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Molecular regulation of the bovine endothelial cell nitric oxide synthase by transforming growth factor-beta 1.

Inoue N, Venema RC, Sayegh HS, Ohara Y, Murphy TJ, Harrison DG
Arterioscler Thromb Vasc Biol. 1995 15 (8): 1255-61

PMID: 7543000 · DOI:10.1161/01.atv.15.8.1255

The promoter region of the endothelial cell nitric oxide synthase (ecNOS) gene contains potential response elements for transforming growth factor-beta 1 (TGF beta 1). TGF beta 1 plays an important role in the pathogenesis of atherosclerosis, vascular hypertrophy, and angiogenesis. We therefore sought to determine whether TGF beta 1 might modulate ecNOS expression in bovine aortic endothelial cells (BAEC). TGF beta 1 increased ecNOS mRNA in a dose-dependent manner. TGF beta 1 also increased ecNOS protein content. The production of nitrogen oxides (NOx), assessed by chemiluminescence, and nitric oxide synthase activity, assessed by arginine/citrulline conversion were increased in TGF beta 1-treated cells. Transcriptional activity of the 5'-flanking promoter region of the ecNOS gene was increased by TGF beta 1, as assessed by transfection with promoter/luciferase constructs. Deletion analysis suggested that the TGF beta 1-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 in a sequence-specific manner. Mutation of the putative NF-1 binding site in the promoter/luciferase construct significantly decreased the responsiveness to TGF beta 1. In conclusion, TGF beta 1 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.

MeSH Terms (22)

Amino Acid Oxidoreductases Animals Base Sequence Binding Sites Cattle Cell Division Cell Nucleus Cells, Cultured DNA-Binding Proteins Endothelium, Vascular In Vitro Techniques Molecular Sequence Data Mutagenesis, Site-Directed NFI Transcription Factors Nitric Oxide Nitric Oxide Synthase Oligodeoxyribonucleotides Promoter Regions, Genetic Structure-Activity Relationship Transcription, Genetic Transcription Factors Transforming Growth Factor beta

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