Targeting of HIF-alpha to the von Hippel-Lindau ubiquitylation complex by O2-regulated prolyl hydroxylation

P Jaakkola; D R Mole; Y M Tian; M I Wilson; J Gielbert; S J Gaskell; A von Kriegsheim; H F Hebestreit; M Mukherji; C J Schofield; P H Maxwell; C W Pugh; P J Ratcliffe

doi:10.1126/science.1059796

Targeting of HIF-alpha to the von Hippel-Lindau ubiquitylation complex by O2-regulated prolyl hydroxylation

Science. 2001 Apr 20;292(5516):468-72. doi: 10.1126/science.1059796. Epub 2001 Apr 5.

Authors

P Jaakkola¹, D R Mole, Y M Tian, M I Wilson, J Gielbert, S J Gaskell, A von Kriegsheim, H F Hebestreit, M Mukherji, C J Schofield, P H Maxwell, C W Pugh, P J Ratcliffe

Affiliation

¹ The Henry Wellcome Building of Genomic Medicine, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK.

PMID: 11292861
DOI: 10.1126/science.1059796

Abstract

Hypoxia-inducible factor (HIF) is a transcriptional complex that plays a central role in the regulation of gene expression by oxygen. In oxygenated and iron replete cells, HIF-alpha subunits are rapidly destroyed by a mechanism that involves ubiquitylation by the von Hippel-Lindau tumor suppressor (pVHL) E3 ligase complex. This process is suppressed by hypoxia and iron chelation, allowing transcriptional activation. Here we show that the interaction between human pVHL and a specific domain of the HIF-1alpha subunit is regulated through hydroxylation of a proline residue (HIF-1alpha P564) by an enzyme we have termed HIF-alpha prolyl-hydroxylase (HIF-PH). An absolute requirement for dioxygen as a cosubstrate and iron as cofactor suggests that HIF-PH functions directly as a cellular oxygen sensor.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Ascorbic Acid / pharmacology
Cell Hypoxia
DNA-Binding Proteins / chemistry
DNA-Binding Proteins / metabolism*
Deferoxamine / pharmacology
Ferrous Compounds / pharmacology
Humans
Hydroxylation
Hydroxyproline / metabolism*
Hypoxia-Inducible Factor 1
Hypoxia-Inducible Factor 1, alpha Subunit
Ligases*
Molecular Sequence Data
Nuclear Proteins / chemistry
Nuclear Proteins / metabolism*
Oxygen / physiology*
Point Mutation
Procollagen-Proline Dioxygenase / antagonists & inhibitors
Procollagen-Proline Dioxygenase / metabolism*
Protein Structure, Tertiary
Proteins / metabolism*
Recombinant Fusion Proteins / metabolism
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Transcription Factors / chemistry
Transcription Factors / metabolism*
Tumor Cells, Cultured
Tumor Suppressor Proteins*
Ubiquitin-Protein Ligases*
Ubiquitins / metabolism
Von Hippel-Lindau Tumor Suppressor Protein

Substances

DNA-Binding Proteins
Ferrous Compounds
HIF1A protein, human
Hypoxia-Inducible Factor 1
Hypoxia-Inducible Factor 1, alpha Subunit
Nuclear Proteins
Proteins
Recombinant Fusion Proteins
Transcription Factors
Tumor Suppressor Proteins
Ubiquitins
Procollagen-Proline Dioxygenase
Ubiquitin-Protein Ligases
Von Hippel-Lindau Tumor Suppressor Protein
Ligases
VHL protein, human
Deferoxamine
Ascorbic Acid
Hydroxyproline
ferrous chloride
Oxygen