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Molecular imaging of hypoxia-inducible factor 1 alpha and von Hippel-Lindau interaction in mice.

Tumor hypoxia plays a crucial role in tumorigenesis. Under hypoxia, hypoxia-inducible factor 1 alpha (HIF-1 alpha) regulates activation of genes promoting malignant progression. Under normoxia, HIF-1 alpha is hydroxylated on prolines 402 and 564 and is targeted for ubiquitin-mediated degradation by interacting with the von Hippel-Lindau protein complex (pVHL). We have developed a novel method of studying the interaction between HIF-1 alpha and pVHL using the split firefly luciferase complementation-based bioluminescence system in which HIF-1 alpha and pVHL are fused to amino-terminal and carboxy-terminal fragments of the luciferase, respectively. We demonstrate that hydroxylation-dependent interaction between the HIF-1 alpha and pVHL leads to complementation of the two luciferase fragments, resulting in bioluminescence in vitro and in vivo. Complementation-based bioluminescence is diminished when mutant pVHLs with decreased affinity for binding HIF-1 alpha are used. This method represents a new approach for studying interaction of proteins involved in the regulation of protein degradation.

Pubmed ID: 19123984

Authors

  • Choi CY
  • Chan DA
  • Paulmurugan R
  • Sutphin PD
  • Le QT
  • Koong AC
  • Zundel W
  • Gambhir SS
  • Giaccia AJ

Journal

Molecular imaging

Publication Data

January 6, 2009

Associated Grants

  • Agency: NCI NIH HHS, Id: CA123823
  • Agency: NCI NIH HHS, Id: CA124435
  • Agency: NCI NIH HHS, Id: CA67166
  • Agency: NCI NIH HHS, Id: P30 CA124435

Mesh Terms

  • Animals
  • Cell Line, Tumor
  • Diagnostic Imaging
  • HCT116 Cells
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Luciferases, Firefly
  • Luminescent Measurements
  • Mice
  • Mice, Nude
  • Models, Molecular
  • Mutation
  • Proline
  • Recombinant Fusion Proteins
  • Von Hippel-Lindau Tumor Suppressor Protein
  • Whole Body Imaging