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Circulation Research. 2003;92:856-864
Published online before print March 27, 2003, doi: 10.1161/01.RES.0000068405.49081.09
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(Circulation Research. 2003;92:856.)
© 2003 American Heart Association, Inc.


Molecular Medicine

Myocardin Is a Key Regulator of CArG-Dependent Transcription of Multiple Smooth Muscle Marker Genes

Tadashi Yoshida, Sanjay Sinha, Frédéric Dandré, Brian R. Wamhoff, Mark H. Hoofnagle, Brandon E. Kremer, Da-Zhi Wang, Eric N. Olson, Gary K. Owens

From the Departments of Molecular Physiology and Biological Physics (T.Y., S.S., F.D., B.R.W., M.H.H., G.K.O.) and Microbiology (B.E.K.), University of Virginia, Charlottesville, Va; and the Department of Molecular Biology (D.-Z.W., E.N.O.), University of Texas Southwestern Medical Center, Dallas, Tex.

Correspondence to Gary K. Owens, PhD, Department of Molecular Physiology and Biological Physics, University of Virginia, PO Box 800736, Charlottesville, VA 22908-0736. E-mail gko{at}virginia.edu

The interactions between serum response factor (SRF) and CArG elements are critical for smooth muscle cell (SMC) marker gene transcription. However, the mechanisms whereby SRF, which is expressed ubiquitously, contributes to SMC-specific transcription are unknown. Myocardin was recently cloned as a coactivator of SRF in the heart, but its role in regulating CArG-dependent expression of SMC differentiation marker genes has not been clearly elucidated. In this study, we examined the expression and the function of myocardin in SMCs. In adult mice, myocardin mRNA was expressed in multiple smooth muscle (SM) tissues including the aorta, bladder, stomach, intestine, and colon, as well as the heart. Myocardin was also expressed in cultured rat aortic SMCs and A404 SMC precursor cells. Of particular interest, expression of myocardin was induced during differentiation of A404 cells, although it was not expressed in parental P19 cells from which A404 cells were derived. Cotransfection studies in SMCs revealed that myocardin induced the activity of multiple SMC marker gene promoters including SM {alpha}-actin, SM-myosin heavy chain, and SM22{alpha} by 9- to 60-fold in a CArG-dependent manner, whereas myocardin short interfering RNA markedly decreased activity of these promoters. Moreover, adenovirus-mediated overexpression of a dominant-negative form of myocardin significantly suppressed expression of endogenous SMC marker genes, whereas adenovirus-mediated overexpression of wild-type myocardin increased expression. Taken together, results provide compelling evidence that myocardin plays a key role as a transcriptional coactivator of SMC marker genes through CArG-dependent mechanisms.


Key Words: smooth muscle cells • transcriptional coactivator • serum response factor • CArG element




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T. Yoshida and G. K. Owens
Molecular Determinants of Vascular Smooth Muscle Cell Diversity
Circ. Res., February 18, 2005; 96(3): 280 - 291.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
J. W. Streb and J. M. Miano
AKAP12{alpha}, an Atypical Serum Response Factor-dependent Target Gene
J. Biol. Chem., February 11, 2005; 280(6): 4125 - 4134.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
F. Yin and B. P. Herring
GATA-6 Can Act as a Positive or Negative Regulator of Smooth Muscle-specific Gene Expression
J. Biol. Chem., February 11, 2005; 280(6): 4745 - 4752.
[Abstract] [Full Text] [PDF]


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Genes Dev.Home page
S. Drori, G. D. Girnun, L. Tou, J. D. Szwaya, E. Mueller, X. Kia, R. A. Shivdasani, and B. M. Spiegelman
Hic-5 regulates an epithelial program mediated by PPAR{gamma}
Genes & Dev., February 1, 2005; 19(3): 362 - 375.
[Abstract] [Full Text] [PDF]


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Arterioscler. Thromb. Vasc. Bio.Home page
J. J. Lepore, T. P. Cappola, P. A. Mericko, E. E. Morrisey, and M. S. Parmacek
GATA-6 Regulates Genes Promoting Synthetic Functions in Vascular Smooth Muscle Cells
Arterioscler. Thromb. Vasc. Biol., February 1, 2005; 25(2): 309 - 314.
[Abstract] [Full Text] [PDF]


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Proc. Natl. Acad. Sci. USAHome page
S. Li, M. P. Czubryt, J. McAnally, R. Bassel-Duby, J. A. Richardson, F. F. Wiebel, A. Nordheim, and E. N. Olson
Requirement for serum response factor for skeletal muscle growth and maturation revealed by tissue-specific gene deletion in mice
PNAS, January 25, 2005; 102(4): 1082 - 1087.
[Abstract] [Full Text] [PDF]


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Mol. Cell. Biol.Home page
D. Cao, Z. Wang, C.-L. Zhang, J. Oh, W. Xing, S. Li, J. A. Richardson, D.-Z. Wang, and E. N. Olson
Modulation of Smooth Muscle Gene Expression by Association of Histone Acetyltransferases and Deacetylases with Myocardin
Mol. Cell. Biol., January 1, 2005; 25(1): 364 - 376.
[Abstract] [Full Text] [PDF]


