Article
Regulation of human Polλ by ATM-mediated phosphorylation during Non-Homologous End Joining
Author/s | Sastre Moreno, Guillermo
Pryor, John M. Moreno Oñate, Marta Herrero Ruiz, Andrés M. Cortés Ledesma, Felipe Ruiz Pérez, José Francisco |
Department | Universidad de Sevilla. Departamento de Bioquímica Vegetal y Biología Molecular |
Publication Date | 2018 |
Deposit Date | 2018-09-20 |
Published in |
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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 ... 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 (Polλ), as a target for both ATM and DNA-PKcs in human cells. We show that Polλ 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 Polλ, and establish that its phosphorylation may facilitate the repair of a subset of IR-induced DSBs and the efficient Polλ-mediated gap-filling during NHEJ. Molecular evidence suggests that Polλ phosphorylation might favor Polλ interaction with the DNA-PK complex at DSBs. Altogether, our work provides the first demonstration of how Polλ is regulated by phosphorylation to connect with the NHEJ core machinery during DSB repair in human cells. |
Funding agencies | Ministerio de Economía y Competitividad (MINECO). España European Commission (EC). Fondo Europeo de Desarrollo Regional (FEDER) |
Project ID. | RYC-2011-08752
BFU2013-44343-P SAF2014-55532-R |
Citation | Sastre Moreno, G., Pryor, J.M., Moreno Oñate, M., Herrero Ruiz, A.M., Cortés Ledesma, F. y Ruiz Pérez, J.F. (2018). Regulation of human Polλ by ATM-mediated phosphorylation during Non-Homologous End Joining. DNA Repair, 51, 31-45. |
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