Viral interaction with host eIF2alpha kinases

病毒与宿主 eIF2α 激酶的相互作用

基本信息

批准号：
10459361
负责人：
Katherine R. Spindler
金额：
$ 59.51万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-24 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10459361
关键词：
Address Adenovirus Infections Adenoviruses Affect Amino Acids Antiviral Response Binding Binding Sites Biological Assay Biology Black race Cells Cellular Stress Color DNA Viruses Data Development Disease Double-Stranded RNA Enzymes Eukaryotic Initiation Factor-2 Family Family Picornaviridae Gene Expression Goals Growth HIV-1 Host Defense Human In Vitro Infection Infection Control Knowledge Laboratories Messenger RNA Mission Molecular Molecular Virology Murid herpesvirus 1 Mus Nucleic Acids Orthobunyavirus Pathogenesis Phosphorylation Phosphotransferases Proteasome Inhibitor Protein Analysis Protein Biosynthesis RNA RNA Polymerase III Reporting Research Role Sindbis Virus System Testing Time Transcript Translating Translations Ubiquitination United States National Institutes of Health Untranslated RNA Viral Viral Genes Viral Pathogenesis Viral Proteins Virus Virus Diseases Virus Replication base cytokine deprivation disorder control experimental study human disease in vivo in vivo Model innovation macromolecule macrophage mortality mouse model multicatalytic endopeptidase complex mutant prevent protein activation protein kinase R response virology virus host interaction

项目摘要

Summary Viruses use many strategies to translate their mRNAs in the face of antiviral responses that would otherwise block viral protein synthesis. The most prominent antiviral mechanism affecting protein synthesis is phosphorylation of eukaryotic initation factor 2 (eIF2 by protein kinase R (PKR). Human adenoviruses (hAds) overcome this via virus-associated (VA) RNAs that block PKR activation. However, mouse adenovirus type 1 (MAV-1) does not encode VA RNAs; how MAV-1 successfully overcomes PKR is unknown. The role of another eIF2 kinase, GCN2, in antiviral responses is less well appreciated, but it is antiviral against MAV-1. There is a critical need to understand how PKR and GCN2 impact viral replication as part of the antiviral response. The long-term goal is to increase fundamental knowledge about adenovirus-host interactions that contribute to disease. The overall objective of this application is to determine how MAV-1 modulates host eIF2 kinases and successfully replicates in the presence of these host antiviral defenses. The central hypothesis is that MAV-1 overcomes the host antiviral response by reducing PKR levels, whereas MAV-1 activates but does not evade GCN2 in macrophages, target cells of viral replication. This hypothesis is based on preliminary data that PKR is depleted in MAV-1 infections, GCN2-deficient macrophages produce more virus, and GCN2-deficient mice are more susceptible and express more proinflammatory cytokines upon MAV-1 infection than control mice. The rationale is that knowing how MAV-1 induces and counteracts the host antiviral responses will enable manipulation of these responses and testing their effects in a powerful in vivo model, which will increase understanding of disease. The specific aims are to determine 1) how MAV-1 counters the host PKR response, and 2) how the host GCN2 response protects mice from MAV-1 infection. These aims will use assays of MAV-1 pathogenesis and molecular virology established in the applicant's laboratory. In Aim 1, preliminary data showing that during infection PKR is degraded via the proteasome and PKR translation is inhibited will be validated and extended to identify the mechanisms and whether specific viral genes are responsible. Approaches will use viral mutants, PKR-/- mice and cells. Aim 2 will address how the lack of GCN2 leads to higher mouse mortality and proinflammatory responses, and whether and how MAV-1 induces GCN2 activation, using viral mutants, and wild type and GCN2-/- mice and cells. The approach is innovative because it will elucidate for the first time how a DNA virus induces depletion of PKR, and it will broaden the focus on eIF2 kinase GCN2 as a host antiviral response. The research is significant because it is expected to have fundamental relevance to virus-host interactions and human disease.

概括病毒使用许多策略来转化其mRNA，面对抗病毒反应阻断病毒蛋白合成。影响蛋白质合成的最突出的抗病毒机制是真核因子因子2的磷酸化（蛋白激酶R（PKR）的EIF2。人腺病毒（HADS）通过与病毒相关的（VA）RNA（阻断PKR激活）克服了这一点。但是，小鼠腺病毒类型1（MAV-1）不编码VA RNA； MAV-1如何成功克服PKR是未知的。的作用在抗病毒反应中的另一种EIF2激酶GCN2不太受欢迎，但它是对MAV-1的抗病毒。迫切需要了解PKR和GCN2如何影响病毒复制作为抗病毒的一部分回复。长期目标是增加有关腺病毒宿主相互作用的基本知识有助于疾病。该应用程序的总体目的是确定MAV-1如何调节主机 EIF2激酶并在存在这些宿主抗病毒防御的情况下成功复制。中央假设MAV-1通过降低PKR水平克服了宿主抗病毒反应，而MAV-1 激活但不会在巨噬细胞，病毒复制的靶细胞中逃避GCN2。该假设是基于关于PKR在MAV-1感染中耗尽的初步数据，GCN2缺乏巨噬细胞产生了更多病毒和GCN2缺乏小鼠更容易敏感，并在上表达更多促炎细胞因子 MAV-1感染比对照小鼠。理由是知道MAV-1如何影响和抵消主机抗病毒反应将使这些反应能够操纵这些反应并在强大的体内测试其效果模型，这将增加对疾病的理解。具体目的是确定1）MAV-1如何计算宿主PKR响应，以及2）宿主GCN2反应如何保护小鼠免受MAV-1感染的影响。这些目标将使用申请人中建立的MAV-1发病机理和分子病毒学的测定实验室。在AIM 1中，初步数据表明，在感染期间，PKR通过蛋白酶体和抑制PKR翻译将得到验证并扩展，以识别机制以及是否具体病毒基因是负责的。方法将使用病毒突变体，PKR - / - 小鼠和细胞。 AIM 2将解决如何缺乏GCN2会导致小鼠死亡率和促炎反应，以及是否以及如何以及如何 MAV-1使用病毒突变体以及野生型和GCN2 - / - 小鼠和细胞诱导GCN2激活。方法具有创新性，因为它将首次阐明DNA病毒如何诱导PKR的耗竭，并且它将扩大对EIF2激酶GCN2作为宿主抗病毒反应的关注。这项研究很重要，因为它是预计将与病毒 - 宿主相互作用和人类疾病具有基本相关性。