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的致病药物），预计将涉及感染宿主细胞的蛋白质因子，这些因子提供了未编码病毒基因组中的活性。该项目着重于病毒选择3'至5'RNA解旋酶以合成负链RNA的宿主RNA的需求，该RNA用作复制阳性基因组的模板。最近的蛋白质组学筛选确定了与SARS-COV-2蛋白相互作用的候选宿主解旋酶DDX10。该研究将采用一套实验方法来表征该酶，而SARS-COV-2基因组复制也实现了另外两个宿主RNA解旋酶。关于宿主蛋白如何有助于病毒复制的详细知识将为治疗干预提供新的靶标，重要的是，以帮助通过病毒突变来规避耐药性的方式。该项目还将支持培训以冠状病毒相关的主题为中心的新本科生物物理学课程的培训。Biochemical和Biophysical技术（X射线晶体学，冷冻em和单分子光谱）将适用于与触发性的速度相关性，包括X射线晶体学，冷冻晶体和单分子光谱的速度，以及如何处理速度，并将其相关性。病毒NSP7/8起启动酶会影响基因组复制。 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值得通过基金会的智力优点和更广泛的影响审查标准来通过评估来支持。