title: The von Hippel-Lindau tumour suppressor gene: uncovering the expression of the pVHL172 isoform.
abstract: BACKGROUND
  The von Hippel-Lindau (VHL) gene encodes two mRNA variants. Variant 1 encodes two protein isoforms, pVHL213 and pVHL160, that have been extensively documented in the literature. Variant 2 is produced by alternative splicing of exon 2 and encodes a pVHL isoform of 172 amino acids with a theoretical molecular weight of 19 kDa (pVHL172), the expression of which has never been demonstrated so far due to the absence of suitable antibodies.
  
  
  METHODS
  We have generated an anti-pVHL monoclonal antibody (JD-1956) using pVHL172 recombinant protein. We tested the antibody against exogenous or endogenous expressed proteins in different cell lines. We identified the pVHL172 using a silencing RNA strategy. The epitope of the antibody was mapped using a peptide array.
  
  
  RESULTS
  We efficiently detected the three different isoforms of pVHL in cell lines and tumorigenic tissues by western blotting and immunohistochemistry and confirmed for the first time the endogenous expression of pVHL172.
  
  
  CONCLUSIONS
  The endogenous expression of the three isoforms and particularly the pVHL172 has never been shown before due to a lack of a highly specific antibody since none of the available commercial antibodies distinguish the three isoforms of pVHL in cells or in both normal and cancerous human tissues. Evidence of pVHL172 expression emphasises the need to further study its implication in renal tumorigenesis and VHL disease.
authors.affiliation: CNRS-UMR 6290 (IGDR)-Université Rennes 1-BIOSIT, 2 Avenue du Professeur L Bernard, 35042 Rennes, France.
authors.forename: F
authors.initials: F
authors.lastname: Chesnel
authors.affiliation: CNRS-UMR 6290 (IGDR)-Université Rennes 1-BIOSIT, 2 Avenue du Professeur L Bernard, 35042 Rennes, France.
authors.forename: P
authors.initials: P
authors.lastname: Hascoet
authors.affiliation: Centre de Recherche du CHU de Québec-Pavillon CHUL-Faculté de Médecine, Université Laval, 2705 Boulevard Laurier, Québec, G1V 4G2 QC, Canada.
authors.forename: J P
authors.initials: JP
authors.lastname: Gagné
authors.affiliation: CNRS-UMR 6290 (IGDR)-Université Rennes 1-BIOSIT, 2 Avenue du Professeur L Bernard, 35042 Rennes, France.
authors.forename: A
authors.initials: A
authors.lastname: Couturier
authors.affiliation: CNRS-UMR 6290 (IGDR)-Université Rennes 1-BIOSIT, 2 Avenue du Professeur L Bernard, 35042 Rennes, France.
authors.forename: F
authors.initials: F
authors.lastname: Jouan
authors.affiliation: Centre de Recherche du CHU de Québec-Pavillon CHUL-Faculté de Médecine, Université Laval, 2705 Boulevard Laurier, Québec, G1V 4G2 QC, Canada.
authors.forename: G G
authors.initials: GG
authors.lastname: Poirier
authors.affiliation: CNRS-UMR 6290 (IGDR)-Université Rennes 1-BIOSIT, 2 Avenue du Professeur L Bernard, 35042 Rennes, France.
authors.forename: C
authors.initials: C
authors.lastname: Le Goff
authors.affiliation: CNRS-UMR 6290 (IGDR)-Université Rennes 1-BIOSIT, 2 Avenue du Professeur L Bernard, 35042 Rennes, France.
authors.forename: C
authors.initials: C
authors.lastname: Vigneau
authors.affiliation: EFS-LNPRM-Etablissement français du sang Rennes, Rue Pierre Jean Gineste, BP 91614, 35016 Rennes, France.
authors.forename: Y
authors.initials: Y
authors.lastname: Danger
authors.affiliation: EFS-LNPRM-Etablissement français du sang Rennes, Rue Pierre Jean Gineste, BP 91614, 35016 Rennes, France.
authors.forename: F
authors.initials: F
authors.lastname: Verite
authors.affiliation: CNRS-UMR 6290 (IGDR)-Université Rennes 1-BIOSIT, 2 Avenue du Professeur L Bernard, 35042 Rennes, France.
authors.forename: X
authors.initials: X
authors.lastname: Le Goff
authors.affiliation: CNRS-UMR 6290 (IGDR)-Université Rennes 1-BIOSIT, 2 Avenue du Professeur L Bernard, 35042 Rennes, France.
authors.forename: Y
authors.initials: Y
authors.lastname: Arlot-Bonnemains
mesh_terms: D000595:Amino Acid Sequence; D000918:Antibody Specificity; D015153:Blotting, Western; D045744:Cell Line, Tumor; D016147:Genes, Tumor Suppressor; D006801:Humans; D007150:Immunohistochemistry; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000032:analysis / Q000737:chemistry / Q000235:genetics*
chemical_list: D020033:Protein Isoforms; D051794:Von Hippel-Lindau Tumor Suppressor Protein; C497446:VHL protein, human