Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Biochem J
2004 Nov 01;383Pt. 3:429-37. doi: 10.1042/BJ20040735.
Show Gene links
Show Anatomy links
Disruption of dimerization and substrate phosphorylation inhibit factor inhibiting hypoxia-inducible factor (FIH) activity.
Lancaster DE
,
McNeill LA
,
McDonough MA
,
Aplin RT
,
Hewitson KS
,
Pugh CW
,
Ratcliffe PJ
,
Schofield CJ
.
???displayArticle.abstract???
HIF (hypoxia-inducible factor) is an alphabeta transcription factor that modulates the hypoxic response in many animals. The cellular abundance and activity of HIF-alpha are regulated by its post-translational hydroxylation. The hydroxylation of HIF is catalysed by PHD (prolyl hydroxylase domain) enzymes and FIH (factorinhibiting HIF), all of which are 2-oxoglutarate- and Fe(II)-dependent dioxygenases. FIH hydroxylates a conserved asparagine residue in HIF-alpha (Asn-803), which blocks the binding of HIF to the transcriptional co-activator p300, preventing transcription of hypoxia-regulated genes under normoxic conditions. In the present paper, we report studies on possible mechanisms for the regulation of FIH activity. Recently solved crystal structures of FIH indicate that it is homodimeric. Site-directed mutants of FIH at residues Leu-340 and Ile-344, designed to disrupt dimerization, were generated in order to examine the importance of the dimeric state in determining FIH activity. A single point mutant, L340R (Leu-340-->Arg), was shown to be predominantly monomeric and to have lost catalytic activity as measured by assays monitoring 2-oxoglutarate turnover and asparagine hydroxylation. In contrast, the I344R (Ile-344-->Arg) mutant was predominantly dimeric and catalytically active. The results imply that the homodimeric form of FIH is required for productive substrate binding. The structural data also revealed a hydrophobic interaction formed between FIH and a conserved leucine residue (Leu-795) on the HIF substrate, which is close to the dimer interface. A recent report has revealed that phosphorylation of Thr-796, which is adjacent to Leu-795, enhances the transcriptional response in hypoxia. Consistent with this, we show that phosphorylation of Thr-796 prevents the hydroxylation of Asn-803 by FIH.
Aravind,
The DNA-repair protein AlkB, EGL-9, and leprecan define new families of 2-oxoglutarate- and iron-dependent dioxygenases.
2001, Pubmed
Aravind,
The DNA-repair protein AlkB, EGL-9, and leprecan define new families of 2-oxoglutarate- and iron-dependent dioxygenases.
2001,
Pubmed
Bruick,
A conserved family of prolyl-4-hydroxylases that modify HIF.
2001,
Pubmed
Chang,
Interdimer processing mechanism of procaspase-8 activation.
2003,
Pubmed
Clifton,
Crystal structure of carbapenem synthase (CarC).
2003,
Pubmed
Cockman,
Hypoxia inducible factor-alpha binding and ubiquitylation by the von Hippel-Lindau tumor suppressor protein.
2000,
Pubmed
Dames,
Structural basis for Hif-1 alpha /CBP recognition in the cellular hypoxic response.
2002,
Pubmed
Dann,
Structure of factor-inhibiting hypoxia-inducible factor 1: An asparaginyl hydroxylase involved in the hypoxic response pathway.
2002,
Pubmed
Datta,
Protein kinase C zeta transactivates hypoxia-inducible factor alpha by promoting its association with p300 in renal cancer.
2004,
Pubmed
Eichhorn,
Characterization of alpha-ketoglutarate-dependent taurine dioxygenase from Escherichia coli.
1997,
Pubmed
Elkins,
Structure of factor-inhibiting hypoxia-inducible factor (HIF) reveals mechanism of oxidative modification of HIF-1 alpha.
2003,
Pubmed
Epstein,
C. elegans EGL-9 and mammalian homologs define a family of dioxygenases that regulate HIF by prolyl hydroxylation.
2001,
Pubmed
Freedman,
Structural basis for negative regulation of hypoxia-inducible factor-1alpha by CITED2.
2003,
Pubmed
Freedman,
Structural basis for recruitment of CBP/p300 by hypoxia-inducible factor-1 alpha.
2002,
Pubmed
Gradin,
The transcriptional activation function of the HIF-like factor requires phosphorylation at a conserved threonine.
