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.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 大量病毒，包括许多人类病原体（登革热、黄热病、西尼罗河病毒等）都是通过昆虫传播的。 该提案的长期目标是研究果蝇抗病毒防御系统的机制。 RNA干扰（RNAi）是昆虫中一种古老且保守的基于核酸的免疫系统。 因此，昆虫病毒进化出了控制这种防御机制的策略。 例如，蟋蟀麻痹病毒（一种双顺反子病毒）编码一种通过与核酸内切酶 Ago2 特异性相互作用来抑制 RNAi 的蛋白质。 更一般地说，我们对 RNA 病毒与无脊椎动物宿主相互作用的了解非常有限，但仍需要紧急关注。 病毒感染的结果是病毒与宿主因素多维集合相互作用的结果。 事实上，许多病毒和宿主蛋白作为复合体发挥作用，而不是作为单个分子来调节大量细胞过程。 对源自病毒感染细胞的纯化复合物进行化学交联质谱分析是一种有用的技术，可用于绘制复合物中两个相互作用伙伴之间的连接位点，并提供有关蛋白质复合物三维功能结构的有价值的信息。 此外，细胞信号通路的翻译后修饰事件（例如磷酸化）对于调节抗病毒机制至关重要，并且通常在感染时发生，导致细胞蛋白表达发生变化。 目前的研究计划旨在利用果蝇作为模型生物以及交联质谱和磷酸蛋白质组学方法来解决 RNA 病毒（特别是双顺反子病毒）如何与宿主机器相互作用以发挥其优势。