Coronavirus Genome Replication

冠状病毒基因组复制

基本信息

批准号：
10372189
负责人：
Jamie Jon Arnold
金额：
$ 67.6万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-15 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10372189
关键词：
2019-nCoV 5&apos -exoribonuclease Address Bacteriophages Biochemical Cells Clinical Complex Core Assembly Coronavirus DNA Data Development Disease Disease Outbreaks Dissociation Drug resistance Enzymatic Biochemistry Enzymes Equilibrium Event Excision Exons Exonuclease Exoribonucleases Frequencies Genetic Genetic Transcription Genetic study Genome Germany Intervention Kinetics Knowledge Laboratories Lead Length Location Mass Spectrum Analysis Measures Middle East Respiratory Syndrome Coronavirus Morbidity - disease rate Multienzyme Complexes Mutation Nonstructural Protein Nucleotides Pathway interactions Population Prodrugs Prokaryotic Cells Proteins Proteome RNA RNA Helicase RNA Viruses RNA-Directed RNA Polymerase Research Resistance Ribonucleotides Role SARS coronavirus SARS-CoV-2 infection Scourge Site Specificity Structure Substrate Specificity System Therapeutic Universities Viral Genome Virus anti-viral efficacy antiviral drug development base crosslink drug action inhibitor insight mortality nucleotide analog pandemic disease prevent remdesivir replicase single molecule stoichiometry structural biology tripolyphosphate viral RNA

项目摘要

ABSTRACT The world is currently in the midst of a global pandemic caused by the second severe acute respiratory syndrome coronavirus (SARS2). In spite of the foreshadowing of such a pandemic by the emergence of SARS1 in 2002 and Middle East respiratory syndrome coronavirus (MERS) in 2012, we were ill equipped to address this scourge. Each of these early outbreaks yielded a substantial body of knowledge on the structures of coronavirus proteins. As with previous outbreaks, we are witnessing a redoubled coronavirus research effort. Structural biology continues to lead the way; however, our laboratory is now pledging a sustained commitment to elucidation of the fundamental enzymology and corresponding mechanisms of coronavirus genome replication. The SARS2 replisome has emerged as a clinically tractable target for development antiviral therapeutics. Remdesivir is a nucleotide analog prodrug, metabolized to the triphosphate in cells, and incorporated by the SARS2 replisome without excision by its proofreading exonuclease, ExoN. The mechanism of action of remdesivir is unclear, and the mechanism of escape from ExoN is unknown. This circumstance reflects the absence of a quantitative, mechanistic perspective of the SARS2 replisome. Such a perspective will be essential to elucidation of the mechanism of drug action and the mechanism of drug resistance. We have demonstrated the feasibility of elucidating the principles governing the dynamics and function of the SARS2 core replicase using state-of-the-art ensemble and single-molecule approaches. We will exploit these advances to pursue the following specific aims: study assembly and function of the SARS2 core replicase and its sub-assemblies (Aim 1); study utilization of incorrect nucleotides and nucleotide analogues by the SARS2 core replicase (Aim 2); and study the mechanism of error correction by the SARS2 exoribonuclease (Aim 3). Completion of these studies will represent the first, deep dive into the mechanistic enzymology of the coronavirus replisome.

抽象的目前，世界正处于第二个严重急性呼吸系统引起的全球大流行综合征冠状病毒（SARS2）。尽管出现了这种大流行的预示 SARS1在2002年和中东呼吸道综合症冠状病毒（MERS）2012年，我们的装备不足解决此祸害。这些早期爆发中的每一个都在结构上产生了大量知识冠状病毒蛋白。与以前的暴发一样，我们目睹了一项加倍的冠状病毒研究努力。结构生物学继续引导。但是，我们的实验室现在正在保证致力于阐明基本酶学和冠状病毒的相应机制基因组复制。 SARS2重建体已成为临床上可用于开发的目标抗病毒疗法。 remdesivir是一种核苷酸类似物前药，代谢为细胞中的三磷酸，并由SARS2重新组合并通过校对外核酸外显酶外显子的切除而掺入。这 Remdesivir的作用机理尚不清楚，从外显子逃脱的机理尚不清楚。这情况反映了SARS2重新构体的定量，机械透视。这样的透视图对于阐明药物作用机理和药物机制至关重要反抗。我们已经证明了阐明控制动态的原则的可行性 SARS2核心复制酶的功能使用最新的集合和单分子方法。我们将利用这些进步来追求以下特定目标：SARS2核心的研究组装和功能复制酶及其子组件（AIM 1）；研究利用不正确的核苷酸和核苷酸类似物 SARS2核心复制酶（AIM 2）；并研究SARS2的误差校正机理 Exoriboneclease（AIM 3）。这些研究的完成将代表第一次深入研究机械冠状病毒的酶学。