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J. Biol. Chem., Vol. 282, Issue 24, 17335-17339, June 15, 2007

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Regulation of GREB1 Transcription by Estrogen Receptor through a Multipartite Enhancer Spread Over 20 kb of Upstream Flanking Sequences^*

Julie Deschênes¹², Véronique Bourdeau¹³, John H. White^¶4, and Sylvie Mader^¶5

From the Institute for Research in Immunology and Cancer and Department of Biochemistry, Université de Montréal, Montréal, Québec H3C 3J7, Canada and the Departments of Physiology and ^¶Medicine, McGill University, Montréal, Québec H3A 1A4, Canada

Estrogen receptors activate transcription in part through direct interactions with specific DNA motifs, called estrogen response elements (EREs). Here we show that the strong and sustained induction of the gene regulated in breast cancer 1 (GREB1), a gene of unknown function that has been previously suggested to play a role in the effects of estradiol on breast cancer cell proliferation (Rae, J. M., Johnson, M. D., Scheys, J. O., Cordero, K. E., Larios, J. M., and Lippman, M. E. (2005) Breast Cancer Res. Treat 92, 141–149), is mediated by binding of estrogen receptor (ER) to three consensus EREs spread over 20 kb of upstream flanking sequences. In addition to ER, coactivator SRC-3, acetylated histones and phosphorylated RNA polymerase II (P-polII) were detected on all three EREs in the presence of estrogen, while basal recruitment of ER and P-polII was observed only on the proximal element. Chromatin loops were formed between each ERE and the GREB1 transcriptional start site in the presence of estrogen but not of a total antiestrogen. Furthermore, estradiol induced physical association between EREs, suggesting that these elements function as a potent multipartite enhancer to regulate GREB1 transcription.

Received for publication, February 21, 2007 , and in revised form, April 24, 2007.

^* This work was supported in part by Canadian Institutes of Health Research (CIHR) Grant MOP-13147 (to S. M.). 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.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. 1–3 and Refs. 34–39.

¹ These authors contributed equally to this work.

² Recipient of studentships from the Fonds de la Recherche en Santé du Québec (FRSQ) and the Faculté desÉtudes Supérieures de l'Université de Montréal.

³ Holder of a post-doctoral fellowship from the CIHR.

⁴ Chercheur-Boursier National of the FRSQ.

⁵ Chercheur-Boursier National of the FRSQ. Holder of the Canadian Imperial Bank of Commerce Breast Cancer Research Chair at Université de Montréal. To whom correspondence should be addressed: IRIC, Université de Montréal, C.P. 6128, Succursale Centre-Ville, Montréal, Quebec H3C 3J7, Canada. Tel.: 514-343-7166; Fax: 514-343-7780; E-mail: sylvie.mader{at}umontreal.ca.

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