Activation of HIF1α ubiquitination by a reconstituted von Hippel-Lindau (VHL) tumor suppressor complex

• Takumi Kamura

  Program in Molecular and Cell Biology, Oklahoma Medical Research Foundation, 825 Northeast 13th Street, Oklahoma City, OK 73104; Department of Molecular and System Biology, Graduate School of Biostudies, Kyoto University, Kyoto, 606-8501, Japan; Department of Molecular and Cellular Physiology, University of Cincinnati College of Medicine, Cincinnati, OH 45267-0576; Howard Hughes Medical Institute, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104; and Department of Biochemistry and...

• Shigeo Sato

  Program in Molecular and Cell Biology, Oklahoma Medical Research Foundation, 825 Northeast 13th Street, Oklahoma City, OK 73104; Department of Molecular and System Biology, Graduate School of Biostudies, Kyoto University, Kyoto, 606-8501, Japan; Department of Molecular and Cellular Physiology, University of Cincinnati College of Medicine, Cincinnati, OH 45267-0576; Howard Hughes Medical Institute, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104; and Department of Biochemistry and...

• Kazuhiro Iwai

  Program in Molecular and Cell Biology, Oklahoma Medical Research Foundation, 825 Northeast 13th Street, Oklahoma City, OK 73104; Department of Molecular and System Biology, Graduate School of Biostudies, Kyoto University, Kyoto, 606-8501, Japan; Department of Molecular and Cellular Physiology, University of Cincinnati College of Medicine, Cincinnati, OH 45267-0576; Howard Hughes Medical Institute, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104; and Department of Biochemistry and...

• Maria Czyzyk-Krzeska

  Program in Molecular and Cell Biology, Oklahoma Medical Research Foundation, 825 Northeast 13th Street, Oklahoma City, OK 73104; Department of Molecular and System Biology, Graduate School of Biostudies, Kyoto University, Kyoto, 606-8501, Japan; Department of Molecular and Cellular Physiology, University of Cincinnati College of Medicine, Cincinnati, OH 45267-0576; Howard Hughes Medical Institute, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104; and Department of Biochemistry and...

• Ronald C. Conaway

  Program in Molecular and Cell Biology, Oklahoma Medical Research Foundation, 825 Northeast 13th Street, Oklahoma City, OK 73104; Department of Molecular and System Biology, Graduate School of Biostudies, Kyoto University, Kyoto, 606-8501, Japan; Department of Molecular and Cellular Physiology, University of Cincinnati College of Medicine, Cincinnati, OH 45267-0576; Howard Hughes Medical Institute, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104; and Department of Biochemistry and...

• Joan Weliky Conaway

  Program in Molecular and Cell Biology, Oklahoma Medical Research Foundation, 825 Northeast 13th Street, Oklahoma City, OK 73104; Department of Molecular and System Biology, Graduate School of Biostudies, Kyoto University, Kyoto, 606-8501, Japan; Department of Molecular and Cellular Physiology, University of Cincinnati College of Medicine, Cincinnati, OH 45267-0576; Howard Hughes Medical Institute, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104; and Department of Biochemistry and...

抄録

Mutations in the VHL tumor suppressor gene result in constitutive expression of many hypoxia-inducible genes, at least in part because of increases in the cellular level of hypoxia-inducible transcription factor HIF1α, which in normal cells is rapidly ubiquitinated and degraded by the proteasome under normoxic conditions. The recent observation that the VHL protein is a subunit of an Skp1-Cul1/Cdc53-F-box (SCF)-like E3 ubiquitin ligase raised the possibility that VHL may be directly responsible for regulating cellular levels of HIF1α by targeting it for ubiquitination and proteolysis. In this report, we test this hypothesis directly. We report development of methods for production of the purified recombinant VHL complex and present direct biochemical evidence that it can function with an E1 ubiquitin-activating enzyme and E2 ubiquitin-conjugating enzyme to activate HIF1α ubiquitination in vitro . Our findings provide new insight into the function of the VHL tumor suppressor protein, and they provide a foundation for future investigations of the mechanisms underlying VHL regulation of oxygen-dependent gene expression.

収録刊行物

• Proceedings of the National Academy of Sciences

  Proceedings of the National Academy of Sciences 97 (19), 10430-10435, 2000-09-05

  Proceedings of the National Academy of Sciences

キーワード

• Multidisciplinary

詳細情報

• CRID: 1360855570743968640
• NII論文ID: 80012258757
• DOI: 10.1073/pnas.190332597
• ISSN: 10916490
• ISSN: 00278424
• Web Site: https://pnas.org/doi/pdf/10.1073/pnas.190332597
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