PDF(Pubmed)
Abstract：
Understanding the spatial organization of nucleoporins (Nups) with intrinsically disordered domains within the nuclear pore complex (NPC) is crucial for deciphering eukaryotic nucleocytoplasmic transport. Leveraging high-speed 2D single-molecule tracking and virtual 3D super-resolution microscopy in live HeLa cells, we investigated the spatial distribution of all eleven phenylalanine-glycine (FG)-rich Nups within individual NPCs. Our study reveals a nuanced landscape of FG-Nup conformations and arrangements. Five FG-Nups are steadfastly anchored at the NPC scaffold, collectively shaping a central doughnut-shaped channel, while six others exhibit heightened flexibility, extending towards the cytoplasmic and nucleoplasmic regions. Intriguingly, Nup214 and Nup153 contribute to cap-like structures that dynamically alternate between open and closed states along the nucleocytoplasmic transport axis, impacting the cytoplasmic and nuclear sides, respectively. Furthermore, Nup98, concentrated at the scaffold region, extends throughout the entire NPC while overlapping with other FG-Nups. Together, these eleven FG-Nups compose a versatile, capped trichoid channel spanning approximately 270 nm across the nuclear envelope. This adaptable trichoid channel facilitates a spectrum of pathways for passive diffusion and facilitated nucleocytoplasmic transport. Our comprehensive mapping of FG-Nup organization within live NPCs offers a unifying mechanism accommodating multiple transport pathways, thereby advancing our understanding of cellular transport processes.
摘要：
了解核孔蛋白（Nups）在核孔复合物（NPC）中具有内在无序结构域的空间组织对于破译真核核质运输至关重要。在活的HeLa细胞中利用高速2D单分子跟踪和虚拟3D超分辨率显微镜，我们调查了单个NPC中所有11个富含苯丙氨酸-甘氨酸（FG）的Nups的空间分布。我们的研究揭示了FG-Nup构象和排列的细微差别。五个FG-Nups坚定地固定在NPC支架上，共同形成一个中央甜甜圈形通道，而其他六个则表现出更高的灵活性，向细胞质和核质区延伸。有趣的是，Nup214和Nup153有助于帽状结构，它们沿着核质运输轴在开放状态和封闭状态之间动态交替，影响细胞质和细胞核，分别。此外，Nup98，集中在脚手架区域，在整个NPC中延伸，同时与其他FG-Nups重叠。一起,这11个FG-Nups组成了一个多才多艺的，覆盖的毛状通道跨越核包膜约270nm。这种适应性的毛状通道促进了被动扩散和促进的核质转运的一系列途径。我们的FG-Nup组织在实时NPC中的全面映射提供了一种统一的机制，可容纳多种运输途径，从而促进我们对细胞运输过程的理解。
