EAGER: Structural Basis for Assembly and Replication of Coronavirus

分子模型提供了基本的见解，为阻止病毒感染和繁殖的治疗策略提供了信息。全世界已经动员了一项努力，通过实验和计算技术来了解所有29种SARS-CoronaVirus-2蛋白质的性质。该项目中的核磁共振实验将为建立病毒组装和病毒基因组复制模型的工作提供信息，并为病毒生命周期的关键阶段提供重要的见解。这些研究的数据将通过公共储存库迅速传播，科学界可以在那里获取这些数据，以最大限度地利用这些数据。该项目将支持研究生和博士后在核磁共振和协作科学方面的培训，以满足紧迫的社会需求。该项目的方法和数据还将通过设在麦迪逊的国家核磁共振设施(NMRFAM)通过其研讨会、网站和其他外展努力传播，以告知更广泛的科学界和公众。这项建议利用NMRFAM现有的合作和仪器，启动对两种SARS冠状病毒2蛋白质：nsp8和膜(M)蛋白质的结构和动力学的核磁共振研究。NSP8是参与大冠状病毒基因组有效复制和翻译的几个复合体的一部分，在已知结构中以多种构象观察到。拟议的研究使用溶液核磁共振来研究在没有任何结合伙伴的情况下NSP8的构象集成，以了解NSP8如何在这些不同的复合体的组装中作为相互作用中心。M蛋白足以形成膜泡，并与其他结构和非结构SARS-CoV-2蛋白相互作用，组装病毒包膜，排除宿主蛋白，并将核衣壳整合到萌芽病毒中。这项拟议的研究使用固态核磁共振和体外分析来检测M的结构、动力学和膜相互作用，以了解它是如何诱导冠状病毒膜膜的萌发和组装的。这项急切的奖项是由分子和细胞生物科学部的分子生物物理计划颁发的，资金来自冠状病毒援助、救济和经济安全(CARE)法案。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Molecular models provide essential insights that inform therapeutic strategies to thwart viral infection and reproduction. A worldwide effort has been mobilized to understand the properties of all 29 SARS-Coronavirus-2 proteins with experimental and computational techniques. The nuclear magnetic resonance (NMR) experiments in this project will inform efforts to model viral assembly and genome replication of the virus and provide vital insights into critical stages of the viral life cycle. Data from these studies will be rapidly disseminated through public repositories where it can be accessed by the scientific community to leverage for maximal benefit. This project will support graduate student and post-doctoral training in NMR and collaborative science to address pressing societal needs. Methods and data resulting from the project will also be disseminated through the National Magnetic Resonance Facility at Madison (NMRFAM) via its workshops, web-site and other outreach efforts to inform the broader scientific community and public.This proposal leverages existing collaborations and instrumentation at NMRFAM to initiate NMR studies of the structure and dynamics of two SARS-coronavirus-2 proteins: nsp8 and membrane (M) protein. Nsp8 is part of several complexes involved in efficient replication and translation of the large coronavirus genome and is observed in multiple conformations in known structures. The proposed research uses solution NMR to examine the ensemble of nsp8 conformations in the absence of any binding partners to understand how nsp8 acts as interaction hub in assembly of these different complexes. The M protein is sufficient for membrane vesicle formation and interacts with other structural and non-structural SARS-CoV-2 proteins to assemble the viral envelope, exclude host proteins, and incorporate nucleocapsid into the budding virus. The proposed research uses solid-state NMR and in vitro assays to examine the structure, dynamics and membrane interaction of M, in true lipid bilayers with functional lipid composition, to provide insight into how it induces budding and assembly of the coronavirus membrane envelope.This EAGER award is made by the Molecular Biophysics Program in the Division of Molecular and Cellular Biosciences, using funds from the Coronavirus Aid, Relief, and Economic Security (CARES) ActThis 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.