{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T11:41:19Z","timestamp":1790768479647,"version":"4.1.0"},"reference-count":44,"publisher":"Public Library of Science (PLoS)","issue":"8","license":[{"start":{"date-parts":[[2022,8,2]],"date-time":"2022-08-02T00:00:00Z","timestamp":1659398400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["R01GM122887"],"award-info":[{"award-number":["R01GM122887"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100000357","name":"North West Cancer Research","doi-asserted-by":"publisher","award":["CR782"],"award-info":[{"award-number":["CR782"]}],"id":[{"id":"10.13039\/501100000357","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p><jats:underline>A<\/jats:underline>taxia <jats:underline>T<\/jats:underline>elangiectasia mutated and <jats:underline>R<\/jats:underline>AD3-related (ATR) kinase is activated by DNA replication stress and also by various forms of DNA damage, including DNA double-strand breaks (DSBs). Recruitment to sites of damage is insufficient for ATR activation as one of two known ATR activators, either topoisomerase II-binding protein (TOPBP1) or Ewing\u2019s tumor-associated antigen 1, must also be present for signaling to initiate. Here, we employ our recently established <jats:underline>D<\/jats:underline>SB-<jats:underline>m<\/jats:underline>ediated <jats:underline>A<\/jats:underline>TR activation in <jats:underline><jats:italic>X<\/jats:italic><\/jats:underline><jats:italic>enopus<\/jats:italic> egg extract (DMAX) system to examine how TOPBP1 is recruited to DSBs, so that it may activate ATR. We report that TOPBP1 is only transiently present at DSBs, with a half-life of less than 10 minutes. We also examined the relationship between TOPBP1 and the MRE11-RAD50-NBS1 (MRN), CtBP interacting protein (CtIP), and <jats:underline>A<\/jats:underline>taxia <jats:underline>T<\/jats:underline>elangiectasia mutated (ATM) network of proteins. Loss of MRN prevents CtIP recruitment to DSBs, and partially inhibits TOPBP1 recruitment. Loss of CtIP has no impact on either MRN or TOPBP1 recruitment. Loss of ATM kinase activity prevents CtIP recruitment and enhances MRN and TOPBP1 recruitment. These findings demonstrate that there are MRN-dependent and independent pathways that recruit TOPBP1 to DSBs for ATR activation. Lastly, we find that both the 9-1-1 complex and MDC1 are dispensable for TOPBP1 recruitment to DSBs.<\/jats:p>","DOI":"10.1371\/journal.pone.0271905","type":"journal-article","created":{"date-parts":[[2022,8,2]],"date-time":"2022-08-02T19:03:37Z","timestamp":1659467017000},"page":"e0271905","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":9,"title":["MRN-dependent and independent pathways for recruitment of TOPBP1 to DNA double-strand breaks"],"prefix":"10.1371","volume":"17","author":[{"given":"Katrina","family":"Montales","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kenna","family":"Ruis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9102-4881","authenticated-orcid":true,"given":"Howard","family":"Lindsay","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8535-5796","authenticated-orcid":true,"given":"W. 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