Structure-Function Mapping of the Nuclear Pore Complex

 描述(申请人提供)：NPC是一个嵌入核膜的大型圆柱形组件，在两个相关的水平上为核功能提供中心。首先，作为运输的监管者，全国人大控制着获取DNA的信号和来自DNA的遗传信息的传递。其次，NPC是基因的重要调节器，通过结合染色质及其调节器来控制表达状态，这一现象在分子水平上知之甚少。这些在所有真核细胞中的关键作用涉及数十条相互作用的途径 几乎影响细胞功能的方方面面。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，尽管核运输机制是一个有效和强大的药物靶点，但NPC和核运输机制并不是治疗策略的重要组成部分。可以说，出现这种情况的根本原因有两个：(I)我们对NPC的结构缺乏足够的了解，无法预测其控制；(Ii)核运输机制影响了一系列令人眼花缭乱的细胞功能--因此，即使对结构有深刻的了解，我们也无法预测有针对性地扰乱运输途径关键成分的结果。我们提出了两个以协同方式相互告知的具体目标。首先，我们将对与疾病相关的NUP复合体进行结构映射，重点放在与致癌和发育缺陷有关的细胞质细丝和内环的成分上。我们将使用我们已经成功部署的方法的增强版本来生成这两个区域及其附属地点的高分辨率地图。这项研究完成后，我们将绘制出全国人大的大部分地图，使这两个地区能够在整个全国人大大会的背景下看到。第二，同时，我们将绘制与疾病相关的NUP复合体的功能图。我们将剖析与目标NUP复合体相关的功能，并确定与它们的改变相关的缺陷-检验这些NUP与疾病有关的假设，因为它们的 干扰以不同于其他核孔蛋白的方式改变了关键的基因表达模式。实现这些目标将产生史无前例的NPC结构-功能图，这些图对于理解NPC的不同部分如何共同行动来确定其功能至关重要。这个项目将阐明与以下疾病相关的许多疾病的性质 人类鼻咽癌功能障碍；最终目的是将核运输机制开放给理性和预测性的药物设计。

 DESCRIPTION (provided by applicant): The 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 signaling 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 control; (ii) the nuclear transport machinery impacts a bewildering array of cellular functions - thus even with a deep understanding of structure, we cannot 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 filaments 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, 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.