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

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 对与结构和功能相关的构象变化知之甚少 冠状病毒病毒粒子蛋白。除了为病毒粒子提供框架外，四个 保守的结构蛋白S、E、M和N在组装、RNA包装、 病毒的出口和入口。我们建议将两个-和三个- 病毒粒子内冠状病毒结构蛋白复合体的空间结构。 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暴发的冠状病毒 以高死亡率和高发病率为特点。这种病毒的宿主还没有被 已经成立了。了解病毒蛋白在病毒粒子内和过程中的作用 与宿主的相互作用将被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阿拉伯病毒的结构蛋白复合体，它们是 电子冷冻显微镜下可见直径为40-500 nm的多形性包膜颗粒。 SARS结构蛋白的初步二维和三维图像在 精致。该项目的NRAMM部分将由成像粒子组成 来自三个冠状病毒家族：猫冠状病毒(组I)、小鼠冠状病毒 (II组)和禽传染性支气管炎病毒(III组)与SARS的比较 冠状病毒(与第II组有远亲关系或可能被归类为 第IV组)。ArenaVirus冷冻-EM研究的结果正在准备出版。 几个蛋白质样本被制备成悬浮在孔洞上的玻璃冰层。 碳衬底。已经从几种冠状病毒变种中获得了数据集 利用莱吉农。调查人员正在对数据进行分析。这项工作应该是 在下个月内完成，然后将被写出来出版。

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.