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（由适用提供）：该提案的总体目标是识别和表征与病毒出血热病毒结合的细胞蛋白。具体来说，我们集中精力 关于四个家庭的RNA病毒：埃博拉病毒和马堡病毒；体育症病毒Junin病毒和Lassa发烧病毒；以及邦雅利亚病毒克里米亚 - 隆戈出血性发烧病毒。这些新兴病毒对人类健康和国家安全构成了重大风险。尽管编码的蛋白质类型的每种差异，但它们具有相似的疾病结果，在人类中引起严重的，经常致命的感染。所有这些都被归类为NIAID类别的优先病原体。由于与其中许多合作所需的高水平遏制，因此我们对这些病毒的理解存在很大的差距，目前没有特定的，批准的疗法。像所有病毒一样，这些新兴病毒依赖于宿主细胞蛋白来复制。这种依赖性代表了病毒中潜在的“阿喀琉斯高跟” 可以探索的生命周期来开发新的治疗病毒感染方法。但是，在这项研究中，仅报道了该病毒的病毒宿主宿主细胞蛋白相互作用，并且没有对新兴病毒宿主宿主细胞蛋白相互作用进行系统分析。在这个项目中，我们开发和评估新兴病毒人类蛋白相互作用的高质量图。在AIM 1中，我们使用完整的酵母双杂交筛查和共亲密纯化以及质谱法来鉴定与新兴病毒结合并从同一家族结合的细胞蛋白。在AIM 2中，我们确定了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。我们使用直源蛋白 - 蛋白质相互作用评估来识别每个病毒家族中共享和病毒的特定相互作用。为了对这些结果产生信心，每个测定法都针对人类正面和随机参考集进行基准测试。然后，我们的员工提供了信息，以发现通过新兴病毒优先针对的途径，发展组织特异性病毒宿主宿主细胞蛋白相互作用网络，并鉴定出可抑制病毒复制的小分子激动剂或拮抗剂。在AIM 3中，我们测试通过生物信息学分析产生的预测。通过RNA干扰对多种病毒靶向的高息细胞蛋白的一部分将受到实验询问。然后聘请最先进的分子技术来确定它们有助于病毒复制的机制。评估病毒和组织特异性相互作用对病毒复制的贡献及其对细胞的影响 信号通路。最后，评估了我们网络模型中鉴定出的小分子抑制剂对细胞培养中病毒复制的影响。从该过程中产生的数据将提高我们对单个病毒蛋白功能的理解，从而深入了解病毒出血热病毒与宿主细胞接触的整体策略，这可能会导致发现新的靶标治疗新兴病毒。