X-linked recessive Menkes disease: identification of partial gene deletions in affected males

• Published in: Clinical genetics 2002-12, Vol.62 (6), p.449-457
• Main Authors: Poulsen, L, Horn, N, Heilstrup, H, Lund, C, Tümer, Z, Møller, LB
• Format: Article
• Language: English
• Subjects: Adenosine Triphosphatases - genetics; Base Sequence; Biological and medical sciences; Cation Transport Proteins - genetics; Copper-transporting ATPases; Deletion; DNA Mutational Analysis - methods; DNA Mutational Analysis - standards; DNA Primers; Gene Deletion; Humans; Male; Medical sciences; Menkes disease; Menkes Kinky Hair Syndrome - diagnosis; Menkes Kinky Hair Syndrome - genetics; Mental Retardation, X-Linked - diagnosis; Mental Retardation, X-Linked - genetics; Metabolic diseases; Metals (hemochromatosis...); multiplex PCR; Other metabolic disorders; Polymerase Chain Reaction - methods; Polymerase Chain Reaction - standards; Recombinant Fusion Proteins; RT-PCR; spanning
• ISSN: 0009-9163; 1399-0004
• DOI: 10.1034/j.1399-0004.2002.620605.x

Poulsen L, Horn N, Heilstrup H, Lund C, Tümer Z, Møller LB. X‐linked recessive Menkes disease: identification of partial gene deletions in affected males. 
Clin Genet 2002: 62: 449–457. © Blackwell Munksgaard, 2002 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.