Molecular determinants of host-cell interactions required for arenavirus replicat

沙粒病毒复制所需的宿主细胞相互作用的分子决定因素

• 批准号: 8424219
• 负责人: Jack H Nunberg
• 金额: $ 16.63万
• 依托单位: UNIVERSITY OF MONTANA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-13 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8424219
• 关键词: Acute; Address; Affinity; Affinity Chromatography; Antiviral Agents; Arenavirus; Arenavirus Infections; Binding; Biochemical; Biology; Categories; Cell Communication; Cell physiology; Cells; Cellular Membrane; Cellular Stress; Chemistry; Complex; Confocal Microscopy; Cytoplasm; Cytoplasmic Granules; Cytoplasmic Structures; Data; Detection; Development; Electron Microscopy; Elements; Environment; Genetic; Genetic Transcription; Genome; Goals; Human; Image; Immune; Infection; Junin virus; Knowledge; Life Cycle Stages; Light; Mediating; Membrane; Messenger RNA; Microscopic; Molecular; Nucleoproteins; Pathway interactions; Peptide Initiation Factors; Population; Property; Protein Biosynthesis; Proteins; Public Health; RNA; RNA Cap-Binding Proteins; RNA Viruses; Recruitment Activity; Regulation; Research Infrastructure; Resistance; Ribosomes; Rodent; Role; Scaffolding Protein; Site; Stress; Structure; Surveys; Techniques; Technology; Transcript; Translating; Translation Initiation; Translations; Two-Dimensional Gel Electrophoresis; Vero Cells; Viral; Viral Hemorrhagic Fevers; Viral Physiology; Viral Proteins; Virus; Virus Diseases; Virus-Cell Membrane Interaction; base; biodefense; drug development; drug discovery; effective therapy; inhibitor/antagonist; innovation; insight; mortality; novel; pathogen; phosphatidylinositol 4-phosphate; polypeptide; small molecule; treatment strategy; viral RNA

DESCRIPTION (provided by applicant): Arenaviruses cause acute hemorrhagic fevers with high mortality worldwide. Jun¿n virus is recognized as a category A pathogen and Material Threat to the US population. Effective therapies against arenaviral hemorrhagic fevers are urgently needed to address ongoing public health and biodefense concerns. Our long- term goal is to elucidate the molecular mechanisms by which Jun¿n virus appropriates specific cellular functions to implement its RNA-based replicative cycle in the host-cell cytoplasm. We have recently identified unique cytoplasmic structures that serve as compartments for arenavirus replication and transcription (RTCs) and may also be involved in viral translation. Our overarching hypothesis is that novel activities of the virus nucleoprotein are essential for induction of these membrane-associated RTCs and for preferential recruitment of ribosomes to viral mRNAs. In this proposal, we will define determinants in the multifunctional nucleoprotein that interface with the cellular infrastructure to organize an environment permissive for viral replication. This knowledge will increase our understanding of arenavirus biology and suggest novel antiviral strategies for the treatment of acute hemorrhagic fevers. Our Specific Aims are to: (1) Investigate the origins, structure and function of RTCs by systematically identifying co-localizing cellular components. Our preliminary results suggest that RTCs comprise a unique combination of cellular elements from both membrane-remodeling and translational-control pathways. Microscopic and biochemical analysis of the co-opted proteins and pathways will establish a framework for understanding fundamental virus- cell interactions required for arenavirus replication. (2) Characterize the functional role of nucleoprotein in ribosome recruitment and viral translation. Confocal microscopy studies reveal that RTCs contain various translation initiation factors and ribosomes, but are specifically lacking the cap-binding protein eIF4E. We posit that N acts as a surrogate to bind m7G-capped viral mRNA and recruit the eIF4G scaffold protein, in order to gain privileged access to ribosomes. We will employ biochemical and pharmacological approaches to determine the functional requirement for nucleoprotein in viral translation. Knowledge gained from these studies will be important in guiding the development of small-molecule compounds to inhibit this novel activity. We anticipate that essential virus-cell interactions can be targeted to generate highly specific antiviral effects that are resistant to genetic escape.

描述（由申请方提供）：沙粒病毒引起急性出血热，在全球范围内死亡率很高。Junn病毒被认为是A类病原体，对美国人口构成重大威胁。迫切需要有效的治疗沙粒病毒出血热，以解决持续的公共卫生和生物防御问题。我们的长期目标是阐明Junn病毒利用特定细胞功能在宿主细胞质中实现其基于RNA的复制周期的分子机制。我们最近发现了独特的细胞质结构，作为沙粒病毒复制和转录（RTCs）的隔室，也可能参与病毒翻译。我们的总体假设是，病毒核蛋白的新活动是必不可少的诱导这些膜相关的RTC和优先招聘的核糖体的病毒mRNA。在这个提议中，我们将定义多功能核蛋白中的决定因素，这些决定因素与细胞基础结构相结合，以组织允许病毒复制的环境。这些知识将增加我们对沙粒病毒生物学的理解，并为治疗急性出血热提出新的抗病毒策略。我们的具体目标是：（1）通过系统鉴定共定位的细胞成分，研究RTCs的起源、结构和功能。我们的初步研究结果表明，RTC包括一个独特的组合，从膜重塑和免疫控制途径的细胞元素。对增选蛋白和途径的显微镜和生物化学分析将建立一个框架，用于理解沙粒病毒复制所需的基本病毒-细胞相互作用。(2)描述核蛋白在核糖体募集和病毒翻译中的功能作用。共聚焦显微镜研究表明，RTCs含有各种翻译起始因子和核糖体，但特别缺乏帽结合蛋白eIF 4 E。我们证实N作为替代物结合m7 G加帽的病毒mRNA并招募eIF 4G支架蛋白，以获得优先进入核糖体的机会。我们将采用生物化学和药理学的方法来确定核蛋白在病毒翻译中的功能需求。从这些研究中获得的知识对于指导小分子化合物的开发以抑制这种新活性将是重要的。我们预计，基本的病毒-细胞相互作用可以靶向产生高度特异性的抗病毒作用，抵抗遗传逃逸。