title: Functional analysis of a large set of BRCA2 exon 7 variants highlights the predictive value of hexamer scores in detecting alterations of exonic splicing regulatory elements.
abstract: Exonic variants can alter pre-mRNA splicing either by changing splice sites or by modifying splicing regulatory elements. Often these effects are difficult to predict and are only detected by performing RNA analyses. Here, we analyzed, in a minigene assay, 26 variants identified in the exon 7 of BRCA2, a cancer predisposition gene. Our results revealed eight new exon skipping mutations in this exon: one directly altering the 5' splice site and seven affecting potential regulatory elements. This brings the number of splicing regulatory mutations detected in BRCA2 exon 7 to a total of 11, a remarkably high number considering the total number of variants reported in this exon (n = 36), all tested in our minigene assay. We then exploited this large set of splicing data to test the predictive value of splicing regulator hexamers' scores recently established by Ke et al. (). Comparisons of hexamer-based predictions with our experimental data revealed high sensitivity in detecting variants that increased exon skipping, an important feature for prescreening variants before RNA analysis. In conclusion, hexamer scores represent a promising tool for predicting the biological consequences of exonic variants and may have important applications for the interpretation of variants detected by high-throughput sequencing.
authors: Inserm U1079, University of Rouen, Institute for Research and Innovation in Biomedicine, Rouen, France; Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, Italy.
authors: Daniela
authors: D
authors: Di Giacomo
authors: Pascaline
authors: P
authors: Gaildrat
authors: Anna
authors: A
authors: Abuli
authors: Julie
authors: J
authors: Abdat
authors: Thierry
authors: T
authors: Frébourg
authors: Mario
authors: M
authors: Tosi
authors: Alexandra
authors: A
authors: Martins
mesh_terms: D019943:Amino Acid Substitution; D024682:BRCA2 Protein / Q000235:genetics*; D001483:Base Sequence; D002874:Chromosome Mapping; D019295:Computational Biology / Q000379:methods; D005091:Exons; D023061:Gene Order; D014644:Genetic Variation; D006801:Humans; D009154:Mutation; D012322:RNA Precursors / Q000235:genetics; D022821:RNA Splice Sites; D012326:RNA Splicing; D012045:Regulatory Sequences, Nucleic Acid
chemical_list: D024682:BRCA2 Protein; D012322:RNA Precursors; D022821:RNA Splice Sites
keywords: BRCA2; RNA splicing regulation; bioinformatics predictions; hereditary breast and ovarian cancer