 1 Rue Calmette, 59019 Lille, France

The Rev-erb and retinoic acid-related orphan receptors (ROR) are two related families of orphan nuclear receptors that recognize similar response elements but have opposite effects on transcription. Recently, the Rev-erbalpha gene promoter has been characterized and shown to harbor a functional Rev-erbalpha -binding site known as Rev-DR2, responsible for negative feedback down-regulation of promoter activity by Rev-erbalpha itself. The present study aimed to investigate whether Rev-erbalpha gene expression is regulated by RORalpha . Gel shift analysis demonstrated that in vitro translated hRORalpha 1 protein binds to the Rev-DR2 site, both as monomer and dimer. Chromatin immunoprecipitation assays demonstrated that binding of RORalpha to this site also occurred in vivo in human hepatoma HepG2 cells. The Rev-DR2 site was further shown to be functional as it conferred hRORalpha 1 responsiveness to a heterologous promoter and to the natural human Rev-erbalpha gene promoter in these cells. Mutation of this site in the context of the natural Rev-erbalpha gene promoter abolished its activation by RORalpha , indicating that this site plays a key role in hRORalpha 1 action. Finally, adenoviral overexpression of hRORalpha 1 in HepG2 cells led to enhanced hRev-erbalpha mRNA accumulation, further confirming the physiological importance of RORalpha 1 in the regulation of Rev-erbalpha expression.

* This work was supported in part by grants from INSERM, FEDER-Conseil Regional Nord-Pas de-Calais (Génopole 01360124) and Merck-Lipha.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.

Supported by a fellowship from the Association pour la Recherche sur le Cancer. Supported by Grants from the Fondation de France and from the Comité d'Aide à la Recherche Fournier.

Dagger Dagger To whom correspondence should be addressed: UR 545 INSERM, Institut Pasteur de Lille, 1 Rue du Pr. Calmette, 59019, Lille, France. Tel.: 33-3-20-87-73-88; Fax: 33-3-20-87-71-98; E-mail: bart.staels@pasteur-lille.fr.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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