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Am J Physiol Cell Physiol 283: C178-C186, 2002. First published February 20, 2002; doi:10.1152/ajpcell.00381.2001
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Vol. 283, Issue 1, C178-C186, July 2002

Role of nitric oxide in the regulation of HIF-1alpha expression during hypoxia

Faton H. Agani, Michelle Puchowicz, Juan Carlos Chavez, Paola Pichiule, and Joseph LaManna

Department of Anatomy, School of Medicine, Case Western Reserve University, Cleveland, Ohio 44106-4938

Hypoxia-inducible factor-1 (HIF-1), a heterodimeric transcription factor consisting of HIF-1alpha and HIF-1beta subunits, controls the expression of a large number of genes involved in the regulation of cellular responses to reduced oxygen availability. The oxygen-regulated subunit, HIF-1alpha , is stabilized in cells exposed to hypoxia. The regulation of hypoxic responses by nitric oxide (NO) is believed to have wide pathophysiological relevance, thus we investigated whether NO affects HIF-1 activation in hypoxic cells. Here we show that NO generated from NO donors prevented HIF-1alpha hypoxic accumulation in Hep 3B and PC-12 cells. Addition of a glutathione analog or peroxynitrite scavengers prevented the NO-induced inhibition of HIF-1alpha accumulation in both cell lines. Exposure to NO was associated with inhibition of mitochondrial electron transport and compensatory glycolysis, which maintained normal cellular ATP content. Succinate, a Krebs cycle intermediate and respiratory chain substrate, restored HIF-1alpha hypoxic induction in the cells, suggesting involvement of mitochondria in regulation of HIF-1alpha accumulation during hypoxia. Regulation of HIF-1alpha by NO is an additional important mechanism by which NO might modulate cellular responses to hypoxia in mammalian cells.

hypoxia-inducible factor-1; mitochondria; oxygen sensing


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