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(Received for publication, August 6, 1996, and in revised form, December 13, 1996)
From the Department of Molecular Physiology and Biological Physics,
University of Virginia Health Sciences Center,
Charlottesville, Virginia 22908
The goal of the present study was to determine
the molecular mechanism whereby transforming growth factor
Volume 272, Number 16,
Issue of April 18, 1997
pp. 10948-10956
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
(TGF) Control Element Drives
TGF-induced Stimulation of Smooth Muscle 
-Actin Gene Expression
in Concert with Two CArG Elements
(TGF)
increases smooth muscle (SM) 
-actin expression. Confluent,
growth-arrested rat aortic smooth muscle cells (SMC) were transiently
transfected with various SM -actin promoter/chloramphenicol
acetyltransferase deletion mutants and stimulated with TGF (2.5 ng/ml). Results demonstrated that the first 125 base pairs of the SM
-actin promoter were sufficient to confer TGF responsiveness.
Three cis elements were shown to be required for TGF
inducibility: two highly conserved CArG boxes, designated A (
62) and
B (112) and a novel TGF control element (TCE) (42). Mutation of
any one of these elements completely abolished TGF-induced reporter
activity. Results of electrophoretic mobility shift assays demonstrated
that nuclear extracts from TGF-treated SMC enhanced binding activity
of serum response factor to the CArG elements and binding of an as yet
unidentified factor to the TCE. Northern analysis showed that TGF
also stimulated transcription of two other SM (SM myosin heavy chain)
differentiation marker genes, SM myosin heavy chain and h1
calponin, whose promoters also contained a TCE-like element. In
summary, we identified a TGF response element in the SM -actin
promoter that may contribute to coordinate regulation of expression of
multiple cell-type specific proteins during SMC differentiation.
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J. SOLWAY, S. M. FORSYTHE, A. J. HALAYKO, J. E. VIEIRA, M. B. HERSHENSON, and B. CAMORETTI-MERCADO Transcriptional Regulation of Smooth Muscle Contractile Apparatus Expression Am. J. Respir. Crit. Care Med., November 1, 1998; 158(2007): S100 - S108. [Abstract] [Full Text] [PDF]
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C. Garat, V. Van Putten, Z. A. Refaat, C. Dessev, S.-Y. Han, and R. A. Nemenoff Induction of Smooth Muscle alpha -Actin in Vascular Smooth Muscle Cells by Arginine Vasopressin Is Mediated by c-Jun Amino-terminal Kinases and p38 Mitogen-activated Protein Kinase J. Biol. Chem., July 14, 2000; 275(29): 22537 - 22543. [Abstract] [Full Text] [PDF]
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P. J. Adam, C. P. Regan, M. B. Hautmann, and G. K. Owens Positive- and Negative-acting Kruppel-like Transcription Factors Bind a Transforming Growth Factor beta Control Element Required for Expression of the Smooth Muscle Cell Differentiation Marker SM22alpha in Vivo J. Biol. Chem., November 22, 2000; 275(48): 37798 - 37806. [Abstract] [Full Text] [PDF]
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N. A. Becker, R. J. Kelm Jr., J. A. Vrana, M. J. Getz, and L. J. Maher III Altered Sensitivity to Single-strand-specific Reagents Associated with the Genomic Vascular Smooth Muscle alpha -Actin Promoter during Myofibroblast Differentiation J. Biol. Chem., May 12, 2000; 275(20): 15384 - 15391. [Abstract] [Full Text] [PDF]
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