Contribution of serum response factor and myocardin to transcriptional regulation of smoothelins

Sander S.M. Rensen, Petra M.G. Niessen, Xiaochun Long, Pieter A. Doevendans, Joseph M. Miano, Guillaume J.J.M. Van Eys

Research output: Contribution to journalArticle

Abstract

Objective: Smoothelin-A and -B isoforms are highly restricted to contractile smooth muscle cells (SMCs). Serum response factor (SRF) and myocardin are essential for contractile SMC differentiation. We evaluated the contribution of SRF/myocardin to transcriptional regulation of smoothelins. Methods: Rat vascular SMCs were transfected with smoothelin-A and smoothelin-B promoter reporter constructs and promoter activity was analyzed. The effects of mutations in the smoothelin-A promoter CArG-boxes and co-transfections with a myocardin expression plasmid were assessed. Electrophoretic mobility shift assays and chromatin immunoprecipitations were performed to investigate SRF-binding to the smoothelin-A CArG-boxes. Results: Smoothelin promoter activity was detected in vascular SMCs. Comparative sequence analysis revealed two conserved CArG elements in the smoothelin-A promoter that bind SRF as shown by chromatin immunoprecipitation. The proximal CArG-near bound SRF stronger than CArG-far in gel shift assays. Mutagenesis studies also indicated that CArG-near is more important than CArG-far in regulating smoothelin-A promoter activity. Myocardin augmented smoothelin-A promoter activity 2.5-fold in a CArG-near-dependent manner. In contrast, myocardin had little effect on the smoothelin-B promoter. Conclusion: Smoothelin-A expression is controlled by an intragenic promoter whose activity is, in part, dependent on two CArG boxes that bind SRF. Our data show a role for SRF/myocardin in regulating smoothelin-A whereas the higher smoothelin-B expression appears to be SRF/myocardin- independent.

Original languageEnglish (US)
Pages (from-to)136-145
Number of pages10
JournalCardiovascular Research
Volume70
Issue number1
DOIs
StatePublished - Apr 1 2006
Externally publishedYes

Fingerprint

Serum Response Factor
Smooth Muscle Myocytes
Chromatin Immunoprecipitation
Vascular Smooth Muscle
myocardin
Electrophoretic Mobility Shift Assay
Mutagenesis
Transfection
Sequence Analysis
Cell Differentiation
Protein Isoforms
Plasmids
Gels
Mutation

Keywords

  • Gene expression
  • Myocardin
  • Serum response factor
  • Smooth muscle cell differentiation
  • Smoothelins

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

Cite this

Rensen, S. S. M., Niessen, P. M. G., Long, X., Doevendans, P. A., Miano, J. M., & Van Eys, G. J. J. M. (2006). Contribution of serum response factor and myocardin to transcriptional regulation of smoothelins. Cardiovascular Research, 70(1), 136-145. https://doi.org/10.1016/j.cardiores.2005.12.018

Contribution of serum response factor and myocardin to transcriptional regulation of smoothelins. / Rensen, Sander S.M.; Niessen, Petra M.G.; Long, Xiaochun; Doevendans, Pieter A.; Miano, Joseph M.; Van Eys, Guillaume J.J.M.

In: Cardiovascular Research, Vol. 70, No. 1, 01.04.2006, p. 136-145.

Research output: Contribution to journalArticle

Rensen, SSM, Niessen, PMG, Long, X, Doevendans, PA, Miano, JM & Van Eys, GJJM 2006, 'Contribution of serum response factor and myocardin to transcriptional regulation of smoothelins', Cardiovascular Research, vol. 70, no. 1, pp. 136-145. https://doi.org/10.1016/j.cardiores.2005.12.018
Rensen, Sander S.M. ; Niessen, Petra M.G. ; Long, Xiaochun ; Doevendans, Pieter A. ; Miano, Joseph M. ; Van Eys, Guillaume J.J.M. / Contribution of serum response factor and myocardin to transcriptional regulation of smoothelins. In: Cardiovascular Research. 2006 ; Vol. 70, No. 1. pp. 136-145.
@article{963c2b39f90f43cdafabada2dee7f0cb,
title = "Contribution of serum response factor and myocardin to transcriptional regulation of smoothelins",
abstract = "Objective: Smoothelin-A and -B isoforms are highly restricted to contractile smooth muscle cells (SMCs). Serum response factor (SRF) and myocardin are essential for contractile SMC differentiation. We evaluated the contribution of SRF/myocardin to transcriptional regulation of smoothelins. Methods: Rat vascular SMCs were transfected with smoothelin-A and smoothelin-B promoter reporter constructs and promoter activity was analyzed. The effects of mutations in the smoothelin-A promoter CArG-boxes and co-transfections with a myocardin expression plasmid were assessed. Electrophoretic mobility shift assays and chromatin immunoprecipitations were performed to investigate SRF-binding to the smoothelin-A CArG-boxes. Results: Smoothelin promoter activity was detected in vascular SMCs. Comparative sequence analysis revealed two conserved CArG elements in the smoothelin-A promoter that bind SRF as shown by chromatin immunoprecipitation. The proximal CArG-near bound SRF stronger than CArG-far in gel shift assays. Mutagenesis studies also indicated that CArG-near is more important than CArG-far in regulating smoothelin-A promoter activity. Myocardin augmented smoothelin-A promoter activity 2.5-fold in a CArG-near-dependent manner. In contrast, myocardin had little effect on the smoothelin-B promoter. Conclusion: Smoothelin-A expression is controlled by an intragenic promoter whose activity is, in part, dependent on two CArG boxes that bind SRF. Our data show a role for SRF/myocardin in regulating smoothelin-A whereas the higher smoothelin-B expression appears to be SRF/myocardin- independent.",
keywords = "Gene expression, Myocardin, Serum response factor, Smooth muscle cell differentiation, Smoothelins",
author = "Rensen, {Sander S.M.} and Niessen, {Petra M.G.} and Xiaochun Long and Doevendans, {Pieter A.} and Miano, {Joseph M.} and {Van Eys}, {Guillaume J.J.M.}",
year = "2006",
month = "4",
day = "1",
doi = "10.1016/j.cardiores.2005.12.018",
language = "English (US)",
volume = "70",
pages = "136--145",
journal = "Cardiovascular Research",
issn = "0008-6363",
publisher = "Oxford University Press",
number = "1",

