TY - JOUR
T1 - XLF and APLF bind Ku80 at two remote sites to ensure DNA repair by non-homologous end joining
AU - Nemoz, Clement
AU - Ropars, Virginie
AU - Frit, Philippe
AU - Gontier, Amandine
AU - Drevet, Pascal
AU - Yu, Jinchao
AU - Guerois, Raphaël
AU - Pitois, Aurelien
AU - Comte, Audrey
AU - Delteil, Christine
AU - Barboule, Nadia
AU - Legrand, Pierre
AU - Baconnais, Sonia
AU - Yin, Yandong
AU - Tadi, Satish
AU - Barbet-Massin, Emeline
AU - Berger, Imre
AU - Le Cam, Eric
AU - Modesti, Mauro
AU - Rothenberg, Eli
AU - Calsou, Patrick
AU - Charbonnier, Jean Baptiste
N1 - Publisher Copyright:
© 2018, The Author(s), under exclusive licence to Springer Nature America, Inc.
PY - 2018/10/1
Y1 - 2018/10/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85054458313&partnerID=8YFLogxK
U2 - 10.1038/s41594-018-0133-6
DO - 10.1038/s41594-018-0133-6
M3 - Article
C2 - 30291363
AN - SCOPUS:85054458313
SN - 1545-9993
VL - 25
SP - 971
EP - 980
JO - Nature Structural and Molecular Biology
JF - Nature Structural and Molecular Biology
IS - 10
ER -