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Originally published In Press as doi:10.1074/jbc.M004625200 on November 28, 2000

J. Biol. Chem., Vol. 276, Issue 10, 7575-7585, March 9, 2001
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Control of Cardiac-specific Transcription by p300 through Myocyte Enhancer Factor-2D*

Tatiana I. SlepakDagger , Keith A. WebsterDagger , Jie ZangDagger , Howard Prentice§, Ann O'Dowd§, Martin N. Hicks, and Nanette H. BishopricDagger ||

From the Dagger  Department of Molecular and Cellular Pharmacology, University of Miami, Miami, Florida 33101 and the Departments of § Molecular Genetics and  Medical Cardiology, Glasgow Royal Infirmary, University of Glasgow, Glasgow G11 6 NU, United Kingdom

The transcriptional integrator p300 regulates gene expression by interaction with sequence-specific DNA-binding proteins and local remodeling of chromatin. p300 is required for cardiac-specific gene transcription, but the molecular basis of this requirement is unknown. Here we report that the MADS (MCM-1, agamous, deficiens, serum response factor) box transcription factor myocyte enhancer factor-2D (MEF-2D) acts as the principal conduit for cardiac transcriptional activation by p300. p300 activation of the native 2130-base pair human skeletal alpha -actin promoter required a single hybrid MEF-2/GATA-4 DNA motif centered at -1256 base pairs. Maximal expression of the promoter in cultured myocytes and in vivo correlated with binding of both MEF-2 and p300, but not GATA-4, to this AT-rich motif. p300 and MEF-2 were coprecipitated from cardiac nuclear extracts by an oligomer containing this element. p300 was found exclusively in a complex with MEF-2D at this and related sites in other cardiac-restricted promoters. MEF-2D, but not other MEFs, significantly potentiated cardiac-specific transcription by p300. No physical or functional interaction was observed between p300 and other factors implicated in skeletal actin transcription, including GATA-4, TEF-1, or SRF. These results show that, in the intact cell, p300 interactions with its protein targets are highly selective and that MEF-2D is the preferred channel for p300-mediated transcriptional control in the heart.

* This work was supported by National Institutes of Health Grants HL49891 (to N. H. B.) and HL44578 (to K. A. W.), by an Established Investigator grant from the American Heart Association (to N.H.B.), by a grant from the Miami Heart Research Institute (to N. H. B.), by a grant from the British Heart Foundation (to H. P.), and by a Wellcome Trust Foundation collaborative travel grant (to H. P., K. A. W., and N. H. B.).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.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF288779.

|| To whom correspondence should be addressed: Dept. of Molecular and Cellular Pharmacology, University of Miami, P.O. Box 106189 (R-189), Miami, FL 33101. Tel.: 305-243-6775; Fax: 305-243-6082; E-mail: nhb@chroma.med.miami.edu.

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