Local activation of mammalian separase in interphase promotes double‐strand break repair and prevents oncogenic transformation

Abstract Separase halves eukaryotic chromosomes in M‐phase by cleaving cohesin complexes holding sister chromatids together. Whether this essential protease functions also in interphase and/or impacts carcinogenesis remains largely unknown. Here, we show that mammalian separase is recruited to DNA double‐strand breaks (DSBs) where it is activated to locally cleave cohesin and facilitate homology‐directed repair (HDR). Inactivating phosphorylation of its NES, arginine methylation of its RG‐repeats, and sumoylation redirect separase from the cytosol to DSBs. In vitro assays suggest that DNA damage response‐relevant ATM, PRMT1, and Mms21 represent the corresponding kinase, methyltransferase, and SUMO ligase, respectively. SEPARASE heterozygosity not only debilitates HDR but also predisposes primary embryonic fibroblasts to neoplasia and mice to chemically induced skin cancer. Thus, tethering of separase to DSBs and confined cohesin cleavage promote DSB repair in G2 cells. Importantly, this conserved interphase function of separase protects mammalian cells from oncogenic transformation.