Identification of Novel Arenavirus Protein-Host Cellular Protein Interactions

新型沙粒病毒蛋白-宿主细胞蛋白相互作用的鉴定

• 批准号: 8077445
• 负责人: Jason W. Botten
• 金额: $ 18.86万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-15 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8077445
• 关键词: Address; Affinity Chromatography; Antiviral Agents; Applications Grants; Arenavirus; Aseptic Meningitis; Basic Science; Biology; Biomedical Research; Categories; Cell Line; Clinical Research; Complex; DNA-Directed RNA Polymerase; Development; Disease; Exploratory/Developmental Grant; FDA approved; Family; Future; Glycoproteins; Goals; Hantavirus; Human; Immune Plasma; Infection; Junin virus; Lead; Libraries; Lymphocytic choriomeningitis virus; Mammalian Cell; Maps; Mass Spectrum Analysis; Measures; Mission; National Institute of Allergy and Infectious Disease; Nature; Nucleoproteins; Open Reading Frames; Pathogenesis; Pilot Projects; Plasmid Cloning Vector; Prevention; Proteins; Proteome; Proteomics; RNA Viruses; Reagent; Research Project Grants; Research Proposals; Ribavirin; Role; Syndrome; Tacaribe Complex Viruses; Technology; Testing; Therapeutic; Vaccines; Viral; Viral Hemorrhagic Fevers; Viral Pathogenesis; Viral Proteins; Virus; Work; base; human RBX1 protein; human disease; innovation; knock-down; novel; pathogen; plasma protein Z; prevent; protein protein interaction; public health relevance; therapeutic vaccine; viral RNA; virus host interaction

DESCRIPTION (provided by applicant): The arenaviruses are a family of negative-sense RNA viruses that cause severe human disease ranging from aseptic meningitis to hemorrhagic fever syndromes. The arenavirus proteome consists of only four proteins: the small RING finger protein Z, nucleoprotein, viral RNA polymerase, and glycoprotein precursor, which is post-translationally modified to yield the heterodimeric envelope glycoproteins GP1 and GP2. While the functional role of these proteins for viral replication is well defined, their interactions with host cellular proteins, and the importance of these interactions for viral replication and disease pathogenesis, is largely unknown. Identification of novel arenavirus protein-host protein interactions that are critical for viral replication and/or viral pathogenesis would advance our understanding of the basic biology of these NIAID Category A viruses and provide new targets for the development of antivirals. Accordingly, we will use a cutting edge proteomics approach that features affinity purification of viral proteins in complex with host cellular proteins and multi-dimensional mass spectrometry protein identification technology (MudPIT) to identify host protein partners that interact with the proteomes of selected Old World (lymphocytic choriomeningitis virus) and New World (Junin virus) pathogenic arenaviruses. Our group has recently utilized this approach to successfully identify several host proteins that interact with viral proteins, including interactions that are conserved among both hantaviruses and arenaviruses. In addition, we have already generated the necessary reagents required for this work, namely a library of plasmid vectors that express, in mammalian cells, each of the ORFs encoded by six arenaviruses that are pathogenic for humans. The arenavirus protein-host protein interactions identified through this work will provide the basis for future grant applications to characterize these interactions more fully, particularly their impact on viral replication and pathogenesis. As an initial means to determine which interactions would be most relevant to pursue in future studies, we will evaluate the importance of the identified arenavirus protein-host protein interactions for viral replication by measuring viral replication in permissive cell lines following selective knock-down of host protein partners. PUBLIC HEALTH RELEVANCE: The arenaviruses cause severe human disease ranging from aseptic meningitis to hemorrhagic fever syndromes. These viruses produce four proteins. The goal of this project is to identify the human host proteins that these arenavirus proteins interact with during infection so that we can better understand how these virus protein-host protein interactions impact both the ability of the virus to replicate and cause human disease. Identification of important interactions will provide new targets for the development of antivirals.

描述（由申请方提供）：沙粒病毒属是一个负义RNA病毒家族，可引起从无菌性脑膜炎到出血热综合征的严重人类疾病。沙粒病毒蛋白质组仅由四种蛋白质组成：小环指蛋白Z、核蛋白、病毒RNA聚合酶和糖蛋白前体，糖蛋白前体经后修饰产生异二聚体包膜糖蛋白GP 1和GP 2。虽然这些蛋白质对病毒复制的功能作用是明确的，但它们与宿主细胞蛋白质的相互作用以及这些相互作用对病毒复制和疾病发病机制的重要性在很大程度上是未知的。对病毒复制和/或病毒发病机制至关重要的新型沙粒病毒蛋白-宿主蛋白相互作用的鉴定将促进我们对这些NIAID A类病毒的基础生物学的理解，并为抗病毒药物的开发提供新的靶点。因此，我们将使用尖端的蛋白质组学方法，其特征在于与宿主细胞蛋白复合的病毒蛋白的亲和纯化和多维质谱蛋白质鉴定技术（MudPIT），以鉴定与选定的旧世界（淋巴细胞性脉络丛脑膜炎病毒）和新世界（朱宁病毒）致病性沙粒病毒的蛋白质组相互作用的宿主蛋白质伴侣。我们小组最近利用这种方法成功地鉴定了几种与病毒蛋白相互作用的宿主蛋白，包括汉坦病毒和沙粒病毒之间保守的相互作用。此外，我们已经产生了这项工作所需的必要试剂，即在哺乳动物细胞中表达由六种对人类致病的沙粒病毒编码的每个ORF的质粒载体库。通过这项工作确定的沙粒病毒蛋白-宿主蛋白相互作用将为未来的拨款申请提供基础，以更充分地表征这些相互作用，特别是它们对病毒复制和发病机制的影响。作为确定在未来研究中最相关的相互作用的初始方法，我们将通过在选择性敲低宿主蛋白伴侣后测量允许细胞系中的病毒复制来评估所鉴定的沙粒病毒蛋白-宿主蛋白相互作用对病毒复制的重要性。 公共卫生相关性：沙粒病毒引起严重的人类疾病，从无菌性脑膜炎到出血热综合征。这些病毒产生四种蛋白质。该项目的目标是确定这些沙粒病毒蛋白在感染过程中与之相互作用的人类宿主蛋白，以便我们能够更好地了解这些病毒蛋白-宿主蛋白相互作用如何影响病毒复制和引起人类疾病的能力。重要相互作用的鉴定将为抗病毒药物的开发提供新的靶点。