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Proc. Natl. Acad. Sci. USAHome page
J. M. Miano, N. Ramanan, M. A. Georger, K. L. de Mesy Bentley, R. L. Emerson, R. O. Balza Jr., Q. Xiao, H. Weiler, D. D. Ginty, and R. P. Misra
Restricted inactivation of serum response factor to the cardiovascular system
PNAS, December 7, 2004; 101(49): 17132 - 17137.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Cell Physiol.Home page
S. Sinha, M. H. Hoofnagle, P. A. Kingston, M. E. McCanna, and G. K. Owens
Transforming growth factor-{beta}1 signaling contributes to development of smooth muscle cells from embryonic stem cells
Am J Physiol Cell Physiol, December 1, 2004; 287(6): C1560 - C1568.
[Abstract] [Full Text] [PDF]


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Physiol. GenomicsHome page
J. M. Spin, S. Nallamshetty, R. Tabibiazar, E. A. Ashley, J. Y. King, M. Chen, P. S. Tsao, and T. Quertermous
Transcriptional profiling of in vitro smooth muscle cell differentiation identifies specific patterns of gene and pathway activation
Physiol Genomics, November 17, 2004; 19(3): 292 - 302.
[Abstract] [Full Text] [PDF]


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Circ. Res.Home page
B.R. Wamhoff, M.H. Hoofnagle, A. Burns, S. Sinha, O.G. McDonald, and G.K. Owens
A G/C Element Mediates Repression of the SM22{alpha} Promoter Within Phenotypically Modulated Smooth Muscle Cells in Experimental Atherosclerosis
Circ. Res., November 12, 2004; 95(10): 981 - 988.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
K. Lockman, J. S. Hinson, M. D. Medlin, D. Morris, J. M. Taylor, and C. P. Mack
Sphingosine 1-Phosphate Stimulates Smooth Muscle Cell Differentiation and Proliferation by Activating Separate Serum Response Factor Co-factors
J. Biol. Chem., October 8, 2004; 279(41): 42422 - 42430.
[Abstract] [Full Text] [PDF]


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Mol. Cell. Biol.Home page
J. Oh, Z. Wang, D.-Z. Wang, C.-L. Lien, W. Xing, and E. N. Olson
Target Gene-Specific Modulation of Myocardin Activity by GATA Transcription Factors
Mol. Cell. Biol., October 1, 2004; 24(19): 8519 - 8528.
[Abstract] [Full Text] [PDF]


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Arterioscler. Thromb. Vasc. Bio.Home page
M. S. Parmacek
Myocardin--Not Quite MyoD
Arterioscler. Thromb. Vasc. Biol., September 1, 2004; 24(9): 1535 - 1537.
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J. Am. Soc. Nephrol.Home page
M. Takeji, N. Kawada, T. Moriyama, K. Nagatoya, S. Oseto, S. Akira, M. Hori, E. Imai, and T. Miwa
CCAAT/Enhancer-Binding Protein {delta} Contributes to Myofibroblast Transdifferentiation and Renal Disease Progression
J. Am. Soc. Nephrol., September 1, 2004; 15(9): 2383 - 2390.
[Abstract] [Full Text] [PDF]


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Arterioscler. Thromb. Vasc. Bio.Home page
T. Yoshida, K. Kawai-Kowase, and G. K. Owens
Forced Expression of Myocardin Is Not Sufficient for Induction of Smooth Muscle Differentiation in Multipotential Embryonic Cells
Arterioscler. Thromb. Vasc. Biol., September 1, 2004; 24(9): 1596 - 1601.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
P. R. Reynolds, M. L. Mucenski, T. D. Le Cras, W. C. Nichols, and J. A. Whitsett
Midkine Is Regulated by Hypoxia and Causes Pulmonary Vascular Remodeling
J. Biol. Chem., August 27, 2004; 279(35): 37124 - 37132.
[Abstract] [Full Text] [PDF]


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Proc. Natl. Acad. Sci. USAHome page
Z. Han, X. Li, J. Wu, and E. N. Olson
A myocardin-related transcription factor regulates activity of serum response factor in Drosophila
PNAS, August 24, 2004; 101(34): 12567 - 12572.
[Abstract] [Full Text] [PDF]


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Circ. Res.Home page
J. M. Miano
Channeling to Myocardin
Circ. Res., August 20, 2004; 95(4): 340 - 342.
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Circ. Res.Home page
B.R. Wamhoff, D.K. Bowles, O.G. McDonald, S. Sinha, A.P. Somlyo, A.V. Somlyo, and G.K. Owens
L-type Voltage-Gated Ca2+ Channels Modulate Expression of Smooth Muscle Differentiation Marker Genes via a Rho Kinase/Myocardin/SRF-Dependent Mechanism
Circ. Res., August 20, 2004; 95(4): 406 - 414.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
S. Albinsson, I. Nordstrom, and P. Hellstrand
Stretch of the Vascular Wall Induces Smooth Muscle Differentiation by Promoting Actin Polymerization
J. Biol. Chem., August 13, 2004; 279(33): 34849 - 34855.
[Abstract] [Full Text] [PDF]


