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。研究得最好的之一 RNA 质量控制途径是无义介导的 mRNA 衰减 (NMD)，一种核糖体相关的监视 使用提前终止密码子 (PTC) 识别和降解细胞转录本的机制。最初 被确定为一种清除细胞中有缺陷 mRNA 的机制，现在已知 NMD 还调节表达 5-10% 的细胞转录物在细胞分化、体内平衡、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通路的许多蛋白都具有功能 细胞自主防御 RNA 病毒，包括人类致病性甲病毒、冠状病毒 和黄病毒。作为专性细胞内寄生虫，这些病毒与细胞 mRNA 加工密切相关 通过了解该机器如何重新调整用途，我们可以对 NMD 功能获得新的见解 对抗病毒感染。目前我们对病毒感染过程中 NMD 功能的了解还非常不完整。许多 病毒缺乏 PTC 或其他 NMD 诱导特征，目前尚不清楚 NMD 蛋白是否识别病毒 RNA 以类似于典型 NMD 的方式，或通过新的和不寻常的相互作用或机制。而且， NMD 在病毒感染过程中通常会在全球范围内下调，这表明病毒具有重新连接的机制 抗病毒 NMD 网络。使用黄病毒寨卡病毒（ZIKV）作为模型 RNA 病毒，我们的目标是了解 NMD 机制与病毒感染之间的分子相互作用。我们将剖析 NMD-ZIKV 蛋白质-蛋白质和蛋白质-RNA相互作用网络，采用结合病毒学的多学科方法 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 本研究解决的问题是：1) NMD 蛋白如何相互作用来对抗病毒感染？ 2）有吗 病毒 RNA 中触发 NMD 的特定特征，如果是的话，它们是如何被 NMD 机制识别的？ 3）病毒蛋白如何抑制NMD？ 4) 重要蚊子的抗病毒 NMD 功能是否保守 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药物提供新的见解 防御网。此外，通过研究 NMD 在病毒感染过程中受到调节的不寻常方式，我们 希望发现 NMD 机制本身的新细胞功能。