Insight into Structure and Assembly of the Nuclear Pore Complex by Utilizing the Genome of a Eukaryotic Thermophile
Insight into Structure and Assembly of the Nuclear Pore Complex by Utilizing the Genome of a Eukaryotic Thermophile
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DOI：
10.1016/j.cell.2011.06.039
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发表时间：
2011-07-22
期刊：
影响因子：
64.5
通讯作者：
Hurt, Ed
中科院分区：
文献类型：
--
作者：
Amlacher, Stefan;Sarges, Phillip;Hurt, Ed
尽管经过数十年的研究，核孔复合体 (NPC) 由类似 30 个核孔蛋白 (Nups) 组成，但其结构和组装仍然难以捉摸。在这里，我们报告了嗜热真菌嗜热毛壳菌 (CT) 的基因组，并鉴定了其中 Nups 的完整库。与嗜温蛋白相比，嗜热蛋白在结构和生化研究中表现出更好的特性，纯化的 ctNups 能够重建内孔环模块，该模块横跨 NPC 从锚定膜到中央运输通道的宽度。该模块由两个大Nups Nup192 和Nup170 组成，它们通过由连接子Nups Nic96 和Nup53 组成的短线性基序灵活桥接。该组装说明了 Nup 相互作用如何在 NPC 支架内产生结构可塑性。因此，我们的研究结果证明了嗜热真核生物基因组在研究复杂分子机器方面的效用。
Despite decades of research, the structure and assembly of the nuclear pore complex (NPC), which is composed of similar to 30 nucleoporins (Nups), remain elusive. Here, we report the genome of the thermophilic fungus Chaetomium thermophilum (ct) and identify the complete repertoire of Nups therein. The thermophilic proteins show improved properties for structural and biochemical studies compared to their mesophilic counterparts, and purified ctNups enabled the reconstitution of the inner pore ring module that spans the width of the NPC from the anchoring membrane to the central transport channel. This module is composed of two large Nups, Nup192 and Nup170, which are flexibly bridged by short linear motifs made up of linker Nups, Nic96 and Nup53. This assembly illustrates how Nup interactions can generate structural plasticity within the NPC scaffold. Our findings therefore demonstrate the utility of the genome of a thermophilic eukaryote for studying complex molecular machines.