Reactive oxygen species activate the HIF-1alpha promoter via a functional NFkappaB site

Arterioscler Thromb Vasc Biol. 2007 Apr;27(4):755-61. doi: 10.1161/01.ATV.0000258979.92828.bc. Epub 2007 Feb 1.


Objective: Reactive oxygen species have been implicated as signaling molecules modulating the activity of redox-sensitive transcription factors such as nuclear factor kappa B (NF-kappaB). Recently, the transcription factor hypoxia-inducible factor-1 (HIF-1), known to mediate gene expression by hypoxia, has been found to be also activated by nonhypoxic factors in a redox-sensitive manner. We therefore aimed to elucidate the link between these 2 important redox-sensitive transcription factors.

Methods and results: In pulmonary artery smooth muscle cells, reactive oxygen species generated either by exogenous H2O2 or by a NOX4-containing NADPH oxidase stimulated by thrombin activated or induced NF-kappaB and HIF-1alpha. The reactive oxygen species-mediated HIF-1alpha induction occurred on the transcriptional level and was dependent on NF-kappaB. Transfection experiments with wild-type or mutant HIF-1alpha promoter constructs revealed the presence of a yet unidentified NF-kappaB binding element. Gel shift analyses and chromatin immunoprecipitation verified binding of NF-kappaB to this site. Furthermore, reactive oxygen species enhanced expression of plasminogen activator inhibitor-1, which was prevented by dominant-negative IkappaB or mutation of the HIF-1 binding site within the plasminogen activator inhibitor-1 promoter.

Conclusion: These findings show for the first time to our knowledge that reactive oxygen species directly link HIF-1alpha and NF-kappaB, implicating an important pathophysiological role of this novel pathway in disorders associated with elevated levels of reactive oxygen species.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Cells, Cultured
  • Gene Expression Regulation
  • Humans
  • Hydrogen Peroxide / pharmacology
  • Hypoxia-Inducible Factor 1, alpha Subunit / genetics*
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism
  • Myocytes, Smooth Muscle / metabolism
  • NF-kappa B / metabolism*
  • NF-kappa B / physiology
  • Plasminogen Activator Inhibitor 1 / genetics
  • Promoter Regions, Genetic / physiology*
  • Pulmonary Artery / cytology
  • RNA, Messenger / metabolism
  • Reactive Oxygen Species / metabolism*
  • Thrombin / pharmacology
  • Transcription, Genetic / physiology


  • Hypoxia-Inducible Factor 1, alpha Subunit
  • NF-kappa B
  • Plasminogen Activator Inhibitor 1
  • RNA, Messenger
  • Reactive Oxygen Species
  • Hydrogen Peroxide
  • Thrombin