Single-molecule studies of nucleocytoplasmic transport: from one dimension to three dimensions.
Single-molecule studies of nucleocytoplasmic transport: from one dimension to three dimensions.
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DOI：
10.1039/c1ib00041a
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发表时间：
2012-01
期刊：
影响因子：
--
通讯作者：
Yang W
中科院分区：
文献类型：
--
作者：
Goryaynov A;Ma J;Yang W
在真核细胞中，蛋白质和遗传物质通过双膜核膜的双向运输是由核孔复合物（nuclear pore complex，NPC）介导的。NPC中苯丙氨酸-甘氨酸（FG）-核孔蛋白（Nup）形成的高度选择性屏障允许两种转运模式：被动扩散和转运受体促进的转运。核质转运的严格调节对于细胞存活、分化、生长和其他基本活动至关重要。然而，由于有限的知识的天然配置的FG-Nup屏障和之间的相互作用的转运分子和NPC的屏障，精确的核质转运机制仍然没有得到解决。为了改进运输机制，单分子荧光显微镜方法已被用来获得单个荧光分子通过NPC的运输动力学，并绘制过境分子和FG-Nup屏障之间的相互作用。核质运输的重要特征，如运输时间，运输效率和空间分布的单个转运分子在NPC中，已经得到，不能测量的合奏平均方法或传统的电子显微镜。在这篇评论中，我们讨论了各种单分子技术的发展及其在体外和体内核质转运中的应用。特别是，我们强调了最近的进展，从一维到三维的单分子表征的运输通过NPC和提出了一个全面的了解核质运输机制，通过这种新的技术发展（105参考文献）。
In eukaryotic cells, the bidirectional trafficking of proteins and genetic materials across the double-membrane nuclear envelope is mediated by nuclear pore complexes (NPCs). A highly selective barrier formed by the phenylalanine–glycine (FG)-nucleoporin (Nup) in the NPC allows for two transport modes: passive diffusion and transport receptor-facilitated translocation. Strict regulation of nucleocytoplasmic transport is crucial for cell survival, differentiation, growth and other essential activities. However, due to the limited knowledge of the native configuration of the FG-Nup barrier and the interactions between the transiting molecules and the barrier in the NPC, the precise nucleocytoplasmic transport mechanism remains unresolved. To refine the transport mechanism, single-molecule fluorescence microscopy methods have been employed to obtain the transport kinetics of individual fluorescent molecules through the NPC and to map the interactions between transiting molecules and the FG-Nup barrier. Important characteristics of nucleocytoplasmic transport, such as transport time, transport efficiency and spatial distribution of single transiting molecules in the NPC, have been obtained that could not be measured by either ensemble average methods or conventional electron microscopy. In this critical review, we discuss the development of various single-molecule techniques and their application to nucleocytoplasmic transport in vitro and in vivo. In particular, we highlight a recent advance from one-dimensional to three-dimensional single-molecule characterization of transport through the NPC and present a comprehensive understanding of the nucleocytoplasmic transport mechanism obtained by this new technical development (105 references).
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影响因子：
7.8
作者：
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通讯作者：
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DOI：
10.1073/pnas.0506345103
发表时间：
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作者：
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DOI：
10.1083/jcb.200806173
发表时间：
2008-10-06
期刊：
The Journal of cell biology
影响因子：
--
作者：
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通讯作者：
Kubitscheck U