CryoEM and image reconstruction of viruses

病毒的冷冻电镜和图像重建

• 批准号: 6340730
• 负责人: Timothy S Baker
• 金额: $ 16.62万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6340730
• 关键词: Flaviviridae; Rubivirus; Togaviridae; bioimaging /biomedical imaging; cryoelectron microscopy; hepatitis C virus; human papillomavirus; image enhancement; plant virus; structural biology; virus assembly; virus infection mechanism

Viruses are parasites of their hosts. Hence, the life cycle of any virus is inextricably tied to that of the host cell. Despite this dependence, all viruses share a number of essential tasks which they must accomplish for survival. A virus must be able to find and recognize a cell in which it can replicate, release its genome into the cell, generate new viral components by transcription and translation and assembly these components into precursors that mature into a stable progeny virion which is released from the host cell and transmitted to encounter a new host. Different viruses accomplish these tasks in different ways as a result of adaptation to different cellular environments. Each of these tasks involves interactions between components within the context of the whole virion and hence require the visualization of the entire structure at which the techniques of cryo-transmission electron microscopy (cryoTEM) and three-dimensional (3D) image reconstruction ('cryo-reconstruction') excel. We have exploited cryo-reconstruction techniques to study a diverse range of viruses, including those that infect mammals as well as insects, bacteria, and plants (including fungi and algae). Several studies funded by the current PPG have illustrated the structural response of different viruses to the common tasks of the viral life cycle. This proposal involves several new cryo-reconstruction studies aimed at exploring the structural basis for key aspects of viral infection. Work with the Kuhn and Rossmann laboratories will continue analyses of the assembly of several enveloped viruses of the alphavirus gene (Togavirus family). We will also initiate new studies of the assembly of three important human pathogenesis: rubella virus (rubrivirus genus) and two members of the closely related Flavivirus family of enveloped viruses, yellow fever virus and Hepatitis C virus. A collaboration with Smith's laboratory will address issues related to viral transmission in two plant viruses, cucumber mosaic virus and zucchini yellow mosaic virus. A collaboration with Friedman's laboratory will define viral epitopes on human papilloma viruses, including the carcinogenic, HPV serotype 16.

病毒是宿主的寄生虫。因此，任何病毒的生命周期都与宿主细胞的生命周期密不可分。尽管存在这种依赖性，所有病毒都有许多为了生存而必须完成的基本任务。病毒必须能够找到并识别其可以在其中复制的细胞，将其基因组释放到细胞中，通过转录和翻译产生新的病毒组分，并将这些组分组装成成熟为稳定的子代病毒体的前体，所述子代病毒体从宿主细胞释放并被传递以遇到新的宿主。不同的病毒以不同的方式完成这些任务，这是适应不同细胞环境的结果。这些任务中的每一个都涉及整个病毒体的背景下的组件之间的相互作用，因此需要整个结构的可视化，其中冷冻透射电子显微镜（cryoTEM）和三维（3D）图像重建（“cryo-reconstruction”）的技术优于。我们已经利用冷冻重建技术来研究各种病毒，包括感染哺乳动物以及昆虫，细菌和植物（包括真菌和藻类）的病毒。目前PPG资助的几项研究已经说明了不同病毒对病毒生命周期共同任务的结构反应。该提案涉及几项新的冷冻重建研究，旨在探索病毒感染关键方面的结构基础。与Kuhn和Rossmann实验室的合作将继续分析甲病毒基因（披膜病毒家族）的几种包膜病毒的组装。我们还将启动三个重要的人类发病机制的新研究：风疹病毒（风疹病毒属）和两个密切相关的黄病毒家族成员的包膜病毒，黄热病病毒和丙型肝炎病毒。与史密斯实验室的合作将解决与两种植物病毒，黄瓜花叶病毒和西葫芦黄色花叶病毒的病毒传播有关的问题。与弗里德曼实验室的合作将确定人类乳头状瘤病毒的病毒表位，包括致癌的HPV血清型16。