title: Molecular analysis of dihydropteridine reductase deficiency: identification of two novel mutations in Japanese patients.
abstract: Mutations in the dihydropteridine reductase (DHPR) gene result in hyperphenylalaninaemia and deficiency of various neurotransmitters in the central nervous system, causing severe neurological symptoms. We studied two Japanese patients with DHPR deficiency and identified a missense and a splicing error mutation, respectively. A homozygous missense mutation (tryptophan36-to-arginine) was detected in patient 1. The mutation abolished DHPR activity according to in vitro expression studies. The DHPR mRNA in patient 2 was markedly decreased. Reverse transcription-polymerase chain reaction of the mRNA generated a cDNA fragment with a 152-bp insertion. The inserted sequence contained a termination codon, which was likely to affect the stability of the mRNA. Analysis of genomic DNA showed that the insertion was derived from putative intron 3 of the DHPR gene, and an intronic A-to-G substitution was present adjacent to the 3'-end of the inserted sequence. The nucleotide change generated a sequence similar to an RNA splice donor site and probably activated an upstream cryptic acceptor site, thus producing an abnormal extra exon.
authors.affiliation: Department of Biochemical Genetics, Tohoku University School of Medicine, Sendai, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Ikeda
authors.forename: Y
authors.initials: Y
authors.lastname: Matsubara
authors.forename: H
authors.initials: H
authors.lastname: Mikami
authors.forename: S
authors.initials: S
authors.lastname: Kure
authors.forename: M
authors.initials: M
authors.lastname: Owada
authors.forename: T
authors.initials: T
authors.lastname: Gough
authors.forename: P M
authors.initials: PM
authors.lastname: Smooker
authors.forename: M
authors.initials: M
authors.lastname: Dobbs
authors.forename: H H
authors.initials: HH
authors.lastname: Dahl
authors.forename: R G
authors.initials: RG
authors.lastname: Cotton
authors.forename: K
authors.initials: K
authors.lastname: Narisawa
mesh_terms: D000592:Amino Acid Metabolism, Inborn Errors / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D004252:DNA Mutational Analysis; D018076:DNA, Complementary / Q000235:genetics; D004093:Dihydropteridine Reductase / Q000235:genetics*; D005260:Female; D005796:Genes / Q000235:genetics; D006801:Humans; D007231:Infant, Newborn; D007438:Introns / Q000235:genetics; D007564:Japan; D008297:Male; D008969:Molecular Sequence Data; D010649:Phenylalanine / Q000097:blood; D010661:Phenylketonurias; D017354:Point Mutation / Q000235:genetics*; D012326:RNA Splicing / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger; D010649:Phenylalanine; D004093:Dihydropteridine Reductase