title: Molecular characterization of large deletions in the von Hippel-Lindau (VHL) gene by quantitative real-time PCR: the hypothesis of an alu-mediated mechanism underlying VHL gene rearrangements.
abstract: BACKGROUND
  Mutations of the von Hippel-Lindau (VHL) gene are responsible for VHL disease. This is a familial autosomal-dominant syndrome, predisposing to the development of benign and malignant tumors, including CNS and retinal hemangioblastomas, pheochromocytomas, and clear cell renal carcinomas. At least 30% of the disease-causing mutations in the VHL gene involve large alterations. Identification of these mutations is not possible using PCR-based mutational scanning methods. Quantitative Southern blot analysis has been traditionally employed for the detection of complete or partial deletions and more complex rearrangements of the gene.
  
  
  METHODS
  An alternative quantitative method was developed using a combination of quantitative Southern blot analysis and real-time PCR. With this approach, we studied 24 large VHL gene alterations to determine the exact nature of the mutations and to possibly characterize the boundaries of the deleted regions.
  
  
  RESULTS
  This combined molecular approach showed that all the VHL alterations studied were due to deletions, from which the position in the gene could be more precisely mapped. One of the samples that was completely characterized was found to carry an intragenic 2.2kb deletion with both 5' and 3' breakpoints located within Alu-repeat sequences.
  
  
  CONCLUSIONS
  This is the first report on the molecular analysis of large VHL alterations. The results of our study and the complete characterization of a large deletion lead to the hypothesis that an Alu-mediated mechanism may be responsible for the common occurrence of large alterations in the VHL gene.
authors.affiliation: Department of Pediatrics, University of Padua, Padua, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Casarin
authors.forename: Maddalena
authors.initials: M
authors.lastname: Martella
authors.forename: Roberta
authors.initials: R
authors.lastname: Polli
authors.forename: Emanuela
authors.initials: E
authors.lastname: Leonardi
authors.forename: Laura
authors.initials: L
authors.lastname: Anesi
authors.forename: Alessandra
authors.initials: A
authors.lastname: Murgia
mesh_terms: D020087:Alu Elements / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D017384:Sequence Deletion; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000235:genetics*; D006623:von Hippel-Lindau Disease / Q000235:genetics*
chemical_list: D051794:Von Hippel-Lindau Tumor Suppressor Protein; C497446:VHL protein, human