Serum response factor-dependent regulation of the smooth muscle calponin gene

Joseph M. Miano, Michael J. Carlson, Jeffrey A. Spencer, Ravi P. Misra

Research output: Contribution to journalArticle

Abstract

Smooth muscle calponin is a multifunctional, thin filament-associated protein whose expression is restricted to smooth muscle cell lineages in developing and postnatal tissues. Although the physiology of smooth muscle calponin has been studied extensively, the cis-elements governing its restricted pattern of expression have yet to be identified. Here we report on smooth muscle-specific enhancer activity within the first intron of smooth muscle calponin. Sequence analysis revealed a proximal consensus intronic CArG box and two distal intronic CArG-like elements, each of which bound recombinant serum response factor (SRF) as well as immunoreactive SRF from smooth muscle nuclear extracts. Site-directed mutagenesis studies suggested that the consensus CArG box mediates much of the intronic enhancer activity; mutating all three CArG elements abolished the ability of SRF to confer enhancer activity on the smooth muscle calponin promoter. Cotransfecting a dominant-negative SRF construct attenuated smooth muscle-specific enhancer activity, and transducing smooth muscle cells with adenovirus harboring the dominant-negative SRF construct selectively reduced steady-state expression of endogenous smooth muscle calponin. These results demonstrate an important role for intronic CArG boxes and the SRF protein in the transcriptional control of smooth muscle calponin in vitro.

Original languageEnglish (US)
Pages (from-to)9814-9822
Number of pages9
JournalJournal of Biological Chemistry
Volume275
Issue number13
DOIs
StatePublished - Mar 31 2000
Externally publishedYes

Fingerprint

Serum Response Factor
Smooth Muscle
Muscle
Genes
Smooth Muscle Myocytes
calponin
Cells
Cell Lineage
Site-Directed Mutagenesis
Mutagenesis
Adenoviridae
Introns
Physiology
Sequence Analysis
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Serum response factor-dependent regulation of the smooth muscle calponin gene. / Miano, Joseph M.; Carlson, Michael J.; Spencer, Jeffrey A.; Misra, Ravi P.

In: Journal of Biological Chemistry, Vol. 275, No. 13, 31.03.2000, p. 9814-9822.

Research output: Contribution to journalArticle

Miano, Joseph M. ; Carlson, Michael J. ; Spencer, Jeffrey A. ; Misra, Ravi P. / Serum response factor-dependent regulation of the smooth muscle calponin gene. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 13. pp. 9814-9822.
@article{12b12725ac6740e284744f1f11679ce0,
title = "Serum response factor-dependent regulation of the smooth muscle calponin gene",
abstract = "Smooth muscle calponin is a multifunctional, thin filament-associated protein whose expression is restricted to smooth muscle cell lineages in developing and postnatal tissues. Although the physiology of smooth muscle calponin has been studied extensively, the cis-elements governing its restricted pattern of expression have yet to be identified. Here we report on smooth muscle-specific enhancer activity within the first intron of smooth muscle calponin. Sequence analysis revealed a proximal consensus intronic CArG box and two distal intronic CArG-like elements, each of which bound recombinant serum response factor (SRF) as well as immunoreactive SRF from smooth muscle nuclear extracts. Site-directed mutagenesis studies suggested that the consensus CArG box mediates much of the intronic enhancer activity; mutating all three CArG elements abolished the ability of SRF to confer enhancer activity on the smooth muscle calponin promoter. Cotransfecting a dominant-negative SRF construct attenuated smooth muscle-specific enhancer activity, and transducing smooth muscle cells with adenovirus harboring the dominant-negative SRF construct selectively reduced steady-state expression of endogenous smooth muscle calponin. These results demonstrate an important role for intronic CArG boxes and the SRF protein in the transcriptional control of smooth muscle calponin in vitro.",
author = "Miano, {Joseph M.} and Carlson, {Michael J.} and Spencer, {Jeffrey A.} and Misra, {Ravi P.}",
year = "2000",
month = "3",
day = "31",
doi = "10.1074/jbc.275.13.9814",
language = "English (US)",
volume = "275",
pages = "9814--9822",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "13",

}

TY - JOUR

T1 - Serum response factor-dependent regulation of the smooth muscle calponin gene

AU - Miano, Joseph M.

AU - Carlson, Michael J.

AU - Spencer, Jeffrey A.

AU - Misra, Ravi P.

PY - 2000/3/31

Y1 - 2000/3/31

N2 - Smooth muscle calponin is a multifunctional, thin filament-associated protein whose expression is restricted to smooth muscle cell lineages in developing and postnatal tissues. Although the physiology of smooth muscle calponin has been studied extensively, the cis-elements governing its restricted pattern of expression have yet to be identified. Here we report on smooth muscle-specific enhancer activity within the first intron of smooth muscle calponin. Sequence analysis revealed a proximal consensus intronic CArG box and two distal intronic CArG-like elements, each of which bound recombinant serum response factor (SRF) as well as immunoreactive SRF from smooth muscle nuclear extracts. Site-directed mutagenesis studies suggested that the consensus CArG box mediates much of the intronic enhancer activity; mutating all three CArG elements abolished the ability of SRF to confer enhancer activity on the smooth muscle calponin promoter. Cotransfecting a dominant-negative SRF construct attenuated smooth muscle-specific enhancer activity, and transducing smooth muscle cells with adenovirus harboring the dominant-negative SRF construct selectively reduced steady-state expression of endogenous smooth muscle calponin. These results demonstrate an important role for intronic CArG boxes and the SRF protein in the transcriptional control of smooth muscle calponin in vitro.

AB - Smooth muscle calponin is a multifunctional, thin filament-associated protein whose expression is restricted to smooth muscle cell lineages in developing and postnatal tissues. Although the physiology of smooth muscle calponin has been studied extensively, the cis-elements governing its restricted pattern of expression have yet to be identified. Here we report on smooth muscle-specific enhancer activity within the first intron of smooth muscle calponin. Sequence analysis revealed a proximal consensus intronic CArG box and two distal intronic CArG-like elements, each of which bound recombinant serum response factor (SRF) as well as immunoreactive SRF from smooth muscle nuclear extracts. Site-directed mutagenesis studies suggested that the consensus CArG box mediates much of the intronic enhancer activity; mutating all three CArG elements abolished the ability of SRF to confer enhancer activity on the smooth muscle calponin promoter. Cotransfecting a dominant-negative SRF construct attenuated smooth muscle-specific enhancer activity, and transducing smooth muscle cells with adenovirus harboring the dominant-negative SRF construct selectively reduced steady-state expression of endogenous smooth muscle calponin. These results demonstrate an important role for intronic CArG boxes and the SRF protein in the transcriptional control of smooth muscle calponin in vitro.

UR - http://www.scopus.com/inward/record.url?scp=0034737603&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034737603&partnerID=8YFLogxK

U2 - 10.1074/jbc.275.13.9814

DO - 10.1074/jbc.275.13.9814

M3 - Article

C2 - 10734136

AN - SCOPUS:0034737603

VL - 275

SP - 9814

EP - 9822

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 13

ER -