Flap endonucleases (FENs) play a vital role in DNA replication, repair and recombination in all living cells. Aspects of how FENs locate their branched DNA substrates and the conformational changes associated with DNA hydrolysis remain unclear. In my project, we are trying to address these questions using a combination of direct atomic force microscopy (AFM) imaging with conventional biochemical and structural approaches. Development of imaging of DNA to major and minor groove resolution using approaches initially proven for membrane protein imaging, has provided the potential to directly visualize local strain within a single molecule during an interaction with a bound protein. Using AFM we aim to explore how FENs engage their substrate using imaging of DNA-FEN complexes to characterize their hydrolysis. |
The Team
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