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Ubiquitination of hypoxia-inducible factor requires direct binding to the β-domain of the von Hippel–Lindau protein

Nature Cell Biology volume 2, pages 423–427 (2000)Cite this article

Abstract

von Hippel–Lindau (VHL) disease is a hereditary cancer syndrome that is characterized by the development of multiple vascular tumors and is caused by inactivation of the von Hippel–Lindau protein (pVHL). Here we show that pVHL, through its β-domain, binds directly to hypoxia-inducible factor (HIF), thereby targeting HIF for ubiquitination in an α-domain-dependent manner. This is the first function to be ascribed to the pVHL β-domain. Furthermore, we provide the first direct evidence that pVHL has a function analogous to that of an F-box protein, namely, to recruit substrates to a ubiquitination machine. These results strengthen the link between overaccumulation of HIF and development of VHL disease.

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Figure 1: The pVHL β-domain binds to a region of HIF implicated in oxygen-dependent proteolysis.
Figure 2: pVHL binds directly to HIF.
Figure 3: Ubiquitination of the HIF pVHL-binding domain.
Figure 4: pVHL-dependent ubiquitination of HIF.
Figure 5: Ubiquitination of HIF requires the pVHL α- and β-domains.
Figure 6: Ubiquitination of HIF inhibited by a hypoxia mimetic.

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References

  1. Stebbins, C. E., Kaelin, W. G. J. & Pavletich, N. P. Structure of the VHL–elonginC–elonginB complex: implications for VHL tumor suppressor function. Science 284, 455–461 ( 1999).

    Article  CAS  Google Scholar 

  2. Kaelin, W. G. Cancer. Many vessels, faulty gene. Nature 399, 203–204 (1999).

    Article  CAS  Google Scholar 

  3. Lisztwan, J., Imbert, G., Wirbelauer, C., Gstaiger, M. & Krek, W. The von Hippel–Lindau tumor suppressor protein is a component of an E3 ubiquitin-protein ligase activity. Genes Dev. 13, 1822–1833 (1999).

    Article  CAS  Google Scholar 

  4. Iwai, K. et al. Identification of the von Hippel–Lindau tumor-suppressor protein as part of an active E3 ubiquitin ligase complex. Proc. Natl Acad. Sci. USA 96, 12436–12441 (1999).

    Article  CAS  Google Scholar 

  5. Maxwell, P. et al. The von Hippel–Lindau gene product is necessary for oxgyen-dependent proteolysis of hypoxia-inducible factor α subunits . Nature 399, 271–275 (1999).

    Article  CAS  Google Scholar 

  6. Huang, L. E., Gu, J., Schau, M. & Bunn, H. F. Regulation of hypoxia-inducible factor 1 alpha is mediated by an O2-dependent degradation domain via the ubiquitin-proteasome pathway. Proc. Natl Acad. Sci. USA 95, 7987–7992 ( 1998).

    Article  CAS  Google Scholar 

  7. Kallio, P., Wilson, W., O’Brien, S., Makino, Y. & Poellinger, L. Regulation of the hypoxia-inducible transcription factor 1 alpha by the ubiquitin-proteasome pathway. J. Biol. Chem. 274, 6519–6525 (1999).

    Article  CAS  Google Scholar 

  8. Pugh, C., O’Rourke, J., Nagao, M., Gleadle, J. & Ratcliffe, P. Activation of hypoxia-inducible factor-1: definition of regulatory domains within the alpha subunit. J. Biol. Chem. 272, 11205–11214 (1997).

    Article  CAS  Google Scholar 

  9. Ohh, M. & Kaelin, W. G. J. The von Hippel–Lindau tumour suppressor protein: new perspectives. Mol. Med. Today 5, 257–263 (1999).

    Article  CAS  Google Scholar 

  10. Kamura, T. et al. The Elongin BC complex interacts with the conserved SOCS-box motif present in members of the SOCS, ras, WD-40 repeat, and ankyrin repeat families. Genes Dev. 12, 3872– 3881 (1998).

