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Previously, ATM activity was implicated in promoting the non-homologous end joining (NHEJ) pathway to repair a subset of DNA double-stranded breaks (DSBs), but how ATM performs this function is still unclear. In this study, we identified that ATM phosphorylates the DNA-dependent protein kinase catalytic subunit (DNA-PKcs), a core NHEJ factor, at its extreme C-terminus at threonine 4102 (T4102) in response to DSBs. Ablating phosphorylation at T4102 attenuates DNA-PKcs kinase activity and this destabilizes the interaction between DNA-PKcs and the Ku-DNA complex, resulting in decreased assembly and stabilization of the NHEJ machinery at DSBs. Phosphorylation at T4102 promotes NHEJ, radioresistance, and increases genomic stability following DSB induction. Collectively, these findings establish a key role for ATM in NHEJ-dependent repair of DSBs through positive regulation of DNA-PKcs.<\/jats:p>","DOI":"10.1093\/nar\/gkad505","type":"journal-article","created":{"date-parts":[[2023,5,31]],"date-time":"2023-05-31T23:12:13Z","timestamp":1685574733000},"page":"6770-6783","source":"Crossref","is-referenced-by-count":18,"title":["ATM phosphorylates the FATC domain of DNA-PKcs at threonine 4102 to promote non-homologous end joining"],"prefix":"10.1093","volume":"51","author":[{"given":"Huiming","family":"Lu","sequence":"first","affiliation":[{"name":"Department of Radiation Oncology, UT Southwestern Medical Center , Dallas , TX 75390 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qin","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Radiation Oncology, UT Southwestern Medical Center , Dallas , TX 75390 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4529-2302","authenticated-orcid":false,"given":"Daniel J","family":"Laverty","sequence":"additional","affiliation":[{"name":"Department of Environmental Health, Harvard T.H. Chan School of Public Health , Boston , MA 02115 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew C","family":"Puncheon","sequence":"additional","affiliation":[{"name":"Department of Radiation Oncology, UT Southwestern Medical Center , Dallas , TX 75390 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mathew\u00a0M","family":"Augustine","sequence":"additional","affiliation":[{"name":"Division of Surgical Oncology, Department of Surgery, UT Southwestern Medical Center , Dallas , TX 75390 , USA"},{"name":"Department of Surgery, North Texas VA Medical Center , Dallas , TX 75216 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gareth\u00a0J","family":"Williams","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Robson DNA Science Centre, Charbonneau Cancer Institute, Cumming School of Medicine, University of Calgary , Calgary , AB , Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zachary\u00a0D","family":"Nagel","sequence":"additional","affiliation":[{"name":"Department of Environmental Health, Harvard T.H. Chan School of Public Health , Boston , MA 02115 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6958-8816","authenticated-orcid":false,"given":"Benjamin\u00a0P C","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Radiation Oncology, UT Southwestern Medical Center , Dallas , TX 75390 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0184-3651","authenticated-orcid":false,"given":"Anthony\u00a0J","family":"Davis","sequence":"additional","affiliation":[{"name":"Department of Radiation Oncology, UT Southwestern Medical Center , Dallas , TX 75390 , USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2023,6,13]]},"reference":[{"key":"2023072104342484300_B1","doi-asserted-by":"crossref","first-page":"644","DOI":"10.1016\/j.cell.2017.01.002","article-title":"Endogenous DNA damage as a source of genomic instability in cancer","volume":"168","author":"Tubbs","year":"2017","journal-title":"Cell"},{"key":"2023072104342484300_B2","doi-asserted-by":"crossref","first-page":"801","DOI":"10.1016\/j.molcel.2017.05.015","article-title":"ATM, ATR, and DNA-PK: the trinity at the heart of the DNA damage response","volume":"66","author":"Blackford","year":"2017","journal-title":"Mol. 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