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

Research output: Contribution to journalArticle

276 Citations (Scopus)

Abstract

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 α-actin, SM-myosin heavy chain, and SM22α 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.

Original languageEnglish (US)
Pages (from-to)856-864
Number of pages9
JournalCirculation Research
Volume92
Issue number8
DOIs
StatePublished - May 2 2003

Fingerprint

Smooth Muscle
Smooth Muscle Myocytes
Genes
Serum Response Factor
Adenoviridae
Cell Differentiation
myocardin
Smooth Muscle Myosins
Myosin Heavy Chains
Differentiation Antigens
Small Interfering RNA
Intestines
Aorta
Actins
Stomach
Colon
Urinary Bladder
Muscles
Messenger RNA

Keywords

  • CArG element
  • Serum response factor
  • Smooth muscle cells
  • Transcriptional coactivator

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Yoshida, T., Sinha, S., Dandré, F., Wamhoff, B. R., Hoofnagle, M. H., Kremer, B. E., ... Owens, G. K. (2003). Myocardin is a key regulator of CArG-dependent transcription of multiple smooth muscle marker genes. Circulation Research, 92(8), 856-864. https://doi.org/10.1161/01.RES.0000068405.49081.09

Myocardin is a key regulator of CArG-dependent transcription of multiple smooth muscle marker genes. / Yoshida, Tadashi; Sinha, Sanjay; Dandré, Frédéric; Wamhoff, Brian R.; Hoofnagle, Mark H.; Kremer, Brandon E.; Wang, Da Zhi; Olson, Eric N.; Owens, Gary K.

In: Circulation Research, Vol. 92, No. 8, 02.05.2003, p. 856-864.

Research output: Contribution to journalArticle

Yoshida, T, Sinha, S, Dandré, F, Wamhoff, BR, Hoofnagle, MH, Kremer, BE, Wang, DZ, Olson, EN & Owens, GK 2003, 'Myocardin is a key regulator of CArG-dependent transcription of multiple smooth muscle marker genes', Circulation Research, vol. 92, no. 8, pp. 856-864. https://doi.org/10.1161/01.RES.0000068405.49081.09
Yoshida, Tadashi ; Sinha, Sanjay ; Dandré, Frédéric ; Wamhoff, Brian R. ; Hoofnagle, Mark H. ; Kremer, Brandon E. ; Wang, Da Zhi ; Olson, Eric N. ; Owens, Gary K. / Myocardin is a key regulator of CArG-dependent transcription of multiple smooth muscle marker genes. In: Circulation Research. 2003 ; Vol. 92, No. 8. pp. 856-864.
@article{bbfef2fe4fd046cfa45d908e156c1d72,
title = "Myocardin is a key regulator of CArG-dependent transcription of multiple smooth muscle marker genes",
abstract = "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 α-actin, SM-myosin heavy chain, and SM22α 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.",
keywords = "CArG element, Serum response factor, Smooth muscle cells, Transcriptional coactivator",
author = "Tadashi Yoshida and Sanjay Sinha and Fr{\'e}d{\'e}ric Dandr{\'e} and Wamhoff, {Brian R.} and Hoofnagle, {Mark H.} and Kremer, {Brandon E.} and Wang, {Da Zhi} and Olson, {Eric N.} and Owens, {Gary K.}",
year = "2003",
month = "5",
day = "2",
doi = "10.1161/01.RES.0000068405.49081.09",
language = "English (US)",
volume = "92",
pages = "856--864",
journal = "Circulation Research",
issn = "0009-7330",
publisher = "Lippincott Williams and Wilkins",
number = "8",

}

TY - JOUR

T1 - Myocardin is a key regulator of CArG-dependent transcription of multiple smooth muscle marker genes

AU - Yoshida, Tadashi

AU - Sinha, Sanjay

AU - Dandré, Frédéric

AU - Wamhoff, Brian R.

AU - Hoofnagle, Mark H.

AU - Kremer, Brandon E.

AU - Wang, Da Zhi

AU - Olson, Eric N.

AU - Owens, Gary K.

PY - 2003/5/2

Y1 - 2003/5/2

N2 - 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 α-actin, SM-myosin heavy chain, and SM22α 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.

AB - 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 α-actin, SM-myosin heavy chain, and SM22α 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.

KW - CArG element

KW - Serum response factor

KW - Smooth muscle cells

KW - Transcriptional coactivator

UR - http://www.scopus.com/inward/record.url?scp=0037968290&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037968290&partnerID=8YFLogxK

U2 - 10.1161/01.RES.0000068405.49081.09

DO - 10.1161/01.RES.0000068405.49081.09

M3 - Article

C2 - 12663482

AN - SCOPUS:0037968290

VL - 92

SP - 856

EP - 864

JO - Circulation Research

JF - Circulation Research

SN - 0009-7330

IS - 8

ER -