High-throughput DNA repair monitoring in Saccharomyces cerevisiae suggests 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 chromosome 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 chromosome reconfiguration for both single- and double-strand breaks following switching induction. Our findings provide new observation of the correlation between chromosome folding and single-strand breaks. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1038/s41598-025-17538-5.