The role of mammarenavirus defective interfering particles in protecting host fitness and the host-driven post-translational modifications that regulate their formation and function

哺乳动物病毒缺陷干扰颗粒在保护宿主健康中的作用以及调节其形成和功能的宿主驱动的翻译后修饰

• 批准号: 10687000
• 负责人: Jason W. Botten
• 金额: $ 54.09万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-19 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10687000
• 关键词: Address; Animals; Arenavirus; Binding; Bunyavirales; Cell physiology; Cells; Defective Viruses; Disease; Ecology; Engineering; Ensure; Environmental Impact; Family; Goals; Human; Immune response; Immunity; Infection; Integration Host Factors; Investigation; Knowledge; Ligase; Link; Lymphocytic choriomeningitis virus; Maintenance; Maps; Modification; Molecular; Mus; Nature; Pathogenesis; Pathway interactions; Phosphorylation; Phosphotransferases; Polymerase; Post-Translational Protein Processing; Production; Property; Protein Tyrosine Kinase; Proteins; Proteome; RNA Viruses; Reagent; Recombinants; Regulation; Resolution; Rodent; Role; Site; Study models; System; Testing; Time; Ubiquitination; Viral; Viral Matrix Proteins; Viral Pathogenesis; Viral Proteins; Virion; Virus; Virus Diseases; Work; Zoonoses; acute infection; chronic infection; fitness; human pathogen; in vivo; innovation; novel; particle; pathogen; pathogenic virus; plasma protein Z; recruit; reverse genetics; tool; trafficking; transmission process; ubiquitin-protein ligase; vector; virology; virus host interaction

The pivotal discovery of RNA viruses, like the bi-segmented, single-stranded arenavirus family, producing defective interfering particles (DIPs) over 50 years ago prompted investigations on their contribution to viral pathogenesis and host immunity. However, many basic questions remain, including the molecular basis for how DIPs are regulated and the contribution of DIPs to virus–host interactions, including the persistence of zoonotic pathogens in their reservoir host. Our overall goal is to identify the cellular machinery responsible for regulating arenavirus DIP production and interrogate the impact DIPs have on reservoir fitness. A major constraint to understanding the true role of DIPs in viral pathogenesis and disease ecology is the lack of tools to modulate DIP levels in experimental systems. We recently discovered that LCMV uses divergent cellular pathways to produce standard virus particles versus DIPs. Using reverse genetic systems, we identified powerful host-driven post-translational modifications (PTMs) that dynamically regulate the production of infectious versus defective viral particles. These studies enabled us to engineer, for the first time, recombinant arenaviruses that no long produce DIPs. Our specific objectives will be to use these innovative approaches and tools to 1) define the mechanism by which host tyrosine kinases and NEDD4 Family E3 ubiquitin ligases regulate DIP formation and function, 2) expand our global map of arenavirus PTMs that may influence DIP production, and 3) connect these molecular findings to pathogenesis studies modeling both persistent infection of the rodent reservoir and acute infection of the rodent or incidental human host. Most RNA viruses that infect animals produce DIPs, which suggests they are fundamentally important for the maintenance of these viruses in nature. Our findings will be broadly applicable beyond the Bunyavirales order. Collectively, the completion of these studies will provide greater resolution on the regulation of DIP production and answer, for the first time, the role of DIP in viral persistence.

RNA 病毒的关键发现，如双节段、单链沙粒病毒家族，产生 50 多年前的缺陷干扰粒子 (DIP) 促使人们研究其对病毒的贡献 发病机制和宿主免疫。然而，许多基本问题仍然存在，包括分子基础 DIP 的监管方式以及 DIP 对病毒与宿主相互作用的贡献，包括持久性 人畜共患病原体在其储存宿主中。我们的总体目标是确定负责的细胞机制 调节沙粒病毒 DIP 的产生并探讨 DIP 对储存库适应性的影响。一个专业 缺乏工具是理解 DIP 在病毒发病机制和疾病生态学中真正作用的限制因素 调节实验系统中的 DIP 水平。我们最近发现 LCMV 使用不同的细胞 生产标准病毒颗粒与 DIP 的途径。使用反向遗传系统，我们确定了 强大的宿主驱动的翻译后修饰 (PTM)，可动态调节 传染性病毒颗粒与有缺陷的病毒颗粒。这些研究使我们能够首次设计重组 不再产生 DIP 的沙粒病毒。我们的具体目标是使用这些创新方法 和工具 1) 定义宿主酪氨酸激酶和 NEDD4 家族 E3 泛素连接酶的机制 调节 DIP 的形成和功能，2) 扩展可能影响 DIP 的沙粒病毒 PTM 的全球图谱 生产，3) 将这些分子发现与建模持续感染的发病机制研究联系起来 啮齿动物宿主的感染和啮齿动物或偶然人类宿主的急性感染。大多数感染的RNA病毒 动物产生 DIP，这表明它们对于维持这些病毒至关重要 在大自然中。我们的研究结果将广泛适用于布尼亚病毒目以外的领域。总的来说，完成 这些研究将为 DIP 生产的监管提供更大的解决方案，并首次回答： DIP 在病毒持久性中的作用。