In cells, enzymes are able to locate and sequence specifically bind to short regions of DNA in a compacted genome of many millions of bases. This fellowship will investigate the mechanism by which they achieve this remarkable feat by examining the behavior of single enzymes diffusing on single DNA molecules. The fellowship will focus on two main objectives. The first is to exploit novel DNA-labeling technology to investigate, for the first time at the single molecule level, the effect of DNA topology on the diffusion of enzymes. Since genetic DNA is highly compacted and adopts plectonemic structures, the study of enzymatic diffusion on supercoiled DNA molecules is critical to our understanding of how enzymes interact with DNA in the cell. The second objective is to investigate the relationship between enzymatic structure and the method used by an enzyme to move on the DNA duplex. This will be done by directly visualizing structurally well-characterized enzymes as they move on a single DNA molecule. The fellow brings a background expertise of fluorescence-based techniques and molecular biology to the fellowship. The fellowship aims to train the fellow in the application of single molecule imaging to the biophysical systems that he has studied in the past. Such knowledge will significantly enhance the ability of the fellow to initiate novel research in the area of biophysical systems in the future.
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