Smooth muscle γ-actin promoter regulation by RhoA and serum response factor signaling

James Carson, Donald E. Culberson, Raymond W. Thompson, Rebecca A. Fillmore, Warren Zimmer

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Smooth muscle γ-actin (SMGA) is both an early marker of smooth muscle cell differentiation, which demonstrates an expression pattern restricted to smooth muscle and the post meiotic spermatocyte. Serum response factor (SRF) DNA-binding is an important regulator of muscle differentiation, including SMGA expression during smooth muscle cell differentiation. RhoA, a low molecular weight GTPase protein, can regulate cardiac, skeletal, and smooth muscle differentiation through SRF-dependent mechanisms. This study's purpose was to examine RhoA expression during smooth muscle cell development, and determine if the SMGA promoter activity is sensitive to RhoA-mediated signaling through SRF. Additionally, the study identified the promoter regulation modifying SMGA expression by RhoA signaling. Western blot analysis of embryonic chick gizzard whole protein extracts during 5 to 14 days of development demonstrated a large induction of RhoA (10-fold) and β1 integrin expression at day 8, which corresponds to the time SMGA expression and differentiation are occurring. Transient transfections in CV-1 fibroblast cells demonstrated that co-overexpression of SRF and RhoA could induce a 40-fold induction of -176 bp SMGA promoter activity. Mutational analysis demonstrated that serum response element (SRE)-1, but not SRE2, was necessary for RhoA/SRF activation of the SMGA promoter. Deletion analysis revealed that although SRE1 was necessary for SMGA promoter activation by RhoA and SRF, it was not sufficient, implicating a possible obligatory role of additional promoter sequences in the response. Overexpression of a mutated SRF protein that was unable to bind DNA demonstrated that the 40-fold RhoA/SRF activation was largely dependent on SRF binding to the SMGA promoter. Thus, as the SMGA promoter appears to be a target of RhoA-mediated transcriptional regulation, the uncovering of these signaling mechanisms effecting SMGA promoter activity should provide a regulatory paradigm that can then be examined during the regulation of other smooth muscle genes.

Original languageEnglish (US)
Pages (from-to)133-139
Number of pages7
JournalBiochimica et Biophysica Acta - Gene Structure and Expression
Volume1628
Issue number2
DOIs
StatePublished - Jul 28 2003
Externally publishedYes

Fingerprint

Serum Response Factor
Smooth Muscle
Muscle
Actins
Cells
Smooth Muscle Myocytes
Chemical activation
Cell Differentiation
Serum Response Element
Proteins
Spermatocytes
Muscle Development
GTP Phosphohydrolases
DNA
Fibroblasts
Integrins
Transfection
Myocardium
Skeletal Muscle
Genes

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Biophysics
  • Biochemistry
  • Genetics

Cite this

Smooth muscle γ-actin promoter regulation by RhoA and serum response factor signaling. / Carson, James; Culberson, Donald E.; Thompson, Raymond W.; Fillmore, Rebecca A.; Zimmer, Warren.

In: Biochimica et Biophysica Acta - Gene Structure and Expression, Vol. 1628, No. 2, 28.07.2003, p. 133-139.

Research output: Contribution to journalArticle

Carson, James ; Culberson, Donald E. ; Thompson, Raymond W. ; Fillmore, Rebecca A. ; Zimmer, Warren. / Smooth muscle γ-actin promoter regulation by RhoA and serum response factor signaling. In: Biochimica et Biophysica Acta - Gene Structure and Expression. 2003 ; Vol. 1628, No. 2. pp. 133-139.
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