Structure-Function Mapping of the Nuclear Pore Complex

项目摘要(摘要) 核孔复合体(NPC)是嵌入核膜中的一个大的圆柱形组件，是 核功能在两个相关的层面上。首先，作为交通监管机构，全国人大控制着进入 DNA和来自DNA的遗传信息的传递。其次，全国人大是基因的重要调节者，通过 结合染色质及其调节剂来控制表达状态，这一现象在 分子水平。这些在所有真核细胞中的关键作用涉及数十个相互作用的途径，影响 细胞功能的几乎所有方面。其结果是，全国人大会议的中断导致了许多人类疾病。 尽管如此，尽管核运输机械是一个有效和强大的毒品目标，但全国人大和 核运输机械并不是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：(一)我们对全国人大的结构还不够了解，无法预测 它的行为；(二)核运输机械影响了一系列令人困惑的细胞功能--因此即使在 深入了解它的结构，我们仍然需要互补的功能信息才能预测 有针对性地扰乱运输途径的关键要素的结果。我们提出了两个具体目标 以一种协同的方式相互通知。首先，我们将执行与疾病相关的结构图 NUP复合体，侧重于细胞质输出平台的组件和已被 与致癌和发育缺陷有关。我们将使用已有方法的增强版本 成功部署以生成这两个区域及其附着点的高分辨率地图。在……上面 这项研究完成后，我们将以高精度绘制出全国人大的大部分地区，使这两个地区能够 在整个全国人大大会的背景下来看。第二，同时，我们将描绘疾病的功能-- 相关的NUP复合体。我们将剖析与目标NUP复合体相关的功能，以及 确定与它们的更改相关的缺陷-测试这些核素与之相关的假设 疾病，因为它们的干扰以一种不同于其他疾病的方式改变了关键基因表达模式 核孔素。实现这些目标将以前所未有的细节生成NPC结构-功能图，其中 对于理解NPC的不同部分如何共同行动以确定其功能是至关重要的。这 该项目将阐明与人类鼻咽癌功能障碍相关的许多疾病的性质；旨在 最终开放核运输机制，进行合理和可预测的药物设计。

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.