Area of interest
My lab has been using atomic force microscopy (AFM) and single-molecule fluorescence coupled with bulk biophysical and biochemical methods to characterize the structure function properties of the protein-DNA complexes involved in DNA repair for over fifteen years. In particular relevance to this proposal, we have been using and developing single-molecule fluorescence methods to characterize the conformations of protein-DNA complexes involved in mismatch repair. We have a longterm collaboration with Keith Weninger (co-sponsor) at North Carolina State University, who is an expert in single-molecule fluorescence. This collaboration coupled with my background in physical chemistry and kinetics has enabled my lab to rapidly come to the forefront of single-molecule fluorescence. We have developed methods for the analysis of DNA bending using single molecule FRET (smFRET) and methods to simultaneously monitor protein (MutSα) binding and DNA bending using non-covalent fluorescent tags and three-color wide-field imaging. In addition, we developed a method to extract kinetic mechanisms out of very complex smFRET data. My lab has also developed methods to overlay AFM and fluorescence images with high-accuracy, which will make it possible to identify different proteins in multiprotein complexes. Our AFM and smFRET studies of DNA mismatch repair have made significant impact on the field, suggesting a novel mechanism of mismatch recognition by MutS and elucidating the role of nucleotides in controlling the conformational states of MutS and MutLα, which are essential for signaling repair.
Awards and Honors
Co-organizer of the 2010 Post-Initiation Transcription Meeting, Mountain Lake, VA
Plenary session speaker at 23rd Annual Symposium of the Protein Society, Boston MA
Co-organizer of the 2008 Post-Initiation Transcription Meeting, Mountain Lake, VA
Co-organizer 2006 Post-Initiation Transcription Meeting, Mountain Lake, VA (11/2-5/06)
Co-chair of the 2002 Biopolymers Gordon Conference
Chair of the Transcription Elongation session at the 2000 Post-Initiation Transcription Meeting, Mountain
Lake, VA (10/27-29/00)
Chair of a Symposium at the 1998 Biophysical Society Meeting titled “The Devil is in the Details: Structural
and Mechanistic Aspects of Transcription,” Kansas City, MO (2/98)
Junior Faculty Development Award, University of North Carolina (1995)
NIH Post-doctoral Fellowship (11/88-11/91).
Stanley Mandeles Graduate Research Award at Rutgers University (1988).