The contribution of VHL substrate binding and HIF1-alpha to the phenotype of VHL loss in renal cell carcinoma

Jodi K Maranchie; James R Vasselli; Joseph Riss; Juan S Bonifacino; W Marston Linehan; Richard D Klausner

doi:10.1016/s1535-6108(02)00044-2

The contribution of VHL substrate binding and HIF1-alpha to the phenotype of VHL loss in renal cell carcinoma

Cancer Cell. 2002 Apr;1(3):247-55. doi: 10.1016/s1535-6108(02)00044-2.

Authors

Jodi K Maranchie¹, James R Vasselli, Joseph Riss, Juan S Bonifacino, W Marston Linehan, Richard D Klausner

Affiliation

¹ Urologic Oncology Branch, National Institutes of Health, Bethesda, MD 20892, USA.

PMID: 12086861
DOI: 10.1016/s1535-6108(02)00044-2

Abstract

Clear-cell renal carcinoma is associated with inactivation of the von Hippel-Lindau (VHL) tumor suppressor gene. VHL is the substrate recognition subunit of an E3 ligase, known to target the alpha subunits of the HIF heterodimeric transcription factor for ubiquitin-mediated degradation under normoxic conditions. We demonstrate that competitive inhibition of the VHL substrate recognition site with a peptide derived from the oxygen degradation domain of HIF1alpha recapitulates the tumorigenic phenotype of VHL-deficient tumor cells. These studies prove that VHL substrate recognition is essential to the tumor suppressor function of VHL. We further demonstrate that normoxic stabilization of HIF1alpha alone, while capable of mimicking some aspects of VHL loss, is not sufficient to reproduce tumorigenesis, indicating that it is not the critical oncogenic substrate of VHL.

MeSH terms

Animals
Binding Sites
Blotting, Western
Carcinoma, Renal Cell / genetics
Carcinoma, Renal Cell / metabolism*
Carcinoma, Renal Cell / pathology
Female
Genes, Tumor Suppressor
Glucose Transporter Type 1
Green Fluorescent Proteins
HeLa Cells
Humans
Hypoxia-Inducible Factor 1, alpha Subunit
Immunoenzyme Techniques
Kidney Neoplasms / genetics
Kidney Neoplasms / metabolism*
Kidney Neoplasms / pathology
Ligases / metabolism*
Luciferases / metabolism
Luminescent Proteins / metabolism
Mice
Mice, SCID
Monosaccharide Transport Proteins / metabolism
Phenotype
Plasmids
Recombinant Fusion Proteins
Transcription Factors / metabolism*
Transcription, Genetic
Transfection
Tumor Suppressor Proteins*
Ubiquitin-Protein Ligases
Von Hippel-Lindau Tumor Suppressor Protein

Substances

Glucose Transporter Type 1
HIF1A protein, human
Hypoxia-Inducible Factor 1, alpha Subunit
Luminescent Proteins
Monosaccharide Transport Proteins
Recombinant Fusion Proteins
SLC2A1 protein, human
Transcription Factors
Tumor Suppressor Proteins
Green Fluorescent Proteins
Luciferases
Ubiquitin-Protein Ligases
Von Hippel-Lindau Tumor Suppressor Protein
Ligases
VHL protein, human