RNA-Protein Interactions in the Saccharomyces Cerevisiae Virus
Jeremy Bruenn Principal Investigator
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Many of the major problems currently being addressed by molecular biology involve the interactions of proteins and nucleic acids. Viruses provide a convenient means of addressing some of these questions. In particular, viruses with their own packaged polymerases provide self-contained systems in which the interaction of both enzymatic and structural proteins with nucleic acids can be studied in an easily purified and isolated form. Among such viruses, the double-stranded RNA viruses are unique in a number of ways. The substrate for their transcriptase is an entirely double- stranded RNA. The transcriptase and replicase are located in the same polypeptide. The substrate for packaging and replication is then viral plus strand. They are multi- segmented and in some double-stranded RNA viruses, package their RNAs in an equimolar ratio in one particle. Some of these viruses replicate semi-conservatively and some conservatively. The Saccharomyces cerevisiae virus is a simple version of such viruses, with one essential RNA of known sequence, one major capsid polypeptide, only one segment of RNA per particle, and only two open reading frames, the second ready by frameshifting. There are extensive genetics of virus-host interaction. This is therefore a useful model system studying packaging in double- stranded RNA virus. The PI proposes to focus on the determination of the optimal viral binding site and its relationship to viral interference and packing; and identification of the specificity determinants of viral binding and packaging.