The RNF138 E3 ligase displaces Ku to promote DNA end resection and regulate DNA repair pathway choice

DNA double-strand breaks (DSBs) are repaired mainly by non-homologous end joining or homologous recombination (HR). Cell cycle stage and DNA end resection are believed to regulate the commitment to HR repair. Here we identify RNF138 as a ubiquitin E3 ligase that regulates the HR pathway. RNF138 is recruited to DNA damage sites through zinc fingers that have a strong preference for DNA with 5′- or 3′-single-stranded overhangs. RNF138 stimulates DNA end resection and promotes ATR-dependent signalling and DSB repair by HR, thereby contributing to cell survival on exposure to DSB-inducing agents. Finally, we establish that RNF138-dependent Ku removal from DNA breaks is one mechanism whereby RNF138 can promote HR. These results establish RNF138 as an important regulator of DSB repair pathway choice. Jackson and colleagues and Hendzel and colleagues reveal that the E3 ligase RNF138 functions in the repair of double-strand breaks by promoting CtIP accumulation and displacement of DNA-PK subunit Ku.