Angiotensin II-induced stimulation of smooth muscle α-actin expression by serum response factor and the homeodomain transcription factor MHox

Martina B. Hautmann, Maria M. Thompson, Ellen A. Swartz, Eric N. Olson, Gary K. Owens

Research output: Contribution to journalArticlepeer-review

105 Scopus citations

Abstract

The objective of the present study was to examine the molecular mechanisms whereby angiotensin II (Ang II) stimulates smooth muscle (SM) α- actin expression in rat aortic smooth muscle cells (SMCs). Nuclear run-on analysis and transfection studies indicated that the effects of Ang II on SM α-actin were mediated at least in part at the transcriptional level. Transfection of various rat SM α-actin promoter/chloramphenicol acetyltransferase (CAT) constructs into SMCs demonstrated that the first 155 bp of the SM α-actin promoter was sufficient to confer maximal Ang II responsiveness, conferring an ≃4-fold increase in reporter activities in these SMCs compared with vehicle-treated SMCs. Mutation of either of two highly conserved CArG elements, designated A (-62) and B (-112), completely abolished Ang II-induced increases in reporter activity, whereas mutation of a homeodomain-like binding sequence at -145 (ATTA) reduced reporter activity by half. Results of EMSAs showed that nuclear extracts from Ang II-treated SMCs exhibited enhanced binding activity of serum response factor (SRF) to the CArG elements and of a homeo-domain factor, MHox, to the ATTA element Northern analyses showed that Ang II also stimulated marked increases in MHox mRNA levels. Western analyses demonstrated that Ang II-induced increases in SRF binding were not due to increased SRF protein expression. Recombinant MHox markedly enhanced binding activity of SRF in EMSAs. Finally, MHox overexpression transactivated a SM α-actin promoter/CAT reporter construct by ≃3.5-fold in transient cotransfection studies. These results provide evidence for involvement of a homeodomain transcription factor, MHox, in Ang II-mediated stimulation of SM α-actin via a CArG/SRF-dependent mechanism.

Original languageEnglish (US)
Pages (from-to)600-610
Number of pages11
JournalCirculation research
Volume81
Issue number4
DOIs
StatePublished - 1997

Keywords

  • Angiotensin II
  • MHox
  • Smooth muscle α-actin promoter
  • Vascular smooth muscle cell

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

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