Structure-Function Mapping of the Nuclear Pore Complex

核孔复合体的结构-功能图谱

• 批准号: 10394295
• 负责人: JOHN D. AITCHISON
• 金额: $ 66.62万
• 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 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10394295
• 关键词: Behavior; Binding; Cell Nucleus; Cell physiology; Chromatin; Communities; Complex; Cytoplasm; DNA; Defect; Development; Disease; Dissection; Drug Design; Drug Targeting; Elements; Epigenetic Process; Eukaryotic Cell; Foundations; Functional disorder; Future; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Human; Knowledge; Lead; Light; Link; Maps; Mediating; Messenger RNA; Methods; Molecular; Mutation; Nature; Nuclear; Nuclear Envelope; Nuclear Pore Complex; Nuclear Pore Complex Proteins; Nuclear Structure; Oncogenic; Orthologous Gene; Pathway interactions; Phenotype; Process; Protein Import; Proteins; Regulation; Regulator Genes; Resolution; Role; Signal Transduction; Site; Structure; Testing; Therapeutic; Viral; Work; Yeasts; design; empowered; genetic information; insight; mRNA Export; nucleocytoplasmic transport; outcome prediction; pleiotropism; virtual

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.

项目摘要（摘要） 核孔复合物（NPC）是嵌入核包膜中的大圆柱组件，中心 两个相关水平的核功能。首先，作为运输的调节器，NPC控制着信号访问到达 DNA和遗传信息从DNA传递。其次，NPC是基因的重要调节因子 结合染色质及其调节剂控制表达状态，这种现象在 分子水平。这些在所有真核细胞中的关键作用涉及数十个影响的相互作用的途径 实际上，细胞功能的所有方面。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。可以说有两个 这样就是这样的基本原因：（i）我们对NPC的结构不太了解 它的行为； （ii）核运输机制会影响一系列令人困惑的细胞功能 - 因此 对其结构有深刻的了解，我们仍然需要互补的功能信息才能预测 运输途径关键要素的目标破坏的结果。我们提出了两个具体目标 这是以协同的方式互相告知的。首先，我们将进行与疾病相关的结构映射 NUP复合物，重点关注细胞质出口平台的组件和内部环 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件站点的高分辨率图。在 完成这项研究的完成，我们将以高精度绘制大部分NPC，从而使两个区域成为 在整个NPC组件的上下文中看到。第二，同时，我们将绘制疾病的功能 - 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 确定与它们的变化相关的缺陷 - 测试这些NUP链接到的假设 疾病是因为它们的破坏以不同于其他的方式改变了关键基因表达模式 核孔蛋白。意识到这些目标将以前所未有的细节生成NPC结构功能地图以及哪些 对于了解NPC的不同部分如何共同确定其功能至关重要。这 项目将阐明与人类NPC功能障碍相关的众多疾病的性质；瞄准 最终，将核运输机制开放到理性和预测药物设计。