Imaging nuclear organization and DNA repair at the single molecule level
Our genome is constantly damaged by a variety of exogenous andendogenous agents. Among the various forms of DNA damage, double-strandbreaks (DSBs) are the most cytotoxic and genotoxic for the cell.Eukaryotic organisms use several mechanisms to repair DSBs among themnon-homologous end-joining (NHEJ) and homologous recombination (HR). Here, here, I will present our recent work to investigatethe molecular mechanisms of HR proteins inside cells at the singlemolecule level. In response to DSB, repair proteins colocalize from adiffuse distribution to repair foci located at the damaged DNA site. Anenduring question in the DNA damage field is how do repair proteins findtheir correct target, accumulate within repair foci and disassemble withthe proper time-window? Despite their functional importance, thephysical nature of repair sub-compartments remains unclear. To answerthese questions, we use single particle tracking (SPT) and PALM (PhotoActivable Localization Microscopy) to assess the physical propertiesunderlying repair foci formation and the internal dynamics of…