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Structural characterization of altered nucleoporin Nup153 expression in human cells by thin-section electron microscopy

机译：薄层电子显微镜在人细胞中改变核仁蛋白Nup153表达的结构表征

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Nuclear pore complexes (NPCs) span the 2 membranes of the nuclear envelope (NE) and facilitate nucleocytoplasmic exchange of macromolecules. NPCs have a roughly tripartite structural organization with the so-called nuclear basket emanating from the NPC scaffold into the nucleoplasm. The nuclear basket is composed of the 3 nucleoporins Nup153, Nup50, and Tpr, but their specific role for the structural organization of this NPC substructure is, however, not well established. In this study, we have used thin-section transmission electron microscopy to determine the structural consequences of altering the expression of Nup153 in human cells. We show that the assembly and integrity of the nuclear basket is not affected by Nup153 depletion, whereas its integrity is perturbed in cells expressing high concentrations of the zinc-finger domain of Nup153. Moreover, even mild over-expression of Nup153 is coinciding with massive changes in nuclear organization and it is the excess of the zinc-finger domain of Nup153 that is sufficient to induce these rearrangements. Our data indicate a central function of Nup153 in the organization of the nucleus, not only at the periphery, but throughout the entire nuclear interior.
机译：核孔复合物（NPC）跨越核被膜（NE）的2个膜，并促进大分子的核质交换。 NPC具有大致三方的结构组织，所谓的核篮从NPC支架散发到核质中。核篮由3个核孔蛋白Nup153，Nup50和Tpr组成，但是，尚不清楚它们在该NPC亚结构的结构组织中的具体作用。在这项研究中，我们使用了薄层透射电子显微镜来确定改变人细胞中Nup153表达的结构后果。我们显示核篮的组装和完整性不受Nup153耗尽的影响，而其完整性在表达高浓度Nup153锌指结构域的细胞中受到干扰。此外，即使Nup153的轻度过度表达也与核组织的巨大变化相吻合，正是Nup153的锌指结构域的过量足以诱导这些重排。我们的数据表明Nup153在核组织中的核心功能，不仅在外围，而且在整个核内部。

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