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Distinct, but not completely separate spatial transport routes in the nuclear pore complex

机译：核孔复合物中不同但不完全分开的空间运输途径

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The nuclear pore complex (NPC), which provides the permeable and selective transport path between the nucleus and cytoplasm of eukaryotic cells, allows both the passive diffusion of small molecules in a signal-independent manner and the transport receptor-facilitated translocation of cargo molecules in a signal-dependent manner. However, the spatial and functional relationships between these two transport pathways, which represent critical information for unraveling the fundamental nucleocytoplasmic transport mechanism, remain in dispute. The direct experimental examination of passive and facilitated transport with a high spatiotemporal resolution under real-time trafficking conditions in native NPCs is still difficult. To address this issue and further define these transport mechanisms, we recently developed single-point edge-excitation sub-diffraction (SPEED) microscopy and a deconvolution algorithm to directly map both passive and facilitated transport routes in three dimensions (3D) in native NPCs. Our findings revealed that passive and facilitated transport occur through spatially distinct transport routes. Signal-independent small molecules exhibit a high probability of passively diffusing through an axial central viscous channel, while transport receptors and their cargo complexes preferentially travel through the periphery, around this central channel, after interacting with phenylalanine-glycine (FG) filaments. Strikingly, these two distinct transport zones are not completely separate either spatially or functionally. Instead, their conformations are closely correlated and simultaneously regulated. In this review, we will specifically highlight a detailed procedure for 3D mapping of passive and facilitated transport routes, demonstrate the correlation between these two distinct pathways, and finally, speculate regarding the regulation of the transport pathways driven by the conformational changes of FG filaments in NPCs.
机译：核孔复合物（NPC）提供了真核细胞核与细胞质之间的渗透性和选择性转运途径，既允许小分子以信号独立的方式进行被动扩散，又允许转运受体促进货物分子的易位依赖信号的方式。然而，这两个运输途径之间的空间和功能关系，代表着揭示基本核质运输机制的关键信息，仍然存在争议。在本地NPC上，在实时贩运条件下以高时空分辨率对被动运输和便利运输进行直接实验检查仍然很困难。为了解决此问题并进一步定义这些传输机制，我们最近开发了单点边缘激发亚衍射（SPEED）显微镜和去卷积算法，以直接在本机NPC中的三个维度（3D）上绘制被动和便利的运输路线。我们的发现表明，被动运输和便利运输通过空间上不同的运输路线发生。与信号无关的小分子在轴向中央粘性通道中被动扩散的可能性很高，而转运受体及其货物配合物在与苯丙氨酸-甘氨酸（FG）细丝相互作用后，优先穿过该中央通道的外围。令人惊讶的是，这两个不同的运输区域在空间或功能上都没有完全分开。相反，它们的构象紧密相关并且同时受到调节。在这篇综述中，我们将重点介绍用于被动和便利运输路径的3D映射的详细过程，演示这两个不同路径之间的相关性，最后推测由FG细丝构象变化驱动的运输路径的调控。 NPC。

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