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Arterioscler. Thromb. Vasc. Bio.Home page
K. Kawai-Kowase, H. Sato, Y. Oyama, H. Kanai, M. Sato, H. Doi, and M. Kurabayashi
Basic Fibroblast Growth Factor Antagonizes Transforming Growth Factor-{beta}1-Induced Smooth Muscle Gene Expression Through Extracellular Signal-Regulated Kinase 1/2 Signaling Pathway Activation
Arterioscler. Thromb. Vasc. Biol., August 1, 2004; 24(8): 1384 - 1390.
[Abstract] [Full Text] [PDF]


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Physiol. Rev.Home page
G. K. Owens, M. S. Kumar, and B. R. Wamhoff
Molecular Regulation of Vascular Smooth Muscle Cell Differentiation in Development and Disease
Physiol Rev, July 1, 2004; 84(3): 767 - 801.
[Abstract] [Full Text] [PDF]


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Circ. Res.Home page
S. Brunelli, E. Tagliafico, F. G. De Angelis, R. Tonlorenzi, S. Baesso, S. Ferrari, M. Niinobe, K. Yoshikawa, R. J. Schwartz, I. Bozzoni, et al.
Msx2 and Necdin Combined Activities Are Required for Smooth Muscle Differentiation in Mesoangioblast Stem Cells
Circ. Res., June 25, 2004; 94(12): 1571 - 1578.
[Abstract] [Full Text] [PDF]


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Mol. Cell. Biol.Home page
J. P. Anderson, E. Dodou, A. B. Heidt, S. J. De Val, E. J. Jaehnig, S. B. Greene, E. N. Olson, and B. L. Black
HRC Is a Direct Transcriptional Target of MEF2 during Cardiac, Skeletal, and Arterial Smooth Muscle Development In Vivo
Mol. Cell. Biol., May 1, 2004; 24(9): 3757 - 3768.
[Abstract] [Full Text] [PDF]


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Circ. Res.Home page
T. Yoshida, M. H. Hoofnagle, and G. K. Owens
Myocardin and Prx1 Contribute to Angiotensin II-Induced Expression of Smooth Muscle {alpha}-Actin
Circ. Res., April 30, 2004; 94(8): 1075 - 1082.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
K. L. Du, M. Chen, J. Li, J. J. Lepore, P. Mericko, and M. S. Parmacek
Megakaryoblastic Leukemia Factor-1 Transduces Cytoskeletal Signals and Induces Smooth Muscle Cell Differentiation from Undifferentiated Embryonic Stem Cells
J. Biol. Chem., April 23, 2004; 279(17): 17578 - 17586.
[Abstract] [Full Text] [PDF]


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Circ. Res.Home page
R. B. Pilz and D. E. Casteel
Regulation of Gene Expression by Cyclic GMP
Circ. Res., November 28, 2003; 93(11): 1034 - 1046.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
A. Selvaraj and R. Prywes
Megakaryoblastic Leukemia-1/2, a Transcriptional Co-activator of Serum Response Factor, Is Required for Skeletal Myogenic Differentiation
J. Biol. Chem., October 24, 2003; 278(43): 41977 - 41987.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
R. J. Kelm Jr., S.-X. Wang, J. A. Polikandriotis, and A. R. Strauch
Structure/Function Analysis of Mouse Pur{beta}, a Single-stranded DNA-binding Repressor of Vascular Smooth Muscle {alpha}-Actin Gene Transcription
J. Biol. Chem., October 3, 2003; 278(40): 38749 - 38757.
[Abstract] [Full Text] [PDF]


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Genes Dev.Home page
E. N. Olson and M. D. Schneider
Sizing up the heart: development redux in disease
Genes & Dev., August 15, 2003; 17(16): 1937 - 1956.
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Proc. Natl. Acad. Sci. USAHome page
S. Li, D.-Z. Wang, Z. Wang, J. A. Richardson, and E. N. Olson
The serum response factor coactivator myocardin is required for vascular smooth muscle development
PNAS, August 5, 2003; 100(16): 9366 - 9370.
[Abstract] [Full Text] [PDF]


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Proc. Natl. Acad. Sci. USAHome page
Z. Wang, D.-Z. Wang, G. C. T. Pipes, and E. N. Olson
Myocardin is a master regulator of smooth muscle gene expression
PNAS, June 10, 2003; 100(12): 7129 - 7134.
[Abstract] [Full Text] [PDF]


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Sci Aging Knowl EnvironHome page
M. Beckman
Rise to Power
Sci. Aging Knowl. Environ., May 28, 2003; 2003(21): nw75 - 75.
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Circ. Res.Home page
M. W. Majesky
Decisions, Decisions ... SRF Coactivators and Smooth Muscle Myogenesis
Circ. Res., May 2, 2003; 92(8): 824 - 826.
[Full Text] [PDF]