In this CAREER proposal the PI proposes to study the molecular mechanisms of DNA mismatch repair. The experimental approach will use single molecule experimental techniques. Specifically the PI will use single molecule fluorescence resonant energy transfer (smFRET) to observe the real time dynamics of the conformational changes taking place in the DNA molecule at an angstrom spatial resolution. It is hypothesized that these conformational changes are important for damage recognition by the repair proteins. A novel combination of force and smFRET spectroscopies is proposed to determine the effect of external force and torque in the protein-DNA interactions. The PI hypothesizes that these external loads can mimic the effect of physical constraints imposed in vivo on the DNA by multiple protein binding and rotational constraints. The focus of the proposal is on studies of DNA mismatch damage recognition. The assays and methodologies developed will allow extending the studies to downstream signaling events in the mismatch repair process resulting from interaction of multiple protein complexes. The PI will develop a new course "Introduction to Biological Physics" at the advanced undergraduate level. Undergraduate and graduate students will develop research projects and the PI will create awareness of emerging interdisciplinary fields and research opportunities for undergraduate students. Special attention is focused on the participation of undergraduate students from underrepresented groups in science in summer research projects.
