Understanding the antiviral roles of acellular RNA quality control pathway

了解非细胞 RNA 质量控制途径的抗病毒作用

• 批准号: 10714668
• 负责人: Anna-Lena Steckelberg
• 金额: $ 41.13万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-15 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10714668
• 关键词: Address; Alphavirus; Binding; Cell Differentiation process; Cell physiology; Cells; Cellular Stress; Coronavirus; Culicidae; Ensure; Eukaryotic Cell; Flavivirus; Gene Expression; Gene Expression Regulation; Goals; Homeostasis; Human; Infection; Link; Mediating; Messenger RNA; Modeling; Pathogenicity; Pathway interactions; Phenotype; Proteins; Quality Control; RNA; RNA Biochemistry; RNA Viruses; RNA-Protein Interaction; Ribosomes; Role; Terminator Codon; Transcript; Viral; Viral Proteins; Virus; Virus Diseases; Zika Virus; fighting; high throughput screening; insight; interdisciplinary approach; mRNA Decay; obligate intracellular parasite; premature; viral RNA; virology

Abstract Eukaryotic cells use multi-layered strategies to ensure the fidelity of gene expression. One of the best-studied RNA quality control pathways is Nonsense-mediated mRNA decay (NMD), a ribosome-associated surveillance machinery that recognizes and degrades cellular transcripts with premature termination codons (PTC). Initially identified as a mechanism to rid the cell of faulty mRNAs, it is now known that NMD also regulates the expression of 5-10% of cellular transcripts with important functions in cell differentiation, homeostasis, cellular stress responses, and more. In addition, recent studies have shown that many proteins of the NMD pathway function in the cell-autonomous defense against RNA viruses, including human-pathogenic alphaviruses, coronaviruses and flaviviruses. As obligate intracellular parasites, these viruses interface closely with cellular mRNA processing pathways, and we can gain new insight to NMD functions by understanding how the machinery is repurposed to fight viral infection. We currently have a very incomplete picture of NMD functions during viral infection. Many viruses lack PTCs or other NMD-inducing features, and it is not known whether NMD proteins recognize viral RNA in manners similar to canonical NMD, or through new and unusual interactions or mechanisms. Moreover, NMD is often globally downregulated during virus infection, suggesting that viruses have mechanisms to rewire the antiviral NMD network. Using the flavivirus Zika virus (ZIKV) as a model RNA virus, we aim to understand the molecular interactions between the NMD machinery and viral infection. We will dissect the NMD-ZIKV protein-protein and protein-RNA interaction network with a multi-disciplinary approach that combines virology with RNA biochemistry and high-throughput assays to link molecular interactions to cellular phenotypes. Specific questions addressed within this study are: 1) How do NMD proteins interact to fight viral infection? 2) Are there specific features in the viral RNA that trigger NMD, and if so, how are they recognized by the NMD machinery? 3) How do viral proteins inhibit NMD? And 4) Are antiviral NMD functions conserved in the important mosquito host of flaviviruses? Collectively, our study will provide new insight into an important cell-autonomous antiviral defense network. In addition, by studying the unusual ways in which NMD is regulated during virus infection, we hope to discover new cellular functions of the NMD machinery itself.

抽象的 真核细胞使用多层策略来确保基因表达的保真度。研究最佳的人之一 RNA质量控制途径是胡说八道介导的mRNA衰减（NMD），这是核糖体相关的监视 识别并降低用过早终止密码子（PTC）的细胞转录本的机械。最初 被确定为消除故障mRNA细胞的机制，现在知道NMD还调节了表达 5-10％的细胞转录本在细胞分化，稳态，细胞应激中具有重要功能 回应等等。此外，最近的研究表明，NMD途径功能的许多蛋白质 在针对RNA病毒的细胞自主防御中，包括人类病毒α病毒，冠状病毒 和黄病毒。作为强制性细胞内寄生虫，这些病毒与细胞mRNA处理紧密接触 途径，我们可以通过了解如何将机械重新使用到NMD功能来获得新的洞察力 对抗病毒感染。目前，我们对病毒感染期间的NMD功能的情况非常不完整。许多 病毒缺乏PTC或其他NMD诱导特征，尚不清楚NMD蛋白是否识别病毒 与规范NMD相似的举止，或通过新的和不寻常的相互作用或机制的RNA。而且， NMD通常在病毒感染期间全球下调，这表明病毒具有重新布线的机制 抗病毒NMD网络。使用黄病毒寨卡病毒（ZIKV）作为模型RNA病毒，我们旨在理解 NMD机械与病毒感染之间的分子相互作用。我们将剖析NMD-ZIKV 蛋白质 - 蛋白质和蛋白-RNA相互作用网络具有多学科方法，该方法结合了病毒学 通过RNA生物化学和高通量测定，将分子相互作用与细胞表型联系起来。具体的 这项研究中解决的问题是：1）NMD蛋白如何相互作用以抵抗病毒感染？ 2）在那里 触发NMD的病毒RNA中的特定特征，如果是，NMD机械如何识别它们？ 3）病毒蛋白如何抑制NMD？和4）是在重要蚊子中保守的抗病毒NMD功能 黄病毒的宿主？总的来说，我们的研究将为重要的细胞自主抗病毒提供新的见解 国防网络。此外，通过研究病毒感染期间NMD的异常方式，我们 希望发现NMD机械本身的新细胞功能。