Viral RNA replication is a fundamental process in viral pathogenesis. This laboratory studies brome mosaic virus (BMV), a model (+)-strand RNA virus, in order to characterize the interactions between RNA-dependent RNA polymerase (RdRp) subunits and the mechanism of RNA synthesis. Three projects will be carried out simultaneously in this research. The first is designed to dissect the requirements for the interaction of the proteins found in the BMV replicase through chemical mutagenesis, selection for stable mutants that have lost protein-protein interaction in the two-hybrid assay, and sequencing the mutations. The second project is designed to elucidate the sequences that contribute to the initiation of (-)-strand viral RNA synthesis and to examine how these sequences affect steps in (-)-strand RNA synthesis. The third project examines genomic (+)-strand RNA synthesis by identifying the minimal functional promoter and constructing promoters with nucleotide substitutions, to identify RNA sequences that affect the various steps in viral RNA synthesis.
Knowledge gained from the execution of this project will contribute to the basic understanding of how viruses replicate. The use of a plant virus, brome mosaic virus, as an experimental model, will directly increase the understanding of less well-studied viruses, some of which cause disease. The detailed understanding of the different steps in viral RNA replication could ultimately uncover targets for the design of highly specific antiviral compounds.
