Importance of Species-Specific Interactions of PKR with Poxvirus Inhibitors for Virus Replication and Host Range

PKR 与痘病毒抑制剂的物种特异性相互作用对于病毒复制和宿主范围的重要性

• 批准号: 8965294
• 负责人: Stefan Rothenburg
• 金额: $ 36.7万
• 依托单位: KANSAS STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-24 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8965294
• 关键词: Address; Affect; Animal Model; Animals; Antiviral Agents; Antiviral Response; Binding; Biological Assay; Cell Line; Cells; Data; Double-Stranded RNA; Effectiveness; Emerging Communicable Diseases; European; Evolution; Foundations; Future; Genes; Goals; Hamsters; Health; Herpesvirus 1; Human; Immune system; Individual; Infection; Influenza A virus; Knowledge; Lead; Modeling; Molecular; Mouse Protein; Mus; Mutagenesis; Myxoma virus; Oncolytic; Orthologous Gene; Oryctolagus cuniculus; Outcome; Peptide Initiation Factors; Population; Poxviridae; Predisposition; Prokaryotic Initiation Factor-2; Protein Synthesis Inhibition; Proteins; Public Health; Relative (related person); Research; Resistance; Role; Testing; Textbooks; Vaccines; Vaccinia virus; Variant; Vertebrates; Viral; Viral Proteins; Virulence; Virus; Virus Diseases; Virus Inhibitors; Virus Replication; Zoonotic Infection; attenuation; base; disorder control; eIF-2 Kinase; improved; inhibitor/antagonist; innovation; insight; member; mutant; novel; novel strategies; pathogen; protein kinase R; public health relevance; research study; resistance mechanism; sensor; tool; transmission process; viral RNA; viral detection; virus virus interaction; weapons

 DESCRIPTION (provided by applicant): Zoonotic viral infections are responsible for a large portion of emerging infectious diseases in humans. However, for many viruses the molecular basis that determines host range, virulence and the potential for cross-species transmission is poorly understood. The long-term goal is to delineate key factors that contribute to viral host range and virulence by analyzing the interaction of viruses with the host innate immune system. A major focus are model poxviruses, which display extreme differences in host range and virulence and include major human and animal pathogens. The objective of this application is to determine the contribution of species-specific interactions of the host antiviral protein kinase R (PKR) with virus inhibitors for virus replication, virulence and host range. The central hypothesis is that both viral host range and virus replication are directly influenced by the interaction of PR and its viral inhibitors. The rationale of this project is that an improved understanding of the mechanisms that determine virus host range can ultimately result in a better prediction of host switches and changes in virulence and also might lead to new alternative strategies for disease control. Based on strong preliminary data, the main hypothesis will be tested by pursuing three specific aims: 1) Characterize host-specific PKR inhibition by poxvirus inhibitors; 2) Elucidate the molecular basis for PKR sensitivity to viral inhibitors; and 3) Evaluate the effects and functional consequences of naturally occurring variants of rabbit PKR and myxoma virus inhibitors. Under the first aim, the interactions between PKR proteins from different host species with PKR inhibitors from prototypic poxviruses will be studied and their functional relevance in cell line infection assays tested. In the second aim, the molecular basis for species-specific PKR-inhibitor interactions will be determined by combining mutagenesis, binding and infection assays and the potential of PKR to resist PKR inhibitors from other viruses will be tested. Under the third aim, naturally evolved variants of PKR from rabbits and from myxoma virus PKR inhibitors, as well as mutants that were generated in artificial evolution experiments, will be analyzed for effects in interaction and infection assays to test the PKR-inhibitor interactions in textbook example for host-virus coevolution. The proposed research is significant because it will lead to the better understanding of the molecular basis for poxvirus host range, yield novel insights into the mechanisms of resistance to viruses and how they evolve, and might lead to a better prediction of which viruses have a strong potential for cross-species transmission. This project is innovative because it introduces new strategies and novel tools to elucidate the importance of host-virus interactions for virus replication and host range and explores the novel concept that the outcome of cross- species virus transmission is strongly influenced by chance, but might be predicted by experimental analyses. Ultimately, the knowledge gained during this project will lead to the better understanding of the molecular mechanisms behind virus host range and resistance and is expected to propel the field forward.

 描述（由申请方提供）：人畜共患病毒感染是人类大部分新发传染病的原因。然而，对于许多病毒，决定宿主范围、毒力和跨物种传播潜力的分子基础知之甚少。长期目标是通过分析病毒与宿主先天免疫系统的相互作用来描述影响病毒宿主范围和毒力的关键因素。一个主要的焦点是模型痘病毒，其在宿主范围和毒力方面显示出极大的差异，并且包括主要的人类和动物病原体。本申请的目的是确定宿主抗病毒蛋白激酶R（PKR）与病毒抑制剂的种属特异性相互作用对病毒复制、毒力和宿主范围的贡献。核心假设 病毒宿主范围和病毒复制都直接受到PR及其病毒抑制剂相互作用的影响。该项目的基本原理是，提高对决定病毒宿主范围的机制的理解，最终可以更好地预测宿主转换和毒力变化，也可能导致新的疾病控制替代策略。基于强有力的初步数据，将通过追求三个特定目标来检验主要假设：1）表征痘病毒抑制剂对宿主特异性PKR的抑制; 2）阐明PKR对病毒抑制剂敏感性的分子基础; 3）评价兔PKR和粘液瘤病毒抑制剂的天然变体的作用和功能后果。在第一个目标下，将研究来自不同宿主物种的PKR蛋白与来自原型痘病毒的PKR抑制剂之间的相互作用，并测试它们在细胞系感染测定中的功能相关性。在第二个目标中，物种特异性PKR-抑制剂相互作用的分子基础将通过结合诱变、结合和感染测定来确定，并且将测试PKR抵抗来自其他病毒的PKR抑制剂的潜力。在第三个目标下，将分析来自兔和粘液瘤病毒PKR抑制剂的PKR的自然进化变体，以及在人工进化实验中产生的突变体在相互作用和感染测定中的作用，以测试PKR-抑制剂相互作用在宿主-病毒共进化的教科书示例中。拟议的研究是重要的，因为它将导致更好地了解痘病毒宿主范围的分子基础，产生对病毒抗性机制及其演变的新见解，并可能导致更好地预测哪些病毒具有跨物种传播的强大潜力。该项目具有创新性，因为它引入了新的策略和新的工具来阐明宿主-病毒相互作用对病毒复制和宿主范围的重要性，并探索了跨物种病毒传播的结果受到偶然性的强烈影响，但可以通过实验分析预测的新概念。最终，在该项目中获得的知识将导致更好地理解病毒宿主范围和抗性背后的分子机制，并有望推动该领域的发展。