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dc.contributor.authorLancey, Claudia
dc.contributor.authorTehseen, Muhammad
dc.contributor.authorBakshi, Souvika
dc.contributor.authorPercival, Matthew
dc.contributor.authorTakahashi, Masateru
dc.contributor.authorSobhy, Mohamed Abdelmaboud
dc.contributor.authorRaducanu, Vlad-Stefan
dc.contributor.authorBlair, Kerry
dc.contributor.authorMuskett, Frederick W.
dc.contributor.authorRagan, Timothy J.
dc.contributor.authorCrehuet, Ramon
dc.contributor.authorHamdan, Samir
dc.contributor.authorDe Biasio, Alfredo
dc.date.accessioned2021-10-20T12:12:01Z
dc.date.available2021-10-20T12:12:01Z
dc.date.issued2021-10-19
dc.date.submitted2020-07-23
dc.identifier.citationLancey, C., Tehseen, M., Bakshi, S., Percival, M., Takahashi, M., Sobhy, M. A., … De Biasio, A. (2021). Cryo-EM structure of human Pol κ bound to DNA and mono-ubiquitylated PCNA. Nature Communications, 12(1). doi:10.1038/s41467-021-26251-6
dc.identifier.issn2041-1723
dc.identifier.doi10.1038/s41467-021-26251-6
dc.identifier.urihttp://hdl.handle.net/10754/672905
dc.description.abstractAbstractY-family DNA polymerase κ (Pol κ) can replicate damaged DNA templates to rescue stalled replication forks. Access of Pol κ to DNA damage sites is facilitated by its interaction with the processivity clamp PCNA and is regulated by PCNA mono-ubiquitylation. Here, we present cryo-EM reconstructions of human Pol κ bound to DNA, an incoming nucleotide, and wild type or mono-ubiquitylated PCNA (Ub-PCNA). In both reconstructions, the internal PIP-box adjacent to the Pol κ Polymerase-Associated Domain (PAD) docks the catalytic core to one PCNA protomer in an angled orientation, bending the DNA exiting the Pol κ active site through PCNA, while Pol κ C-terminal domain containing two Ubiquitin Binding Zinc Fingers (UBZs) is invisible, in agreement with disorder predictions. The ubiquitin moieties are partly flexible and extend radially away from PCNA, with the ubiquitin at the Pol κ-bound protomer appearing more rigid. Activity assays suggest that, when the internal PIP-box interaction is lost, Pol κ is retained on DNA by a secondary interaction between the UBZs and the ubiquitins flexibly conjugated to PCNA. Our data provide a structural basis for the recruitment of a Y-family TLS polymerase to sites of DNA damage.
dc.description.sponsorshipThis research was supported by King Abdullah University of Science and Technology through core funding (to S.M.H.) and the Competitive Research Award Grant CRG8 URF/1/4036-01-01 (to S.M.H. and A.D.B.), and by the Wellcome Trust (to A.D.B.). R.C. acknowledges funding from the MINECO (CTQ2016-78636-P) and to AGAUR, (2017 SGR 324). The MD project has been carried out using CSUC resources. We acknowledge The Midlands Regional Cryo-EM Facility at the Leicester Institute of Structural and Chemical Biology (LISCB), major funding from MRC (MC_PC_17136). We thank Christos Savva (LISCB, University of Leicester) for his help in cryo-EM data collection and advice on data processing
dc.publisherSpringer Science and Business Media LLC
dc.relation.urlhttps://www.nature.com/articles/s41467-021-26251-6
dc.rightsThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.titleCryo-EM structure of human Pol κ bound to DNA and mono-ubiquitylated PCNA
dc.typeArticle
dc.contributor.departmentBiological and Environmental Science and Engineering (BESE) Division
dc.contributor.departmentBioscience Program
dc.contributor.departmentLaboratory of DNA Replication and Recombination
dc.identifier.journalNature Communications
dc.eprint.versionPublisher's Version/PDF
dc.contributor.institutionLeicester Institute of Structural & Chemical Biology and Department of Molecular & Cell Biology, University of Leicester, Lancaster Rd, Leicester LE1 7HB, UK.
dc.contributor.institutionThese authors contributed equally: Claudia Lancey, Muhammad Tehseen.
dc.contributor.institutionCSIC-Institute for Advanced Chemistry of Catalonia (IQAC) C/ Jordi Girona 18-26, 08034 Barcelona, Spain.
dc.identifier.volume12
dc.identifier.issue1
kaust.personTehseen, Muhammad
kaust.personTakahashi, Masateru
kaust.personSobhy, Mohamed Abdelmaboud
kaust.personRaducanu, Vlad-Stefan
kaust.personHamdan, Samir
kaust.personDe Biasio, Alfredo
kaust.grant.numberCRG8 URF/1/4036-01-01
dc.date.accepted2021-09-22
refterms.dateFOA2021-10-20T12:14:01Z
dc.date.published-online2021-10-19
dc.date.published-print2021-12


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This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.
Except where otherwise noted, this item's license is described as This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.