Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA.
Nanobiology Institute, Yale University, West Haven, CT, USA.
Sci Adv. 2024 Nov 15;10(46):eadq8773. doi: 10.1126/sciadv.adq8773. Epub 2024 Nov 13.
Nucleoporins (nups) in the nuclear pore complex (NPC) form a selective barrier that suppresses the diffusion of most macromolecules while enabling rapid transport of nuclear transport receptor (NTR)-bound cargos. Recent studies have shown that the NPC may dilate and constrict, but how altering the NPC diameter affects its selective barrier properties remains unclear. Here, we build DNA nanopores with programmable diameters and nup arrangements to model the constricted and dilated NPCs. We find that Nup62 proteins form a dynamic cross-channel barrier impermeable to hepatitis B virus (HBV) capsids when grafted inside 60-nm-wide nanopores but not in 79-nm pores, where Nup62 cluster locally. Furthermore, importin-β1 substantially changes the dynamics of Nup62 assemblies and facilitates the passage of HBV capsids through the 60-nm NPC mimics containing Nup62 and Nup153. Our study shows that transport channel width is critical to the permeability of nup barriers and underscores NTRs' role in dynamically remodeling nup assemblies and mediating the nuclear entry of viruses.
核孔复合体（NPC）中的核孔蛋白（nups）形成一个选择性屏障，抑制大多数大分子的扩散，同时允许核转运受体（NTR）结合的货物快速运输。最近的研究表明，NPC 可能会扩张和收缩，但改变 NPC 直径如何影响其选择性屏障特性尚不清楚。在这里，我们构建了具有可编程直径和 nups 排列的 DNA 纳米孔，以模拟收缩和扩张的 NPC。我们发现，当 Nup62 蛋白被嫁接在 60nm 宽的纳米孔内时，它们会形成一个动态的跨通道屏障，阻止乙型肝炎病毒（HBV）衣壳通过，但在 79nm 的孔内不会，因为 Nup62 会在那里局部聚集。此外，importin-β1 会极大地改变 Nup62 组装体的动力学，并促进 HBV 衣壳通过含有 Nup62 和 Nup153 的 60nm NPC 模拟物的传递。我们的研究表明，运输通道的宽度对 nups 屏障的通透性至关重要，并强调了 NTR 在动态重塑 nups 组装体和介导病毒进入核内方面的作用。
