title: Functional analysis of splicing mutations in MYO7A and USH2A genes.
abstract: Usher syndrome is defined by the association of sensorineural hearing loss, retinitis pigmentosa and variable vestibular dysfunction. Many disease-causative mutations have been identified in MYO7A and USH2A genes, which play a major role in Usher syndrome type I and type II, respectively. The pathogenic nature of mutations that lead to premature stop codons is not questioned; nevertheless, additional studies are needed to verify the pathogenicity of some changes such as those putatively involved in the splice process. Five putative splice-site variants were detected in our cohort of patients: c.2283-1G>T and c.5856G>A in MYO7A and c.1841-2A>G, c.2167+5G>A and c.5298+1G>C in the USH2A gene. In this study, we analyze these changes with bioinformatic tools and investigate the expression of MYO7A and USH2A transcripts through hybrid minigene assays. Our study showed that all five mutations abolished the consensus splice site producing the skipping of involved exons. In addition, for variant c.2167+5G>A, a new donor splice site was observed. Our data reveal the pathogenic nature of the analyzed variants. The fact that splicing mutations led to in-frame or out-of-frame alterations cannot explain phenotypic differences, thus, genotype-phenotype correlations cannot be inferred.
authors.affiliation: Unidad de Genética y Diagnóstico Prenatal, Hospital Universitario La Fe, Avenida Campanar 21, Valencia, Spain.
authors.forename: T
authors.initials: T
authors.lastname: Jaijo
authors.forename: E
authors.initials: E
authors.lastname: Aller
authors.forename: M J
authors.initials: MJ
authors.lastname: Aparisi
authors.forename: G
authors.initials: G
authors.lastname: García-García
authors.forename: I
authors.initials: I
authors.lastname: Hernan
authors.forename: M J
authors.initials: MJ
authors.lastname: Gamundi
authors.forename: C
authors.initials: C
authors.lastname: Nájera
authors.forename: M
authors.initials: M
authors.lastname: Carballo
authors.forename: J M
authors.initials: JM
authors.lastname: Millán
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D016326:Extracellular Matrix Proteins / Q000235:genetics* / Q000378:metabolism; D005260:Female; D023061:Gene Order; D005838:Genotype; D006801:Humans; D008297:Male; D009154:Mutation; D000081412:Myosin VIIa; D009218:Myosins / Q000235:genetics* / Q000378:metabolism; D022821:RNA Splice Sites; D012326:RNA Splicing / Q000235:genetics*; D052245:Usher Syndromes / Q000235:genetics
chemical_list: D016326:Extracellular Matrix Proteins; C000631455:MYO7A protein, human; D000081412:Myosin VIIa; D022821:RNA Splice Sites; C112735:USH2A protein, human; D009218:Myosins