Regulation of human pol? by ATM-mediated phosphorylation during non-homologous end joining.

ABSTRACT: DNA double strand breaks (DSBs) trigger a variety of cellular signaling processes, collectively termed the DNA-damage response (DDR), that are primarily regulated by protein kinase ataxia-telangiectasia mutated (ATM). Among DDR activated processes, the repair of DSBs by non-homologous end joining (NHEJ) is essential. The proper coordination of NHEJ factors is mainly achieved through phosphorylation by an ATM-related kinase, the DNA-dependent protein kinase catalytic subunit (DNA-PKcs), although the molecular basis for this regulation has yet to be fully elucidated. In this study we identify the major NHEJ DNA polymerase, DNA polymerase lambda (Pol?), as a target for both ATM and DNA-PKcs in human cells. We show that Pol? is efficiently phosphorylated by DNA-PKcs in vitro and predominantly by ATM after DSB induction with ionizing radiation (IR) in vivo. We identify threonine 204 (T204) as a main target for ATM/DNA-PKcs phosphorylation on human Pol?, and establish that its phosphorylation may facilitate the repair of a subset of IR-induced DSBs and the efficient Pol?-mediated gap-filling during NHEJ. Molecular evidence suggests that Pol? phosphorylation might favor Pol? interaction with the DNA-PK complex at DSBs. Altogether, our work provides the first demonstration of how Pol? is regulated by phosphorylation to connect with the NHEJ core machinery during DSB repair in human cells.

SUBMITTER: Sastre-Moreno G 

PROVIDER: S-EPMC5444907 | biostudies-literature | 2017 Mar

REPOSITORIES: biostudies-literature