Molecular regulation of the bovine endothelial cell nitric oxide synthase by transforming growth factor-β1

Nobutaka Inoue, Richard C. Venema, Hassan S. Sayegh, Yuichi Ohara, T. J. Murphy, David G. Harrison

Research output: Contribution to journalArticlepeer-review

176 Scopus citations

Abstract

The promoter region of the endothelial cell nitric oxide synthase (ecNOS) gene contains potential response elements for transforming growth factor- β1 (TGFβ1). TGFβ1 plays an important role in the pathogenesis of atherosclerosis, vascular hypertrophy, and angiogenesis. We therefore sought to determine whether TGFβ1 might modulate ecNOS expression in bovine aortic endothelial cells (BAEC). TGFβ1 increased ecNOS mRNA in a dose-dependent manner. TGFβ1 also increased ecNOS protein content. The production of nitrogen oxides (NO(x)), assessed by chemiluminescence, and nitric oxide synthase activity, assessed by arginine/citrulline conversion, were increased in TGFβ1-treated cells. Transcriptional activity of the 5'-flanking promoter region of the ecNOS gene was increased by TGFβ1, as assessed by transfection with promoter/luciferase constructs. Deletion analysis suggested that the TGFβ1-response element was present between nucleotides -1269 and - 935 from the first transcription start site, in which a putative nuclear factor-1 (NF-1) binding site existed. Gel shift assays showed that nuclear protein(s), immunologically similar to CCAAT transcription factor/NF-1, bound to the putative NF-1 binding site a sequence-specific manner. Mutation of the putative NF-1 binding site in the promoter/luciferase construct significantly decreased the responsiveness to TGFβ1. In conclusion, TGFβ1 increases ecNOS expression associated with an increase in production of NO in BAEC. This response is probably mediated by transcriptional activation of the ecNOS gene promoter.

Original languageEnglish (US)
Pages (from-to)1255-1261
Number of pages7
JournalArteriosclerosis, thrombosis, and vascular biology
Volume15
Issue number8
DOIs
StatePublished - Aug 1995

Keywords

  • endothelium
  • nitric oxide synthase
  • nuclear factor-1
  • transforming growth factor- β

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

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