Multiple roles for Mre11 at uncapped telomeres

Progressive telomere attrition or uncapping of the shelterin complex elicits a DNA damage response (DDR) as a result of a cell’s inability to distinguish dysfunctional telomeric ends from DNA double-strand breaks (DSBs)1. Telomere deprotection activates both ataxia telangiectasia mutated (ATM) and telangiectasia and Rad3-related (ATR) kinase dependent DDR pathways and promotes efficient non-homologous end-joining (NHEJ) of dysfunctional telomeres2–5. The mammalian Mre11-Rad50-NBS1 (MRN) complex interacts with ATM to sense chromosomal DSBs and coordinate global DNA damage responses6, 7. While the MRN complex accumulates at dysfunctional telomeres, it is not known whether mammalian MRN promotes repair at these sites. Here we address this question by utilizing mouse alleles that either inactivate the entire MRN complex or eliminate only the nuclease activities of Mre118. Cells lacking MRN do not activate ATM when telomeric repeat binding factor 2 (TRF2) is removed from telomeres, and Ligase 4 (Lig4) dependent chromosome end-to-end fusions are markedly reduced. Residual chromatid fusions involve only telomeres generated by leading strand synthesis. Strikingly, while cells deficient for Mre11 nuclease activity efficiently activate ATM and recruit 53BP1 to deprotected telomeres, the 3’ telomeric overhang persists to prevent NHEJ-mediated chromosomal fusions. Removal of shelterin proteins that protect the 3’ overhang in the setting of Mre11 nuclease deficiency restores Lig4 dependent chromosome fusions. Our data suggest a critical role for the MRN complex in sensing dysfunctional telomeres, with Mre11 nuclease activity required to remove the 3’ telomeric overhang to promote chromosome fusion. Mre11 is also required to protect newly replicated leading strand telomeres from engaging the NHEJ pathway, likely by promoting 5’ strand resection to generate Pot1a-TPP1 bound 3’ overhangs that prevents NHEJ.