Non-NF-κ-B elements are required for full induction of the rat type II nitric oxide synthase in vascular smooth muscle cells

Hanfang Zhang, Xingwu Teng, Connie Snead, John D. Catravas

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

1. We have investigated the role of the NF-κB binding sites and other promoter elements beyond NF-κB in iNOS induction in rat vascular smooth muscle cells (SMC). 2. Rat aortic SMC transfected with iNOS promoter constructs with either mutation or deletion of the downstream NF-κB site exhibited about 50% reduction in promoter activity in response to a cytokine mixture, whereas either mutation or deletion of the upstream NF-κB site reduced promoter activity by 90%, suggesting that the latter site is the most important, and that co-existence of two NF-κB sites is necessary for iNOS induction. 3. Nuclear NF-κB activity was robustly induced by TNF-α. However, TNF-α alone did not induce iNOS promoter activity, protein expression, or nitrite production, indicating that NF-κB activation alone is not sufficient for iNOS induction. 4. The construct up to -890 bp, containing the downstream NF-κB site, exhibited little response to cytokines. The construct up to -1.0 kb, containing the two NF-κB sites exhibited only 22% of full promoter activity. The regions -1001 to -1368 bp and -2 to -2.5 kb contributed an additional 43 and 22% promoter activity, respectively. 5. Internal deletion or reversal of the orientation of -1001 to -1368 bp in the full promoter resulted in 40% reduction in promoter activity. 6. These data suggest that the co-existence of two NF-κB sites is essential for core promoter activity, but that full induction of the rat SMC iNOS gene requires other elements located between -1.0 to -1.37 and -2.0 to -2.5 kb of the promoter.

Original languageEnglish (US)
Pages (from-to)270-278
Number of pages9
JournalBritish Journal of Pharmacology
Volume130
Issue number2
DOIs
StatePublished - Jan 1 2000

Fingerprint

Nitric Oxide Synthase Type II
Vascular Smooth Muscle
Smooth Muscle Myocytes
Sequence Deletion
Cytokines
Nitrites
Binding Sites
Genes
Proteins

Keywords

  • Deletion
  • Mutation
  • NF-κB
  • Rat iNOS promoter
  • Smooth muscle cells
  • Transfection
  • iNOS gene induction

ASJC Scopus subject areas

  • Pharmacology

Cite this

Non-NF-κ-B elements are required for full induction of the rat type II nitric oxide synthase in vascular smooth muscle cells. / Zhang, Hanfang; Teng, Xingwu; Snead, Connie; Catravas, John D.

In: British Journal of Pharmacology, Vol. 130, No. 2, 01.01.2000, p. 270-278.

Research output: Contribution to journalArticle

Zhang, Hanfang ; Teng, Xingwu ; Snead, Connie ; Catravas, John D. / Non-NF-κ-B elements are required for full induction of the rat type II nitric oxide synthase in vascular smooth muscle cells. In: British Journal of Pharmacology. 2000 ; Vol. 130, No. 2. pp. 270-278.
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