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Vol. 273, Issue 3, 1511-1518, January 16, 1998

MEF2B Is a Component of a Smooth Muscle-specific Complex That Binds an A/T-rich Element Important for Smooth Muscle Myosin Heavy Chain Gene Expression

Youichi KatohDagger , Jeffery D. Molkentin§, Vrushank DaveDagger , Eric N. Olson§, and Muthu PeriasamyDagger

From the Dagger  Section of Molecular Cardiology, Division of Cardiology and Cardiovascular Research Center, University of Cincinnati, Cincinnati, Ohio 45267 and the § Department of Molecular Biology and Oncology, University of Texas Southwestern Medical Center, Dallas, Texas 75235

To understand smooth muscle-specific gene expression, we have focused our studies on the smooth muscle myosin heavy chain (SMHC) gene, a smooth muscle-specific marker. In this study, we demonstrate that the SMHC promoter region (-1594 to -1462 base pairs) containing the A/T-rich element can activate the heterologous thymidine kinase promoter in smooth muscle cells, but not in fibroblasts. Mutations of this A/T-rich element decreased SMHC promoter activity significantly. Both gel mobility shift assays and DNase I footprinting revealed that this region binds to specific protein complexes from smooth muscle nuclear extracts, whereas nuclear extracts from skeletal muscle and fibroblasts produced a different binding pattern. We also demonstrate that the protein complex obtained from smooth muscle nuclear extract reacts with MEF2B-specific antibody, but not with antibodies specific to MEF2A, MEF2C, or MEF2D, suggesting that only MEF2B protein binds to the A/T-rich element. Furthermore, MEF2B overexpression in smooth muscle cells up-regulated the SMHC promoter, suggesting that MEF2B is important for SMHC gene regulation. This is the first report demonstrating a role for MEF2 factors in smooth muscle-specific gene expression.

Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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