Hypoxia-inducible Factor (HIF) Asparagine Hydroxylase Is Identical to Factor Inhibiting HIF (FIH) and Is Related to the Cupin Structural Family*

By K. Hewitson, Luke A. McNeill, Madeline V Riordan, Ya-min Tian, A. Bullock, R. Welford, J. Elkins, N. Oldham, S. Bhattacharya, J. Gleadle and 3 more. Journal of Biological Chemistry, 2002

716 citations recorded by the source corpus.

Activity of the hypoxia-inducible factor (HIF) complex is controlled by oxygen-dependent hydroxylation of prolyl and asparaginyl residues. Hydroxylation of specific prolyl residues by 2-oxoglutarate (2-OG)-dependent oxygenases mediates ubiquitinylation and proteasomal destruction of HIF-α. Hydroxylation of an asparagine residue in the C-terminal transactivation domain (CAD) of HIF-α abrogates interaction with p300, preventing transcriptional activation. Yeast two-hybrid assays recently identified factor inhibiting HIF (FIH) as a protein that associates with the CAD region of HIF-α. Since FIH contains certain motifs present in iron- and 2-OG-dependent oxygenases we investigated whether FIH was the HIF asparaginyl hydroxylase. Assays using recombinant FIH and HIF-α fragments revealed that FIH is the enzyme that hydroxylates the CAD asparagine residue, that the activity is directly inhibited by cobalt(II) and limited by hypoxia, and that the oxygen in the alcohol of the hydroxyasparagine residue is directly derived from dioxygen. Sequence analyses involving FIH link the 2-OG oxygenases with members of the cupin superfamily, including Zn(II)-utilizing phosphomannose isomerase, revealing structural and evolutionary links between these metal-binding proteins that share common motifs.

Subfields: Cancer Research, Cell Biology, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/e1a7bbd152f4980f6c5274d761604918e9b9ab1e

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In Cancer Research

1. G. Semenza
2. Craig B. Thompson
3. M. C. Simon
4. Eyal Gottlieb
5. J. Chiche
6. P. Ratcliffe
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8. J. Pouysségur

In Cell Biology

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2. Eyal Gottlieb
3. J. Chiche
4. M. Brahimi-Horn
5. P. Ratcliffe
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In Molecular Biology

1. G. Semenza
2. M. C. Simon
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4. Robert P. Hausinger
5. P. Ratcliffe
6. J. Chiche
7. Jessica A. Bertout
8. Eyal Gottlieb

Citations in the corpus

• Oxygen sensing by HIF hydroxylases (2004, background): “Like prolyl hydroxylation, HIF asparaginyl hydroxylation is also catalysed by a member of the 2-oxoglutarate-dependent-oxygenase superfamily, but one that is significantly different in sequence to the PHD”
• Tumour hypoxia induces a metabolic shift causing acidosis: a common feature in cancer (2009, background): “This post-translational modification impairs the interaction between the C-TAD and the coactivator p300/CREB binding protein (CBP) [97, 98].”
• Fe(II)/α-Ketoglutarate-Dependent Hydroxylases and Related Enzymes (2004, methodology): “Crystal structures also are reported for Fe(II)-bound proline 3-hydroxylase (Clifton et al., 2001) and FIH (Dann et al., 2002).”
• Hypoxia-Inducible Factor-1 (HIF-1) (2006, neutral): “Under normal oxygen tension, hydroxylation of the asparagines residue 803 (Asn803) in the C-TAD of HIF-1 (Asn851 in HIF-2 ) by factor inhibiting HIF-1 (FIH-1) prevented the interaction of HIF-1 with CBP/p300 (Hewitson et al., 2002; Lando et al., 2002b; Sang et al., 2002).”
• Succinate links TCA cycle dysfunction to oncogenesis by inhibiting HIF-alpha prolyl hydroxylase. (2005, background): “For studying reduced and total glutathione levels, cell lysates were regulate HIF activity by modulating another (cid:1) -ketoglutarate de-prepared using a lysis buffer (Cell Signaling) freshly supplemented with pendent dioxygenase, FIH, thereby altering the ability of HIF-1 (cid:1) protease inhibit”