VHL frameshift mutation as target of nonsense-mediated mRNA decay in Drosophila melanogaster and human HEK293 cell line

J Biomed Biotechnol. 2009:2009:860761. doi: 10.1155/2009/860761. Epub 2010 Jan 21.

Abstract

There are many well-studied examples of human phenotypes resulting from nonsense or frameshift mutations that are modulated by Nonsense-Mediated mRNA Decay (NMD), a process that typically degrades transcripts containing premature termination codons (PTCs) in order to prevent translation of unnecessary or aberrant transcripts. Different types of germline mutations in the VHL gene cause the von Hippel-Lindau disease, a dominantly inherited familial cancer syndrome with a marked phenotypic variability and age-dependent penetrance. By generating the Drosophila UAS:Upf1(D45B) line we showed the possible involvement of NMD mechanism in the modulation of the c.172delG frameshift mutation located in the exon 1 of Vhl gene. Further, by Quantitative Real-time PCR (QPCR) we demonstrated that the corresponding c.163delG human mutation is targeted by NMD in human HEK 293 cells. The UAS:Upf1(D45B) line represents a useful system to identify novel substrates of NMD pathway in Drosophila melanogaster. Finally, we suggest the possible role of NMD on the regulation of VHL mutations.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alcohol Dehydrogenase / genetics
  • Alcohol Dehydrogenase / metabolism
  • Animals
  • Animals, Genetically Modified
  • Cell Line
  • Drosophila Proteins / genetics
  • Drosophila Proteins / metabolism
  • Drosophila melanogaster / genetics*
  • Female
  • Frameshift Mutation*
  • Humans
  • Insect Proteins / genetics
  • Insect Proteins / metabolism
  • Male
  • Ovary / chemistry
  • Ovary / metabolism
  • Polymerase Chain Reaction
  • RNA Stability / physiology*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Testis / chemistry
  • Testis / metabolism
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Von Hippel-Lindau Tumor Suppressor Protein / genetics*
  • Von Hippel-Lindau Tumor Suppressor Protein / metabolism

Substances

  • Drosophila Proteins
  • Insect Proteins
  • Transcription Factors
  • ADH protein, Drosophila
  • Alcohol Dehydrogenase
  • Von Hippel-Lindau Tumor Suppressor Protein
  • VHL protein, human