Determining mechanisms of innate immune modulation by ADP-ribosylation

通过 ADP-核糖基化确定先天免疫调节机制

• 批准号: 10386112
• 负责人: Anthony R Fehr
• 金额: $ 3万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10386112
• 关键词: ADP Ribose Transferases; ADP ribosylation; Address; Adenosine Diphosphate Ribose; Affect; Antiviral Agents; Antiviral Response; Antiviral Therapy; Automobile Driving; Biological Process; Biology; Cell Culture Techniques; Cells; Chiroptera; Communicable Diseases; Coronavirus; Family; Foundations; Goals; Hydrolase; Hypersensitivity; Immune response; Impairment; Infection; Innate Immune Response; Interferons; Knock-out; Knowledge; Lead; Malignant Neoplasms; Mediating; Mission; Modeling; Murine hepatitis virus; National Institute of General Medical Sciences; Pathway interactions; Phenotype; Phosphorylation; Post-Translational Protein Processing; Process; Proteins; RNA replication; Research; Research Personnel; Role; System; Togaviridae; Translations; Viral; Viral hepatitis; Virus; Virus Diseases; Virus Replication; Work; human disease; immunoregulation; inhibitor/antagonist; innate immune mechanisms; innovation; mutant; novel; pathogenic virus; protein function; response

PROJECT SUMMARY ADP-ribosylation is an important post-translational modification that directly influences several biological processes including cancer, allergy, and infectious disease. ADP-ribose can be added to proteins as one or more consecutive units by ADP-ribosyltransferases, also termed PARPs, resulting in mono- or poly- ADP-ribosylation (mAR or pAR). Most mARylating PARPs are upregulated by interferon (IFN) upon virus infection and several are predicted to have antiviral functions. In addition, several viral families, including the Coronaviridae and Togaviridae, encode for macrodomain proteins that have mono-ADP-ribosyl hydrolase (ARH) activity. This activity allows these viruses the ability to specifically counteract the effects of mAR, further implicating mAR in the mammalian antiviral response. Despite these findings, there are only a few known examples where mAR is known to inhibit virus replication. This is largely due to the lack of cell culture models of virus infections where the mAR status of a cell can be specifically controlled, such as models using mutant viruses or PARP knockout cells that have significant phenotypes. Importantly, the PI has established a virus infection system using a model coronavirus, Murine Hepatitis Virus (MHV), where virus lacking ARH activity is i) significantly impaired in virus replication and ii) independently induces a robust IFN response. These phenotypes are reversed by PARP inhibitors, establishing mAR as a key factor driving this anti-viral response. The investigator’s long-term goal is to determine mechanistically how mAR inhibits virus replication and enhances the innate immune response following virus infection. This gap in knowledge will be resolved by answering the following questions: 1) How does mAR inhibit MHV infection? Does it inhibit the entry, RNA replication, protein translation, assembly, or release of MHV? 2) What step(s) of the IFN induction pathway is enhanced by mAR, and does mAR also affect the IFN response in bats, which are known to harbor many highly pathogenic viruses? 3) What proteins are modified by PARPs following virus infection and which substrates are relevant for specific phenotypes? The rationale for this research is that it will enhance our understanding of mAR, including its ability to modulate protein function and will uncover novel cellular proteins or processes that mediate virus replication. The work is innovative because: i) it will bridge a significant knowledge gap between ADP-ribose biology, the innate immune response, and virus replication; ii) it utilizes unique models of infection utilizing both mutant viruses and PARP knockout cells; and iii) will be the first to address the role of mAR in bats. Finally, these projects are significant and relevant to the NIGMS mission because they will provide a thorough understanding of how mAR impacts the anti-viral response that could lay the foundation for advances in the treatment of virus infections or other human diseases impacted by mAR.

项目摘要 ADP-核糖基化是一种重要的翻译后修饰，直接影响几种 包括癌症，过敏和传染病在内的生物过程。可以将ADP-核糖添加到蛋白质中 通过ADP-核糖基转移酶（也称为PARPS）连续的一个或多个单位，导致单或聚 ADP-核糖基（MAR或PAR）。大多数Marylating Parps由Interferon（IFN）对病毒进行更新 预计感染和一些具有抗病毒功能。此外，包括几个病毒家庭，包括 冠状病和togaviridae，编码具有单ADP-核糖基水解酶的大域蛋白 （ARH）活动。该活动使这些病毒能够特别抵消MAR的影响，进一步 哺乳动物抗病毒反应中的隐性MAR。尽管有这些发现，但只有少数已知 已知MAR抑制病毒复制的示例。这主要是由于缺乏细胞培养模型 病毒感染可以特异性控制的病毒感染，例如使用突变体的模型 具有重要表型的病毒或PARP基因敲除细胞。重要的是，PI已经建立了病毒 使用模型冠状病毒，鼠肝炎病毒（MHV）的感染系统，缺乏ARH活性的病毒为 i）在病毒复制中显着损害，ii）独立诱导了强大的IFN反应。这些 表型被PARP抑制剂逆转，确立了MAR作为推动这种抗病毒反应的关键因素。 研究者的长期目标是从机械上确定MAR如何抑制病毒复制和 病毒感染后增强先天免疫反应。知识差距将由 回答以下问题：1）MAR如何抑制MHV感染？它会抑制入口，RNA吗 MHV的复制，蛋白质翻译，组装或释放？ 2）IFN感应途径的哪一步 MAR增强，并且MAR也会影响蝙蝠中的IFN反应，蝙蝠已知许多 高致病的病毒？ 3）病毒感染后，哪种蛋白质通过PARP修饰，哪些蛋白质是 底物与特定表型相关吗？这项研究的理由是它将增强我们的 了解MAR，包括调节蛋白质功能的能力，并将发现新型细胞蛋白 或介导病毒复制的过程。这项工作是创新的，因为：i）它将桥梁 ADP-核糖生物学，先天免疫反应和病毒复制之间的知识差距； ii）它利用 使用突变病毒和PARP基因敲除细胞的独特感染模型； iii）将是第一个 解决MAR在蝙蝠中的作用。最后，这些项目很重要，并且与NIGMS任务相关 因为他们将对可能产生的抗病毒反应有透彻的理解 治疗病毒感染或其他人类疾病的基础。