Transport of large cargoes, such as ribosomal subunits, through the permeability barrier of nuclear pore complexes (P08)

具有嵌入的核孔复合物（NPC）的核膜将真核细胞分成核和胞质区室。NPC通常会限制惰性大分子的通过。然而，穿梭核转运受体（NTR）赋予与核孔蛋白FG重复的相互作用，从而介导所选货物的促进的NPC通过。在这个项目中，我们使用核糖体亚基作为模型系统来研究大（MDA大小）货物的运输。通过生成针对成熟核糖体组分的纳米抗体，将其与核转运信号融合并将其与NTR复合，我们将分析使成熟核糖体亚基具有转运能力所需的相互作用的数量，分布和性质。作为一种补充的方法，我们将表征内源性的，有出口能力的核糖体前体，并研究其通过NPC和通过重建的渗透性屏障易位所需的功能。

The nuclear envelope with embedded nuclear pore complexes (NPCs) divides eukaryotic cells into a nuclear and a cytoplasmic compartment. NPCs usually restrict the passage of inert macromolecules. Shuttling nuclear transport receptors (NTRs), however, confer interactions with nucleoporin FG repeats and thereby mediate facilitated NPC-passage of selected cargoes. In this project, we are studying the transport of large (MDa-sized) cargoes using ribosomal subunits as a model system. By generating nanobodies against components of mature ribosomes, fusing them with nuclear transport signals and complexing them with NTRs, we will analyze the number, distribution, and nature of interactions that are required for making mature ribosomal subunits transport-competent. As a complementary approach, we will characterize endogenous, export-competent ribosomal precursors, and study the features needed for their translocation through NPCs and through reconstituted permeability barriers.