Dissecting ADP-ribosylation as an innate immune response countering influenza virus replication

摘要 病毒感染操纵不同的翻译后修饰(PTM)，改变现有的蛋白质景观 以创造有利于复制的蜂窝环境。PTM允许主机环境的快速调制 通过改变蛋白质的丰度、定位和活性。由于它们的动态性质，PTM是理想的典型 免疫反应效应器或启动者。我们的数据确定了ADP-核糖化，蛋白质的修饰 腺苷二磷酸核糖(ADPr)，作为一种来自感染细胞的快速免疫样反应，构成了抗病毒反应 对抗流感病毒感染的药物。使用尖端的ADPr特定质谱学方法，我们 ADP-核糖体在流感病毒感染和感染过程中具有单一氨基酸拆分的特征 确定了对病毒和宿主蛋白质的数千种修饰。ADP-核糖化与 对多种病毒的抗病毒反应。然而，关于病毒感染是如何引发这种情况的知之甚少。 腺苷二磷酸核糖基化和多聚(ADPR)聚合酶的反应或激活机制(S) 催化ADPr加成到蛋白质上。在此，我们提出了流感病毒过程中ADP核糖化的研究。 感染。我们将确定特定ADPR修改的功能后果，并在我们独特的 ADP-核糖体数据集，研究ADP-核糖化如何改变特定病毒和宿主蛋白的功能。 我们将询问负责启动ADP-核糖化反应的细胞通路，定义 在流感病毒感染期间被激活的分子触发器和PAP。这些实验将 阐明ADPr介导的病毒抑制的机制，触发这种反应的因素，以及如何 病毒对其进行对抗，将ADP-核糖化确立为细胞抗病毒反应的一个关键方面，并可能作为 一个完全独立的抗病毒防御系统。

ABSTRACT Viral infections manipulate diverse post-translational modifications (PTMs), altering existing protein landscapes to create cellular environments favorable to replication. PTMs allow for rapid modulation of host environments by altering protein abundance, localization, and activity. Because of their dynamic nature, PTMs typify ideal immune response effectors or initiators. Our data identifies ADP-ribosylation, the modification of proteins with ADP-ribose (ADPr), as a rapid, immune-like response from infected cells that constitutes an antiviral response that counters influenza virus infection. Using cutting-edge ADPr-specific mass spectrometry approaches, we have characterized with single amino acid resolution the ADP-ribosylome during influenza virus infection and identified thousands of modifications on viral and host proteins. ADP-ribosylation has been associated with antiviral responses against multiple viruses. However, little is known about how viral infections trigger this response or the activiral mechanism(s) of ADP-ribosylation and poly(ADPr)-polymerases (PARPs), the enzymes that catalyze addition of ADPr to proteins. Here, we propose studies of ADP-ribosylation during influenza virus infection. We will identify the functional consequences of specific ADPr modifications, and building on our unique ADP-ribosylome dataset, investigate how ADP-ribosylation alters the function of specific viral and host proteins. We will interrogate the cellular pathways responsible for initiating ADP-ribosylation responses, defining the molecular triggers and PARPs that are activated during influenza virus infection. These experiments will elucidate the mechanisms of ADPr-mediated viral inhibition, the factors that trigger this response, and how viruses counter it, establishing ADP-ribosylation as a key aspect of cellular antiviral responses and perhaps as an entirely independent arm of antiviral defenses.