High-throughput DNA repair monitoring in Saccharomyces cerevisiae reveals SSB- and DSB-induced chromatin reconfiguration

DNA repair is critical for cellular function and genomic stability across organisms. Yeast mating-type switching serves as an established model for studying DNA break repair and chromatin dynamics. However, real-time tracking of mating-type switching in live cells remains challenging due to resolution limitations of existing techniques. Here, we use high-throughput methods, including three-dimensional imaging, to follow the dynamics of DNA damage and repair and to quantify mating-type switching occurrences at the single live cell level, with unprecedented resolution. We reveal chromatin reconfiguration for both single- and double-strand breaks following switching induction. Our findings provide new observation of the correlation between chromatin folding and single-strand breaks.