Non-coding RNA and ADP-ribosylation in Antiviral Defense

项目概要/摘要 了解宿主如何防御病毒的分子基础是未来研究的基本要求。 抗病毒治疗的发展。聚ADP核糖聚合酶（PARP）家族成员 抗病毒防御蛋白的典型特征包括干扰素诱导、加速进化和 抑制病毒感染的能力。PARP的定义特征是ADP-核糖基化的酶活性。 尽管迫切需要了解抗病毒反应的机制， 我们还不知道在抗病毒反应期间是什么激活了PARP酶活性，也不知道这种活性是如何激活的。 促进抗病毒状态。有一个关键的需要，以填补这一空白，因为了解PARP- 介导的抗病毒反应开辟了药物干预的新策略， 病毒性疾病我们的长期目标是确定病毒和宿主非蛋白编码RNA （ncRNA）调节病毒感染和疾病。根据这一目标，我们的总体目标是 确定RNA酶L，抗病毒反应的典型成分，如何执行新描述的 产生诱导PARP活性的ncRNA的活性以及这种增加的PARP活性如何抗病毒。我们 中心假设是RNase L活性产生的ncRNA增加PARP活性，并产生一个 通过转录后机制增强抗病毒状态。我们的假设是建立在我们的初步 数据表明RNase L活性是诱导可扩散PARP的必要和充分条件。 依赖性抗病毒因子这项研究的基本原理是，了解RNase L 产品激活抗病毒PARP可能会识别出关键的新靶点，用于治疗尼古丁驱动的疾病和病毒性疾病， 并提高我们对抗病毒反应的三个已建立/新兴领域的整体理解： RNase L、ncRNA生物学和PARP。我们计划测试我们的中心假设并完成目标 通过以下两个具体目标概述了本提案：1）确定RNase L如何诱导抗病毒 ADP-核糖基化，和2）确定PARP活性如何产生抗病毒状态。我们的贡献是 预期将详细了解ADP-核糖基化是如何由RNase L-产物ncRNA产生的， 这是如何影响抗病毒反应的。这一贡献将是巨大的，因为预计 在预防和治疗一系列由尼古丁引起的疾病中具有转化重要性。的 我们认为，本申请中提出的研究是创新的，因为它代表了一种新的实质性的 通过关注PARP活性的调节作为改变抗病毒的手段， 反应和细胞因子水平。

PROJECT SUMMARY/ABSTRACT Understanding the molecular basis for how hosts defend against viruses is an essential requirement for future development of antiviral therapeutics. Members of the poly ADP ribose polymerase (PARP) family display hallmarks typical of antiviral defense proteins including interferon-inducibility, accelerated evolution, and the ability to inhibit virus infection. A defining feature of PARPs is the enzymatic activity of ADP-ribosylation. Despite the imperative for understanding the mechanisms of the antiviral response, there is almost nothing known about what activates PARP enzymatic activity during the antiviral response, nor how this activity promotes an antiviral state. There is a critical need to fill in this gap because understanding the PARP- mediated antiviral response opens up new strategies of pharmacological intervention for cytokine-driven and viral diseases. Our long-term goal is to define the mechanisms by which viral and host non-protein-coding-RNA (ncRNA) regulates virus infection and disease. Consistent with this goal, our overall objective here is to determine how RNase L, a canonical component of the antiviral response, performs the newly described activity of generating ncRNA that induces PARP activity and how this increased PARP activity is antiviral. Our central hypothesis is that ncRNA generated by RNase L activity increases PARP activity and creates a heightened antiviral state by post-transcriptional mechanisms. Our hypothesis is formulated on our preliminary data demonstrating that RNase L activity is both necessary and sufficient to induce a diffusible PARP- dependent antiviral factor. The rationale for this proposed research is that understanding how RNase L products activate antiviral PARPs may identify critical new targets for cytokine-driven and viral diseases, as well as improve our overall understanding of three established/emerging areas of the antiviral response: RNase L, ncRNA biology and PARPs. We plan to test our central hypothesis and complete the objectives outlined in this proposal via the following two specific aims: 1) Determine how RNase L induces antiviral ADP-ribosylation, and 2) Determine how PARP activity creates an antiviral state. Our contribution here is expected to be a detailed understanding of how ADP-ribosylation is generated by RNase L-product ncRNA and how this contributes to the antiviral response. This contribution will be significant because it is expected to have translational importance in the prevention and treatment of a range of cytokine-driven diseases. The research proposed in this application is innovative, in our opinion, because it represents a new and substantive departure from the status quo by focusing on the modulation of PARP activity as a means to alter antiviral response and cytokine levels.