EAGER: Structural Basis for Assembly and Replication of Coronavirus

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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.