Osmani Aysha H, Davies Jonathan, Liu Hui-Lin, Nile Aaron, Osmani Stephen A
Department of Molecular Genetics, The Ohio State University, Columbus, OH 43210, USA.
Mol Biol Cell. 2006 Dec;17(12):4946-61. doi: 10.1091/mbc.e06-07-0657. Epub 2006 Sep 20.
To define the extent of the modification of the nuclear pore complex (NPC) during Aspergillus nidulans closed mitosis, a systematic analysis of nuclear transport genes has been completed. Thirty genes have been deleted defining 12 nonessential and 18 essential genes. Several of the nonessential deletions caused conditional phenotypes and self-sterility, whereas deletion of some essential genes caused defects in nuclear structure. Live cell imaging of endogenously tagged NPC proteins (Nups) revealed that during mitosis 14 predicted peripheral Nups, including all FG repeat Nups, disperse throughout the cell. A core mitotic NPC structure consisting of membrane Nups, all components of the An-Nup84 subcomplex, An-Nup170, and surprisingly, An-Gle1 remained throughout mitosis. We propose this minimal mitotic NPC core provides a conduit across the nuclear envelope and acts as a scaffold to which dispersed Nups return during mitotic exit. Further, unlike other dispersed Nups, An-Nup2 locates exclusively to mitotic chromatin, suggesting it may have a novel mitotic role in addition to its nuclear transport functions. Importantly, its deletion causes lethality and defects in DNA segregation. This work defines the dramatic changes in NPC composition during A. nidulans mitosis and provides insight into how NPC disassembly may be integrated with mitosis.
为了确定构巢曲霉封闭有丝分裂期间核孔复合体(NPC)的修饰程度,已完成对核转运基因的系统分析。已删除30个基因,其中12个为非必需基因,18个为必需基因。一些非必需基因的缺失导致了条件性表型和自我不育,而一些必需基因的缺失则导致了核结构缺陷。对内源标记的NPC蛋白(核孔蛋白)进行活细胞成像显示,在有丝分裂期间,14个预测的外周核孔蛋白,包括所有FG重复核孔蛋白,分散在整个细胞中。一个由膜核孔蛋白、An-Nup84亚复合体的所有成分、An-Nup170以及令人惊讶的是An-Gle1组成的核心有丝分裂NPC结构在整个有丝分裂过程中都存在。我们提出,这个最小的有丝分裂NPC核心提供了一个穿过核膜的通道,并作为一个支架,在有丝分裂后期分散的核孔蛋白会回到该支架上。此外,与其他分散的核孔蛋白不同,An-Nup2仅定位于有丝分裂染色质,这表明它除了具有核转运功能外,可能还具有新的有丝分裂作用。重要的是,它的缺失会导致致死性和DNA分离缺陷。这项工作定义了构巢曲霉有丝分裂期间NPC组成的巨大变化,并深入了解了NPC拆卸如何与有丝分裂整合。