Gepner J, Li M, Ludmann S, Kortas C, Boylan K, Iyadurai S J, McGrail M, Hays T S
Department of Genetics and Cell Biology, University of Minnesota, St. Paul 55108, USA.
Genetics. 1996 Mar;142(3):865-78. doi: 10.1093/genetics/142.3.865.
The microtubule motor cytoplasmic dynein has been implicated in a variety of intracellular transport processes. We previously identified and characterized the Drosophila gene Dhc64C, which encodes a cytoplasmic dynein heavy chain. To investigate the function of the cytoplasmic dynein motor, we initiated a mutational analysis of the Dhc64C dynein gene. A small deletion that removes the chromosomal region containing the heavy chain gene was used to isolate EMS-induced lethal mutations that define at least eight essential genes in the region. Germline transformation with a Dhc64C transgene rescued 16 mutant alleles in the single complementation group that identifies the dynein heavy chain gene. All 16 alleles were hemizygous lethal, which demonstrates that the cytoplasmic dynein heavy chain gene Dhc64C is essential for Drosophila development. Furthermore, our failure to recover somatic clones of cells homozygous for a Dhc64C mutation indicates that cytoplasmic dynein function is required for cell viability in several Drosophila tissues. The intragenic complementation of dynein alleles reveals multiple mutant phenotypes including male and/or female sterility, bristle defects, and defects in eye development.
微管马达胞质动力蛋白已被证明参与多种细胞内运输过程。我们之前鉴定并表征了果蝇基因Dhc64C,它编码一种胞质动力蛋白重链。为了研究胞质动力蛋白马达的功能,我们启动了对Dhc64C动力蛋白基因的突变分析。利用一个小缺失来去除包含重链基因的染色体区域,以分离出经乙基亚硝基脲(EMS)诱导的致死突变,这些突变定义了该区域至少八个必需基因。用Dhc64C转基因进行种系转化挽救了单一互补组中的16个突变等位基因,该互补组确定了动力蛋白重链基因。所有16个等位基因半合子致死,这表明胞质动力蛋白重链基因Dhc64C对果蝇发育至关重要。此外,我们未能获得Dhc64C突变纯合的体细胞克隆,这表明胞质动力蛋白功能在果蝇的多个组织中对细胞活力是必需的。动力蛋白等位基因的基因内互补揭示了多种突变表型,包括雄性和/或雌性不育、刚毛缺陷以及眼睛发育缺陷。