Structural Characterization of SARS-CoV-2 Genome Organization by NMR

通过 NMR 表征 SARS-CoV-2 基因组组织的结构

• 批准号: 2231099
• 负责人: Tatyana Polenova
• 金额: $ 120万
• 依托单位: University of Delaware
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2231099&HistoricalAwards=false
• 关键词: Structural; Characterization; SARS; CoV; Genome

The goal of this project is to structurally characterize the nucleocapsid (N)-mediated packaging of the genome of the SARS-CoV-2 virus, the causative agent of the deadly COVID-19 pandemic. The proposed investigations and the general methodological framework to be developed will have broad applications since they will provide 1) biological insights that open new strategies for interfering with viral genome packaging, and 2) integrative magnetic resonance-based protocols that can be used by other practitioners (biochemists, biophysicists, molecular biologists). As such, the proposed research will impact several areas of science, including structural biology, biochemistry, physical chemistry, magnetic resonance spectroscopy, biotechnology and pharmacology. The proposed innovative research program will provide unique training for students at all levels in state-of-the-art experimental methods. Importantly, an educational program within this project will include embedding opportunities for members of the laboratories of the PI, Co-PI and the industrial collaborator that will greatly enhance the training of the STEM workforce. The goal of this project is to structurally characterize how the nucleocapsid (N) packages the genome in the SARS-CoV-2 virus, by integrating experimental magic angle spinning (MAS) and dynamic nuclear polarization (DNP) nuclear magnetic resonance (NMR) methods. This new methodological framework, established through the proposed research, will overcome the current challenges in studying complex heterogeneous viral genome ribonuclear particles that exhibit different degrees of disorder and dynamics. The proposed research will provide atomic-level knowledge on the structure and dynamics of the SARS-CoV-2 N protein as well as the structural organization of the N protein in assemblies with genomic RNA. Employing high magnetic fields together with fast and ultrafast 1H- and 19F- detected MAS and DNP-enhanced MAS NMR will enable studies into very large virus systems, requiring only a fraction of the material used in conventional experiments. The general methodological framework that will be developed will be broadly applicable to investigate packaged genomes of other viruses and can be used by a wide range of practitioners of these methodologies (biochemists, biophysicists, molecular biologists). As such, the planned research will impact several areas of science, including structural biology, biochemistry, physical chemistry, magnetic resonance spectroscopy, biotechnology and pharmacology.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.

该项目的目标是从结构上表征 SARS-CoV-2 病毒基因组的核衣壳 (N) 介导的包装，该病毒是致命的 COVID-19 大流行的病原体。拟议的研究和待开发的通用方法框架将具有广泛的应用，因为它们将提供 1) 生物学见解，开辟干扰病毒基因组包装的新策略，以及 2) 可供其他从业者使用的基于综合磁共振的方案（生物化学家、生物物理学家、分子生物学家）。因此，拟议的研究将影响多个科学领域，包括结构生物学、生物化学、物理化学、磁共振波谱、生物技术和药理学。拟议的创新研究计划将为各级学生提供最先进实验方法的独特培训。重要的是，该项目中的教育计划将包括为 PI、Co-PI 和工业合作者的实验室成员提供机会，这将大大加强 STEM 劳动力的培训。该项目的目标是通过整合实验魔角旋转 (MAS) 和动态核极化 (DNP) 核磁共振 (NMR) 方法，从结构上表征 SARS-CoV-2 病毒中核衣壳 (N) 如何包装基因组。通过拟议的研究建立的这种新的方法框架将克服当前研究表现出不同程度的无序和动态的复杂异质病毒基因组核糖核颗粒的挑战。拟议的研究将提供有关 SARS-CoV-2 N 蛋白的结构和动力学以及 N 蛋白与基因组 RNA 组装的结构组织的原子级知识。采用高磁场以及快速和超快的 1H 和 19F 检测 MAS 以及 DNP 增强的 MAS NMR 将使研究非常大的病毒系统成为可能，而只需要传统实验中使用的材料的一小部分。将开发的通用方法框架将广泛适用于研究其他病毒的包装基因组，并且可供这些方法的广泛实践者（生物化学家、生物物理学家、分子生物学家）使用。因此，计划中的研究将影响多个科学领域，包括结构生物学、生物化学、物理化学、磁共振波谱、生物技术和药理学。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值进行评估，被认为值得支持以及更广泛的影响审查标准。