XLF and APLF bind Ku80 at two remote sites to ensure DNA repair by non-homologous end joining.

Nature structural & molecular biology 2018 Oct

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The Ku70-Ku80 (Ku) heterodimer binds rapidly and tightly to the ends of DNA double-strand breaks and recruits factors of the non-homologous end-joining (NHEJ) repair pathway through molecular interactions that remain unclear. We have determined crystal structures of the Ku-binding motifs (KBM) of the NHEJ proteins APLF (A-KBM) and XLF (X-KBM) bound to a Ku-DNA complex. The two KBM motifs bind remote sites of the Ku80 α/β domain. The X-KBM occupies an internal pocket formed by an unprecedented large outward rotation of the Ku80 α/β domain. We observe independent recruitment of the APLF-interacting protein XRCC4 and of XLF to laser-irradiated sites via binding of A- and X-KBMs, respectively, to Ku80. Finally, we show that mutation of the X-KBM and A-KBM binding sites in Ku80 compromises both the efficiency and accuracy of end joining and cellular radiosensitivity. A- and X-KBMs may represent two initial anchor points to build the intricate interaction network required for NHEJ.

Citation

Clement Nemoz, Virginie Ropars, Philippe Frit, Amandine Gontier, Pascal Drevet, Jinchao Yu, Raphaël Guerois, Aurelien Pitois, Audrey Comte, Christine Delteil, Nadia Barboule, Pierre Legrand, Sonia Baconnais, Yandong Yin, Satish Tadi, Emeline Barbet-Massin, Imre Berger, Eric Le Cam, Mauro Modesti, Eli Rothenberg, Patrick Calsou, Jean Baptiste Charbonnier. XLF and APLF bind Ku80 at two remote sites to ensure DNA repair by non-homologous end joining. Nature structural & molecular biology. 2018 Oct;25(10):971-980