CRYO-EM STRUCTURAL STUDIES OF FELINE, MURINE, AVIAN AND SARS CORONA VIRUSES

猫科动物、鼠科动物、禽类和 SARS 冠状病毒的冷冻电镜结构研究

• 批准号: 8362481
• 负责人: Michael Joseph Buchmeier
• 金额: $ 1.29万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362481
• 关键词: Acute; Antiviral Agents; Arenavirus; Birds; Caliber; Carbon; Coronavirus; Cryoelectron Microscopy; Data; Data Set; Disease Outbreaks; Drug Design; Family; Family Felidae; Feline Coronavirus; Funding; Grant; Ice; Image; Infectious bronchitis virus; Microscopy; Molecular; Morbidity - disease rate; Mus; National Center for Research Resources; Principal Investigator; Protein S; Proteins; Publications; RNA; Research; Research Infrastructure; Research Personnel; Resources; SARS coronavirus; Severe Acute Respiratory Syndrome; Source; Specimen; Structural Protein; Structure; Syndrome; United States National Institutes of Health; Viral Proteins; Virion; Virus; Work; Writing; cost; member; mortality; particle; protein complex; protein function; three dimensional structure; two-dimensional

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. Little is known about structure and function-associated conformational changes in coronavirus virion proteins. In addition to providing the framework for the virion, the four conserved structural proteins S, E, M and N are essential in assembly, RNA packaging, egress and ingress of the virion. We propose to characterize the two- and three- dimensional structure of the coronavirus structural protein complexes within the virion. Changes that occur during fusion activation will also be studied. The coronavirus that causes severe acute resporatory syndrome outbreak of 2002-2003 was marked by high mortality and morbidity. The reservoir of this virus has not been established. Understanding of how the viral proteins function within the virion and during interaction with the host would prove very useful in rational antiviral drug design. We have reconstructed the structural protein complexes of three arenaviruses, which are pleomorphic enveloped particles 40-500 nm in diameter, using electron cryomicroscopy. Preliminary two- and three-dimensional images of the SARS structural proteins are in refinement. The NRAMM component of this project would consist of imaging particles from the three coronavirus families: feline coronavirus (group I), murine coronavirus (group II) and avian infectious bronchitis virus (group III) for comparison with SARS coronavirus (distantly related to group II or perhaps assigned as the only member of group IV). Results of the arenavirus cryo-EM studies are being prepared for publication. Several protein specimens have been prepared as vitreous ice layers suspended over holey carbon substrates. Datasets have been acquired from several varieties of corona virus using Leginon. The data is being analyzed by the investigator. This work should be completed within the next month and will then be written up for publication.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 关于结构和功能相关的构象变化， 冠状病毒病毒粒子蛋白。 除了为病毒体提供框架外， 保守的结构蛋白S、E、M和N在装配，RNA包装， 病毒粒子的进出 我们建议把两个和三个- 病毒粒子内冠状病毒结构蛋白复合物的三维结构。 还将研究融合激活过程中发生的变化。 2002-2003年引起严重急性呼吸综合征爆发的冠状病毒 以高死亡率和发病率为特点。 这种病毒的宿主 确立了习 了解病毒蛋白质在病毒体内和病毒感染过程中的功能 与宿主的相互作用将证明在合理的抗病毒药物设计中非常有用。 我们已经重建了三种沙粒病毒的结构蛋白复合物， 使用电子低温显微镜，观察到直径为40-500 nm的多形包膜颗粒。 SARS结构蛋白的初步二维和三维图像在 精致。 该项目的NRAMM组件将包括成像粒子 来自三个冠状病毒家族：猫冠状病毒（I组）、鼠冠状病毒 鸡传染性支气管炎病毒（Ⅲ组）与SARS病毒的比较 冠状病毒（与II组有远亲关系，或者可能是唯一的成员） 第四组）。 沙粒病毒冷冻电镜研究的结果正在准备发表。 几种蛋白质标本被制备成悬浮在多孔表面的玻璃状冰层， 碳基质 数据集已经从几种冠状病毒中获得 用的是雷吉农 研究者正在分析数据。 这项工作应当 下个月内完成，然后准备出版。