Bacterial effectors as probes to study (+) RNA virus-host cell biology

细菌效应子作为研究 ( ) RNA 病毒-宿主细胞生物学的探针

• 批准号: 9089955
• 负责人: Brett D. Lindenbach
• 金额: $ 20.81万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-15 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9089955
• 关键词: Address; Affect; Autophagocytosis; Award; Bacteria; Bacterial Infections; Biochemical; Biological; Cell Communication; Cells; Cellular biology; Collaborations; Collection; Coxiella; Coxiella burnetii; Cytosol; Data; Dengue Virus; Development; Enzymes; Eukaryotic Cell; Family; Flaviviridae; Follow-Up Studies; Funding; Future; Genes; Genetic; Golgi Apparatus; Health; Hepatitis C virus; Immune response; Integration Host Factors; Legionella pneumophila; Legionnaires' Disease; Mammalian Cell; Maps; Membrane Protein Traffic; Methods; Operative Surgical Procedures; Outcome Study; Pathogenesis; Pathway interactions; Pharmacopoeias; Phenotype; Proteins; Proteomics; Qualifying; RNA Interference; RNA Viruses; RNA interference screen; Reagent; Reproducibility; Research; Research Proposals; Role; Specificity; Staging; Transmembrane Transport; United States National Institutes of Health; Virus; Virus Replication; Work; Yellow fever virus; base; cellular targeting; follow-up; genome-wide; genome-wide analysis; high throughput screening; innovation; insight; interest; knock-down; member; novel strategies; pathogen; prevent; protein protein interaction; research study; screening; success; tool; virus host interaction; yeast two hybrid system

 DESCRIPTION (provided by applicant): All viruses depend on their host cells for replication. However, many aspects of virus-host cell interaction - such as the role of membrane trafficking in (+) RNA virus replication - are still poorly understood, in part due to a lack of sufficiently precise and powerful tools. This Exploratory and Developmental Research Proposal explores a new strategy to identify virus-host cell interaction by employing a large collection of bacterial effector proteins as cell biological probes, based on the observation that these proteins frequently target the same cellular pathways used by (+) RNA viruses. Because bacterial effector proteins are genetically encoded enzymes that have evolved to manipulate host cell biology with high specificity and activity, they have distinct advantages over other reagents for probing virus-host interaction, such as RNAi or pharmacological agents. By using a high-throughput screening approach, we will express a panel of 435 bacterial effector proteins to manipulate host cell biology, and examine how these perturbations affect the replication of hepatitis C virus, dengue virus, and yellow fever virus, three representative members of the Flaviviridae. Hits will then be prioritized for further mechanistic characterization and follow up studies. Our preliminary data demonstrate feasibility of this approach and provide interesting leads for future experiments. The outcome of these studies will provide proof of concept for our strategy, will generate a set of unique tools to study an important group of (+) RNA viruses, and will collaboratively synergize with ongoing research into bacterial effector mechanisms. We envision a future experimental niche wherein virologists and cell biologists will be able to utiliz a toolkit of bacterial effector proteins to probe many different biological questions.

 描述（由适用提供）：所有病毒都取决于其宿主细胞进行复制。然而，病毒宿主宿主相互作用的许多方面（例如膜运输在（+）RNA病毒复制中的作用）仍然很少了解，部分原因是缺乏足够精确和强大的工具。这项探索性和发育研究提案探索了一种新策略，通过使用大量细菌效应蛋白作为细胞生物学问题来鉴定病毒宿主 - 宿主相互作用，这是基于这样的观察结果，即这些蛋白经常针对（+）RNA病毒使用相同的细胞途径。由于细菌效应蛋白是遗传编码的酶，这些酶已经演变成具有高特异性和活性的宿主细胞生物学，因此它们比其他试剂具有明显的优势，用于探测病毒宿主相互作用，例如RNAI或药理剂。通过使用高通量筛选方法，我们将表达一组435种细菌效应蛋白来操纵宿主细胞生物学，并研究这些扰动如何影响丙型肝炎病毒，登革热病毒和黄热病病毒的复制，这是三个代表性的flaviviviridae。然后将优先命中命中，以进一步的机械表征和后续研究。我们的初步数据证明了这种方法的可行性，并为将来的实验提供了有趣的潜在客户。这些研究的结果将为我们的策略提供概念验证，将生成一组独特的工具来研究一组（+）RNA病毒的重要组，并将随着对细菌效应器机制的持续研究进行协作协作。我们设想了一个未来的实验生态位，其中病毒学家和细胞生物学家将能够使用细菌效应蛋白的工具包来探测许多不同的生物学问题。