Structure-function studies of alpha-and flaviviruses

描述（由申请方提供）：将对α病毒和黄病毒蛋白进行全面分析，以详细了解这些蛋白的结构和功能及其与病毒生命周期的关系。这两类病毒都是重要的人类病原体，其中几种已被确定为可能的生物武器。 我们将采用高通量的方法来检查来自两个病毒组的代表性成员的每种病毒蛋白。这将使用X射线晶体学（Chen，Rossmann，Smith）来检查病毒蛋白或蛋白复合物的原子结构。将采用冷冻电子显微镜（cryo-EM）检查单个病毒及其组装和进入过程中的中间体的结构（Baker，Rossmann）。从结构研究中获得的知识将应用于使用病毒或复制子的分子遗传学和生物化学方法（Kuhn，Strauss）。将开发体外试验，以检查复制周期的各个方面，如RNA复制、蛋白质加工、颗粒组装和病毒融合（Baker、Chen、Kuhn、Rossmann、Smith、Strauss）。该提案的长期目标是在原子水平上了解这两种病毒群的生命周期，并将这些信息转化为新的抗病毒（Chen，Kuhn，Rossmann，Smith）和疫苗（Baker，Kuhn，Rossmann，Strauss）方法来控制人类感染。 各个实验室位于同一建筑物的同一楼层，可以交流晶体学（Chen，Rossmann，Smith）和冷冻EM（Baker，Rossmann）专业知识以及病毒学和分子生物学技术（Kuhn）。关于甲病毒和黄病毒的合作已经在PI的一个子集之间进行了十多年（Baker，Kuhn，Rossmann），并已扩展到包括新聘请的教员（Chen）以及高级晶体学研究员（Smith）。近年来，施特劳斯实验室和普渡大学结构病毒学小组之间的合作不断发展，这反映在施特劳斯实验室虽然脱离了普渡大学的环境，但仍将为该项目的长期成功做出重大贡献。

DESCRIPTION (provided by applicant): A comprehensive analysis of alpha- and flavivirus proteins will be undertaken to obtain detailed knowledge of the structure and function of these proteins and their relationship to their virus life cycles. Viruses from both groups are significant human pathogens and several have been identified as possible biological weapons. We will employ a high-throughput approach to examine each of the viral proteins from representative members of the two virus groups. This will be accomplished using X-ray crystallography (Chen, Rossmann, Smith) to examine the atomic structure of the viral proteins or complexes of proteins. Cryo-electron microscopy (cryo-EM) will be employed to examine the structure of the individual viruses and their intermediates in assembly and entry (Baker, Rossmann). Knowledge obtained from structural studies will be applied to molecular genetic and biochemical approaches using viruses or replicons (Kuhn, Strauss). In vitro assays will be developed to examine various aspects of the replication cycle such as RNA replication, protein processing, particle assembly, and virus fusion (Baker, Chen, Kuhn, Rossmann, Smith, Strauss). The long-term goal of this proposal is to understand the life cycles of these two virus groups at the atomic level and to translate this information into novel antiviral (Chen, Kuhn, Rossmann, Smith) and vaccine (Baker, Kuhn, Rossmann, Strauss) approaches to control human infections. The proximity of the individual laboratories on the same floor of one building permits exchange of crystallographic (Chen, Rossmann, Smith) and cryo-EM (Baker, Rossmann) expertise as well as virological and molecular biological techniques (Kuhn). Collaborations on alphaviruses and flaviviruses have been ongoing between a subset of the PIs for more than a decade (Baker, Kuhn, Rossmann) and have expanded to include a newly hired faculty member (Chen) as well as a senior crystallographic investigator (Smith). Collaborations between the Strauss laboratory and the Purdue Structural Virology Group have grown in recent years and this is reflected in the conviction that the Strauss laboratory, although removed from the Purdue environment, will contribute significantly to the long-term successes of the project.