DNA sequence analysis for structure/function and mutation studies in Becker muscular dystrophy

• Published in: Clinical genetics 2005-07, Vol.68 (1), p.69-79
• Main Authors: Hamed, SA, Sutherland-Smith, AJ, Gorospe, JRM, Kendrick-Jones, J, Hoffman, EP
• Format: Article
• Language: English
• Subjects: 5' Untranslated Regions; actin-binding domain; Adult; Amino Acid Sequence; Base Sequence; Becker muscular dystrophy; Biological and medical sciences; Child; Child, Preschool; Diseases of striated muscles. Neuromuscular diseases; dystrophin; Dystrophin - chemistry; Dystrophin - genetics; Dystrophin - metabolism; dystrophin point mutations; General aspects. Genetic counseling; Humans; Infant; Male; Medical genetics; Medical sciences; Middle Aged; Molecular Sequence Data; Molecular Weight; Muscular dystrophy; Muscular Dystrophy, Duchenne - genetics; Mutation; Neurology; Pedigree; Polymerase Chain Reaction - methods; quantitative multiplex fluorescence polymerase chain reaction; Ribonucleic acid; RNA; RNA, Messenger - analysis; Sequence Analysis, DNA - methods; Structure-Activity Relationship
• ISSN: 0009-9163; 1399-0004
• DOI: 10.1111/j.1399-0004.2005.00455.x

We systematically screened the whole coding region of 18 male muscular dystrophy patients whose clinical, histological and laboratory findings suggest Becker muscular dystrophy (present but abnormal dystrophin). No systematic mutation study of a cohort of patients with dystrophin of normal quality but abnormal quantity has been published. The complete coding sequence of the dystrophin gene (11 kb) of each patient was subjected to an automated sequence analysis by using muscle biopsy RNA; 535 bp of the gene promoter and 5′UTR were likewise sequenced. We identified seven disease‐causing mutations (40%). Six were novel, including missense, nonsense, small deletion and splice site mutations. Sixty percent (11/18) of patients with decreased quantities of normal molecular weight dystrophin showed no mutation, but most of them had a family history highly suggestive of X‐linked inheritance, suggesting transcription or translational deleterious affection, i.e. outside what was screened. Quantitative multiplex fluorescence polymerase chain studies of mutation‐negative patients showed normal levels of dystrophin mRNA. In three patients, there was some reduction of the transcript suggesting a deleterious undetected gene change resulted in the reduction of RNA levels. Our data address important structure/function and genotype/phenotype correlations and it suggests that dystrophin protein studies must be interpreted with caution in deletion‐negative male muscular dystrophy patients.