}

TY - JOUR

T1 - Contribution of serum response factor and myocardin to transcriptional regulation of smoothelins

AU - Rensen, Sander S.M.

AU - Niessen, Petra M.G.

AU - Long, Xiaochun

AU - Doevendans, Pieter A.

AU - Miano, Joseph M.

AU - Van Eys, Guillaume J.J.M.

PY - 2006/4/1

Y1 - 2006/4/1

N2 - Objective: Smoothelin-A and -B isoforms are highly restricted to contractile smooth muscle cells (SMCs). Serum response factor (SRF) and myocardin are essential for contractile SMC differentiation. We evaluated the contribution of SRF/myocardin to transcriptional regulation of smoothelins. Methods: Rat vascular SMCs were transfected with smoothelin-A and smoothelin-B promoter reporter constructs and promoter activity was analyzed. The effects of mutations in the smoothelin-A promoter CArG-boxes and co-transfections with a myocardin expression plasmid were assessed. Electrophoretic mobility shift assays and chromatin immunoprecipitations were performed to investigate SRF-binding to the smoothelin-A CArG-boxes. Results: Smoothelin promoter activity was detected in vascular SMCs. Comparative sequence analysis revealed two conserved CArG elements in the smoothelin-A promoter that bind SRF as shown by chromatin immunoprecipitation. The proximal CArG-near bound SRF stronger than CArG-far in gel shift assays. Mutagenesis studies also indicated that CArG-near is more important than CArG-far in regulating smoothelin-A promoter activity. Myocardin augmented smoothelin-A promoter activity 2.5-fold in a CArG-near-dependent manner. In contrast, myocardin had little effect on the smoothelin-B promoter. Conclusion: Smoothelin-A expression is controlled by an intragenic promoter whose activity is, in part, dependent on two CArG boxes that bind SRF. Our data show a role for SRF/myocardin in regulating smoothelin-A whereas the higher smoothelin-B expression appears to be SRF/myocardin- independent.

AB - Objective: Smoothelin-A and -B isoforms are highly restricted to contractile smooth muscle cells (SMCs). Serum response factor (SRF) and myocardin are essential for contractile SMC differentiation. We evaluated the contribution of SRF/myocardin to transcriptional regulation of smoothelins. Methods: Rat vascular SMCs were transfected with smoothelin-A and smoothelin-B promoter reporter constructs and promoter activity was analyzed. The effects of mutations in the smoothelin-A promoter CArG-boxes and co-transfections with a myocardin expression plasmid were assessed. Electrophoretic mobility shift assays and chromatin immunoprecipitations were performed to investigate SRF-binding to the smoothelin-A CArG-boxes. Results: Smoothelin promoter activity was detected in vascular SMCs. Comparative sequence analysis revealed two conserved CArG elements in the smoothelin-A promoter that bind SRF as shown by chromatin immunoprecipitation. The proximal CArG-near bound SRF stronger than CArG-far in gel shift assays. Mutagenesis studies also indicated that CArG-near is more important than CArG-far in regulating smoothelin-A promoter activity. Myocardin augmented smoothelin-A promoter activity 2.5-fold in a CArG-near-dependent manner. In contrast, myocardin had little effect on the smoothelin-B promoter. Conclusion: Smoothelin-A expression is controlled by an intragenic promoter whose activity is, in part, dependent on two CArG boxes that bind SRF. Our data show a role for SRF/myocardin in regulating smoothelin-A whereas the higher smoothelin-B expression appears to be SRF/myocardin- independent.

KW - Gene expression

KW - Myocardin

KW - Serum response factor

KW - Smooth muscle cell differentiation

KW - Smoothelins

UR - http://www.scopus.com/inward/record.url?scp=33645057677&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33645057677&partnerID=8YFLogxK

U2 - 10.1016/j.cardiores.2005.12.018

DO - 10.1016/j.cardiores.2005.12.018

M3 - Article

C2 - 16451796

AN - SCOPUS:33645057677

VL - 70

SP - 136

EP - 145

JO - Cardiovascular Research

JF - Cardiovascular Research

SN - 0008-6363

IS - 1

ER -