PSEUDO-ATOMIC STRUCTURE OF THE NUCLEAR PORE COMPLEX (NPC) USING SAXS

使用 SAXS 分析核孔复合体 (NPC) 的伪原子结构

• 批准号: 8362058
• 负责人: HIROTSUGU TSURUTA
• 金额: $ 0.14万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362058
• 关键词: Architecture; Benchmarking; Cell Nucleus; Cell physiology; Data; Eukaryota; Funding; Genetic Transcription; Grant; Head; Homologous Gene; Malignant Neoplasms; N-terminal; National Center for Research Resources; Nuclear Envelope; Nuclear Pore Complex; Nuclear Pore Complex Proteins; Play; Positioning Attribute; Principal Investigator; Proteins; RNA; Radiation; Relative (related person); Research; Research Infrastructure; Resolution; Resources; Solutions; Source; Structure; Tail; United States National Institutes of Health; Universities; Work; Yeasts; base; cost; dimer; human disease; improved; macromolecule; protein complex; restraint; structural biology

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The Nuclear Pore Complex (NPC, about 50 MDa) is the sole passageway for the transport of macromolecules across the nuclear envelope. The pore plays a key role in numerous critical cellular processes such as transcription, and many of its components are implicated in human diseases such as cancer. The recent work by the Sali group provided the first description of the macromolecular architecture of the yeast NPC. This structure defined the relative positions and proximities of its 456 constituent nucleoporin (nup) proteins, based on spatial restraints derived from experimental data. Further elucidation of the evolutionary origin and transport mechanism of the NPC will require higher resolution information. To help improve upon the resolution and accuracy of the NPC structure, we have begun small angle x-ray scattering studies at SSRL. Individial nup proteins and complexes are being expressed, produced and purified by our collaborators at Rockefeller University and Eli Lilly on a regular basis. We selected several nup proteins whose crystal structures have been solved to establish a benchmark. Among them is Nup145 which belongs to a highly conserved group of homologs found throughout the eukaryotes. Nup145N, the autoproteolised N-terminal half of the protein has been implicated in carrying RNA between the nucleus and the NPC by its analogy with Nup98. The crystal structures of Nup145N(443-605) and Nup98N, however, shows different modes of association within their respective crystallographic asymmetric units. Our preliminary solution x-ray scattering results demonstrate that Nup145N(443-605) forms the head-to-tail dimer in solution as observed in the asymmetric units of two different space groups.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 核孔复合物（NPC，约50 MDA）是大分子横跨核包膜的唯一通道。该孔在许多关键细胞过程（例如转录）中起着关键作用，其许多成分与人类疾病（如癌症）有关。 Sali Group最近的工作提供了酵母NPC大分子建筑的首次描述。该结构基于从实验数据得出的空间约束，定义了其456个组成核蛋白（NUP）蛋白的相对位置和接近性。 NPC的进化起源和运输机制的进一步阐明将需要更高的分辨率信息。为了帮助提高NPC结构的分辨率和准确性，我们已经开始在SSRL上进行小角度X射线散射研究。我们的合作者在洛克菲勒大学和伊利·莉莉（Eli Lilly）定期表达，生产和纯化个性化蛋白质和复合物。我们选择了几种NUP蛋白，其晶体结构已被溶解以建立基准。其中包括NUP145，它属于整个真核生物中发现的一组高度保守的同源物。 NUP145N，该蛋白的自蛋白N端一半与NUP98的类比在细胞核和NPC之间携带RNA有关。然而，NUP145N（443-605）和NUP98N的晶体结构在其各自的晶体不对称单元中显示出不同的缔合模式。我们的初步溶液X射线散射结果表明，NUP145N（443-605）在两个不同空间组的不对称单元中观察到的溶液中的头到尾二聚体。