Emerging virus-host cell protein interaction networks

新兴的病毒-宿主细胞蛋白质相互作用网络

• 批准号: 8964885
• 负责人: ROBERT A DAVEY
• 金额: $ 62.46万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-04 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8964885
• 关键词: Address; Affinity Chromatography; Agonist; Antiviral Agents; Arenavirus; Benchmarking; Binding; Biochemical; Bioinformatics; Biological; Biological Assay; Categories; Cell Communication; Cell Culture Techniques; Cells; Complex; Containment; Crimean-Congo Hemorrhagic Fever Virus; Data; Data Set; Dengue Virus; Dependence; Development; Disease Outbreaks; Disease Outcome; Drug effect disorder; Ebola virus; Environment; Family; Filovirus; Frankfurt-Marburg Syndrome Virus; Genotype; Goals; Health; Hepatitis C virus; Human; Human Virus; Individual; Infection; Informatics; Junin virus; Laboratories; Lassa Fever; Lassa fever virus; Lead; Life; Life Cycle Stages; Location; Maps; Mass Spectrum Analysis; Methods; Modeling; Molecular; Mus; National Institute of Allergy and Infectious Disease; National Security; Orthobunyavirus; Pathogenesis; Pathway interactions; Preparation; Principal Investigator; Protein Binding; Proteins; RNA Interference; RNA Viruses; Recruitment Activity; Reporting; Research; Risk; Role; Signal Pathway; Techniques; Technology; Testing; Tissues; Viral Hemorrhagic Fevers; Viral Proteins; Virus; Virus Diseases; Virus Replication; Work; Yeasts; cellular targeting; cofactor; experience; gammaherpesvirus; genome-wide; hemorrhagic fever virus; improved; inhibitor/antagonist; insight; interest; mortality; network models; novel strategies; pathogen; programs; protein protein interaction; public health relevance; screening; small molecule; virus host interaction; yeast two hybrid system

 DESCRIPTION (provided by applicant): The overall goal of this proposal is to identify and characterize cellular proteins that bind to viral hemorrhagic fever viruses. Specifically, we focus on RNA viruses from four families: the filoviruses Ebolavirus and Marburg virus; the Arenaviruses Junin virus and Lassa Fever virus; and the Bunyavirus Crimean-Congo hemorrhagic fever virus. These emerging viruses pose significant risks to human health and national security. While each differs in the types of proteins encoded, they share similar disease outcomes, causing severe, often fatal infections in humans; all are classified as NIAID Category A Priority Pathogens. Due to the high level containment needed to work with many of them, there is a significant gap in our understanding of these viruses and no specific, approved therapies are currently available. Like all viruses, these emerging viruses rely on host cell proteins in order to replicate. This dependence represents a potential "Achilles heel" in the virus life cycle that may be exploited to develop new approaches to treat viral infections. However, only a small number of virus-host cell protein interactions have been reported for the viruses in this study, and no systematic analysis of emerging virus-host cell protein interactions has been performed. In this project we develop and evaluate high quality maps of emerging virus-human protein interactions. In Aim 1 we use complementary yeast two-hybrid screening and co-affinity purification plus mass spectrometry approaches to identify cellular proteins that bind to emerging viruses and closely related non-pathogenic viruses from the same family. In aim 2 we identify shared and virus-specific features of the host cell interaction networks of emerging viruses. We use orthologonal protein-protein interaction assays to identify shared and virus specific interactions within each virus family. To gain confidence in these results, each assay is benchmarked against the human positive and random reference sets. We then employ informatic approaches to discover pathways that targeted preferentially by emerging viruses, develop tissue-specific virus-host cell protein interaction networks, and identify small molecule agonists or antagonists predicted to inhibit virus replication. In aim 3, we test the predictions generated through the bioinformatic analyses. A subset of high-interest cellular proteins targeted by multiple viruses will be experimentally interrogated for their contribution to infectio using RNA interference. State-of-the-art molecular techniques are then employed to determine the mechanisms by which they contribute to virus replication. Virus- and tissue-specific interactions are evaluated for their contribution to virus replication and their effect on cellular signaling pathways. Finally, small molecule inhibitors identified in our network models are evaluated for their effect on virus replication in cell culture. The data generated from this projet will improve our understanding of the functions of individual viral proteins, provide insight into the overall strategies used by viral hemorrhagic fever virus to interface with their host cells, an may lead to the discovery of new targets for treatments of emerging viruses.

 描述（由申请人提供）：该提案的总体目标是鉴定和表征与病毒性出血热病毒结合的细胞蛋白。 四个科的 RNA 病毒：丝状病毒、埃博拉病毒、胡宁病毒和拉沙热病毒，以及布尼亚病毒、克里米亚-刚果出血热病毒，这些新出现的病毒对人类健康和国家安全构成重大风险。由于所编码的蛋白质类型不同，它们具有相似的疾病结果，导致人类严重且往往致命的感染；由于需要高水平的遏制，所有这些病原体都被列为 NIAID A 类优先病原体。与其中许多病毒一起工作，我们对这些病毒的了解存在很大差距，并且目前没有特定的、经批准的治疗方法，与所有病毒一样，这些新兴病毒依赖宿主细胞蛋白进行复制，这种依赖性代表了一种潜在的“。病毒中的“致命弱点” 然而，本研究中仅报道了少量病毒-宿主细胞蛋白相互作用，并且没有对新出现的病毒-宿主细胞蛋白相互作用进行系统分析。在该项目中，我们开发并评估了新兴病毒与人类蛋白质相互作用的高质量图谱。在目标 1 中，我们使用互补酵母双杂交筛选和共亲和纯化以及质谱方法来识别与新兴病毒结合的细胞蛋白质。病毒和密切相关的非致病性在目标 2 中，我们识别新兴病毒的宿主细胞相互作用网络的共享和病毒特异性特征，我们使用正交蛋白质-蛋白质相互作用测定来识别每个病毒家族内的共享和病毒特异性相互作用。在这些结果中，每个测定都以人类阳性和随机参考集为基准，然后采用信息学方法来发现新兴病毒优先靶向的途径，开发组织特异性病毒-宿主细胞蛋白相互作用网络，并识别小分子激动剂或药物。预计拮抗剂会抑制在目标 3 中，我们将使用最先进的分子干扰技术来测试通过生物信息学分析生成的预测，以了解多种病毒靶向的一组高度关注的细胞蛋白对感染的贡献。然后采用技术来确定它们促进病毒复制的机制，评估它们对病毒复制的贡献及其对细胞的影响。 最后，我们的网络模型中鉴定的小分子抑制剂对细胞培养中病毒复制的影响进行了评估，该项目生成的数据将提高我们对单个病毒蛋白功能的理解，提供对所使用的总体策略的深入了解。通过病毒性出血热病毒与其宿主细胞相互作用，可能会发现治疗新兴病毒的新靶点。