HIF-1alpha binding to VHL is regulated by stimulus-sensitive proline hydroxylation.

Hypoxia-inducible factor-1alpha (HIF-1alpha) is a global transcriptional regulator of the hypoxic response. Under normoxic conditions, HIF-1alpha is recognized by the von Hippel-Lindau tumor-suppressor protein (VHL), a component of an E3 ubiquitin ligase complex. This interaction thereby promotes the rapid degradation of HIF-1alpha. Under hypoxic conditions, HIF-1alpha is stabilized. We have ...
previously shown that VHL binds in a hypoxia-sensitive manner to a 27-aa segment of HIF-1alpha, and that this regulation depends on a posttranslational modification of HIF-1alpha. Through a combination of in vivo coimmunoprecipitation assays using VHL and a panel of point mutants of HIF-1alpha in this region, as well as MS and in vitro binding assays, we now provide evidence that this modification, which occurs under normoxic conditions, is hydroxylation of Pro-564 of HIF-1alpha. The data furthermore show that this proline hydroxylation is the primary regulator of VHL binding.
Mesh Terms:
Amino Acid Sequence, Animals, COS Cells, Cell Hypoxia, Cercopithecus aethiops, Cobalt, DNA-Binding Proteins, Hela Cells, Humans, Hydroxylation, Hydroxyproline, Hypoxia-Inducible Factor 1, Hypoxia-Inducible Factor 1, alpha Subunit, Ligases, Macromolecular Substances, Molecular Sequence Data, Nuclear Proteins, Peptide Fragments, Point Mutation, Proline, Protein Binding, Protein Processing, Post-Translational, Proteins, Recombinant Fusion Proteins, Transcription Factors, Transfection, Tumor Suppressor Proteins, Ubiquitin-Protein Ligases, Von Hippel-Lindau Tumor Suppressor Protein
Proc. Natl. Acad. Sci. U.S.A.
Date: Aug. 14, 2001
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