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

基本信息

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
项目类别：
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

项目摘要

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（由申请人提供）：体育症病毒是一个负义RNA病毒家族，引起严重的人类疾病，从无菌性脑膜炎到出血热综合征。体育症病毒蛋白质组仅由四种蛋白质组成：小环蛋白Z，核蛋白，病毒RNA聚合酶和糖蛋白前体，后者在后修饰后修饰，以产生异质二聚体胶片糖蛋白GP1和GP2。尽管这些蛋白质在病毒复制方面的功能作用已很好地定义，但它们与宿主细胞蛋白的相互作用以及这些相互作用对于病毒复制和疾病发病机理的重要性在很大程度上尚不清楚。对病毒复制和/或病毒发病机理至关重要的新型体育经瘤病毒蛋白宿主蛋白相互作用的鉴定将提高我们对这些NIAID类别A病毒的基本生物学的理解，并为抗病毒药的发展提供了新的靶标。因此，我们将使用一种最先进的蛋白质组学方法，该方法具有与宿主细胞蛋白的复合物和多维质谱蛋白鉴定技术（MUDPIT）的复杂性纯度纯化，以鉴定与所选旧世界（淋巴细胞炎性炎性炎性炎）和新世界（ned World）（anden World）（anden World）相互作用的宿主蛋白伴侣。我们的小组最近利用这种方法成功地识别了几种与病毒蛋白相互作用的宿主蛋白，包括在汉坦病毒和体育症病毒中保守的相互作用。此外，我们已经生成了这项工作所需的必要试剂，即在哺乳动物细胞中表达的质粒载体库，每个ORF在每个属于人类致病的体育病毒中编码的ORF。通过这项工作确定的竞技病毒蛋白宿主蛋白相互作用将为未来的赠款应用提供基础，以更充分地表征这些相互作用，尤其是它们对病毒复制和发病机理的影响。作为确定哪些相互作用在未来研究中最相关的一种方法，我们将通过测量宿主蛋白伴侣选择性敲门后，通过测量允许细胞系中的病毒复制来评估已鉴定的体育经瘤病毒蛋白质宿主蛋白相互作用对病毒复制的重要性。公共卫生相关性：体育症病毒会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。该项目的目的是确定这些体育症病毒蛋白在感染过程中与之相互作用的人类宿主蛋白质，以便我们可以更好地了解这些病毒蛋白质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。重要相互作用的识别将为抗病毒药的发展提供新的目标。