Structure-function studies of alpha-and flaviviruses

描述(由申请人提供)：将对甲型和黄病毒蛋白进行全面分析，以获得这些蛋白的结构和功能及其与病毒生命周期的关系的详细知识。这两组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。 我们将使用高通量方法来检查这两个病毒组中具有代表性的成员的每个病毒蛋白。这将使用X射线结晶学(Chen，Rossmann，Smith)来检查病毒蛋白质或蛋白质复合体的原子结构。低温电子显微镜(CRYO-EM)将被用来检查单个病毒及其组装和进入的中间体(Baker，Rossmann)的结构。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和生化方法(Kuhn，Strauss)。将开发体外试验来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)。这项提议的长期目标是在原子水平上了解这两个病毒组的生命周期，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，以控制人类感染。 同一建筑同一楼层的各个实验室的距离很近，因此可以交流结晶学(Chen，Rossmann，Smith)和低温EM(Baker，Rossmann)的专业知识，以及病毒学和分子生物学技术(Kuhn)。十多年来，在甲型病毒和黄病毒方面的合作一直在一群PI(贝克、库恩、罗斯曼)之间进行，并已扩大到包括一名新聘用的教员(陈)以及一名高级结晶学研究员(史密斯)。施特劳斯实验室和普渡结构病毒学小组之间的合作近年来有所增加，这反映在人们的信念上，即施特劳斯实验室虽然从普渡环境中移除，但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.