RAPID: Host-pathogen interactions during genome replication of SARS-CoV2

RAPID：SARS-CoV2 基因组复制过程中宿主与病原体的相互作用

• 批准号: 2031094
• 负责人: Taekjip Ha
• 金额: $ 16.23万
• 依托单位: Johns Hopkins University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2031094&HistoricalAwards=false
• 关键词: RAPID; Host; pathogen; interactions; during

Replication of coronaviruses, including SARS-CoV-2 – the causative agent of COVID-19, is expected to involve protein factors from the infected host cell that provide activities not encoded in the viral genome. This project focuses on the need for the virus to co-opt a host 3’ to 5’ RNA helicase to synthesize the negative-strand RNA that serves as template for copying the positive-strand genome. A recent proteomic screen identified a candidate host helicase, DDX10, that interacts with SARS-CoV-2 proteins. The research will employ a suite of experimental approaches to characterize this enzyme and two other host RNA helicases also implicated in SARS-CoV-2 genome replication. Detailed knowledge of how host proteins contribute to viral replication will provide new targets for therapeutic intervention, importantly, in ways that help circumvent drug resistance through viral mutation. The project will also support training of a postdoctoral scholar and development of a new undergraduate biophysics course centered on coronavirus-related topics.Biochemical and biophysical techniques, such as X-ray crystallography, cryo-EM and single molecule spectroscopy, will be applied to determine structure-function properties that govern the helicase mechanism, including polarity, speed, processivity and associated energetics, and how its interactions with the viral NSP7/8 primase influence genome replication. The outcomes are expected to reveal the workings of host proteins during SARS-CoV-2 replication, and thereby open new avenues and novel drug targets to block its proliferation.This RAPID award is made by the Genetic Mechanisms Program in the Division of Molecular and Cellular Biosciences, using funds from the Coronavirus Aid, Relief, and Economic Security (CARES) Act.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

冠状病毒的复制，包括SARS-CoV-2 -COVID-19的病原体，预计涉及来自受感染宿主细胞的蛋白因子，这些蛋白因子提供病毒基因组中未编码的活性。该项目的重点是病毒需要选择宿主3'至5' RNA解旋酶来合成负链RNA，作为复制正链基因组的模板。最近的蛋白质组学筛选确定了一个候选宿主解旋酶DDX 10，它与SARS-CoV-2蛋白相互作用。这项研究将采用一套实验方法来表征这种酶和其他两种与SARS-CoV-2基因组复制有关的宿主RNA解旋酶。详细了解宿主蛋白质如何促进病毒复制将为治疗干预提供新的靶点，重要的是，通过病毒突变帮助规避耐药性。该项目还将支持培养一名博士后学者，并开发一门以冠状病毒相关主题为中心的新的本科生生物物理课程。生物化学和生物物理技术，如X射线晶体学、冷冻电镜和单分子光谱学，将用于确定决定解旋酶机制的结构-功能特性，包括极性、速度、持续合成能力和相关能量学，以及它与病毒NSP 7/8引发酶的相互作用如何影响基因组复制。这些成果有望揭示SARS-CoV-2复制过程中宿主蛋白的工作机制，从而开辟新的途径和新的药物靶点，以阻止其增殖。该RAPID奖由分子和细胞生物科学部的遗传机制计划颁发，使用冠状病毒援助，救济，该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。