Abstract
During hypoxia, hypoxia-inducible factor-1alpha (HIF-1alpha) is required for induction of a variety of genes including erythropoietin and vascular endothelial growth factor. Hypoxia increases mitochondrial reactive oxygen species (ROS) generation at Complex III, which causes accumulation of HIF-1alpha protein responsible for initiating expression of a luciferase reporter construct under the control of a hypoxic response element. This response is lost in cells depleted of mitochondrial DNA (rho(0) cells). Overexpression of catalase abolishes hypoxic response element-luciferase expression during hypoxia. Exogenous H(2)O(2) stabilizes HIF-1alpha protein during normoxia and activates luciferase expression in wild-type and rho(0) cells. Isolated mitochondria increase ROS generation during hypoxia, as does the bacterium Paracoccus denitrificans. These findings reveal that mitochondria-derived ROS are both required and sufficient to initiate HIF-1alpha stabilization during hypoxia.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Androstadienes / pharmacology
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Animals
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Cell Line
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Cell Nucleus / metabolism
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Chelating Agents / pharmacology
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Cobalt / pharmacology
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Cytosol / chemistry
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DNA-Binding Proteins / metabolism*
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Deferoxamine / pharmacology
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Dose-Response Relationship, Drug
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Electron Transport Complex III / chemistry*
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Electron Transport Complex III / metabolism*
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Electron Transport Complex IV / metabolism
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Enzyme Inhibitors / pharmacology
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Genes, Reporter
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Humans
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Hydrogen Peroxide / metabolism
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Hypoxia*
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Hypoxia-Inducible Factor 1
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Hypoxia-Inducible Factor 1, alpha Subunit
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Immunoblotting
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Marine Toxins
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Mitochondria / enzymology
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Mitochondria / metabolism*
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Mitochondria, Liver / metabolism
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Models, Biological
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Nuclear Proteins / metabolism*
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Oxazoles / pharmacology
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Oxidation-Reduction
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Oxygen / metabolism
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Paracoccus denitrificans / metabolism
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Rats
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Reactive Oxygen Species / metabolism*
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Time Factors
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Transcription Factors*
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Transfection
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Tumor Cells, Cultured
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Wortmannin
Substances
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Androstadienes
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Chelating Agents
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DNA-Binding Proteins
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Enzyme Inhibitors
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HIF1A protein, human
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Hif1a protein, rat
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Hypoxia-Inducible Factor 1
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Hypoxia-Inducible Factor 1, alpha Subunit
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Marine Toxins
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Nuclear Proteins
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Oxazoles
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Reactive Oxygen Species
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Transcription Factors
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Cobalt
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calyculin A
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Hydrogen Peroxide
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Electron Transport Complex IV
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Electron Transport Complex III
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cobaltous chloride
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Deferoxamine
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Oxygen
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Wortmannin