Robinson J T, Wojcik E J, Sanders M A, McGrail M, Hays T S
University of Minnesota, Department of Genetics, Cell Biology and Development, St. Paul, Minnesota 55108-1095, USA.
J Cell Biol. 1999 Aug 9;146(3):597-608. doi: 10.1083/jcb.146.3.597.
Cytoplasmic dynein is a multisubunit minus-end-directed microtubule motor that serves multiple cellular functions. Genetic studies in Drosophila and mouse have demonstrated that dynein function is essential in metazoan organisms. However, whether the essential function of dynein reflects a mitotic requirement, and what specific mitotic tasks require dynein remains controversial. Drosophila is an excellent genetic system in which to analyze dynein function in mitosis, providing excellent cytology in embryonic and somatic cells. We have used previously characterized recessive lethal mutations in the dynein heavy chain gene, Dhc64C, to reveal the contributions of the dynein motor to mitotic centrosome behavior in the syncytial embryo. Embryos lacking wild-type cytoplasmic dynein heavy chain were analyzed by in vivo analysis of rhodamine-labeled microtubules, as well as by immunofluorescence in situ methods. Comparisons between wild-type and Dhc64C mutant embryos reveal that dynein function is required for the attachment and migration of centrosomes along the nuclear envelope during interphase/prophase, and to maintain the attachment of centrosomes to mitotic spindle poles. The disruption of these centrosome attachments in mutant embryos reveals a critical role for dynein function and centrosome positioning in the spatial organization of the syncytial cytoplasm of the developing embryo.
细胞质动力蛋白是一种多亚基的向微管负端移动的分子马达,具有多种细胞功能。对果蝇和小鼠的遗传学研究表明,动力蛋白的功能在多细胞生物中至关重要。然而,动力蛋白的基本功能是否反映了有丝分裂的需求,以及动力蛋白具体参与哪些有丝分裂任务仍存在争议。果蝇是分析有丝分裂中动力蛋白功能的优秀遗传系统,能为胚胎细胞和体细胞提供出色的细胞学研究。我们利用先前鉴定的动力蛋白重链基因Dhc64C中的隐性致死突变,来揭示动力蛋白分子马达对合胞体胚胎中有丝分裂中心体行为的作用。通过对罗丹明标记微管的体内分析以及免疫荧光原位方法,对缺乏野生型细胞质动力蛋白重链的胚胎进行了分析。野生型和Dhc64C突变体胚胎之间的比较表明,在间期/前期,动力蛋白功能对于中心体沿核膜的附着和迁移是必需的,并且对于维持中心体与有丝分裂纺锤体极的附着也是必需的。突变体胚胎中这些中心体附着的破坏揭示了动力蛋白功能和中心体定位在发育中胚胎合胞体细胞质空间组织中的关键作用。