Abstract
Hypoxia-inducible factor 1α (HIF-1α) degradation under normoxia is critical to modulating vascular growth. This degradation is mediated during normoxia by the von Hippel–Lindau tumour suppressor protein (VHL)-E3 ubiquitin ligase in partnership with the E2 enzyme UbcH5. In current models of the functionally similar Skp1, cullin, F-box (SCF)-E3 ligase, the E3 binds the target protein and the E2 catalyses ubiquitin transfer to lysines in an appropriately positioned domain. In the present study, we report that for efficient ubiquitination of HIF-1α to occur, three conserved lysines are required in both the HIF-1α and endothelial Per-ARNT-Sim domain protein (EPAS) sequences. The site of ubiquitin attachment via UbcH5 was mapped, and is shown to involve three HIF-1α lysines, K532, K538 and K547, and the same aligned lysines in EPAS. Only one of these lysines need to be intact for full ubiquitination to occur, analogous to the mechanism of Sic1 ubiquitination by the SCF/Cdc34 complex and further strengthening the functional link between the VHL and SCF-E3 ubiquitin ligases. We also report that lysines can be moved around the HIF-1α sequence with only minor losses in ubiquitination efficiency, thus suggesting HIF-1α and EPAS regulation by hypoxia depends primarily on an interaction with VHL per se, rather than the highly specific positioning of flanking lysine acceptors.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Cardozo T, Pagano M . (2004). Nat Rev Mol Cell Biol 5: 739–751.
Chan DA, Sutphin PD, Yen SE, Giaccia AJ . (2005). Mol Cell Biol 25: 6415–6426.
Deffenbaugh AE, Scaglione KM, Zhang L, Moore JM, Buranda T, Sklar LA et al (2003). Cell 114: 611–622.
Epstein AC, Gleadle JM, McNeill LA, Hewitson KS, O’Rourke J, Mole DR et al (2001). Cell 107: 43–54.
Hatfield PM, Callis J, Vierstra RD . (1990). J Biol Chem 265: 15813–15817.
Hergovich A, Lisztwan J, Barry R, Ballschmieter P, Krek W . (2003). Nat Cell Biol 5: 64–70.
Ivan M, Kondo K, Yang H, Kim W, Valiando J, Ohh M et al (2001). Science 292: 464–468.
Jaakkola P, Mole DR, Tian YM, Wilson MI, Gielbert J, Gaskell SJ et al (2001). Science 292: 468–472.
Jackson PK, Eldridge AG . (2002). Mol Cell 9: 923–925.
Jensen JP, Bates PW, Yang M, Vierstra RD, Weissman AM . (1995). J Biol Chem 270: 30408–30414.
Kondo K, Kaelin Jr WG . (2001). Exp Cell Res 264: 117–125.
Lewis MD, Roberts BJ . (2003). Oncogene 22: 3992–3997.
Lewis MD, Roberts BJ . (2004). Oncogene 23: 2315–2323.
Lonser RR, Glenn GM, Walther M, Chew EY, Libutti SK, Linehan WM et al (2003). Lancet 361: 2059–2067.
Masson N, Willam C, Maxwell PH, Pugh CW, Ratcliffe PJ . (2001). EMBO J 20: 5197–5206.
Maynard MA, Qi H, Chung J, Lee EH, Kondo Y, Hara S et al (2003). J Biol Chem 278: 11032–11040.
Ohh M, Yauch RL, Lonergan KM, Whaley JM, Stemmer-Rachamimov AO, Louis DN et al (1998). Mol Cell 1: 959–968.
Petroski MD, Deshaies RJ . (2003). Mol Cell 11: 1435–1444.
Pugh CW, O’Rourke JF, Nagao M, Gleadle JM, Ratcliffe PJ . (1997). J Biol Chem 272: 11205–11214.
Pugh CW, Ratcliffe PJ . (2003). Semin Cancer Biol 13: 83–89.
Tanimoto K, Makino Y, Pereira T, Poellinger L . (2000). EMBO J 19: 4298–4309.
Yu F, White SB, Zhao Q, Lee FS . (2001). Cancer Res 61: 4136–4142.
Zhou MI, Wang H, Ross JJ, Kuzmin I, Xu C, Cohen HT . (2002). J Biol Chem 277: 39887–39898.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Paltoglou, S., Roberts, B. HIF-1α and EPAS ubiquitination mediated by the VHL tumour suppressor involves flexibility in the ubiquitination mechanism, similar to other RING E3 ligases. Oncogene 26, 604–609 (2007). https://doi.org/10.1038/sj.onc.1209818
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.onc.1209818
Keywords
This article is cited by
-
UBE3B promotes breast cancer progression by antagonizing HIF-2α degradation
Oncogene (2023)
-
Elucidating the role of hypoxia-inducible factor in rheumatoid arthritis
Inflammopharmacology (2022)
-
An oxygen sensitive self-decision making engineered CAR T-cell
Scientific Reports (2017)
-
Parkin targets HIF-1α for ubiquitination and degradation to inhibit breast tumor progression
Nature Communications (2017)
-
Development of an Oxygen-Sensitive Degradable Peptide Probe for the Imaging of Hypoxia-Inducible Factor-1-Active Regions in Tumors
Molecular Imaging and Biology (2013)