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.

摘要 真核细胞使用多层策略来确保基因表达的保真度。研究最多的 RNA质量控制途径是无义介导的mRNA衰变（NMD），一种核糖体相关的监视 识别和降解具有提前终止密码子（PTC）的细胞转录物的机制。最初 NMD被认为是清除细胞中错误mRNA的一种机制，现在已知NMD也调节细胞中 5-10%的细胞转录物在细胞分化、稳态、细胞应激中具有重要功能 答案，还有更多。此外，最近的研究表明，NMD途径的许多蛋白质功能 在细胞自主防御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物，这些病毒与细胞mRNA加工密切相关 途径，我们可以通过了解NMD机制如何被重新利用， 对抗病毒感染。我们目前对NMD在病毒感染期间的功能有一个非常不完整的了解。许多 病毒缺乏PTC或其他NMD诱导特征，并且不知道NMD蛋白是否识别病毒 RNA的方式类似于典型的NMD，或者通过新的和不寻常的相互作用或机制。此外，委员会认为， NMD在病毒感染过程中通常是全局下调的，这表明病毒具有重新连接的机制。 抗病毒国家导弹防御系统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机制和病毒感染之间的分子相互作用。我们将解剖NMD-ZIKV 蛋白质-蛋白质和蛋白质-RNA相互作用网络，结合病毒学的多学科方法 与RNA生物化学和高通量测定，以连接分子相互作用的细胞表型。具体 本研究所要解决的问题是：1）NMD蛋白如何相互作用以对抗病毒感染？2)有 病毒RNA中触发NMD的特定特征，如果是这样，它们是如何被NMD机制识别的？ 3)病毒蛋白如何抑制NMD？抗病毒NMD功能在重要的蚊子中是否保守 黄病毒宿主总的来说，我们的研究将为一种重要的细胞自主抗病毒药物提供新的见解。 防御网络。此外，通过研究NMD在病毒感染过程中的异常调节方式， 希望发现NMD机制本身的新的细胞功能。