Hypoxia Inducible Factor-α Binding and Ubiquitylation by the von Hippel-Lindau Tumor Suppressor Protein*

By M. Cockman, N. Masson, D. Mole, P. Jaakkola, G. Chang, S. Clifford, E. Maher, C. Pugh, P. Ratcliffe, P. Maxwell. Journal of Biological Chemistry, 2000

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The von Hippel-Lindau tumor suppressor protein (pVHL) has emerged as a key factor in cellular responses to oxygen availability, being required for the oxygen-dependent proteolysis of α subunits of hypoxia inducible factor-1 (HIF). Mutations in VHL cause a hereditary cancer syndrome associated with dysregulated angiogenesis, and up-regulation of hypoxia inducible genes. Here we investigate the mechanisms underlying these processes and show that extracts from VHL-deficient renal carcinoma cells have a defect in HIF-α ubiquitylation activity which is complemented by exogenous pVHL. This defect was specific for HIF-α among a range of substrates tested. Furthermore, HIF-α subunits were the only pVHL-associated proteasomal substrates identified by comparison of metabolically labeled anti-pVHL immunoprecipitates from proteosomally inhibited cells and normal cells. Analysis of pVHL/HIF-α interactions defined short sequences of conserved residues within the internal transactivation domains of HIF-α molecules sufficient for recognition by pVHL. In contrast, while full-length pVHL and the p19 variant interact with HIF-α, the association was abrogated by further N-terminal and C-terminal truncations. The interaction was also disrupted by tumor-associated mutations in the β-domain of pVHL and loss of interaction was associated with defective HIF-α ubiquitylation and regulation, defining a mechanism by which these mutations generate a constitutively hypoxic pattern of gene expression promoting angiogenesis. The findings indicate that pVHL regulates HIF-α proteolysis by acting as the recognition component of a ubiquitin ligase complex, and support a model in which its β domain interacts with short recognition sequences in HIF-α subunits.

Subfields: Cancer Research, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/d81ae1d044ef17dccee4c1b4013fdde7a1ddd416

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Citations in the corpus

Hypoxia requires notch signaling to maintain the undifferentiated cell state. (2005, background): “In contrast, no interaction between pected (Cockman et al., 2000; Ohh et al., 2000; Tanimoto et al., 2000), HIF-1α and pVHL interacted (Figure 4B). pVHL did not significantly increase the level of ubiquitylation of Notch 1 ICD, while Sel-10 markedly elevated Notch 1 ICD ubiquitylation (data not…”
FIH-1: a novel protein that interacts with HIF-1alpha and VHL to mediate repression of HIF-1 transcriptional activity. (2001, background): “2000) via binding of the von Hippel-Lindau (VHL) tumor suppressor protein, which is the recognition component of an E3 ubiquitin-protein ligase (Cockman et al. 2000; Kamura et al. 2000; Ohh et al. 2000) that targets HIF-1 for proteasomal degradation (Salceda and Caro 1997).”
Gene mutations in the succinate dehydrogenase subunit SDHB cause susceptibility to familial pheochromocytoma and to familial paraganglioma. (2001, background): “…cancers and in mouse models, results in up-regulation of angiogenic growth factors, such as vascular endothelial growth factor, secondary to a defect in ubiquitylation of the a-subunits of the HIF-1 and the HIF-2 transcription factors (Maxwell et al. 1999; Cockman et al. 2000; Ohh et al. 2000).”
Inhibition of HIF is necessary for tumor suppression by the von Hippel-Lindau protein. (2002, background): “…transcription factor called HIF (hypoxia-inducible factor) in the pres-von Hippel-Lindau Disease is a hereditary cancer syndrome that displays an autosomal dominant pattern of inheritance (Maher ence of oxygen (Cockman et al., 2000; Kamura et al., 2000; Ohh et al., 2000; Tanimoto et al., 2000).”
FIH-1 is an asparaginyl hydroxylase enzyme that regulates the transcriptional activity of hypoxia-inducible factor. (2002, background): “…pVHL is a component of a protein–ubiquitin ligase complex that targets the alpha subunit for degradation by the proteasome (Maxwell et al. 1999; Cockman et al. 2000; Ohh et al. 2000; Tanimoto et al. 2000). pVHL recognition of HIF- is dependent on hydroxylation of proline residues within the…”
Hypoxia-inducible expression of tumor-associated carbonic anhydrases. (2000, background): “The gene product pVHL forms part of a ubiquitin-ligase complex (19, 20) that targets HIF-a subunits for oxygen-dependent proteolysis (13, 21 ).”