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Originally published In Press as doi:10.1074/jbc.M106649200 on December 11, 2001

J. Biol. Chem., Vol. 277, Issue 8, 6287-6295, February 22, 2002
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Transforming Growth Factor-beta Induction of Smooth Muscle Cell Phenotpye Requires Transcriptional and Post-transcriptional Control of Serum Response Factor*

Karen K. HirschiDagger §, Lihua LaiDagger , Narasimhaswamy S. Belaguli, David A. Dean||**, Robert J. SchwartzDagger Dagger , and Warren E. Zimmer||**

From the Dagger  Departments of Pediatrics and Molecular and Cellular Biology, Center for Cell and Gene Therapy and Children's Nutrition Research Center, Baylor College of Medicine, Houston, Texas 77030, the  Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas 77030, and the || Department of Cell Biology and Neuroscience, University of South Alabama, Mobile, Alabama 36688

Transforming growth factor-beta induces a smooth muscle cell phenotype in undifferentiated mesenchymal cells. To elucidate the mechanism(s) of this phenotypic induction, we focused on the molecular regulation of smooth muscle-gamma -actin, whose expression is induced at late stages of smooth muscle differentiation and developmentally restricted to this lineage. Transforming growth factor-beta induced smooth muscle-gamma -actin protein, cytoskeletal localization, and mRNA expression in mesenchymal cells. Smooth muscle-gamma -actin promoter-luciferase reporter activity was enhanced by transforming growth factor-beta , and deletion analysis revealed that CArG box 2 in the promoter was necessary for this transcriptional activation. CArG motifs bind transcriptional activator serum response factor; gel shift analyses revealed increased binding of serum response factor-containing complexes to this site in response to transforming growth factor-beta , paralleled by increased serum response factor protein expression. Serum response factor expression was found to be up-regulated by transforming growth factor-beta via transcriptional activation of the gene and post-transcriptional regulation. Using mesenchymal cells stably transfected with wild type or dominant-negative serum response factor, we demonstrated that its expression is sufficient for induction of a smooth muscle phenotype in mesenchymal cells and is necessary for transforming growth factor-beta -mediated smooth muscle induction.

* 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.

§ Supported by United States Department of Agriculture (USDA) Grant 6250-51000-033, National Institutes of Health (NIH) Grant 1R01-HL61408, and American Heart Association-National Scientist Development Grant 9930054N. To whom correspondence should be addressed: Depts. of Pediatrics & Molecular and Cellular Biology, Centers for Cell and Gene Therapy & Children's Nutrition Research, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030. Tel.: 713-798-7771; Fax: 713-798-1230; khirschi@bcm.tmc.edu.

** Supported by NIH Grant RO1-HL59956.

Dagger Dagger Supported by USDA Grant 6250-51000-037 and NIH Grants RO1-HL50423 and PO1-HL49953.

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