Lis-1 is required for dynein-dependent cell division process in C-elegans embryos
Lis-1 is required for dynein-dependent cell division process in C-elegans embryos
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DOI：
10.1242/jcs.01344
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发表时间：
2004-09-01
影响因子：
4
通讯作者：
Gönczy, P
中科院分区：
文献类型：
--
作者：
Cockell, MM;Baumer, M;Gönczy, P
我们研究了进化保守蛋白Lis 1在秀丽隐杆线虫胚胎细胞分裂过程中的作用。我们鉴定了lis-1的表观无效等位基因，其导致的缺陷与动力蛋白重链dhc-1失活后观察到的缺陷相同，包括中心体分离和纺锤体组装的缺陷。我们提出了针对LIS-1的抗体，并产生了表达功能性GFP-LIS-1的转基因动物。使用间接免疫荧光和旋转盘共聚焦显微镜，我们发现，LIS-1存在于整个细胞质中，并在离散的亚细胞位置，包括细胞皮质，附近的微管星状体，核周边和动粒富集。我们确定，lis-1有助于，但不是必不可少的，DHC-1富集在特定的亚细胞位置。相反，我们发现dhc-1，以及dynactin组件dnc-1（p150（Glued））和dnc-2（p50/dynitin），是必不可少的LIS-1靶向核周边，但不是细胞皮质，也不动粒。这些结果表明，动力蛋白和Lis 1，虽然在相同的过程中发挥作用，有针对性的部分独立于彼此。
We investigated the role of the evolutionarily conserved protein Lis1 in cell division processes of Caenorhabditis elegans embryos. We identified apparent null alleles of lis-1, which result in defects identical to those observed after inactivation of the dynein heavy chain dhc-1, including defects in centrosome separation and spindle assembly. We raised antibodies against LIS-1 and generated transgenic animals expressing functional GFP-LIS-1. Using indirect immunofluorescence and spinning-disk confocal microscopy, we found that LIS-1 is present throughout the cytoplasm and is enriched in discrete subcellular locations, including the cell cortex, the vicinity of microtubule asters, the nuclear periphery and kinetochores. We established that lis-1 contributes to, but is not essential for, DHC-1 enrichment at specific subcellular locations. Conversely, we found that dhc-1, as well as the dynactin components dnc-1 (p150(Glued)) and dnc-2 (p50/dynamitin), are essential for LIS-1 targeting to the nuclear periphery, but not to the cell cortex nor to kinetochores. These results suggest that dynein and Lis1, albeit functioning in identical processes, are targeted partially independently of one another.