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Originally published In Press as doi:10.1074/jbc.M001678200 on July 31, 2000

J. Biol. Chem., Vol. 275, Issue 41, 31722-31732, October 13, 2000
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Transcriptional Activation of the Human ucp1 Gene in a Rodent Cell Line
SYNERGISM OF RETINOIDS, ISOPROTERENOL, AND THIAZOLIDINEDIONE IS MEDIATED BY A MULTIPARTITE RESPONSE ELEMENT*

Maria del Mar Gonzalez-BarrosoDagger , Claire Pecqueur§, Chantal Gelly, Daniel SanchisDagger , Marie-Clotilde Alves-Guerra, Frederic Bouillaud, Daniel Ricquier, and Anne-Marie Cassard-Doulcier

From the Centre de Recherches sur l'Endocrinologie Moléculaire et le Développement, CNRS, 92190 Meudon, France

Uncoupling protein 1 (UCP1) is uniquely expressed in brown adipocytes and generates heat production by uncoupling respiration from ATP synthesis. The activatory effects of norepinephrine and retinoic acid (RA) on rodent ucp1 gene transcription have been well characterized. These effects are mediated by a 211-base pair (bp) enhancer which is also sufficient to restrict expression to brown adipose tissue. The molecular mechanisms controlling the transcription of the human ucp1 gene are unknown. In order to study the transcriptional regulation of the human gene, we set up chloramphenicol acetyltransferase constructs containing the entire or deleted 5' regions upstream of the transcriptional start site of the gene. These constructs were transiently transfected in a mouse cell line. A 350-bp hormone response region showing a significant homology with the rat ucp1 enhancer and located between the BclI polymorphic site and an AatII site (bp -3820/-3470) was detected. This region was sufficient to mediate the stimulation by RA and by combined treatments (RA + isoproterenol (ISO), RA + thiazolidinedione (TZD), or RA + ISO + TZD). The highest stimulation, a 26-fold increase in basal activity, was obtained by RA + ISO + TZD treatment. In contrast to the rodent gene, under our conditions, the effect of ISO and/or TZD is dependent on RA stimulation. Analysis of 105 bp inside the 350-bp element by site-directed mutagenesis and gel retardation experiments demonstrated that a multipartite response element mediates the drug stimulation. This region binds RARs and RXRs nuclear factors, CREB/ATF factors, and also PPARgamma despite the absence of a consensus peroxisome-proliferator response element. The activation of the human ucp1 gene transcription by certain hormones or drugs, and the identification of the cis-elements involved, will help to identify new compounds activating fat oxidation and energy expenditure in humans.

* This work was supported in part by the Centre National de la Recherche Scientifique (CNRS), the Institut National de la Santé et de la Recherche Médicale (INSERM), the Association pour la Recherche contre le Cancer, the Human Frontier Science Program organization (HFSP), and the Institut de Recherches Servier.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.

Dagger Supported by a grant from the European Union (Marie Curie Mobility Training Fellowship).

§ Supported by a Ph.D. thesis fellowship from the Association pour la Recherche contre le Cancer.

To whom correspondence should be addressed: CEREMOD, CNRS, 9 rue Jules Hetzel, 92190 Meudon, France. Tel.: 33-0-1-45-07-57-48; E-mail: doulcier@infobiogen.fr.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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