Viral interaction with host eIF2alpha kinases

病毒与宿主 eIF2α 激酶的相互作用

• 批准号: 9976443
• 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
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-24 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9976443
• 关键词: Address; Adenovirus Infections; Adenoviruses; Affect; Amino Acids; Antiviral Agents; Antiviral Response; Binding; Binding Sites; Biological Assay; Biology; Bunyaviridae; 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; 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

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.

总结 病毒在面对抗病毒反应时使用许多策略来翻译它们的mRNA， 阻断病毒蛋白质合成。影响蛋白质合成的最突出的抗病毒机制是 蛋白激酶R（PKR）对真核细胞启动因子2 β（eIF 2 β）的磷酸化作用。人腺病毒 （hAds）通过阻断PKR活化的病毒相关（VA）RNA克服这一点。然而，小鼠腺病毒 1型（MAV-1）不编码VA RNA; MAV-1如何成功克服PKR尚不清楚。的作用 另一种eIF 2 β激酶GCN 2在抗病毒应答中的作用还不太清楚，但它对MAV-1具有抗病毒作用。 目前迫切需要了解PKR和GCN 2作为抗病毒药物的一部分如何影响病毒复制。 反应长期目标是增加关于腺病毒-宿主相互作用的基础知识， 有助于疾病。本申请的总体目标是确定MAV-1如何调节宿主 eIF 2激酶，并在这些宿主抗病毒防御的存在下成功复制。中央 假设MAV-1通过降低PKR水平克服宿主抗病毒应答，而MAV-1 激活但不逃避巨噬细胞中的GCN 2，巨噬细胞是病毒复制的靶细胞。这个假设是基于 根据初步数据，PKR在MAV-1感染中被耗尽，GCN 2缺陷的巨噬细胞产生更多的PKR， 病毒和GCN 2缺陷小鼠更易感，并表达更多的促炎细胞因子， MAV-1感染比对照小鼠更严重。基本原理是，了解MAV-1如何诱导和抵消宿主 抗病毒反应将使得能够操纵这些反应并在强有力的体内试验中测试它们的作用。 这将增加对疾病的了解。具体目标是确定1）MAV-1如何 对抗宿主PKR应答，以及2）宿主GCN 2应答如何保护小鼠免受MAV-1感染。 这些目的将使用在申请人的专利申请中建立的MAV-1发病机制和分子病毒学的测定。 实验室在目标1中，初步数据显示，在感染期间，PKR通过蛋白酶体降解， PKR翻译被抑制将被验证和扩展，以确定机制和是否特异性 病毒基因是罪魁祸首方法将使用病毒突变体、PKR-/-小鼠和细胞。目标2将解决如何 缺乏GCN 2会导致小鼠死亡率和促炎反应升高， MAV-1诱导GCN 2活化，使用病毒突变体，和野生型和GCN 2-/-小鼠和细胞。的方法 是创新的，因为它将首次阐明DNA病毒如何诱导PKR的消耗， 扩大了对eIF 2 β激酶GCN 2作为宿主抗病毒反应的关注。这项研究意义重大，因为它 预期与病毒-宿主相互作用和人类疾病具有根本相关性。