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In the ATR-Chk1 DDR pathway, it has been proposed that ATR is recruited to RPA-coated single-stranded DNA (ssDNA) by direct ATRIP-RPA interaction. However, it remains elusive how ATRIP is recruited to ssDNA in an RPA-independent manner. Here, we provide evidence that APE1 directly associates ssDNA to recruit ATRIP onto ssDNA in an RPA-independent fashion. The N-terminal motif within APE1 is required and sufficient for the APE1-ATRIP interaction in vitro and the distinct APE1-ATRIP interaction is required for ATRIP recruitment to ssDNA and the ATR-Chk1 DDR pathway activation in\n                    <jats:italic>Xenopus<\/jats:italic>\n                    egg extracts. In addition, APE1 directly associates with RPA70 and RPA32 via two distinct motifs. Taken together, our evidence suggests that APE1 recruits ATRIP onto ssDNA in an RPA-dependent and -independent manner in the ATR DDR pathway.\n                  <\/jats:p>","DOI":"10.7554\/elife.82324","type":"journal-article","created":{"date-parts":[[2023,5,9]],"date-time":"2023-05-09T12:01:57Z","timestamp":1683633717000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.82324","source":"Crossref","is-referenced-by-count":21,"title":["APE1 recruits ATRIP to ssDNA in an RPA-dependent and -independent manner to promote the ATR DNA damage response"],"prefix":"10.7554","volume":"12","author":[{"given":"Yunfeng","family":"Lin","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04dawnj30","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biological Sciences, University of North Carolina at Charlotte","place":["Charlotte, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jia","family":"Li","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04dawnj30","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biological Sciences, University of North Carolina at Charlotte","place":["Charlotte, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haichao","family":"Zhao","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04dawnj30","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biological Sciences, University of North Carolina at Charlotte","place":["Charlotte, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne","family":"McMahon","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04dawnj30","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biological Sciences, University of North Carolina at Charlotte","place":["Charlotte, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kelly","family":"McGhee","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04dawnj30","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biological Sciences, University of North Carolina at Charlotte","place":["Charlotte, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8106-6295","authenticated-orcid":true,"given":"Shan","family":"Yan","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04dawnj30","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biological Sciences, University of North Carolina at Charlotte","place":["Charlotte, United States"]},{"id":[{"id":"https:\/\/ror.org\/04dawnj30","id-type":"ROR","asserted-by":"publisher"}],"name":"School of Data Science, University of North Carolina at Charlotte","place":["Charlotte, United States"]},{"id":[{"id":"https:\/\/ror.org\/04dawnj30","id-type":"ROR","asserted-by":"publisher"}],"name":"Center for Biomedical Engineering and Science, University of North Carolina at Charlotte","place":["Charlotte, United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"4374","published-online":{"date-parts":[[2023,5,22]]},"reference":[{"key":"bib1","doi-asserted-by":"publisher","first-page":"425","DOI":"10.1016\/j.ctrv.2009.12.006","article-title":"Human AP Endonuclease 1 (Ape1): From mechanistic insights to Druggable target in cancer","volume":"36","author":"Abbotts","year":"2010","journal-title":"Cancer Treatment Reviews"},{"key":"bib2","doi-asserted-by":"publisher","first-page":"13124","DOI":"10.1074\/jbc.M116.729194","article-title":"Direct binding to Replicationprotein A (RPA)-Coated single-stranded DNA allows recruitment of the ATR activator Topbp1 to sites of DNA damage","volume":"291","author":"Acevedo","year":"2016","journal-title":"The Journal of Biological Chemistry"},{"key":"bib3","doi-asserted-by":"crossref","first-page":"14","DOI":"10.2174\/1874467211205010014","article-title":"Small-molecule inhibitors of Ape1 DNA repair function: An overview","volume":"5","author":"Al-Safi","year":"2012","journal-title":"Current Molecular Pharmacology"},{"key":"bib4","doi-asserted-by":"publisher","first-page":"2372","DOI":"10.1091\/mbc.e04-11-1006","article-title":"ATRIP binding to replication protein A-single-stranded DNA promotes ATR-ATRIP localization but is Dispensable for Chk1 Phosphorylation","volume":"16","author":"Ball","year":"2005","journal-title":"Molecular Biology of the Cell"},{"key":"bib5","doi-asserted-by":"publisher","first-page":"792","DOI":"10.1038\/nrm1493","article-title":"Checking on DNA damage in S phase","volume":"5","author":"Bartek","year":"2004","journal-title":"Nature Reviews. 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