Chromatography sysem to purify nucleic acid molecules and proteins for biophysical analysis

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A hallmark of many viral infections is the production of foreign nucleic acid molecules generated during the viral life cycle. Cellular proteins that detect invasion through the recognition of viral nucleic acid molecules allow host cells to mount an immune response. At the heart of this recognition event are the molecular features of the protein-nucleic acid complexes that allow for successful interaction to occur. The McKenna laboratory is principally concerned with understanding the interactions between viral nucleic acid molecules and 2'-5' oligoadenylate synthetase (OAS) enzymes, a family of nucleic-acid binding proteins that prevent viral replication by blocking necessary viral protein production. The McKenna laboratory will establish a research program to understand, at the molecular level, the features of naturally occurring viral nucleic acid molecules that are specifically recognized by this family of enzymes. Biophysical and structural biology approaches will be used to characterize OAS-nucleic acid complexes, and help understand how these enzymes distinguish between viral and host-cell nucleic acid molecules. A crucial starting point for this research program is the development of strategies to produce and isolate protein and nucleic acid materials of high yield and purity. The requested Fast Performance Liquid Chromatography system outlined in this proposal will be the central device used to purify large quantities of these reagents, and is required equipment in modern structural biology laboratories. With the acquisition of the requested equipment, the McKenna laboratory will place itself at the cutting edge of the nucleic acid isolation field for use in structural biology applications. Given the important roles that nucleic acid molecules play in biological systems and the biotechnology field, the inclusion of a Fast Performance Liquid Chromatography system into an already multidisciplinary training environment will result in the training of highly qualified personnel who are well equipped for careers in science.