SGER: Structure and Function of the SARS Virus RNA Polymerase

Texas A&M Research Foundation, College Station TX

Investigators

Abstract

The SARS virus, SCoV, is a newly emerged, highly virulent member of the coronavirus family of positive-strand RNA viruses. While coronaviruses have unique aspects of RNA replication and transcription, little biochemical and structural information exists for the key enzyme of these processes, the RNA-dependent RNA polymerase (RdRp). This SGER project will combine the biochemical and structural biology expertise of the two principal investigators to: 1) use oligonucletoides to construct a cDNA to express the RdRp of the Toronto isolate of the SCoV, purify the protein using conventional and affinity chromatography, establish a polymerase assay, identify the catalytic residues, and map the minimal region required for RNA synthesis in vitro; 2) characterize the initiation of RNA synthesis by the SCoV RdRp; and 3) grow high-resolution crystals of the SCoV RdRp and use X-ray crystallography to elucidate the structure of the protein alone and bound to substrate mimics. Results from these studies will establish biochemical assays and solve the structure for the SCoV polymerase. The results will be highly relevant to the mechanism of viral RNA replication and will allow the design of more effective antivirals against a significant class of human and animal pathogens.