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We previously determined the structure of a short motif in the disordered XPA N-terminus bound to the RPA32C domain. However, a second contact between the XPA DNA-binding domain (XPA DBD) and the RPA70AB tandem ssDNA-binding domains, which is likely to influence the orientation of XPA and RPA on the damaged DNA substrate, remains poorly characterized. NMR was used to map the binding interfaces of XPA DBD and RPA70AB. Combining NMR and X-ray scattering data with comprehensive docking and refinement revealed how XPA DBD and RPA70AB orient on model NER DNA substrates. The structural model enabled design of XPA mutations that inhibit the interaction with RPA70AB. These mutations decreased activity in cell-based NER assays, demonstrating the functional importance of XPA DBD\u2013RPA70AB interaction. Our results inform ongoing controversy about where XPA is bound within the NER bubble, provide structural insights into the molecular basis for malfunction of disease-associated XPA missense mutations, and contribute to understanding of the structure and mechanical action of the NER machinery.<\/jats:p>","DOI":"10.1093\/nar\/gkz1231","type":"journal-article","created":{"date-parts":[[2019,12,21]],"date-time":"2019-12-21T07:06:40Z","timestamp":1576912000000},"page":"2173-2188","source":"Crossref","is-referenced-by-count":52,"title":["A key interaction with RPA orients XPA in NER complexes"],"prefix":"10.1093","volume":"48","author":[{"given":"Agnieszka M","family":"Topolska-Wo\u015b","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Vanderbilt University, Nashville, TN 37240-7917, USA"},{"name":"Center for Structural Biology, Vanderbilt University, Nashville, TN 37240-7917, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Norie","family":"Sugitani","sequence":"first","affiliation":[{"name":"Center for Structural Biology, Vanderbilt University, Nashville, TN 37240-7917, USA"},{"name":"Department of Chemistry, Vanderbilt University, Nashville, TN 37240-7917, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John J","family":"Cordoba","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Vanderbilt University, Nashville, TN 37240-7917, USA"},{"name":"Center for Structural Biology, Vanderbilt University, Nashville, TN 37240-7917, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kateryna V","family":"Le\u00a0Meur","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Vanderbilt University, Nashville, TN 37240-7917, USA"},{"name":"Center for Structural Biology, Vanderbilt University, Nashville, TN 37240-7917, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R\u00e9my A","family":"Le\u00a0Meur","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Vanderbilt University, Nashville, TN 37240-7917, USA"},{"name":"Center for Structural Biology, Vanderbilt University, Nashville, TN 37240-7917, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hyun Suk","family":"Kim","sequence":"first","affiliation":[{"name":"Center for Genomic Integrity, Institute for Basic Science, Ulsan,\u00a0Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jung-Eun","family":"Yeo","sequence":"first","affiliation":[{"name":"Center for Genomic Integrity, Institute for Basic Science, Ulsan,\u00a0Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Rosenberg","sequence":"first","affiliation":[{"name":"Molecular Biophysics and Integrated Bioimaging, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michal","family":"Hammel","sequence":"first","affiliation":[{"name":"Molecular Biophysics and Integrated Bioimaging, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Orlando D","family":"Sch\u00e4rer","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Vanderbilt University, Nashville, TN 37240-7917, USA"},{"name":"Center for Genomic Integrity, Institute for Basic Science, Ulsan,\u00a0Republic of Korea"},{"name":"Department of Biological Sciences, School of Life Sciences, Ulsan National Institute of Science and Technology, Ulsan, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2180-0790","authenticated-orcid":false,"given":"Walter J","family":"Chazin","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Vanderbilt University, Nashville, TN 37240-7917, USA"},{"name":"Center for Structural Biology, Vanderbilt University, Nashville, TN 37240-7917, USA"},{"name":"Department of Chemistry, Vanderbilt University, Nashville, TN 37240-7917, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2020,1,11]]},"reference":[{"key":"2020022408401823400_B1","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1021\/cr040483f","article-title":"Molecular mechanisms of mammalian global genome nucleotide excision repair","volume":"106","author":"Gillet","year":"2006","journal-title":"Chem. 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