MASS SPECTROMETRY BASED APPROACHES FOR STUDYING HOST VIRUS INTERACTIONS

用于研究宿主病毒相互作用的基于质谱的方法

• 批准号: 8363841
• 负责人: Raul Andino
• 金额: $ 0.58万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363841
• 关键词: Address; Animal Model; Antiviral Agents; Architecture; Attention; Cell physiology; Cells; Chemicals; Complex; Defense Mechanisms; Dengue; Drosophila genus; Drosophila melanogaster; Event; Funding; Goals; Grant; Human; Immune system; Infection; Insect Viruses; Insecta; Integration Host Factors; Maps; Mass Spectrum Analysis; National Center for Research Resources; Nucleic Acids; Outcome; Post-Translational Protein Processing; Principal Investigator; Proteins; RNA Interference; RNA Viruses; Research; Research Infrastructure; Resources; Signal Pathway; Site; Source; System; Techniques; United States National Institutes of Health; Viral; Virus; Virus Diseases; West Nile virus; Yellow Fever; base; cost; cricket paralysis virus; crosslink; design; endonuclease; invertebrate host; pathogen; protein complex; protein expression; protein function; single molecule; virus host interaction

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. A large number of viruses, including many human pathogens (dengue, yellow fever, west nile virus, etc), are transmitted by insects. The long-term goal of this proposal is to examine the mechanism of the antiviral defense system in Drosophila melanogaster. RNA interference (RNAi) is an ancient and conserved nucleic acid-based immune system in insects. Accordingly, insect viruses evolved strategies to control this defense mechanism. For example, Cricket Paralysis Virus (a Dicistrovirus) encodes a protein that suppresses RNAi by interacting specifically with the endonuclease Ago2. In more general terms, our understanding of the interactions of RNA viruses with the invertebrate hosts is very limited nevertheless requires urgent attention. The outcome of virus infection is the result of multidimensional set interactions of virus and host factors. Indeed, many viral and host proteins function as a complex than acting as a single molecule to regulate a vast number of cellular processes. Chemical crosslinking mass spectrometry of purified complexes derived from virus infected cells is a useful technique for mapping the site of linkage between two interacting partners in a complex and provides valuable information on three dimensional functional architecture of the protein complex. In addition, post translational modification events (e.g. phosporylation) of cellular signaling pathways are critical to regulate the antiviral mechanisms and often takes place upon infection that leads to change in cellular protein expression. The present research plan is designed to address how RNA viruses particularly Dicistroviruses interacts with host machinery to their advantage using Drosophila as a model organism and crosslinking mass spectrometry and phospoproteomics approaches.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 许多病毒，包括许多人类病原体（登革热，黄热病，西尼罗河病毒等），都是由昆虫传播的。 该提案的长期目标是检查果蝇果蝇抗病毒防御系统的机制。 RNA干扰（RNAi）是一种古老而保守的基于核酸的免疫系统。 因此，昆虫病毒发展了控制这种防御机制的策略。 例如，板球瘫痪病毒（一种二菌病毒）编码一种蛋白质，该蛋白质通过与核酸内切酶AGO2进行专门相互作用来抑制RNAi。 从更一般的角度来看，我们对RNA病毒与无脊椎动物宿主的相互作用的理解非常有限，但仍需要紧急关注。 病毒感染的结果是病毒和宿主因素的多维集相互作用的结果。 实际上，许多病毒和宿主蛋白比充当单个分子来调节大量细胞过程。 从病毒感染细胞中得出的纯化复合物的化学交联质谱是一种有用的技术，用于映射复合物中两个相互作用伙伴之间的链接位点，并在蛋白质复合物的三维功能结构上提供有价值的信息。 此外，细胞信号传导途径的翻译后修饰事件（例如荧光素化）对于调节抗病毒机制至关重要，并且经常发生在感染后，导致细胞蛋白表达的变化。 本研究计划旨在解决RNA病毒尤其是Dicistrovires如何与宿主机械相互作用，以使用果蝇作为模型生物和交联的质谱和凤凰蛋白质组学方法，从而使其优势相互作用。