    Article  CAS  Google Scholar 

  11. Duan, D. R. et al. Inhibition of transcriptional elongation by the VHL tumor suppressor protein. Science 269, 1402– 1406 (1995).

    Article  CAS  Google Scholar 

  12. Kibel, A., Iliopoulos, O., DeCaprio, J. D. & Kaelin, W. G. Binding of the von Hippel–Lindau tumor suppressor protein to elongin B and C. Science 269, 1444– 1446 (1995).

    Article  CAS  Google Scholar 

  13. Salceda, S. & Caro, J. Hypoxia-inducible factor 1 alpha (HIF-1 alpha) protein is rapidly degraded by the ubiquitin-proteasome system under normoxic conditions. Its stabilization by hypoxia depends on redox-induced changes. J. Biol. Chem. 272, 22642– 2267 (1997).

    Article  CAS  Google Scholar 

  14. Ohh, M. et al. Synthetic peptides define critical contacts between elongin C, elongin B, and the von Hippel–Lindau protein. J. Clin. Invest. 104, 1583–1591 (1999).

    Article  CAS  Google Scholar 

  15. An, W. et al. Stabilization of wild-type p53 by hypoxia-inducible factor 1 alpha . Nature 392, 405–408 (1998).

    Article  CAS  Google Scholar 

  16. Chandel, N. et al. Mitochondrial reactive oxygen species trigger hypoxia-induced transcription. Proc. Natl Acad. Sci. USA 95, 11715–11720 (1998).

    Article  CAS  Google Scholar 

  17. Ritter, M. et al. Isolated familial pheochromocytoma as a variant of von Hippel–Lindau Disease. J. Clin. Endocrinol. Met. 81, 1035 –1037 (1996).

    CAS  Google Scholar 

  18. Crossey, P. A. et al. Identification of intragenic mutations in the von Hippel–Lindau disease tumor suppressor gene and correlation with disease phenotype. Hum. Mol. Genet. 3, 1303–1308 (1994).

    Article  CAS  Google Scholar 

  19. Iliopoulos, O., Kibel, A., Gray, S. & Kaelin, W. G. Tumor suppression by the human von Hippel–Lindau gene product. Nature Med. 1, 822–826 ( 1995).

    Article  CAS  Google Scholar 

  20. Lonergan, K. M. et al. Regulation of hypoxia-inducible mRNAs by the von Hippel–Lindau protein requires binding to complexes containing elongins B/C and Cul2. Mol. Cell Biol. 18, 732–741 (1998).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank C. Stebbins for purified pVHL/elongin C/elongin B complexes and H. Franklin Bunn and M. Meyerson for critical reading of the manuscript. This work was supported by grants from the NIH and from the Murray Foundation. M.O. is a Fellow of the National Cancer Institute of Canada. W.G.K. is a Howard Hughes Medical Institute assistant investigator.

Correspondence and requests for materials should be addressed to W.G.K.

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Authors and Affiliations

  1. Department of Medicine, Brigham and Womens Hospital and Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts, 02115, USA

    Michael Ohh, Cheol Won Park, Mircea Ivan, Michael A. Hoffman, Tae-You Kim & William G. Kaelin

  2. Howard Hughes Medical Institute,

    Cheol Won Park, Tae-You Kim, Nikola Pavletich & William G. Kaelin

  3. Division of Hematology, Brigham and Womens Hospital, Harvard Medical School, Boston, Massachusetts, 02115, USA

    L. Eric Huang

  4. Memorial Sloan-Kettering Cancer Center, New York, New York, 10021, USA

    Nikola Pavletich

  5. Department of Cellular and Molecular Physiology, Pennsylvania State University School of Medicine, Hershey, Pennsylvania, 17033, USA

    Vincent Chau

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  1. Michael Ohh
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Correspondence to William G. Kaelin.

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Ohh, M., Park, C., Ivan, M. et al. Ubiquitination of hypoxia-inducible factor requires direct binding to the β-domain of the von Hippel–Lindau protein. Nat Cell Biol 2, 423–427 (2000). https://doi.org/10.1038/35017054

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