TY - JOUR
T1 - Zinc finger protein ZNF384 is an adaptor of Ku to DNA during classical non-homologous end-joining
AU - Singh, Jenny Kaur
AU - Smith, Rebecca
AU - Rother, Magdalena B.
AU - de Groot, Anton J. L.
AU - Wiegant, Wouter W.
AU - Vreeken, Kees
AU - D’Augustin, Ostiane
AU - Kim, Robbert Q.
AU - Qian, Haibin
AU - Krawczyk, Przemek M.
AU - González-Prieto, Román
AU - Vertegaal, Alfred C. O.
AU - Lamers, Meindert
AU - Huet, S. bastien
AU - van Attikum, Haico
N1 - Funding Information:
We thank Robin van Schendel and Marcel Tijsterman for the custom Sanger Sequence analyzer and help with the sequence analysis, and Dik van Gent, Stephen Taylor, Geert Kopps, Bernard Lopez, Roger Greenberg, Maria Jasin, Jeremy Stark, Nicholas Lakin, Sylvia Gelpke-Vermeulen, and Karoly Szuhai for kindly providing valuable reagents. We also thank the Microscopy-Rennes Imaging Center (BIOSIT, Université Rennes 1), a member of the national infrastructure France-BioImaging supported by the French National Research Agency (ANR-10-INBS-04), for providing access to their imaging setups, as well as S. Dutertre and X. Pinson for technical assistance with the microscopes. This research was financially supported by the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (ERC-StG 310913 to A.C.O.V.; ERC-CoG 50364 to H.v.A.), the Ligue contre le Cancer du Grand-Ouest (committees 22 and 35), the Fondation ARC pour la recherche sur le cancer (20161204883), the Agence Nationale de la Recherche (PRC-2018 REPAIRCHROM) and the Institut Universitaire de France (all grants to S.H.). R.S. is supported by the Fon-dation ARC pour la recherche sur le cancer (PDF20181208405).
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/12/1
Y1 - 2021/12/1
N2 - DNA double-strand breaks (DSBs) are among the most deleterious types of DNA damage as they can lead to mutations and chromosomal rearrangements, which underlie cancer development. Classical non-homologous end-joining (cNHEJ) is the dominant pathway for DSB repair in human cells, involving the DNA-binding proteins XRCC6 (Ku70) and XRCC5 (Ku80). Other DNA-binding proteins such as Zinc Finger (ZnF) domain-containing proteins have also been implicated in DNA repair, but their role in cNHEJ remained elusive. Here we show that ZNF384, a member of the C2H2 family of ZnF proteins, binds DNA ends in vitro and is recruited to DSBs in vivo. ZNF384 recruitment requires the poly(ADP-ribosyl) polymerase 1 (PARP1)-dependent expansion of damaged chromatin, followed by binding of its C2H2 motifs to the exposed DNA. Moreover, ZNF384 interacts with Ku70/Ku80 via its N-terminus, thereby promoting Ku70/Ku80 assembly and the accrual of downstream cNHEJ factors, including APLF and XRCC4/LIG4, for efficient repair at DSBs. Altogether, our data suggest that ZNF384 acts as a ‘Ku-adaptor’ that binds damaged DNA and Ku70/Ku80 to facilitate the build-up of a cNHEJ repairosome, highlighting a role for ZNF384 in DSB repair and genome maintenance.
AB - DNA double-strand breaks (DSBs) are among the most deleterious types of DNA damage as they can lead to mutations and chromosomal rearrangements, which underlie cancer development. Classical non-homologous end-joining (cNHEJ) is the dominant pathway for DSB repair in human cells, involving the DNA-binding proteins XRCC6 (Ku70) and XRCC5 (Ku80). Other DNA-binding proteins such as Zinc Finger (ZnF) domain-containing proteins have also been implicated in DNA repair, but their role in cNHEJ remained elusive. Here we show that ZNF384, a member of the C2H2 family of ZnF proteins, binds DNA ends in vitro and is recruited to DSBs in vivo. ZNF384 recruitment requires the poly(ADP-ribosyl) polymerase 1 (PARP1)-dependent expansion of damaged chromatin, followed by binding of its C2H2 motifs to the exposed DNA. Moreover, ZNF384 interacts with Ku70/Ku80 via its N-terminus, thereby promoting Ku70/Ku80 assembly and the accrual of downstream cNHEJ factors, including APLF and XRCC4/LIG4, for efficient repair at DSBs. Altogether, our data suggest that ZNF384 acts as a ‘Ku-adaptor’ that binds damaged DNA and Ku70/Ku80 to facilitate the build-up of a cNHEJ repairosome, highlighting a role for ZNF384 in DSB repair and genome maintenance.
UR - https://www.scopus.com/pages/publications/85119019981
U2 - 10.1038/s41467-021-26691-0
DO - 10.1038/s41467-021-26691-0
M3 - Article
C2 - 34772923
SN - 2041-1723
VL - 12
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 6560
ER -
SDG 3 Good Health and Well-being