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Science 2001 Apr 20;292(5516):468-72
Targeting of HIF-alpha to the von Hippel-Lindau ubiquitylation complex by
O2-regulated prolyl hydroxylation.
Jaakkola P, Mole DR, Tian YM, Wilson MI, Gielbert J, Gaskell SJ, Kriegsheim
Av, Hebestreit HF, Mukherji M, Schofield CJ, Maxwell PH, Pugh CW, Ratcliffe
PJ
The Henry Wellcome Building of Genomic Medicine, University of Oxford,
Roosevelt Drive, Oxford OX3 7BN, UK.
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.
PMID: 11292861, UI: 21214630
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Nature 1999 May 20;399(6733):271-5
The tumour suppressor protein VHL targets hypoxia-inducible factors for
oxygen-dependent proteolysis.
Maxwell PH, Wiesener MS, Chang GW, Clifford SC, Vaux EC, Cockman ME, Wykoff
CC, Pugh CW, Maher ER, Ratcliffe PJ
Wellcome Trust Centre for Human Genetics, Oxford, UK.
Hypoxia-inducible factor-1 (HIF-1) has a key role in cellular
responses to hypoxia, including the regulation of genes involved in
energy metabolism, angiogenesis and apoptosis. The alpha subunits of
HIF are rapidly degraded by the proteasome under normal conditions,
but are stabilized by hypoxia. Cobaltous ions or iron chelators mimic
hypoxia, indicating that the stimuli may interact through effects on a
ferroprotein oxygen sensor. Here we demonstrate a critical role for
the von Hippel-Lindau (VHL) tumour suppressor gene product pVHL in
HIF-1 regulation. In VHL-defective cells, HIF alpha-subunits are
constitutively stabilized and HIF-1 is activated. Re-expression of
pVHL restored oxygen-dependent instability. pVHL and HIF
alpha-subunits co-immunoprecipitate, and pVHL is present in the
hypoxic HIF-1 DNA-binding complex. In cells exposed to iron chelation
or cobaltous ions, HIF-1 is dissociated from pVHL. These findings
indicate that the interaction between HIF-1 and pVHL is iron
dependent, and that it is necessary for the oxygen-dependent
degradation of HIF alpha-subunits. Thus, constitutive HIF-1 activation
may underlie the angiogenic phenotype of VHL-associated tumours. The
pVHL/HIF-1 interaction provides a new focus for understanding cellular
oxygen sensing.
PMID: 10353251, UI: 99279691
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