School of Graduate Entry Medicine and Health, University of Nottingham, Royal Derby Hospital, Derby, United Kingdom.
PLoS Genet. 2012 Jan;8(1):e1002471. doi: 10.1371/journal.pgen.1002471. Epub 2012 Jan 12.
Two components of integrin containing attachment complexes, UNC-97/PINCH and UNC-112/MIG-2/Kindlin-2, were recently identified as negative regulators of muscle protein degradation and as having decreased mRNA levels in response to spaceflight. Integrin complexes transmit force between the inside and outside of muscle cells and signal changes in muscle size in response to force and, perhaps, disuse. We therefore investigated the effects of acute decreases in expression of the genes encoding these multi-protein complexes. We find that in fully developed adult Caenorhabditis elegans muscle, RNAi against genes encoding core, and peripheral, members of these complexes induces protein degradation, myofibrillar and mitochondrial dystrophies, and a movement defect. Genetic disruption of Z-line- or M-line-specific complex members is sufficient to induce these defects. We confirmed that defects occur in temperature-sensitive mutants for two of the genes: unc-52, which encodes the extra-cellular ligand Perlecan, and unc-112, which encodes the intracellular component Kindlin-2. These results demonstrate that integrin containing attachment complexes, as a whole, are required for proper maintenance of adult muscle. These defects, and collapse of arrayed attachment complexes into ball like structures, are blocked when DIM-1 levels are reduced. Degradation is also blocked by RNAi or drugs targeting calpains, implying that disruption of integrin containing complexes results in calpain activation. In wild-type animals, either during development or in adults, RNAi against calpain genes results in integrin muscle attachment disruptions and consequent sub-cellular defects. These results demonstrate that calpains are required for proper assembly and maintenance of integrin attachment complexes. Taken together our data provide in vivo evidence that a calpain-based molecular repair mechanism exists for dealing with attachment complex disruption in adult muscle. Since C. elegans lacks satellite cells, this mechanism is intrinsic to the muscles and raises the question if such a mechanism also exists in higher metazoans.
两个整合素连接附着复合物的成分,UNC-97/PINCH 和 UNC-112/MIG-2/Kindlin-2,最近被鉴定为肌肉蛋白降解的负调节剂,并且在响应空间飞行时其 mRNA 水平降低。整合素复合物在肌肉细胞内外传递力,并响应力和(或许)废用而发出肌肉大小变化的信号。因此,我们研究了这些多蛋白复合物编码基因表达急性下降的影响。我们发现,在完全发育的成年秀丽隐杆线虫肌肉中,针对这些复合物核心和外周成分编码基因的 RNAi 会诱导蛋白质降解、肌原纤维和线粒体病变以及运动缺陷。Z 线或 M 线特异性复合物成员的遗传缺失足以诱导这些缺陷。我们证实,在两个基因的温度敏感突变体中会发生缺陷:unc-52,其编码细胞外配体 Perlecan;以及 unc-112,其编码细胞内成分 Kindlin-2。这些结果表明,整合素连接附着复合物作为一个整体,对于维持成年肌肉的正常功能是必需的。这些缺陷以及排列的附着复合物塌陷成球状结构,在 DIM-1 水平降低时被阻断。降解也被针对钙蛋白酶的 RNAi 或药物阻断,这意味着整合素连接复合物的破坏导致钙蛋白酶的激活。在野生型动物中,无论是在发育过程中还是在成年期,钙蛋白酶基因的 RNAi 都会导致整合素肌肉附着的破坏以及随后的亚细胞缺陷。这些结果表明,钙蛋白酶对于整合素附着复合物的正确组装和维持是必需的。总之,我们的数据提供了体内证据,证明存在一种基于钙蛋白酶的分子修复机制来处理成年肌肉中附着复合物的破坏。由于秀丽隐杆线虫缺乏卫星细胞,这种机制是肌肉固有的,并提出了这样一个问题,即在高等后生动物中是否也存在这种机制。