von Hippel–Lindau (VHL) gene inactivation occurs in von Hippel–Lindau (VHL) disease. The protein pVHL functions in a multi-subunit E3 ubiquitin ligase that targets the hypoxia-inducible transcription factor Hif1α for proteasomal degradation during normoxia. We establish that pVHL binds to Tat-binding protein-1 (TBP-1), a component of the 19S regulatory complex of the proteasome. TBP-1 associates with the β-domain of pVHL and complexes with pVHL and Hif1α in vivo. Overexpression of TBP-1 promotes degradation of Hif1α in a pVHL-dependent manner that requires the ATPase domain of TBP-1. Blockade of TBP-1 expression by small interfering RNA (siRNA) causes prolonged degradation kinetics of Hif1α. Several distinct mutations in exon 2 of VHL disrupt binding of pVHL to TBP-1. A pVHL mutant containing a P154L substitution coimmunoprecipitates with Hif1α, but not TBP-1, and does not promote degradation of Hif1α. Thus, the ability of pVHL to degrade Hif1α depends in part on its interaction with TBP-1 and suggests a new mechanism for Hif1α stabilization in some pVHL-deficient tumors.
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The authors thank M. Rountree and K. Bachman for technical assistance with the yeast two-hybrid assay in the initial phase of this study and C. Stolle for clinical information on individuals with VHL disease. This work was supported by grants from the US National Institutes of Health and by the Howard Hughes Medical Institute. W.S.E-D. is an Assistant Investigator of the Howard Hughes Medical Institute.
The authors declare no competing financial interests.
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