DNA-dependent protein kinase promotes DNA end processing by MRN and CtIP.

Deshpande, Rajashree A; Myler, Logan R; Soniat, Michael M; Makharashvili, Nodar; Lee, Linda; Lees-Miller, Susan P; Finkelstein, Ilya J; Paull, Tanya T · Sci Adv · 2020

basic_science · Level V

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Abstract

The repair of DNA double-strand breaks occurs through nonhomologous end joining or homologous recombination in vertebrate cells-a choice that is thought to be decided by a competition between DNA-dependent protein kinase (DNA-PK) and the Mre11/Rad50/Nbs1 (MRN) complex but is not well understood. Using ensemble biochemistry and single-molecule approaches, here, we show that the MRN complex is dependent on DNA-PK and phosphorylated CtIP to perform efficient processing and resection of DNA ends in physiological conditions, thus eliminating the competition model. Endonucleolytic removal of DNA-PK-bound DNA ends is also observed at double-strand break sites in human cells. The involvement of DNA-PK in MRN-mediated end processing promotes an efficient and sequential transition from nonhomologous end joining to homologous recombination by facilitating DNA-PK removal.

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