Analysis of Coronavirus-Host Cell Interactions

冠状病毒-宿主细胞相互作用分析

• 批准号: 8442842
• 负责人: Shinji Makino
• 金额: $ 35.96万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-15 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8442842
• 关键词: 5' Untranslated Regions; Address; Antiviral Agents; Binding; Binding Sites; Biological Models; Biological Process; Cell Communication; Cell Extracts; Cells; China; Chiroptera; Complex; Coronavirus; Coronavirus Infections; Country; Disease Outbreaks; Family Picornaviridae; Foundations; Future; Gene Expression; Genes; Genetic Translation; Genome; Hela Cells; Hepatitis C virus; Human Coronavirus 229E; Integration Host Factors; Interferon Type I; Interferons; Internal Ribosome Entry Site; Knowledge; Lung diseases; Maps; Messenger RNA; Molecular; Murine hepatitis virus; Nonstructural Protein; Oryctolagus cuniculus; Pathogenesis; Pathogenicity; Peptide Initiation Factors; Play; Polyproteins; Process; Production; Protein Biosynthesis; Proteins; RNA; RNA chemical synthesis; Regulation; Resistance; Resources; Reticulocytes; Ribonucleases; Ribosomes; Role; SARS coronavirus; Severe Acute Respiratory Syndrome; System; Testing; Translation Initiation; Translations; Transmissible gastroenteritis virus; Viral; Viral Genes; Viral Genome; Viral Proteins; Virulence Factors; Virus; cricket paralysis virus; eIF-4B; inhibitor/antagonist; mRNA Transcript Degradation; mortality; novel; viral RNA

DESCRIPTION (provided by applicant): The translation of the viral genome begins immediately after coronavirus (CoV) infection to produce two large polyproteins that are processed into 15 or 16 mature, nonstructural proteins. Most of these proteins are involved in viral RNA synthesis, and some have other biological functions. The nsp1 protein of betaCoVs, which includes SARS-CoV (SCoV), bat CoVs, and mouse hepatitis virus (MHV), and of alphaCoVs, which includes transmissible gastroenteritis virus (TGEV) and human CoV 229E, inhibits host gene expression. As past studies have shown that viral proteins that inhibit host gene expression are major virulence factors, nsp1 proteins of CoVs most probably play a critical role in CoV pathogenesis. Consistent with this notion, MHV nsp1 is a major virulence factor, and SCoV nsp1 inhibits the production of type I interferon and interferon-stimulated genes in infected cells. Nsp1 proteins of different CoVs share a common biological function to inhibit host gene expression but use different strategies to exert this function. SCoV nsp1 uses a novel, two-pronged strategy to inhibit host protein synthesis/gene expression. SCoV nsp1 binds to the 40S ribosomal subunit to inhibit mRNA translation and also induces a template-dependent endonucleolytic mRNA cleavage. In contrast to SCoV nsp1, TGEV nsp1 employs a different strategy to inhibit mRNA translation, as it is unable to bind the 40S ribosomal subunit or promote host mRNA degradation. This application aims to delineate the novel mechanisms of CoV nsp1-induced inhibition of gene expression by using SCoV and TGEV nsp1 proteins as model systems. We will uncover the mechanism of SCoV nsp1-induced template-dependent mRNA cleavage. We will also clarify the different mechanisms of SCoV and TGEV nsp1-induced translation inhibition and reveal the strategy that allows robust viral gene expression in SCoV-infected cells under conditions of nsp1-induced translation inhibition. The proposed studies will provide a foundation for understanding the modulation of host gene expression by CoV, expand our knowledge of CoV pathogenicity at the molecular level, and potentially identify a novel mechanism for the regulation of eukaryotic gene expression.

描述（由申请人提供）：病毒基因组的翻译在冠状病毒 (CoV) 感染后立即开始，产生两种大的多蛋白，这些多蛋白被加工成 15 或 16 个成熟的非结构蛋白。这些蛋白质大部分参与病毒RNA合成，有些还具有其他生物学功能。 betaCoV（包括 SARS-CoV (SCoV)、蝙蝠 CoV 和小鼠肝炎病毒 (MHV)）和 alphaCoV（包括传染性胃肠炎病毒 (TGEV) 和人类 CoV 229E）的 nsp1 蛋白抑制宿主基因表达。过去的研究表明，抑制宿主基因表达的病毒蛋白是主要毒力因子，冠状病毒的 nsp1 蛋白很可能在冠状病毒的发病机制中发挥着关键作用。与这一概念一致，MHV nsp1 是主要毒力因子，而 SCoV nsp1 抑制受感染细胞中 I 型干扰素和干扰素刺激基因的产生。不同冠状病毒的Nsp1蛋白具有共同的抑制宿主基因表达的生物学功能，但使用不同的策略来发挥该功能。 SCoV nsp1 使用一种新颖的双管齐下的策略来抑制宿主蛋白质合成/基因表达。 SCoV nsp1 与 40S 核糖体亚基结合，抑制 mRNA 翻译，并诱导模板依赖性核酸内切 mRNA 裂解。与 SCoV nsp1 相比，TGEV nsp1 采用不同的策略来抑制 mRNA 翻译，因为它无法结合 40S 核糖体亚基或促进宿主 mRNA 降解。本申请旨在通过使用 SCoV 和 TGEV nsp1 蛋白作为模型系统来描述 CoV nsp1 诱导的基因表达抑制的新机制。我们将揭示 SCoV nsp1 诱导的模板依赖性 mRNA 裂解的机制。我们还将阐明 SCoV 和 TGEV nsp1 诱导的翻译抑制的不同机制，并揭示在 nsp1 诱导的翻译抑制条件下允许 SCoV 感染的细胞中病毒基因稳健表达的策略。拟议的研究将为了解冠状病毒对宿主基因表达的调节奠定基础，扩大我们对分子水平上冠状病毒致病性的了解，并有可能确定真核基因表达调节的新机制。