WASH interacts with Ku to regulate DNA double-stranded break repair

Tao Wang; Xiao-Hui Du; Yu Hong; Xian Hong; Li Fan; Jian-Wen Zhou; He Sun; Jie Ge; Daniel D Billadeau; Zhi-Hui Deng

doi:10.1016/j.isci.2021.103676

WASH interacts with Ku to regulate DNA double-stranded break repair

iScience. 2021 Dec 25;25(1):103676. doi: 10.1016/j.isci.2021.103676. eCollection 2022 Jan 21.

Authors

Tao Wang¹, Xiao-Hui Du¹, Yu Hong¹, Xian Hong¹, Li Fan¹, Jian-Wen Zhou¹, He Sun¹, Jie Ge², Daniel D Billadeau³, Zhi-Hui Deng¹

Affiliations

¹ Laboratory of Protein Structure and Function, Institute of Medicine and Pharmacy, Qiqihar Medical University, Qiqihar, Heilongjiang 161006, China.
² Department of Epidemiology and Statistics, School of Public Health, Qiqihar Medical University, Qiqihar, Heilongjiang 161006, China.
³ Division of Oncology Research and Schulze Center for Novel Therapeutics, Mayo Clinic College of Medicine, Rochester, MN 55905, USA.

Abstract

The Wiskott-Aldrich syndrome protein and SCAR homolog (WASH), an actin nucleation-promoting factor, is present in the nucleus where it regulates gene transcription and maintains nuclear organization. Here, we show that WASH interacts with core non-homologous end-joining (NHEJ) factors including Ku70/Ku80 and DNA-PKcs, and Ku70/Ku80 is involved in the recruitment of WASH to the sites of DNA double-stranded break (DSB). WASH depletion leads to increased cell sensitivity and impaired DNA repair capacity in response to etoposide-induced DSBs and reduces NHEJ efficiency. Mechanistically, we show that loss of WASH inhibits the phosphorylation of DNA-PKcs, H2AX, and KAP1 after DSB induction and reduces chromatin relaxation and the recruitment of several downstream NHEJ factors to DSBs. Moreover, WASH role in DSB repair depends on its conserved C-terminal VCA domain and Arp2/3 activation. Our findings reveal a function and mechanistic insight for WASH in DNA DSB repair by the NHEJ pathway.

Keywords: Biological sciences; Cell biology; Molecular biology.