title: Identification of novel mutations in the PCCB gene in European propionic acidemia patients. Mutation in brief no. 253. Online.
abstract: Propionyl-CoA carboxylase (PCC) is a biotin-dependent enzyme located in the mitochondrial matrix. Mutations in the PCCA and PCCB genes, which encode the a and b subunits of this heteropolymer, result in propionic acidemia (PA). We report the molecular analysis of b-deficient patients from Spain and Austria. Subjects were screened for defects affecting the PCCB gene by direct sequencing from genomic PCR products, restriction digests and mRNA analysis by RT-PCR. Study by western blot of the presence of immunoreactive b-PCC protein was also performed. A total of four novel sequence variations were found including the point mutations V205D, and M442T, and the frameshift mutation 790-791insG. Additionally, a new point change, L17M, was identified on the same allele as 790-791insG. The missense changes described above were not found in at least 40 control chromosomes analyzed. The Austrian patients were homozygous for V205D. One of the Spanish subjects was heterozygous for M442T and the known mutation c1170insT. The other Spanish patient carried L17M+790-791insG on one allele, and the described mutation E168K on the other mutant chromosome. The mutations V205D and M442T were confirmed at RNA level and also we have detected the presence of immunoreactive b-PCC protein translated from these mutant alleles. The patient having L17M+790-791insG and E168K also presented immunoreactive b-PCC protein. However, no cDNA product was obtained from the chromosome carrying L17M+790-791insG. We propose that 790-791insG, which causes a frameshift and a premature stop codon, is responsible for this finding. In any case, the translation from this mutant cDNA would produce a severity truncated peptide and, in consequence, a non-functional protein. Expression analysis of all these changes will help us to clarify their structural/functional consequences.
authors.affiliation: Departamento de Biología Molecular, Centro de Biología Molecular Severo Ochoa CSIC-UAM, Universidad Autónoma de Madrid, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Muro
authors.forename: P
authors.initials: P
authors.lastname: Rodríguez-Pombo
authors.forename: B
authors.initials: B
authors.lastname: Pérez
authors.forename: C
authors.initials: C
authors.lastname: Pérez-Cerdá
authors.forename: L R
authors.initials: LR
authors.lastname: Desviat
authors.forename: W
authors.initials: W
authors.lastname: Sperl
authors.forename: D
authors.initials: D
authors.lastname: Skladal
authors.forename: J O
authors.initials: JO
authors.lastname: Sass
authors.forename: M
authors.initials: M
authors.lastname: Ugarte
mesh_terms: D000592:Amino Acid Metabolism, Inborn Errors / Q000201:enzymology / Q000235:genetics*; D001317:Austria; D019735:Carbon-Carbon Ligases / Q000172:deficiency* / Q000235:genetics*; D006801:Humans; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D011422:Propionates / Q000097:blood*; D017422:Sequence Analysis, DNA; D013030:Spain
chemical_list: D011422:Propionates; D019735:Carbon-Carbon Ligases; C022458:propionyl CoA carboxylase (ATP-hydrolyzing)