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: 10514041
• 负责人: Jason W. Botten
• 金额: $ 54.39万
• 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/10514041
• 关键词: Address; Alpha Particles; Animals; Arenavirus; Binding; Bunyavirales; Cell physiology; Cells; Defective Viruses; Disease; Ecology; Engineering; Ensure; Environmental Impact; Family; Generations; Goals; Human; Immune response; Immunity; Infection; Integration Host Factors; Investigation; Knowledge; Life; 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，例如双分段的单链体aRANAVIRUS家族，产生 50年前的干扰颗粒（DIPS）有缺陷，促使对病毒的贡献进行了调查 发病机理和宿主免疫。但是，仍然存在许多基本问题，包括 如何调节倾角以及倾角对病毒 - 霍斯特相互作用的贡献，包括 他们的水库主机中的人畜共患病原体。我们的总体目标是确定负责 调节体育症病毒浸入生产并询问DIP对储层健身的影响。专业 限制理解DIP在病毒发病机理和疾病生态学中的真正作用是缺乏工具 调节实验系统中的倾斜水平。我们最近发现LCMV使用不同的细胞 产生标准病毒颗粒与浸入的途径。使用反向遗传系统，我们确定了 强大的主机驱动后翻译后修饰（PTMS），该修饰动态调节的产生 传染性与有缺陷的病毒颗粒。这些研究使我们能够首次设计重组 没有长时间降低的体aver病毒。我们的具体目标是使用这些创新的方法 和1）定义宿主酪氨酸激酶和NEDD4家族E3泛素连接酶的机制 调节倾角的形成和功能，2）扩展我们可能影响DIP的全球轨迹病毒PTM 生产和3）将这些分子发现与两种持续感染建模的发病机理研究联系起来 啮齿动物或偶然的人类宿主的啮齿动物储层和急性感染。大多数感染的RNA病毒 动物产生倾角，这表明它们对于维持这些病毒至关重要 本质。我们的发现将广泛适用于Bunyavirales命令。共同完成 这些研究将首次提供有关调节蘸水和答案的更大分辨率， DIP在病毒持久性中的作用。