In vivo single-molecule studies of DNA repair in E. Coli
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Bacteria use highly dynamic interaction networks to coordinate the activities of individual DNA replication and repair proteins, allowing them to divide rapidly and respond to changing environmental conditions. We are using single-molecule fluorescence microscopy techniques to study activation of the translesion polymerase Pol V, after E. coli cells are damaged by UV light. Unexpectedly, our analysis revealed that a significant portion of Pol V is sequestered at the cell membrane and that it only enters the cytosol once a key post-translational modification has been made. This spatial control limits the (mutagenic) activity of Pol V during the early stages of the DNA damage response, giving error-free repair systems a chance to act first.