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Requirement of the hypoxia-responsive enhancer in the activation of the inducible nitric oxide synthase promoter by desferrioxamine

  1. Author:
    Bosco, M. C.
    Brooks, A.
    Musso, T.
    Taylor, L.
    Cox, G. W.
    Varesio, L.
    Melillo, G.
    1. Year of Conference: 1997
  2. Conference Name: American Association For Cancer Research Annual Meeting
    1. 38
    2. Pages: A3624
  4. Type of Work: Meeting Abstract
  1. Abstract:

    The activation of the inducible nitric oxide synthase (iNOS) promoter by picolinic acid or hypoxia in IFNgamma-treated murine macrophages is mediated by a 19-base pair element of the 5'-flanking region of the iNOS gene containing a sequence homology to the hypoxia-responsive enhancer (HRE) of the human erythropoietin (hEpo) promoter. The iron chelator desferrioxamine (DFX) induces the activity of the hEpo enhancer in Hep3B cells. We investigated the effects of DFX on the activation of the iNOS promoter and iNOS gene expression in ANA-1 murine macrophages. We found that DFX induced DNA-binding activity to the hypoxia-inducible factor 1 consensus sequence of the iNOS promoter and activated the iNOS-HRE. These activities were associated with a synergistic induction of iNOS transcription and iNOS mRNA expression in IFNgamma-treated ANA-1 cells. Functional analysis of the 5'-flanking region of the iNOS gene demonstrated that IFNgamma plus DFX activated the full-length iNOS promoter and that the iNOS-HRE was required for iNOS transcriptional activation by DFX. These data demonstrate that DFX is a costimulus for the transcriptional induction of the iNOS gene in IFNgamma-treated macrophages and that the DFX-dependent activation of the iNOS promoter is mediated by the iNOS-HRE.

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