Novel COL6A1 splicing mutation in a family affected by mild Bethlem myopathy.
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.
Vanegas
Olga Camacho
OC
Department of Internal Medicine, University of Rome "Tor Vergata," Via Tor Vergata 135, 00133 Roma, Italy.
Zhang
Rui-Zhu
RZ
Sabatelli
Patrizia
P
Lattanzi
Giovanna
G
Bencivenga
Paola
P
Giusti
Betti
B
Columbaro
Marta
M
Chu
Mon-Li
ML
Merlini
Luciano
L
Pepe
Guglielmina
G
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
D024142:Collagen Type VI; D012333:RNA, Messenger