title: A nonsense mutation in the 3-hydroxy-3-methylglutaryl-CoA lyase gene produces exon skipping in two patients of different origin with 3-hydroxy-3-methylglutaryl-CoA lyase deficiency.
abstract: A novel nonsense mutation associated with the skipping of constitutive exon 2 of the 3-hydroxy-3-methylglutaryl-CoA lyase gene was found in two patients, from Portugal and Morocco, with 3-hydroxy-3-methylglutaric acidemia. By reverse transcriptase PCR and single-strand conformational polymorphism a G-T transversion was located, at nucleotide 109, of the 3-hydroxy-3-methylglutaryl-CoA lyase cDNA, within exon 2. Two mRNAs were produced as a result of this nonsense mutation: one of the expected size that contains the premature stop codon UAA, and the other with a deletion of 84 bp corresponding to the whole of exon 2. This deletion produced the loss of the last seven amino acids of the leader peptide and the first 21 amino acids of the mature protein. The nonsense mutation was found in a purine-rich GGAAG sequence, which is equal to, or similar to, others reported to be exonic splicing enhancers (ESE). We suggest that the nonsense mutation may affect a possible ESE on exon 2, which would hinder the splice site selection and facilitate an aberrant splice with the skipping of this exon. Determination by quantitative PCR shows that the ratio of mRNA with the nonsense mutation to the mRNA with the deletion is approx. 3:1.
authors.affiliation: Unit of Biochemistry, School of Pharmacy, University of Barcelona, Barcelona, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Pié
authors.forename: N
authors.initials: N
authors.lastname: Casals
authors.forename: C H
authors.initials: CH
authors.lastname: Casale
authors.forename: C
authors.initials: C
authors.lastname: Buesa
authors.forename: C
authors.initials: C
authors.lastname: Mascaró
authors.forename: A
authors.initials: A
authors.lastname: Barceló
authors.forename: M O
authors.initials: MO
authors.lastname: Rolland
authors.forename: T
authors.initials: T
authors.lastname: Zabot
authors.forename: D
authors.initials: D
authors.lastname: Haro
authors.forename: F
authors.initials: F
authors.lastname: Eyskens
authors.forename: P
authors.initials: P
authors.lastname: Divry
authors.forename: F G
authors.initials: FG
authors.lastname: Hegardt
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D005091:Exons; D005260:Female; D006801:Humans; D007223:Infant; D007438:Introns; D008297:Male; D008969:Molecular Sequence Data; D016296:Mutagenesis; D007652:Oxo-Acid-Lyases / Q000172:deficiency* / Q000235:genetics*; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational
chemical_list: D007652:Oxo-Acid-Lyases; C021410:3-hydroxy-3-methylglutaryl-coenzyme A lyase