Interplay between coronaviruses and nonsense-mediated mRNA decay pathway

冠状病毒与无义介导的 mRNA 衰减途径之间的相互作用

• 批准号: 10358595
• 负责人: Shinji Makino
• 金额: $ 30.92万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-11 至 2024-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10358595
• 关键词: 3' Untranslated Regions; Address; Area; Attenuated; Binding; Binding Proteins; Biological; Biological Models; Biology; Cells; Clinical; Common Cold; Coronavirus; Coronavirus nucleocapsid protein; Country; Cytoplasm; Data; Development; Disease; Domestic Animals; Elements; Ensure; Epidemic; Europe; Exhibits; Far East; Gene Expression; Generations; Genetic Diseases; Genome; Host Defense Mechanism; Human; Introns; Knowledge; Lead; Measures; Mediating; Messenger RNA; Middle East; Middle East Respiratory Syndrome; Middle East Respiratory Syndrome Coronavirus; Molecular; Murine hepatitis virus; Nonsense-Mediated Decay; Northern Africa; Open Reading Frames; Pathogenicity; Pathway interactions; Phenotype; Phosphorylation; Physiological; Play; Post-Transcriptional Regulation; Predisposition; Proteins; Public Health; Quality Control; RNA; RNA Viruses; RNA-Binding Proteins; Research; Respiratory System; Role; SARS coronavirus; Saudi Arabia; Severe Acute Respiratory Syndrome; Structure; Symptoms; Terminator Codon; Testing; Therapeutic Agents; Time; Transcript; Vaccines; Viral; Viral Gene Expression Regulation; Viral Genes; Viral N Protein; Viral Proteins; Virus; Virus Assembly; Virus Replication; coronavirus vaccine; economic impact; endonuclease; gene product; human coronavirus; inhibitor; insight; mRNA Decay; mRNA Stability; mRNA Surveillance; mortality; mutant; novel; novel strategies; premature; prevent; recruit; respiratory; vaccine candidate; viral RNA; virology

Coronaviruses (CoVs), which carry a large, single-stranded, positive-sense RNA genome, cause a variety of diseases in humans and domestic animals. Human CoVs (HCoVs) usually infect the respiratory tract and cause a range of symptoms varying from mild, such as the common cold, to more serious respiratory illnesses like severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS), caused by two highly pathogenic HCoVs, SARS-CoV and MERS-CoV. SARS-CoV caused a worldwide epidemic in 2002- 2003, resulting in more than 8,000 cases with an approximate mortality of 10%, while MERS-CoV emerged in Saudi Arabia in 2012 and has been disseminated into other countries in the Middle East, North Africa, Europe, and East Asia. HCoVs represent a major threat to public health and have the potential to cause a significant negative economic impact. Currently, there are no approved vaccines and therapeutic agents against HCoVs. The development of effective control measures against CoVs requires a comprehensive understanding of viral gene expression strategies and host-CoV interactions. A plethora of studies have focused on investigating CoV biology and have significantly contributed to our understanding of CoV replication mechanisms, including the structure-function analyses of viral RNA elements as well as the viral proteins that are involved in viral replication and assembly. However, there are still gaps in our knowledge of the post-transcriptional regulation of viral gene expression, as only a limited number of studies have addressed this area of CoV research. Particularly, very little is known about the cis-acting viral RNA elements and trans-acting host and viral factors that regulate CoV mRNA transcript stability. One newly emerging research area in virology is understanding interactions between viruses and host mRNA surveillance pathways that prevent generation/accumulation of unwanted gene products. We have demonstrated that CoV mRNAs are the targets of the nonsense-mediated mRNA decay (NMD) pathway, one of the host mRNA surveillance pathways, and that viral N protein protects CoV mRNAs from NMD. Our data suggest the importance of N-mediated NMD suppression for efficient virus replication. The present application will study the interplay between the NMD pathway and CoVs by testing the following hypotheses: UPF1, the principal orchestrator of NMD, binds to the 3’ UTR of CoV mRNAs having specific motifs, undergoes phosphorylation, and recruits SMG6, an endonuclease, leading to endonucleolytic RNA cleavage; N binds to the 3’ UTRs of NMD targets and prevents an NMD factor(s) from accessing these targets and/or N interacts with an NMD factor(s) and sequesters it away from the NMD pathway; and CoV mutants having an increased susceptibility to NMD, cannot replicate as efficiently as the parental viruses. The data obtained from these studies will provide mechanistic insights into NMD of CoV mRNAs and N-mediated NMD pathway suppression, and will reveal the feasibility of novel strategies for attenuating CoVs through increased susceptibility to the NMD pathway.

冠状病毒（COVS）携带大型，单链，阳性RNA基因组，引起多种 人类和家畜的疾病。人类COV（HCOV）通常会感染呼吸道和 导致一系列症状从轻度感冒到更严重的呼吸系统疾病不等 像严重的急性呼吸综合征（SARS）和中东呼吸道综合征（MERS）一样，由两个 高致病性HCOV，SARS-COV和MERS-COV。 SARS-COV在2002年引起了全球流行病 - 2003年，导致8,000多例死亡率约为10％，而MERS-COV出现了 沙特阿拉伯在2012年，已被传播到中东，北非，欧洲的其他国家， 和东亚。 HCOVS代表了对公共卫生的主要威胁，并有可能引起重大的 负面的经济影响。目前，尚无针对HCOV的批准疫苗和治疗剂。 针对COV的有效控制措施的制定需要对病毒的全面了解 基因表达策略和宿主-COV相互作用。大量的研究重点是研究COV 生物学，并为我们对COV复制机制的理解做出了重大贡献，包括 病毒RNA元素以及参与病毒的病毒蛋白的结构功能分析 复制和组装。但是，我们对转录后法规的了解仍然存在差距 病毒基因的表达，因为只有有限的研究已经解决了COV研究的这一领域。 特别是，关于顺式作用的病毒RNA元素以及跨作用宿主和病毒因子知之甚少 调节COV mRNA转录本稳定性。病毒学的一个新兴研究领域是理解 病毒与宿主mRNA监视途径之间的相互作用，以防止 不需要的基因产品。我们已经证明COV mRNA是废话介导的目标 mRNA衰变（NMD）途径，宿主mRNA监测途径之一，该病毒N蛋白保护 NMD的COV mRNA。我们的数据表明N介导的NMD抑制对有效病毒的重要性 复制。本应用将通过测试NMD途径与COV之间的相互作用 以下假设：NMD的主要编排UPF1与具有COV mRNA的3’UTR结合具有 特定的基序，经历磷酸化，并募集SMG6，一种内核解释性分析 RNA裂解； n与NMD目标的3’UTR结合，并防止NMD因子访问这些因子 靶标和/或n与NMD因子相互作用，并将其隔离为NMD途径；和Cov 对NMD敏感的突变体不能像父母病毒那样有效地复制。 从这些研究中获得的数据将提供对COV mRNA和N介导的NMD的机械见解 NMD途径抑制，并将揭示通过通过 增加对NMD途径的敏感性。