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Originally published In Press as doi:10.1074/jbc.M202490200 on May 6, 2002

J. Biol. Chem., Vol. 277, Issue 27, 24390-24398, July 5, 2002
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The Transcription Factors GATA4 and dHAND Physically Interact to Synergistically Activate Cardiac Gene Expression through a p300-dependent Mechanism*

Yan-Shan DaiDagger §, Peter Cserjesi, Bruce E. Markham§||, and Jeffery D. MolkentinDagger **

From the Dagger  Department of Pediatrics, University of Cincinnati, Children's Hospital Medical Center, Cincinnati, Ohio 45229-3039, the § Department of Molecular Sciences, Pfizer Global Research and Development, Ann Arbor, Michigan 48105, and the  Department of Cell Biology and Anatomy, Louisiana State University Health Sciences Center, New Orleans, Louisiana 70112

An intricate array of heterogeneous transcription factors participate in programming tissue-specific gene expression through combinatorial interactions that are unique to a given cell-type. The zinc finger-containing transcription factor GATA4, which is widely expressed in mesodermal and endodermal derived tissues, is thought to regulate cardiac myocyte-specific gene expression through combinatorial interactions with other semi-restricted transcription factors such as myocyte enhancer factor 2, nuclear factor of activated T-cells, serum response factor, and Nkx2.5. Here we determined that GATA4 also interacts with the cardiac-expressed basic helix-loop-helix transcription factor dHAND (also known as HAND2). GATA4 and dHAND synergistically activated expression of cardiac-specific promoters from the atrial natriuretic factor gene, the b-type natriuretic peptide gene, and the alpha -myosin heavy chain gene. Using artificial reporter constructs this functional synergy was shown to be GATA site-dependent, but E-box site-independent. A mechanism for the transcriptional synergy was suggested by the observation that the bHLH domain of dHAND physically interacted with the C-terminal zinc finger domain of GATA4 forming a higher order complex. This transcriptional synergy observed between GATA4 and dHAND was associated with p300 recruitment, but not with alterations in DNA binding activity of either factor. Moreover, the bHLH domain of dHAND directly interacted with the CH3 domain of p300 suggesting the existence of a higher order complex between GATA4, dHAND, and p300. Taken together with previous observations, these results suggest the existence of an enhanceosome complex comprised of p300 and multiple semi-restricted transcription factors that together specify tissue-specific gene expression in the heart.

* This work was supported by grants from the National Institutes of Health (to J. D. M. and B. E. M.) and the Pew Charitable Trust Foundation (to J. D. M.).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 may be addressed: Dept. of Molecular Sciences, Pfizer Global Research and Development, 2800 Plymouth Rd., Ann Arbor, MI 48105. E-mail: bruce.markham@pfizer.com.

** To whom correspondence may be addressed: Division of Molecular Cardiovascular Biology, Dept. of Pediatrics, Children's Hospital Medical Center, 3333 Burnet Ave., Cincinnati, OH 45229-3039. E-mail: jeff.molkentin@chmcc.org.

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