Structure-Function Mapping of the Nuclear Pore Complex

项目概要（摘要） 核孔复合体 (NPC) 是嵌入核膜中的大型圆柱形组件，位于核膜的中心 两个相关水平的核功能。首先，全国人大作为交通运输的监管者，控制着交通信号的接入。 DNA 和 DNA 遗传信息的传递。其次，NPC是基因的重要调节者。 结合染色质及其调节因子来控制表达状态，这一现象目前人们知之甚少 分子水平。所有真核细胞中的这些关键作用涉及数十种相互作用的途径，影响 几乎细胞功能的所有方面。因此，NPC 的破坏会导致许多人类疾病。 尽管如此，尽管核运输机制是一个有效且强大的药物目标，但 NPC 和 核转运机制尚未成为治疗策略的重要组成部分。可以说，有两个 造成这种情况的根本原因是：（i）我们对 NPC 的结构了解不够，无法预测 它的行为； (ii) 核运输机制影响着一系列令人眼花缭乱的细胞功能——因此即使 深入了解其结构，我们仍然需要补充功能信息才能预测 有针对性地破坏运输路径关键要素的结果。我们提出两个具体目标 以协同方式相互告知。首先，我们将进行疾病相关的结构图谱 Nup 复合物，重点关注细胞质输出平台和内环的组件 与致癌和发育缺陷有关。我们将使用现有方法的增强版本 成功部署以生成这两个区域及其附着点的高分辨率地图。开 完成这项研究后，我们将以高精度绘制大部分 NPC 的地图，从而使这两个区域能够被 从整个全国人大会议的背景来看。其次，同时，我们将绘制疾病的功能图—— 相关的 Nup 复合物。我们将剖析与目标 Nup 复合物相关的功能，并且 确定与其改变相关的缺陷 - 检验这些 Nups 所关联的假设 疾病，因为它们的破坏以不同于其他疾病的方式改变关键基因表达模式 核孔蛋白。实现这些目标将生成前所未有的详细的 NPC 结构功能图，并且 对于理解 NPC 的不同部分如何共同作用以确定其功能至关重要。这个 该项目将揭示与人类鼻咽癌功能障碍相关的多种疾病的性质；瞄准的 最终打开核运输机制，实现合理和预测性药物设计。

PROJECT SUMMARY (Abstract) The Nuclear Pore Complex (NPC) is a large cylindrical assembly embedded in the nuclear envelope, central for nuclear function at two related levels. First, as a regulator of transport, the NPC controls signalling access to the DNA and the passage of genetic information from DNA. Second, the NPC is an important regulator of genes by binding chromatin and its regulators to control expression states, a phenomenon that is poorly understood at the molecular level. These pivotal roles in all eukaryotic cells involve dozens of interacting pathways influencing virtually all aspects of cellular function. As a consequence, disruption of the NPC leads to many human disorders. Despite this, and though the nuclear transport machinery is a valid and powerful drug target, the NPC and the nuclear transport machinery have not been a significant part of therapeutic strategies. Arguably, there are two fundamental reasons why this is the case: (i) we do not know enough about the structure of the NPC to predict its behavior; (ii) the nuclear transport machinery impacts a bewildering array of cellular functions - thus even with a deep understanding of its structure, we still require complementary functional information to be able to predict the outcome of the targeted disruption of key elements of the transport pathway. We propose two Specific Aims that inform each other in a synergistic fashion. First, we will perform structural mapping of disease-associated Nup complexes, focusing on components of the cytoplasmic export platform and inner rings that have been linked to oncogenic and developmental defects. We will use enhanced versions of the methods we have already successfully deployed to generate high resolution maps of these two regions and their attachment sites. On completion of this study, we will have mapped most of the NPC at high precision, allowing the two regions to be seen in the context of the whole NPC assembly. Second, and in parallel, we will map the functions of disease- associated Nup complexes. We will dissect the functionalities associated with the target Nup complexes, and determine the defects associated with their alteration - testing the hypothesis that these Nups are linked to diseases because their disruption alters critical gene expression patterns in a manner distinct from other nucleoporins. Realizing these aims will generate NPC structure-function maps in unprecedented detail and which are essential to understanding how different parts of the NPC act together to determine its functionality. This project will shed light on the nature of numerous disorders associated with human NPC dysfunction; aimed ultimately to open the nuclear transport machinery to rational and predictive drug design.