Characterization of the VHL Tumor Suppressor Gene Product: Localization, Complex Formation, and the Effect of Natural Inactivating Mutations

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 92; no. 14; pp. 6459 - 6463
• Main Authors: Duan, D R, Humphrey, J S, Chen, D Y, Weng, Y, Sukegawa, J, Lee, S, Gnarra, J R, Linehan, W M, Klausner, R D
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 03.07.1995; National Acad Sciences; National Academy of Sciences
• Subjects: Amino Acid Sequence; Amino acids; Animals; Base Sequence; Biochemistry; carcinoma; Cell Line; Cercopithecus aethiops; Complementary DNA; COS cells; Disease; DNA, Complementary - chemistry; Genes; Genes, Tumor Suppressor; Genetic mutation; HeLa Cells; Humans; Kidney; Ligases; man; Molecular Sequence Data; Mutation; Nuclear Proteins - biosynthesis; Nuclear Proteins - genetics; Nuclear Proteins - isolation & purification; Open reading frames; Proteins; Rats; Recombinant Proteins - biosynthesis; Recombinant Proteins - isolation & purification; Sequence Homology, Amino Acid; Sequence Homology, Nucleic Acid; Subcellular Fractions - metabolism; Transfection; Tumor suppressor genes; Tumor Suppressor Proteins; Tumors; Ubiquitin-Protein Ligases; VHL gene; von Hippel-Lindau disease; von Hippel-Lindau Disease - genetics; Von Hippel-Lindau Tumor Suppressor Protein
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.92.14.6459

The human VHL tumor suppressor gene has been implicated in the inherited disorder von Hippel-Lindau disease and in sporadic renal carcinoma. The homologous rat gene encodes a 185-amino acid protein that is 88% sequence identical to the aligned 213-amino acid human VHL gene product. When expressed in COS-7 cells, both the human and the rat VHL proteins showed predominant nuclear, nuclear and cytosolic, or predominant cytosolic VHL staining by immunofluorescence. A complicated pattern of cellular proteins was seen that could be specifically coimmunoprecipitated with the introduced VHL protein. A complex containing VHL and proteins of apparent molecular masses 16 and 9 kDa was the most consistently observed. Certain naturally occurring VHL missense mutations demonstrated either complete or partial loss of the p16-p9 complex. Thus, the VHL tumor suppressor gene product is a nuclear protein, perhaps capable of specifically translocating between the nucleus and the cytosol. It is likely that VHL executes its functions via formation of specific multiprotein complexes. Identification of these VHL-associated proteins will likely clarify the physiology of this tumor suppressor gene.