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: 8968695
• 负责人: Brett D. Lindenbach
• 金额: $ 24.98万
• 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 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8968695
• 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; 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; Reagent; Reproducibility; Research; Research Proposals; Role; Specificity; Staging; Transmembrane Transport; United States National Institutes of Health; Viral; Virus; Virus Replication; Work; Yellow fever virus; base; cellular targeting; follow-up; genome-wide; genome-wide analysis; high throughput screening; innovation; insight; interest; member; novel strategies; pathogen; prevent; protein protein interaction; public health relevance; 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（由申请人提供）：所有病毒都依赖于其宿主细胞进行复制。然而，病毒与宿主细胞相互作用的许多方面——例如膜运输在（+）RNA病毒复制中的作用——仍然部分了解甚少。由于缺乏足够精确和强大的工具，该探索性和发展研究提案基于对这些蛋白质频繁出现的观察，探索了一种通过使用大量细菌效应蛋白作为细胞生物探针来识别病毒与宿主细胞相互作用的新策略。目标相同(+) RNA 病毒使用的细胞途径。由于细菌效应蛋白是基因编码的酶，已进化为具有高特异性和活性的操纵宿主细胞生物学，因此它们比其他用于探测病毒-宿主相互作用的试剂（例如 RNAi 或 RNAi）具有明显的优势。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，并检查这些扰动如何影响丙型肝炎病毒、登革热病毒和黄热病的复制。我们的初步数据证明了这种方法的可行性，并为未来的实验提供了有趣的线索。这些研究的结果将为未来的实验提供概念证明。我们的战略将产生一套独特的工具来研究一组重要的（+）RNA病毒，并将与正在进行的细菌效应机制研究协同作用，我们设想病毒学家和细胞生物学家将能够在未来实现实验利基。利用细菌效应蛋白工具包来探索许多不同的生物学问题。