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Although the disease\u2010specific mutations are thought to affect different DNA repair pathways, the molecular basis for this is unknown. Here we examine the function of XPF\u2010ERCC1 in DNA interstrand crosslink (ICL) repair. We used\n                    <jats:italic>Xenopus<\/jats:italic>\n                    egg extracts to measure both ICL and nucleotide excision repair, and we identified mutations that are specifically defective in ICL repair. One of these separation\u2010of\u2010function mutations resides in the helicase\u2010like domain of XPF and disrupts binding to SLX4 and recruitment to the ICL. A small deletion in the same domain supports recruitment of XPF to the ICL, but inhibited the unhooking incisions most likely by disrupting a second, transient interaction with SLX4. Finally, mutation of residues in the nuclease domain did not affect localization of XPF\u2010ERCC1 to the ICL but did prevent incisions on the ICL substrate. Our data support a model in which the ICL repair\u2010specific function of XPF\u2010ERCC1 is dependent on recruitment, positioning and substrate recognition.\n                  <\/jats:p>","DOI":"10.15252\/embj.201695223","type":"journal-article","created":{"date-parts":[[2017,3,14]],"date-time":"2017-03-14T20:10:32Z","timestamp":1489522232000},"page":"2034-2046","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":50,"title":["Recruitment and positioning determine the specific role of the XPF\u2010ERCC1 endonuclease in interstrand crosslink repair"],"prefix":"10.1038","volume":"36","author":[{"given":"Daisy","family":"Klein Douwel","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wouter S","family":"Hoogenboom","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rick ACM","family":"Boonen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4198-0132","authenticated-orcid":false,"given":"Puck","family":"Knipscheer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2017,3,14]]},"reference":[{"key":"BFEMBJ201695223_CR1","doi-asserted-by":"crossref","first-page":"e1000871","DOI":"10.1371\/journal.pgen.1000871","volume":"6","author":"A Ahmad","year":"2010","unstructured":"Ahmad A, Enzlin JH, Bhagwat NR, Wijgers N, Raams A, Appledoorn E, Theil AF, Hoeijmakers JH, Vermeulen W, Jaspers NG, Sch\u00e4rer OD, Niedernhofer LJ (2010) Mislocalization of XPF\u2010ERCC1 nuclease contributes to reduced DNA repair in XP\u2010F patients. 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