title: New splicing mutations in propionic acidemia.
abstract: Propionic acidemia results from mutations in either of the two genes, PCCA or PCCB, that encode the two subunits of the propionyl-CoA carboxylase (PCC) enzyme. In this study, we report the identification and analysis of seven novel splicing mutations involving consensus donor and acceptor splice sites. Most of them were identified in patients with a Central Asian origin, and some present in several alleles, probably reflecting founder effects. The functional consequences of the splicing mutations were analyzed in patients' fibroblasts, as well as transcript quantification using real-time PCR methods. In the PCCA gene, two mutations were demonstrated to affect 5' splice sites (c.231+1G>C and c.1209+3A>G) and two 3' acceptor splice sites (c.1210delG and c.1430G>T), all causing skipping of the exons involved, with no detectable levels of normally spliced transcript. In the PCCB gene, all three mutations involved 5' donor splice sites-two affected exon 1 splicing (c.154_183+17del46 and c.183+2T>C), the latter activating a cryptic splice site in intron 1, and the remaining mutation (c.1498+2T>C) resulted in exon 14 skipping. The results highlight the necessity to perform transcript analysis in addition to genomic DNA sequencing to characterize the effect of splicing mutations and add relevant information on the genetic epidemiology of the disease.
authors.affiliation: Centro de Biología Molecular "Severo Ochoa" CSIC-UAM, Universidad Autónoma de Madrid, Cantoblanco, 28049, Madrid, Spain.
authors.forename: Lourdes R
authors.initials: LR
authors.lastname: Desviat
authors.affiliation: Centro de Biología Molecular "Severo Ochoa" CSIC-UAM, Universidad Autónoma de Madrid, Cantoblanco, 28049, Madrid, Spain.
authors.forename: Sonia
authors.initials: S
authors.lastname: Clavero
authors.affiliation: Centro de Biología Molecular "Severo Ochoa" CSIC-UAM, Universidad Autónoma de Madrid, Cantoblanco, 28049, Madrid, Spain.
authors.forename: Celia
authors.initials: C
authors.lastname: Perez-Cerdá
authors.affiliation: Centro de Biología Molecular "Severo Ochoa" CSIC-UAM, Universidad Autónoma de Madrid, Cantoblanco, 28049, Madrid, Spain.
authors.forename: Rosa
authors.initials: R
authors.lastname: Navarrete
authors.affiliation: Centro de Biología Molecular "Severo Ochoa" CSIC-UAM, Universidad Autónoma de Madrid, Cantoblanco, 28049, Madrid, Spain. mugarte@cbm.uam.es.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Ugarte
authors.affiliation: Centro de Biología Molecular "Severo Ochoa" CSIC-UAM, Universidad Autónoma de Madrid, Cantoblanco, 28049, Madrid, Spain.
authors.forename: Belen
authors.initials: B
authors.lastname: Perez
mesh_terms: D017398:Alternative Splicing; D000592:Amino Acid Metabolism, Inborn Errors / Q000453:epidemiology / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D019735:Carbon-Carbon Ligases / Q000235:genetics*; D005091:Exons; D005347:Fibroblasts / Q000378:metabolism; D018703:Founder Effect; D005838:Genotype; D006579:Heterozygote; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D009154:Mutation; D012333:RNA, Messenger / Q000378:metabolism; D017422:Sequence Analysis, DNA
chemical_list: D012333:RNA, Messenger; D019735:Carbon-Carbon Ligases; C022458:propionyl CoA carboxylase (ATP-hydrolyzing)