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

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 impa:-ed 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 deponds on its conserved C-terminal VCA domain and Arp32/3 activation. Our findings reveal a function and mechanistic insight for WASH in DNA DSB repair by the NHEJ pathway.

Automated summary

The Wiskott-Aldrich syndrome protein and SCAR homolog (WASH) is a protein present in the nucleus that regulates gene transcription and maintains nuclear organization. It has been found to interact with core factors involved in DNA repair and play a role in the recruitment of repair machinery to DNA double-stranded break sites. Loss of WASH leads to increased cell sensitivity to DNA damage and impairs DNA repair efficiency.