Structure-Function Mapping of the Nuclear Pore Complex

核孔复合体的结构-功能图谱

• 批准号: 8888675
• 负责人: JOHN D. AITCHISON
• 金额: $ 92.04万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8888675
• 关键词: Binding; Caliber; Cell Nucleus; Cell physiology; Charge; Chromatin; Complex; Cytoplasm; Cytoplasmic Filaments; DNA; Defect; Development; Disease; Drug Design; Drug Targeting; Elements; Epigenetic Process; Eukaryotic Cell; Functional disorder; Gene Expression; Gene Expression Profile; Genes; Health; Human; Knowledge; Lead; Light; Link; Malignant Neoplasms; Maps; Mediating; Membrane; Messenger RNA; Methods; Molecular; Mutation; Nature; Nuclear; Nuclear Envelope; Nuclear Pore Complex; Nuclear Pore Complex Proteins; Oncogenic; Outcome; Pathway interactions; Process; Protein Import; Regulator Genes; Resolution; Role; Signal Transduction; Site; Structure; Testing; Therapeutic; Viral; Virus Diseases; Work; genetic information; glycylphenylalanine; improved; mRNA Export; mRNA Expression; nucleocytoplasmic transport; public health relevance

 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.

 描述（由适用提供）：NPC是嵌入核包膜中的大圆柱组件，在两个相关水平上是核功能的中心。首先，作为运输的调节剂，NPC控制了对DNA的信号传递的访问，并通过DNA传递了遗传信息。其次，NPC是通过结合染色质及其调节剂来控制表达状态的重要调节基因的调节剂，这一现象在分子水平上尚不清楚。这些关键作用在所有真核细胞中都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。结果，NPC的破坏会导致许多人类疾病。尽管如此，尽管核运输机械是有效而有力的药物靶标，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，情况有两个基本原因：（i）我们对NPC的结构不了解以预测其控制； （ii）核运输机制会影响一系列令人困惑的细胞功能 - 因此，即使对结构有深刻的了解，我们也无法预测靶向破坏运输途径的关键要素的结果。我们提出了两个具体的目标，它们以协同的方式互相告知。首先，我们将对疾病相关的NUP复合物进行结构映射，重点关注与致癌和发育缺陷有关的细胞质细丝和内环的组成部分。我们将使用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件站点的高分辨率图。这项研究完成后，我们将绘制NPC的大部分，从而可以在整个NPC组装的背景下看到这两个区域。其次，同时，我们将绘制与疾病相关的NUP复合物的功能。我们将剖析与目标NUP复合物相关的功能，并确定与它们的变化相关的缺陷 - 测试这些NUP与疾病有关的假设，因为它们 破坏以不同于其他核oporin的方式改变关键基因表达模式。意识到这些目标将以前所未有的细节生成NPC结构功能图，这对于了解NPC的不同部分如何共同作用以确定其功能至关重要。该项目将阐明与 人NPC功能障碍；最终旨在将核运输机制开放到理性和预测的药物设计中。