RGD Reference Report - Hypoxia up-regulates prolyl hydroxylase activity: a feedback mechanism that limits HIF-1 responses during reoxygenation. - Chinchilla Research Resource Database
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Hypoxia up-regulates prolyl hydroxylase activity: a feedback mechanism that limits HIF-1 responses during reoxygenation.

Authors: D'Angelo, G  Duplan, E  Boyer, N  Vigne, P  Frelin, C 
Citation: D'Angelo G, etal., J Biol Chem 2003 Oct 3;278(40):38183-7.
CRRD ID: 727754
Pubmed: (View Article at PubMed) PMID:12876291
DOI: Full-text: DOI:10.1074/jbc.M302244200

The mechanism by which hypoxia induces gene transcription is now well established. Hypoxia reduces activity of prolyl hydroxylases (PHD) that hydroxylate specific proline residues in the oxygen-dependent degradation domain (ODD) of hypoxia-inducible factor-1alpha (HIF-1alpha). As a consequence, HIF-1alpha accumulates and promotes hypoxic tolerance by activating gene transcription. This paper identifies the three forms of PHDs in rats and shows that a period of hypoxia selectively increases expression of PHD-2 mRNAs levels. We developed assays for PHD activity that used (i) the peptide-specific conversion of labeled 2-oxoglutarate into succinate and (ii) the binding of the von Hippel-Lindau protein to a glutathione S-transferase-ODD fusion protein. The two assays indicated a low enzymatic activity in normoxic and hypoxic cells and a rapid increase during reoxygenation. We also developed hydroxyproline-specific antibodies that recognized hydroxylated forms of a fusion protein (ODD-green fluorescent protein) that combined the ODD domain of HIF-1alpha and the green fluorescent protein. Using this antibody, we demonstrated that reoxygenation induced a rapid hydroxylation of Pro-564, which was followed by a massive degradation of the proteins. The results suggest that a hypoxic upregulation of PHD (presumably PHD-2) acts as a feedback mechanism to stop hypoxic responses in reoxygenated cells. We propose that proline hydroxylation might play a role in hypoxic preconditioning.


Gene Ontology Annotations    

Biological Process

Molecular Pathway Annotations    
Objects Annotated

Objects referenced in this article
0 Egln3 egl-9 family hypoxia-inducible factor 3 All species
0 Hif1a hypoxia inducible factor 1 subunit alpha All species

Additional Information



RGD is funded by grant HL64541 from the National Heart, Lung, and Blood Institute on behalf of the NIH.