Structural Studies of the Corona Virus Life Cycle

Coronaviruses are a diverse family of viruses infecting many animals including humans. The 21stcentury is now experiencing its third outbreak of a novel pathogenic coronavirus that has crossed from ananimal host into humans for the first time. The outbreak of SARS-CoV-2 is unprecedented among humancoronaviruses in its size and the speed of its spread. Countering this viral outbreak will require a detailedmechanistic understanding of virus protein function. The goals of this project are to gain highly detailedinformation about the SARS-CoV-2 replication complex using single-particle cryo-electron microscopy. We willuse this high-resolution imaging technique to determine structures of the SARS-CoV-2 polymerase complexbound to substrates and small molecule antiviral drugs. We will complement these structural analyses with adetailed biochemical study of protein-protein and protein-RNA interactions and the influence of theseinteractions on SARS-CoV-2 polymerase activity. These studies include mutagenesis of key interfaces in theprotein complex and testing of the recombinant proteins in polymerase activity assays. These studies have thepotential to illuminate the mechanisms used by the SARS-CoV-2 polymerase complex to replicate its viralgenome and provide a mechanistic understanding of antiviral therapeutic action key to the development ofnovel antiviral therapeutics to treat COVID-19.