VINCULIN IS ESSENTIAL FOR MUSCLE FUNCTION IN THE NEMATODE
VINCULIN IS ESSENTIAL FOR MUSCLE FUNCTION IN THE NEMATODE
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DOI：
10.1083/jcb.114.4.715
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发表时间：
1991-08-01
影响因子：
7.8
通讯作者：
WATERSTON, RH
中科院分区：
文献类型：
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作者：
BARSTEAD, RJ;WATERSTON, RH
线虫体壁肌肉中的肌动蛋白细丝通过含有纽蛋白的结构(称为致密小体，Z线类似物)附着在肌膜上。为了研究纽蛋白在体内的功能，我们进行了一项旨在恢复线虫纽蛋白基因Deb-1区域突变的遗传筛选。根据四个独立的标准，其中两个被分离的突变体被证明是由于Deb-1基因的改变。首先，抗体染色显示突变体的纽蛋白水平降低。其次，每个突变基因的序列都与野生型基因的序列发生了变化，其中一个突变改变了保守的剪接序列，另一个突变产生了过早的琥珀终止密码子。第三，琥珀突变体受到超7琥珀抑制基因tRNA的抑制。最后，注射该基因的克隆野生型拷贝拯救了该突变体。缺乏纽蛋白的突变动物抑制了L1幼虫的发育。在这些动物中，胚胎的伸长在两倍的长度处中断，因此突变的动物比同期的野生型动物要短。突变体瘫痪，肌肉组织紊乱，这一表型与纽蛋白对线虫肌肉功能至关重要的观点一致。
Actin filaments in the body wall muscle of the nematode Caenorhabditis elegans are attached to the sarcolemma through vinculin-containing structures called dense bodies, Z-line analogues. To investigate the in vivo function of vinculin, we executed a genetic screen designed to recover mutations in the region of the nematode vinculin gene, deb-1. According to four independent criteria, two of the isolated mutants were shown to be due to alterations in the deb-1 gene. First, antibody staining showed that the mutants had reduced levels of vinculin. Second, the sequence of each mutant gene was altered from that of wild type, with one mutation altering a conserved splice sequence and the other generating a premature amber stop codon. Third, the amber mutant was suppressed by the sup-7 amber suppressor tRNA gene. Finally, injection of a cloned wild type copy of the gene rescued the mutant. Mutant animals lacking vinculin arrested development as L1 larvae. In such animals, embryonic elongation was interrupted at the twofold length, so that the mutants were shorter than wild type animals at the same stage. The mutants were paralyzed and had disorganized muscle, a phenotype consistent with the idea that vinculin is essential for muscle function in the nematode.