2002,
Pubmed
Hewitson,
Hypoxia-inducible factor (HIF) asparagine hydroxylase is identical to factor inhibiting HIF (FIH) and is related to the cupin structural family.
2002,
Pubmed
Hon,
Structural basis for the recognition of hydroxyproline in HIF-1 alpha by pVHL.
2002,
Pubmed
Huang,
Regulation of hypoxia-inducible factor 1alpha is mediated by an O2-dependent degradation domain via the ubiquitin-proteasome pathway.
1998,
Pubmed
Ivan,
HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing.
2001,
Pubmed
Jaakkola,
Targeting of HIF-alpha to the von Hippel-Lindau ubiquitylation complex by O2-regulated prolyl hydroxylation.
2001,
Pubmed
Koivunen,
Catalytic properties of the asparaginyl hydroxylase (FIH) in the oxygen sensing pathway are distinct from those of its prolyl 4-hydroxylases.
2004,
Pubmed
Lando,
FIH-1 is an asparaginyl hydroxylase enzyme that regulates the transcriptional activity of hypoxia-inducible factor.
2002,
Pubmed
Lando,
Oxygen-dependent regulation of hypoxia-inducible factors by prolyl and asparaginyl hydroxylation.
2003,
Pubmed
Lee,
Structure of human FIH-1 reveals a unique active site pocket and interaction sites for HIF-1 and von Hippel-Lindau.
2003,
Pubmed
Linke,
Substrate requirements of the oxygen-sensing asparaginyl hydroxylase factor-inhibiting hypoxia-inducible factor.
2004,
Pubmed
Mahon,
FIH-1: a novel protein that interacts with HIF-1alpha and VHL to mediate repression of HIF-1 transcriptional activity.
2001,
Pubmed
Makino,
Inhibitory PAS domain protein is a negative regulator of hypoxia-inducible gene expression.
2001,
Pubmed
Masson,
Independent function of two destruction domains in hypoxia-inducible factor-alpha chains activated by prolyl hydroxylation.
2001,
Pubmed
Maxwell,
The tumour suppressor protein VHL targets hypoxia-inducible factors for oxygen-dependent proteolysis.
1999,
Pubmed
McNeill,
Hypoxia-inducible factor asparaginyl hydroxylase (FIH-1) catalyses hydroxylation at the beta-carbon of asparagine-803.
2002,
Pubmed
Min,
Structure of an HIF-1alpha -pVHL complex: hydroxyproline recognition in signaling.
2002,
Pubmed
Nicholls,
Protein folding and association: insights from the interfacial and thermodynamic properties of hydrocarbons.
1991,
Pubmed
Pugh,
Hypoxia and oxidative stress in breast cancer. Hypoxia signalling pathways.
2001,
Pubmed
Ratcliffe,
From erythropoietin to oxygen: hypoxia-inducible factor hydroxylases and the hypoxia signal pathway.
2002,
Pubmed
Richard,
p42/p44 mitogen-activated protein kinases phosphorylate hypoxia-inducible factor 1alpha (HIF-1alpha) and enhance the transcriptional activity of HIF-1.
1999,
Pubmed
Roach,
Crystal structure of isopenicillin N synthase is the first from a new structural family of enzymes.
1995,
Pubmed
Sabourin,
Purification and characterization of an alpha-ketoisocaproate oxygenase of rat liver.
1982,
Pubmed
Safran,
HIF hydroxylation and the mammalian oxygen-sensing pathway.
2003,
Pubmed
Schofield,
Oxygen sensing by HIF hydroxylases.
2004,
Pubmed
Shen,
Hormone-sensitive lipase functions as an oligomer.
2000,
Pubmed
Taylor,
Characterization and comparative analysis of the EGLN gene family.
2001,
Pubmed
Veenstra,
Electrospray ionization mass spectrometry in the study of biomolecular non-covalent interactions.
1999,
Pubmed
Wang,
Purification and characterization of hypoxia-inducible factor 1.
1995,
Pubmed
Wang,
Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension.
1995,
Pubmed
Yu,
HIF-1alpha binding to VHL is regulated by stimulus-sensitive proline hydroxylation.
2001,
Pubmed
Zhang,
Structural origins of the selectivity of the trifunctional oxygenase clavaminic acid synthase.
2000,
Pubmed