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dc.contributor.authorAbdullah, Ummi B
dc.contributor.authorMcGouran, Joanna F
dc.contributor.authorBrolih, Sanja
dc.contributor.authorPtchelkine, Denis
dc.contributor.authorEl‐Sagheer, Afaf H
dc.contributor.authorBrown, Tom
dc.contributor.authorMcHugh, Peter J
dc.date.accessioned2017-06-21T06:51:53Z
dc.date.available2017-06-21T06:51:53Z
dc.date.issued2017-06-12
dc.identifier.citationAbdullah UB, McGouran JF, Brolih S, Ptchelkine D, El-Sagheer AH, et al. (2017) RPA activates the XPF-ERCC1 endonuclease to initiate processing of DNA interstrand crosslinks. The EMBO Journal: e201796664. Available: http://dx.doi.org/10.15252/embj.201796664.
dc.identifier.issn0261-4189
dc.identifier.issn1460-2075
dc.identifier.issn0261-4189
dc.identifier.issn1460-2075
dc.identifier.doi10.15252/embj.201796664
dc.identifier.urihttp://hdl.handle.net/10754/625123
dc.description.abstractDuring replication-coupled DNA interstrand crosslink (ICL) repair, the XPF-ERCC1 endonuclease is required for the incisions that release, or “unhook”, ICLs, but the mechanism of ICL unhooking remains largely unknown. Incisions are triggered when the nascent leading strand of a replication fork strikes the ICL. Here, we report that while purified XPF-ERCC1 incises simple ICL-containing model replication fork structures, the presence of a nascent leading strand, modelling the effects of replication arrest, inhibits this activity. Strikingly, the addition of the single-stranded DNA (ssDNA)-binding replication protein A (RPA) selectively restores XPF-ERCC1 endonuclease activity on this structure. The 5′–3′ exonuclease SNM1A can load from the XPF-ERCC1-RPA-induced incisions and digest past the crosslink to quantitatively complete the unhooking reaction. We postulate that these collaborative activities of XPF-ERCC1, RPA and SNM1A might explain how ICL unhooking is achieved in vivo.
dc.description.sponsorshipWe thank our laboratory member Sook Y. Lee and Opher Gileadi (Structural Genomics Consortium, Oxford) for the recombinant SNM1A; Fumiko Esashi (Dunn School of Pathology, University of Oxford) and Marc Wold (Iowa State University, USA) for the RPA and RPA70ΔC442, respectively; Hannah Baddock (WIMM), Hazel Aitkenhead and Joseph A. Newman (Structural Genomics Consortium, University of Oxford) for assistance with fluorescence anisotropy. P.J.M. was supported by MRC (Medical Research Council, grant MR/L007665/1). A.H.E.S. and T.B. were supported by BBSRC (Biotechnology and Biological Sciences Research Council, grant BB/J001694/1) BBSRC: sLOLA grant BB/J001694/1 “Extending the boundaries of nucleic acid chemistry” J.F.M. was supported by King Abdullah University of Science and Technology (KAUST) for a grant awarded to B. Nordén. U.B.A. was supported by the Malaysia's King Scholarship (Biasiswa Yang Di-Pertuan Agong).
dc.publisherEMBO
dc.titleRPA activates the XPF-ERCC1 endonuclease to initiate processing of DNA interstrand crosslinks
dc.typeArticle
dc.identifier.journalThe EMBO Journal
dc.contributor.institutionDepartment of Oncology, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, UK
dc.contributor.institutionDepartment of Chemistry, University of Oxford, Oxford, UK
dc.contributor.institutionDepartment of Chemistry, Trinity College Dublin, Dublin, Ireland
dc.contributor.institutionResearch Complex at Harwell, Rutherford Appleton Laboratory, Oxford, UK
dc.contributor.institutionDepartment of Oncology, University of Oxford, Oxford, UK
dc.date.published-online2017-06-12
dc.date.published-print2017-07-14


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