title: Exon skipping in IVD RNA processing in isovaleric acidemia caused by point mutations in the coding region of the IVD gene.
abstract: Isovaleric acidemia (IVA) is a recessive disorder caused by a deficiency of isovaleryl-CoA dehydrogenase (IVD). We have reported elsewhere nine point mutations in the IVD gene in fibroblasts of patients with IVA, which lead to abnormalities in IVD protein processing and activity. In this report, we describe eight IVD gene mutations identified in seven IVA patients that result in abnormal splicing of IVD RNA. Four mutations in the coding region lead to aberrantly spliced mRNA species in patient fibroblasts. Three of these are amino acid altering point mutations, whereas one is a single-base insertion that leads to a shift in the reading frame of the mRNA. Two of the coding mutations strengthen pre-existing cryptic splice acceptors adjacent to the natural splice junctions and apparently interfere with exon recognition, resulting in exon skipping. This mechanism for missplicing has not been reported elsewhere. Four other mutations alter either the conserved gt or ag dinucleotide splice sites in the IVD gene. Exon skipping and cryptic splicing were confirmed by transfection of these mutations into a Cos-7 cell line model splicing system. Several of the mutations were predicted by individual information analysis to inactivate or significantly weaken adjacent donor or acceptor sites. The high frequency of splicing mutations identified in these patients is unusual, as is the finding of missplicing associated with missense mutations in exons. These results may lead to a better understanding of the phenotypic complexity of IVA, as well as provide insight into those factors important in defining intron/exon boundaries in vivo.
authors.affiliation: Mayo Clinic, Department of Medical Genetics, 200 First Street, SW, Rochester, MN 55905, USA. vockley@mayo.edu
authors.forename: J
authors.initials: J
authors.lastname: Vockley
authors.forename: P K
authors.initials: PK
authors.lastname: Rogan
authors.forename: B D
authors.initials: BD
authors.lastname: Anderson
authors.forename: J
authors.initials: J
authors.lastname: Willard
authors.forename: R S
authors.initials: RS
authors.lastname: Seelan
authors.forename: D I
authors.initials: DI
authors.lastname: Smith
authors.forename: W
authors.initials: W
authors.lastname: Liu
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004795:Enzyme Stability; D005091:Exons / Q000235:genetics*; D005347:Fibroblasts; D005787:Gene Frequency / Q000235:genetics; D045782:Hemiterpenes; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007438:Introns / Q000235:genetics; D050769:Isovaleryl-CoA Dehydrogenase; D008661:Metabolism, Inborn Errors / Q000201:enzymology / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D010088:Oxidoreductases / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D044925:Oxidoreductases Acting on CH-CH Group Donors; D010421:Pentanoic Acids / Q000097:blood*; D017354:Point Mutation / Q000235:genetics*; D011110:Polymorphism, Genetic / Q000235:genetics; D034622:RNA Interference; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D020080:Tandem Repeat Sequences / Q000235:genetics
chemical_list: D045782:Hemiterpenes; D010421:Pentanoic Acids; D012333:RNA, Messenger; C008216:isovaleric acid; D010088:Oxidoreductases; D044925:Oxidoreductases Acting on CH-CH Group Donors; D050769:Isovaleryl-CoA Dehydrogenase