title: Three novel splice mutations in the PCCA gene causing identical exon skipping in propionic acidemia patients.
abstract: Propionyl-CoA carboxylase (PCC) is a mitochondrial, biotin-dependent enzyme involved in the catabolism of branched chain amino acids, odd chain fatty acids, and other metabolites. PCC consists of non-identical subunits, alpha and beta, encoded by the PCCA and PCCB genes, respectively. Inherited deficiency of PCC due to mutations in either the PCCA or the PCCB gene results in propionic acidemia (PA), a clinically heterogeneous disorder with a severe, often lethal, neonatal form, and a mild, later onset form. To characterize PCCA gene mutations responsible for PCC deficiency, we analyzed RT-PCR products obtained from cultured fibroblasts from Spanish PCC-alpha deficient patients. In three patients, smaller than normal PCR products were observed, and sequence analysis revealed the deletion of a 54-bp exon in the cDNA. Sequencing of genomic DNA from these three patients led to the identification of three novel mutations in the PCCA gene, two short deletions and one small insertion, adjacent to short direct repeats, and all of them affecting the consensus splice sites of the skipped exon. These mutations, 1771IVS-2del9, 1824IVS+3del4, and 1824IVS+3insCT, are the cause of the aberrant splicing of the PCCA pre-mRNA and result in an in-frame deletion of 54 nucleotides in the cDNA, probably leading to an unstable protein structure which is responsible for the lack of activity leading to PCC deficiency in these patients.
authors.affiliation: Centro de Biología Molecular Severo Ochoa, CSIC-UAM, Facultad de Ciencias, Universidad Autónoma de Madrid, Spain.
authors.forename: E
authors.initials: E
authors.lastname: Richard
authors.forename: L R
authors.initials: LR
authors.lastname: Desviat
authors.forename: B
authors.initials: B
authors.lastname: Pérez
authors.forename: C
authors.initials: C
authors.lastname: Pérez-Cerdá
authors.forename: M
authors.initials: M
authors.lastname: Ugarte
mesh_terms: D000592:Amino Acid Metabolism, Inborn Errors / Q000201:enzymology* / Q000235:genetics*; D001483:Base Sequence; D002262:Carboxy-Lyases / Q000172:deficiency / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D005091:Exons; D005838:Genotype; D006801:Humans; D043586:Methylmalonyl-CoA Decarboxylase; D009154:Mutation; D010641:Phenotype; D016133:Polymerase Chain Reaction; D011422:Propionates / Q000097:blood*; D012326:RNA Splicing
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D011422:Propionates; D002262:Carboxy-Lyases; D043586:Methylmalonyl-CoA Decarboxylase