Real-Time Interactions Between SV40 T-Antigen and a Stream of Analytes
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The primary goal of the funded research involved using surface plasmon resonance (SPR) for determining real-time interactions between viral SV40 large T-antigen (T-ag), DNA, and a stream of quadruplex DNA-interactive small molecules. We successfully developed an SPR-based assay for monitoring the ability of T-ag to unwind both normal duplex DNA and quadruplex DNA and characterized the ability of several small molecules to inhibit the helicase activity of T-ag. The ability to monitor unwinding of quadruplex DNA in real-time is an advance for investigating the inhibitory effects of quadruplex-interactive agents and this work was published recently in Cell Biochemistry and Biophysics (Feb. 2009). Our results will aid in future characterization of potential therapeutic approaches involving G-quadruplex interactive small molecules and in understanding the structural basis for selective targeting of different families of G-quadruplex DNA helicases. These results form the basis for a recent NIH ARRA grant in collaboration with UT-Austin. In addition, data generated in this research was presented at poster presentations for three national biochemistry meetings. A total of 4 undergraduate students and 1 graduate student were involved in the research.