title: Maple syrup urine disease due to a new large deletion at BCKDHA caused by non-homologous recombination.
abstract: Maple syrup urine disease (MSUD) is a rare disorder of branched-chain amino acid (BCAA) metabolism caused by the defective function of branched-chain α-ketoacid dehydrogenase complex (BCKD). Many MSUD-causing mutations have already been described in genes that encode the complex (BCKDHA, BCKDHB and DBT), but up to now only four large deletions are known, all located in the DBT gene. In a previous study we identified a Portuguese MSUD patient with a homozygous deletion of exons 2, 3 and 4 at the BCKDHA gene; however, the corresponding breakpoints and, consequently, the exact deletion extension were not identified. Here, using long-range PCR and sequencing methodologies we were able to refine the characterization of this gross rearrangement. A genomic DNA loss of about 13.8 kb was detected, starting at intron 1 and ending at intron 4, thus encompassing exons 2, 3 and 4. Molecular characterization showed that the deletion junction contained a short sequence whose motif was CGGG. Since this motif is present in introns 1 and 4 of normal genomic DNA, we have hypothesized that non-homologous recombination was the mechanism underlying the identified large deletion, within which the CGGG could be derived either from intron 1 or from intron 4.
authors.affiliation: IPATIMUP - Institute of Pathology and Molecular Immunology of University of Porto, Rua Dr Roberto Frias s/n, 4200-465, Porto, Portugal. mquental@ipatimup.pt
authors.forename: S
authors.initials: S
authors.lastname: Quental
authors.forename: E
authors.initials: E
authors.lastname: Martins
authors.forename: L
authors.initials: L
authors.lastname: Vilarinho
authors.forename: A
authors.initials: A
authors.lastname: Amorim
authors.forename: M
authors.initials: M
authors.lastname: João Prata
mesh_terms: D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide) / Q000235:genetics*; D000597:Amino Acids, Branched-Chain / Q000097:blood; D001483:Base Sequence; D015415:Biomarkers / Q000097:blood; D004252:DNA Mutational Analysis; D005091:Exons; D020022:Genetic Predisposition to Disease; D006801:Humans; D007231:Infant, Newborn; D007438:Introns; D008297:Male; D008375:Maple Syrup Urine Disease / Q000097:blood / Q000175:diagnosis / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D059372:Nucleotide Motifs; D010641:Phenotype; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D017384:Sequence Deletion; D015854:Up-Regulation
chemical_list: D000597:Amino Acids, Branched-Chain; D015415:Biomarkers; D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)