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MBC in Press, published online ahead of print July 28, 2004
Mol. Biol. Cell 10.1091/mbc.E04-04-0348

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Submitted on April 28, 2004
Revised on July 2, 2004
Accepted on July 20, 2004

Induction of Vascular Smooth Muscle {alpha}-Actin Gene Transcription in TGF{beta}1-Activated Myofibroblasts Mediated by Dynamic Interplay Between the Pur Repressor Proteins and Sp1/Smad Co-activators

Sukanya V. Subramanian*{dagger}, John A. Polikandriotis*{dagger}, Robert J. Kelm Jr.{ddagger}, Jason J. David*, Charles G. Orosz{sect}, and Arthur R. Strauch*||

Departments of *Physiology and Cell Biology and {sect}Surgery, Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, College of Medicine and Public Health, Columbus, OH 43210; {ddagger}Department of Medicine, College of Medicine, University of Vermont, Colchester, VT 05446

Monitoring Editor: Carl-Henrik Heldin

The mouse vascular smooth muscle {alpha}-actin (SMA) gene enhancer is activated in fibroblasts by TGF{beta}1, a potent mediator of myofibroblast differentiation and wound healing. The SMA enhancer contains tandem sites for the Sp1 transcriptional activator protein and Pur{alpha} and {beta} repressor proteins. We have examined dynamic interplay between these divergent proteins to identify checkpoints for possible control of myofibroblast differentiation during chronic inflammatory disease. A novel element in the SMA enhancer named SPUR was responsible for both basal and TGF{beta}1-dependent transcriptional activation in fibroblasts and capable of binding Sp1 and Pur proteins. A novel Sp1:Pur:SPUR complex was dissociated when SMA enhancer activity was increased by TGF{beta}1 or Smad protein overexpression. Physical association of Pur proteins with Smad2/3 was observed as was binding of Smads to an upstream enhancer region that undergoes DNA duplex unwinding in TGF{beta}1-activated myofibroblasts. Pur{beta} repression of the SMA enhancer could not be relieved by TGF{beta}1 whereas repression mediated by Pur{alpha} was partially rescued by TGF{beta}1 or overexpression of Smad proteins. Interplay between Pur repressor isoforms and Sp1 and Smad coactivators may regulate SMA enhancer output in TGF{beta}1-activated myofibroblasts during episodes of wound repair and tissue remodeling.

{dagger}These authors contributed equally to this work.

||Corresponding author. E-mail: strauch1{at}osu.edu




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