title: Rapid and efficient ATM mutation detection by fluorescent chemical cleavage of mismatch: identification of four novel mutations.
abstract: Mutations in the Ataxia Telangiectasia Mutated (ATM) gene are responsible for the autosomal recessive disease Ataxia Telangiectasia (A-T). A wide variety of mutations scattered across the entire coding region (9168bp) of ATM have been found, which presents a challenge in developing an efficient mutation screening strategy for detecting unknown mutations. Fluorescent chemical cleavage of mismatch (FCCM) is an ideal mutation screening method, offering a non-radioactive alternative to other techniques such as restriction endonuclease fingerprinting (REF). Using FCCM, we have developed an efficient, accurate and sensitive mutation detection method for screening RT-PCR products for ATM mutations. We have identified seven ATM mutations in five A-T families, four of which are previously unknown. We quantified ATM protein expression in four of the families and found variable ATM protein expression (0-6.4%), further evidence for mutant ATM protein expression in both classic and variant A-T patients. We conclude that FCCM offers a robust ATM mutation detection method and can be used to screen for ATM mutations in cancer-prone populations.
authors.affiliation: Division of Medical and Molecular Genetics GKT, Guy's Hospital, London, UK. l.izatt@umds.ac.uk
authors.forename: L
authors.initials: L
authors.lastname: Izatt
authors.forename: C
authors.initials: C
authors.lastname: Vessey
authors.forename: S V
authors.initials: SV
authors.lastname: Hodgson
authors.forename: E
authors.initials: E
authors.lastname: Solomon
mesh_terms: D000293:Adolescent; D000328:Adult; D000483:Alleles; D017398:Alternative Splicing; D001260:Ataxia Telangiectasia / Q000235:genetics*; D064007:Ataxia Telangiectasia Mutated Proteins; D015153:Blotting, Western; D018797:Cell Cycle Proteins; D002648:Child; D002675:Child, Preschool; D004268:DNA-Binding Proteins; D005260:Female; D005456:Fluorescent Dyes; D016368:Frameshift Mutation; D006801:Humans; D008297:Male; D009154:Mutation; D020125:Mutation, Missense; D017346:Protein Serine-Threonine Kinases; D011506:Proteins / Q000235:genetics*; D011933:Reagent Kits, Diagnostic; D013997:Time Factors; D025521:Tumor Suppressor Proteins
chemical_list: D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes; D011506:Proteins; D011933:Reagent Kits, Diagnostic; D025521:Tumor Suppressor Proteins; C576324:ATM protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; D017346:Protein Serine-Threonine Kinases