title: Novel COL6A1 splicing mutation in a family affected by mild Bethlem myopathy.
abstract: Bethlem myopathy is an early-onset benign myopathy characterized by proximal muscular weakness and multiple flexion contractures. It is a dominantly inherited disorder associated with mutations in the three COL6 genes encoding type VI collagen. We detected a g-->a substitution at +1 position of COL6A1 intron 3 in a four-generation Italian family affected by a mild form of Bethlem myopathy. The mutation results in the activation of a cryptic splice donor site at the 3' end of exon 3, leading to the loss of 66 nucleotides and an "in-frame" deletion of 22 amino acids in the NH2-domain. Molecular analysis on fibroblasts of the propositus showed that the mutated mRNA was present and stable, but the mutated protein could not be detected. Western blot and immunofluorescence analyses showed a decreased level of collagen VI synthesis and deposition in fibroblasts of the propositus. Together, the results suggest that the mutated protein was highly unstable and rapidly degraded, and that the mild phenotype was caused by a reduced amount of normal collagen VI microfibrils. In addition, we demonstrated that lymphocytes can be used for the first mutation screening analysis of patients with Bethlem myopathy.
authors.affiliation: Department of Internal Medicine, University of Rome "Tor Vergata," Via Tor Vergata 135, 00133 Roma, Italy.
authors.forename: Olga Camacho
authors.initials: OC
authors.lastname: Vanegas
authors.forename: Rui-Zhu
authors.initials: RZ
authors.lastname: Zhang
authors.forename: Patrizia
authors.initials: P
authors.lastname: Sabatelli
authors.forename: Giovanna
authors.initials: G
authors.lastname: Lattanzi
authors.forename: Paola
authors.initials: P
authors.lastname: Bencivenga
authors.forename: Betti
authors.initials: B
authors.lastname: Giusti
authors.forename: Marta
authors.initials: M
authors.lastname: Columbaro
authors.forename: Mon-Li
authors.initials: ML
authors.lastname: Chu
authors.forename: Luciano
authors.initials: L
authors.lastname: Merlini
authors.forename: Guglielmina
authors.initials: G
authors.lastname: Pepe
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D017398:Alternative Splicing / Q000235:genetics*; D024142:Collagen Type VI / Q000096:biosynthesis / Q000235:genetics*; D004252:DNA Mutational Analysis; D005260:Female; D005786:Gene Expression Regulation / Q000235:genetics; D005820:Genetic Testing; D006801:Humans; D008297:Male; D008875:Middle Aged; D018482:Muscle, Skeletal / Q000378:metabolism* / Q000473:pathology; D009135:Muscular Diseases / Q000235:genetics* / Q000473:pathology; D010375:Pedigree; D017354:Point Mutation / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics
chemical_list: D024142:Collagen Type VI; D012333:RNA, Messenger