Structure-Function Mapping of the Nuclear Pore Complex

核孔复合体的结构-功能图谱

• 批准号: 9430429
• 负责人: JOHN D. AITCHISON
• 金额: $ 88.58万
• 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 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9430429
• 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; Nuclear Structure; Oncogenic; 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; outcome prediction; public health relevance; virtual

 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（由申请人提供）：NPC是一个嵌入核被膜中的大型圆柱形组件，在两个相关水平上对核功能起中心作用。首先，作为运输的调节器，NPC控制信号进入DNA和遗传信息从DNA的通过。第二，NPC是一个重要的基因调节因子，通过结合染色质及其调节因子来控制表达状态，这是一种在分子水平上了解甚少的现象。在所有真核细胞中，这些关键作用涉及数十种相互作用的途径 几乎影响细胞功能的所有方面。因此，NPC的破坏导致许多人类疾病。尽管如此，尽管核转运机制是一个有效和强大的药物靶点，NPC和核转运机制并不是治疗策略的重要组成部分。可以说，有两个基本原因导致这种情况：（i）我们对NPC的结构了解不够，无法预测其控制;（ii）核转运机制影响了一系列令人困惑的细胞功能-因此，即使对结构有深入的了解，我们也无法预测转运途径关键元件的靶向破坏的结果。我们提出了两个具体的目标，以协同的方式相互通报。首先，我们将进行疾病相关的Nup复合物的结构定位，重点是与致癌和发育缺陷有关的细胞质细丝和内环的组分。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着位点的高分辨率地图。这项研究完成后，我们将绘制大部分NPC的地图，使这两个地区能够在整个NPC大会的背景下看到。其次，与此同时，我们将绘制疾病相关的Nup复合物的功能。我们将剖析与目标Nup复合物相关的功能，并确定与其改变相关的缺陷-测试这些Nup与疾病相关的假设，因为它们的功能与疾病有关。 破坏以不同于其它核孔蛋白的方式改变关键基因表达模式。实现这些目标将产生前所未有的详细的NPC结构-功能图，这对于理解NPC的不同部分如何共同作用以确定其功能至关重要。这个项目将阐明与之相关的许多疾病的性质。 人类NPC功能障碍;旨在最终打开核转运机制，以进行合理和预测性的药物设计。