Coronavirus Genome Replication

冠状病毒基因组复制

基本信息

批准号：
10585913
负责人：
Jamie Jon Arnold
金额：
$ 67.58万
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 2027-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10585913
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 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 Severe Acute Respiratory Syndrome Site Specificity Structure Substrate Specificity Syndrome System Therapeutic Universities Viral Viral Genome Virus anti-viral efficacy antiviral drug development crosslink drug action inhibitor insight mortality new pandemic nucleotide analog pandemic disease prevent previous outbreak 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，世界正处于由第二次严重急性呼吸道疾病引起的全球大流行之中冠状病毒综合症（SARS2）。尽管出现了这种大流行的预兆 2002 年的 SARS1 和 2012 年的中东呼吸综合征冠状病毒 (MERS)，我们没有能力应对解决这一祸害。每一次早期爆发都产生了大量有关结构的知识冠状病毒蛋白。与之前的疫情爆发一样，我们正在见证对冠状病毒的研究加倍努力。结构生物学继续引领潮流；然而，我们的实验室现在承诺持续致力于阐明冠状病毒的基本酶学和相应机制基因组复制。 SARS2复制体已成为临床上易于开发的靶标抗病毒治疗。瑞德西韦是一种核苷酸类似物前药，在细胞中代谢为三磷酸盐，并且由 SARS2 复制体整合，而不被其校对核酸外切酶 ExoN 切除。这瑞德西韦的作用机制尚不清楚，ExoN 的逃逸机制尚不清楚。这这种情况反映了 SARS2 复制体缺乏定量、机制的观点。这样一个观点对于阐明药物作用机制和药物机制至关重要反抗。我们已经证明了阐明控制动力学和使用最先进的整体和单分子方法研究 SARS2 核心复制酶的功能。我们将利用这些进步来实现以下具体目标：研究 SARS2 核心的组装和功能复制酶及其子组件（目标 1）；研究错误核苷酸和核苷酸类似物的利用 SARS2 核心复制酶（目标 2）；研究SARS2的纠错机制核糖核酸外切酶（目标 3）。这些研究的完成将代表着首次深入研究其机制冠状病毒复制体的酶学。