Interactions between a Nuclear Transporter and a Subset of Nuclear Pore Complex Proteins Depend on Ran GTPase
Interactions between a Nuclear Transporter and a Subset of Nuclear Pore Complex Proteins Depend on Ran GTPase
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核转运蛋白和核孔复合蛋白子集之间的相互作用依赖于 Ran GTPase
DOI：
10.1128/mcb.19.2.1547
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发表时间：
1999
影响因子：
5.3
通讯作者：
P. Silver
中科院分区：
文献类型：
--
作者：
M. Seedorf;M. Damelin;J. Kahana;T. Taura;P. Silver
摘要 被转运到细胞核中的蛋白质被输入核转运蛋白核转运受体家族的成员识别。与核孔复合体 (NPC) 对接后，货物-受体复合体穿过水孔通道。一旦货物放行，进口货物就会通过通道返回，进行新一轮的运输。因此，输入蛋白和输出蛋白（该家族中参与输出的另一个成员）被认为在核内部和细胞质之间不断穿梭。为了了解核转运蛋白如何穿过 NPC，我们在酵母导入蛋白家族的几个成员（包括 Pse1p、Sxm1p、Xpo1p 和 Kap95p）与绿色荧光蛋白 (GFP) 之间构建了功能性蛋白融合。使用高度特异性的抗 GFP 抗体分离含有核转运蛋白的复合物。对 Pse1-GFP 进行了最详细的研究。 Pse1-GFP 与 importin-α 和 -β（酵母细胞中的 Srp1p 和 Kap95p）形成复合物，对 Ran GTPase 的核苷酸结合状态敏感。此外，Pse1p 与核孔蛋白 Nsp1p、Nup159p 和 Nup116p 结合，而 Sxm1p、Xpo1p 和 Kap95p 显示与核孔蛋白相互作用的不同模式。 Pse1p 与核孔蛋白的关联也取决于 Ran 的核苷酸结合状态。当 Ran 处于 GTP 结合状态时，核孔蛋白结合就会丢失。不结合 Ran 的 Pse1p 突变体也无法与核孔蛋白相互作用。这些数据表明，Pse1p 等转运受体以 Ran 依赖性方式与某些核孔蛋白相互作用。当 Pse1p 通过 NPC 进出细胞核时，这些核孔蛋白可能代表 Pse1p 的主要对接位点。
ABSTRACT Proteins to be transported into the nucleus are recognized by members of the importin-karyopherin nuclear transport receptor family. After docking at the nuclear pore complex (NPC), the cargo-receptor complex moves through the aqueous pore channel. Once cargo is released, the importin then moves back through the channel for new rounds of transport. Thus, importin and exportin, another member of this family involved in export, are thought to continuously shuttle between the nuclear interior and the cytoplasm. In order to understand how nuclear transporters traverse the NPC, we constructed functional protein fusions between several members of the yeast importin family, including Pse1p, Sxm1p, Xpo1p, and Kap95p, and the green fluorescent protein (GFP). Complexes containing nuclear transporters were isolated by using highly specific anti-GFP antibodies. Pse1-GFP was studied in the most detail. Pse1-GFP is in a complex with importin-α and -β (Srp1p and Kap95p in yeast cells) that is sensitive to the nucleotide-bound state of the Ran GTPase. In addition, Pse1p associates with the nucleoporins Nsp1p, Nup159p, and Nup116p, while Sxm1p, Xpo1p, and Kap95p show different patterns of interaction with nucleoporins. Association of Pse1p with nucleoporins also depends on the nucleotide-bound state of Ran; when Ran is in the GTP-bound state, the nucleoporin association is lost. A mutant form of Pse1p that does not bind Ran also fails to interact with nucleoporins. These data indicate that transport receptors such as Pse1p interact in a Ran-dependent manner with certain nucleoporins. These nucleoporins may represent major docking sites for Pse1p as it moves in or out of the nucleus via the NPC.
影响因子：
10.5
作者：
Marie A. Vodicka;D. Koepp;P. Silver;M. Emerman
通讯作者：
Marie A. Vodicka;D. Koepp;P. Silver;M. Emerman
Rat7p/Nup159p（酿酒酵母必需核孔蛋白）的结构/功能分析。
影响因子：
4
作者：
DelPriore,V;Heath,C;Snay,C;MacMillan,A;Gorsch,L;Dagher,S;Cole,C
通讯作者：
Cole,C