title: X-linked recessive Menkes disease: identification of partial gene deletions in affected males.
abstract: Menkes disease is an X-linked recessive lethal disorder of copper metabolism, caused by defects in the ATP7A gene. Partial gene deletions comprise about 15% of the mutations causing Menkes disease. We have previously demonstrated identification of partial ATP7A deletions in patients by Southern blot analysis. In the present study, we report the use of three fast and reliable polymerase chain reaction (PCR)-based methods for the identification of partial ATP7A deletions in Menkes disease patients. First we demonstrate the use of multiplex PCR, a fast method for identification and rough localization of partial gene deletions, in which two exons of ATP7A are coamplified. Second, we present PCR amplification of genomic DNA across the deletion junctions, a method enabling identification of the deletion breakpoints and hence the exact size of the deletion. Finally, application of reverse transcription PCR (RT-PCR) for identification and localization of gene deletions at the cDNA level is demonstrated. By studying the mutation at the cDNA level the predicted effect of the mutation on the amino acid sequence and consequently the protein structure and function can be inferred. We demonstrate characterization of partial gene deletions in five patients, and in three of these we were able to determine the breakpoint sequences.
authors.affiliation: The John F. Kennedy Institute, Glostrup, Denmark.
authors.forename: L
authors.initials: L
authors.lastname: Poulsen
authors.forename: N
authors.initials: N
authors.lastname: Horn
authors.forename: H
authors.initials: H
authors.lastname: Heilstrup
authors.forename: C
authors.initials: C
authors.lastname: Lund
authors.forename: Z
authors.initials: Z
authors.lastname: Tümer
authors.forename: L B
authors.initials: LB
authors.lastname: Møller
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics*; D001483:Base Sequence; D027682:Cation Transport Proteins / Q000235:genetics*; D000073840:Copper-Transporting ATPases; D004252:DNA Mutational Analysis / Q000379:methods* / Q000592:standards; D017931:DNA Primers; D017353:Gene Deletion; D006801:Humans; D008297:Male; D007706:Menkes Kinky Hair Syndrome / Q000175:diagnosis* / Q000235:genetics; D038901:X-Linked Intellectual Disability / Q000175:diagnosis / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards; D011993:Recombinant Fusion Proteins
chemical_list: D027682:Cation Transport Proteins; D017931:DNA Primers; D011993:Recombinant Fusion Proteins; D000251:Adenosine Triphosphatases; C081184:ATP7A protein, human; D000073840:Copper-Transporting ATPases