Homotypic association between tumour-associated VHL proteins leads to the restoration of HIF pathway

Oncogene. 2006 May 18;25(21):3079-83. doi: 10.1038/sj.onc.1209328.

Abstract

The von Hippel-Lindau (VHL) tumour suppressor gene encodes a substrate-specifying component of an E3 ubiquitin ligase that targets hypoxia-inducible factor (HIF) alpha subunits for degradation under normoxia. The VHL protein is composed of an N-terminal HIFalpha-binding beta domain and a C-terminal alpha domain, which is necessary and sufficient for the formation of the E3 multiprotein enzyme. A large number of disease-causing mutations in either the alpha or beta domain renders HIFalpha stable irrespective of oxygen tension, leading to the upregulation of numerous HIF-target genes, such as GLUT1 and VEGF. Here, we show that VHL forms a self-associated complex in vivo, but not in vitro, and demonstrate that coexpression of two different VHL missense mutants -- one in the alpha domain and the other in the beta domain -- restores HIF-mediated gene expression profile. These findings indicate that VHL homotypic complexes can function in vivo in a complementary fashion to target HIFalpha for ubiquitin-mediated proteolysis, and potentially explain why VHL-associated tumours with a missense mutation-carrying VHL allele is almost invariably accompanied by a second VHL allele harbouring a gross truncation or deletion.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Bone Neoplasms / pathology
  • Carcinoma, Renal Cell / pathology
  • Cell Line, Tumor
  • Dimerization
  • Elongin
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / physiology*
  • Kidney Neoplasms / pathology
  • Mutation, Missense
  • Neoplasm Proteins / biosynthesis
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / physiology
  • Osteosarcoma / pathology
  • Oxygen / metabolism
  • Protein Interaction Mapping
  • Protein Structure, Tertiary / genetics
  • Structure-Activity Relationship
  • Transcription Factors / metabolism
  • Transfection
  • Ubiquitin-Protein Ligase Complexes / chemistry
  • Ubiquitin-Protein Ligase Complexes / physiology*
  • Ubiquitin-Protein Ligases / physiology*
  • Up-Regulation
  • Von Hippel-Lindau Tumor Suppressor Protein / chemistry
  • Von Hippel-Lindau Tumor Suppressor Protein / genetics
  • Von Hippel-Lindau Tumor Suppressor Protein / physiology*

Substances

  • Elongin
  • HIF1A protein, human
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Neoplasm Proteins
  • Transcription Factors
  • Ubiquitin-Protein Ligase Complexes
  • Ubiquitin-Protein Ligases
  • Von Hippel-Lindau Tumor Suppressor Protein
  • VHL protein, human
  • Oxygen