Myopathy mutations in α-skeletal-muscle actin cause a range of molecular defects

被引:73
作者
Costa, CF
Rommelaere, H
Waterschoot, D
Sethi, KK
Nowak, KJ
Laing, NG
Ampe, C
Machesky, LM [1 ]
机构
[1] Univ Birmingham, Div Mol Cell Biol, Sch Biosci, Birmingham B15 2TT, W Midlands, England
[2] Univ Ghent, Dept Biochem, B-9052 Ghent, Belgium
[3] Flanders Interuniv Inst Biotechnol, B-9052 Ghent, Belgium
[4] Univ Western Australia, Australian Neuromuscular Res Inst, Ctr Neuromuscular & Neurol Disorders, Nedlands, WA 6009, Australia
[5] Western Australian Inst Med Res, Med Res Ctr, QEII Med Ctr, Nedlands, WA 6009, Australia
关键词
actin; nematine myopathy; actin polymerization; protein foldin; myopathy; actin mutations;
D O I
10.1242/jcs.01172
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Mutations in the gene encoding alpha-skeletal-muscle actin, ACTA1, cause congenital myopathies of various phenotypes that have been studied since their discovery in 1999. Although much is now known about the clinical aspects of myopathies resulting from over 60 different ACTA1 mutations, we have very little evidence for how mutations alter the behavior of the actin protein and thus lead to disease. We used a combination of biochemical and cell biological analysis to classify 19 myopathy mutants and found a range of defects in the actin. Using in vitro expression systems, we probed actin folding and actin's capacity to interact with actin-binding proteins and polymerization. Only two mutants failed to fold; these represent recessive alleles, causing severe myopathy, indicating that patients produce nonfunctional actin. Four other mutants bound tightly to cyclase-associated protein, indicating a possible instability in the nucleotide-binding pocket, and formed rods and aggregates in cells. Eleven mutants showed defects in the ability to co-polymerize with wild-type actin. Some of these could incorporate into normal actin structures in NIH 3T3 fibroblasts, but two of the three tested also formed aggregates. Four mutants showed no defect in vitro but two of these formed aggregates in cells, indicating functional defects that we have not yet tested for. Overall, we found a range of defects and behaviors of the mutants in vitro and in cultured cells, paralleling the complexity of actin-based muscle myopathy phenotypes.
引用
收藏
页码:3367 / 3377
页数:11
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