Rafael J A, Townsend E R, Squire S E, Potter A C, Chamberlain J S, Davies K E
Department of Human Anatomy and Genetics, University of Oxford, Oxford OX1 3QX, UK.
Hum Mol Genet. 2000 May 22;9(9):1357-67. doi: 10.1093/hmg/9.9.1357.
The X-linked muscle wasting disease Duchenne muscular dystrophy is caused by the lack of dystrophin in muscle. Protein structure predictions, patient mutations, in vitro binding studies and transgenic and knockout mice suggest that dystrophin plays a mechanical role in skeletal muscle, linking the subsarcolemmal cytoskeleton with the extracellular matrix through its direct interaction with the dystrophin-associated protein complex (DAPC). Although a signaling role for dystrophin has been postulated, definitive data have been lacking. To identify potential non-mechanical roles of dystrophin, we tested the ability of various truncated dystrophin transgenes to prevent any of the skeletal muscle abnormalities associated with the double knockout mouse deficient for both dystrophin and the dystrophin-related protein utrophin. We show that restoration of the DAPC with Dp71 does not prevent the structural abnormalities of the post-synaptic membrane or the abnormal oxidative properties of utrophin/dystrophin-deficient muscle. In marked contrast, a dystrophin protein lacking the cysteine-rich domain, which is unable to prevent dystrophy in the mdx mouse, is able to ameliorate these abnormalities in utrophin/dystrophin-deficient mice. These experiments provide the first direct evidence that in addition to a mechanical role and relocalization of the DAPC, dystrophin and utrophin are able to alter both structural and biochemical properties of skeletal muscle. In addition, these mice provide unique insights into skeletal muscle fiber type composition.
X连锁肌肉萎缩疾病杜氏肌营养不良症是由肌肉中缺乏抗肌萎缩蛋白引起的。蛋白质结构预测、患者突变、体外结合研究以及转基因和基因敲除小鼠研究表明,抗肌萎缩蛋白在骨骼肌中发挥机械作用,通过与抗肌萎缩蛋白相关蛋白复合体(DAPC)直接相互作用,将肌膜下细胞骨架与细胞外基质连接起来。尽管有人推测抗肌萎缩蛋白具有信号传导作用,但一直缺乏确凿的数据。为了确定抗肌萎缩蛋白潜在的非机械作用,我们测试了各种截短的抗肌萎缩蛋白转基因预防与同时缺乏抗肌萎缩蛋白和抗肌萎缩蛋白相关蛋白——肌养蛋白的双基因敲除小鼠相关的任何骨骼肌异常的能力。我们发现,用Dp71恢复DAPC并不能预防突触后膜的结构异常或肌养蛋白/抗肌萎缩蛋白缺陷型肌肉的异常氧化特性。与之形成鲜明对比的是,一种缺乏富含半胱氨酸结构域的抗肌萎缩蛋白,虽然不能预防mdx小鼠的肌营养不良,但却能够改善肌养蛋白/抗肌萎缩蛋白缺陷型小鼠的这些异常。这些实验提供了首个直接证据,表明除了具有机械作用和重新定位DAPC外,抗肌萎缩蛋白和肌养蛋白还能够改变骨骼肌的结构和生化特性。此外,这些小鼠为骨骼肌纤维类型组成提供了独特的见解。