Structure of Factor-inhibiting Hypoxia-inducible Factor (HIF) Reveals Mechanism of Oxidative Modification of HIF-1α
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The activity of the transcription factor hypoxia-inducible factor (HIF) is regulated by oxygen-dependent hydroxylation. Under normoxic conditions, hydroxylation of proline residues triggers destruction of its alpha-subunit while hydroxylation of Asn(803) in the C-terminal transactivation domain of HIF-1 alpha (CAD) prevents its interaction with p300. Here we report crystal structures of the asparagine hydroxylase (factor-inhibiting HIF, FIH) complexed with Fe((II)), 2-oxoglutarate cosubstrate, and CAD fragments, which reveal the structural basis of HIF modification. CAD binding to FIH occurs via an induced fit process at two distinct interaction sites. At the hydroxylation site CAD adopts a loop conformation, contrasting with a helical conformation for the same residues when bound to p300. Asn(803) of CAD is buried and precisely orientated in the active site such that hydroxylation occurs at its beta-carbon. Together with structures with the inhibitors Zn((II)) and N-oxaloylglycine, analysis of the FIH-CAD complexes will assist design of hydroxylase inhibitors with proangiogenic properties. Conserved structural motifs within FIH imply it is one of an extended family of Fe((II)) oxygenases involved in gene regulation.
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Both R-2HG and S-2HG coordinate the metal with the 2-hydroxyl group trans to D201 in FIH or E190 in JMJD2A, and one oxygen of the C1-carboxylate trans to H199 in FIH or H188 in JMJD2A.
“Similarly to 2OG and NOG (Elkins et al, 2003;Ng et al, 2007), both R-and S-2HG coordinate the metal with the 2-hydroxyl group trans to D201 FIH /E190 JMJD2A and one oxygen of the the C1-carboxylate trans to H199 FIH /H188 JMJD2A (Fig 2A,B,D,E).”
From The oncometabolite 2-hydroxyglutarate inhibits histone lysine demethylases (2011) · cites this paper as Elkins et al, 2003
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- Authors
- Elkins, J. M., Hewitson, K. S., McNeill, L. A., Seibel, J. F., Schlemminger, I., Pugh, C. W., Ratcliffe, P. J., & Schofield, C. J.
- Affiliations
- University of Oxford (GB)
- Corresponding author
- Hewitson, Kirsty S.
- Schofield, Christopher J.
- ORCID
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2003-01
- Volume
- 278
- Issue
- 3
- Pages
- 1802-1806
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2068053372
- Cited by
- 391
- References
- 29
- Citations by year
- 2026: 8 · 2025: 12 · 2024: 14 · 2023: 7 · 2022: 12 · 2021: 13 · 2020: 10 · 2019: 14 · 2018: 14 · 2017: 12 · 2016: 16 · 2015: 13 · 2014: 12 · 2013: 19 · 2012: 13
- FWCI
- 11.90
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- Wellcome Trust
- Medical Research Council
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Hypoxia, and Metabolism
- Physical Sciences › Chemistry › Inorganic Chemistry › Metal-Catalyzed Oxygenation Mechanisms
- Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
- Keywords
- factor inhibiting HIF, FIH, hypoxia-inducible factor-1α, HIF-1α, oxidative modification, crystal structure, induced fit, P300, transactivation domain, alpha-ketoglutarate, gene expression regulation
- MeSH terms
- Amino Acid Sequence, DNA-Binding Proteins, Models, Molecular, Molecular Sequence Data, Nuclear Proteins, Oxidation-Reduction, Protein Conformation, Transcription Factors, Sequence Homology, Amino Acid, Crystallography, X-Ray, Hypoxia-Inducible Factor 1
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