Investigating How ADP-ribosylation Impacts Innate Immunity During Coronavirus Infection

研究 ADP-核糖基化如何影响冠状病毒感染期间的先天免疫

• 批准号: 9428897
• 负责人: Anthony R Fehr
• 金额: $ 16.2万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-12-07 至 2020-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9428897
• 关键词: ADP ribosylation; Activation Analysis; Address; Adenosine Diphosphate Ribose; Animals; Antiviral Agents; Biology; Cell physiology; Code; Confocal Microscopy; Coronavirus; Coronavirus Infections; Cytoplasmic Structures; Data; Development; Disease; Enzymes; Epidemic; Faculty; Family; Goals; Human; I-kappa B Proteins; Immune; Immune response; Immune signaling; Infection; Inflammatory; Innate Immune Response; Interferons; K22 Award; Knockout Mice; Label; Life; Literature; Lung diseases; Mammals; Mediating; Medical; Modeling; Murine hepatitis virus; Mus; Mutation; Natural Immunity; Nonstructural Protein; Pathogenesis; Pathogenicity; Pathway interactions; Poly(ADP-ribose) Polymerases; Post-Translational Protein Processing; Process; Property; Proteins; RNA; RNA Interference; Regulation; Resources; SARS coronavirus; Scientist; Signal Pathway; Signaling Protein; Site-Directed Mutagenesis; Societies; Techniques; Testing; Therapeutic; Transcription Factor AP-1; Vaccine Design; Viral; Viral Genome; Viral Pathogenesis; Virulence Factors; Virus; Virus Diseases; Virus Replication; Work; blocking factor; career; combat; cytokine; experimental study; in vivo; insight; interest; knock-down; mutant; pathogen; porcine epidemic diarrhea virus; prevent; public health relevance; respiratory virus; response; reverse genetics; sensor; stress granule; transcription factor

SUMMARY Coronaviruses (CoVs) have proven to be significant pathogens of both veterinary and medical importance since their discovery over 50 years ago and are responsible for two recent epidemics (SARS-CoV and MERS- CoV). The ability of CoVs to establish infection and to cause disease is dependent on their ability to inhibit the host innate immune response. Many questions still remain in this field, such as what viral factors function in vivo to prevent cytokine expression, and whether RNA sensors other than MDA5 can detect CoV RNA. I am interested in exploring both viral factors that block the immune response, as well as host proteins that promote the response and that mediate protection from infection. I have discovered that the conserved CoV macrodomain both suppresses the innate immune response and promotes in vivo replication to facilitate viral pathogenesis. Furthermore, our group and others have shown that the CoV macrodomain is an enzyme that removes ADP-ribose from proteins. These studies indicate that protein ADP-ribosylation is a mechanism used by the host to promote the innate immune response. However, neither the enzymes that catalyze the ADP- ribosylation (PARPs) or the targets of this modification are known. The central objectives in this proposal are to identify the factors that mediate antiviral ADP-ribosylation and enhance our understanding of how ADP-ribosylation impacts the innate immune response to counter CoV infection. In Aim 1 I will screen for the PARP(s) that impacts innate immunity during CoV infection and determine its localization. In Aim 2, I will systematically identify the sensor and specific step of the signaling pathway that is activated in the absence of the macrodomain. Utilizing the results from these two aims, I will then begin the process of identifying the direct protein target of the CoV macrodomain. Taken together, these aims will further define how ADP-ribosylation impacts innate immunity and CoV pathogenesis and will address the mechanism of how macrodomains combat cellular ADP-ribosylation, which remains a significant gap in the literature. A detailed understanding of the interplay between ADP-ribosylation and CoV macrodomains will provide significant insight into mechanisms of the host response and how viruses combat this response.

概括 冠状病毒 (CoV) 已被证明是具有兽医和医学重要性的重要病原体 自 50 多年前发现以来，它们是最近两种流行病（SARS-CoV 和 MERS- 冠状病毒）。冠状病毒建立感染和引起疾病的能力取决于它们抑制病毒的能力 宿主先天免疫反应。该领域仍然存在许多问题，例如病毒因子在其中发挥什么作用？ 体内阻止细胞因子表达，以及除 MDA5 之外的 RNA 传感器是否可以检测 CoV RNA。我是 有兴趣探索阻断免疫反应的病毒因素，以及促进免疫反应的宿主蛋白 反应并介导防止感染的保护。我发现保守的 CoV 宏结构域既抑制先天免疫反应，又促进体内复制以促进病毒 发病。此外，我们的小组和其他人已经证明，冠状病毒宏结构域是一种酶， 从蛋白质中去除 ADP-核糖。这些研究表明蛋白质 ADP-核糖基化是一种使用的机制 由宿主促进先天免疫反应。然而，催化 ADP- 核糖基化（PARP）或这种修饰的目标是已知的。本提案的中心目标 的目的是确定介导抗病毒 ADP-核糖基化的因素并增强我们的理解 ADP-核糖基化如何影响对抗 CoV 感染的先天免疫反应。在目标 1 中我会 筛选在 CoV 感染期间影响先天免疫的 PARP 并确定其定位。在 目标2，我将系统地识别传感器和信号通路中激活的具体步骤 缺少宏结构域。利用这两个目标的结果，我将开始以下过程： 识别 CoV 宏结构域的直接蛋白质靶标。总而言之，这些目标将进一步明确如何 ADP-核糖基化影响先天免疫和 CoV 发病机制，并将解决如何机制 宏结构域对抗细胞 ADP-核糖基化，这在文献中仍然是一个重大空白。详细的 了解 ADP-核糖基化和 CoV 宏结构域之间的相互作用将提供重要的见解 研究宿主反应的机制以及病毒如何对抗这种反应。