title: TOPBP1 regulates RAD51 phosphorylation and chromatin loading and determines PARP inhibitor sensitivity.
abstract: Topoisomerase IIβ-binding protein 1 (TOPBP1) participates in DNA replication and DNA damage response; however, its role in DNA repair and relevance for human cancer remain unclear. Here, through an unbiased small interfering RNA screen, we identified and validated TOPBP1 as a novel determinant whose loss sensitized human cells to olaparib, an inhibitor of poly(ADP-ribose) polymerase. We show that TOPBP1 acts in homologous recombination (HR) repair, impacts olaparib response, and exhibits aberrant patterns in subsets of human ovarian carcinomas. TOPBP1 depletion abrogated RAD51 loading to chromatin and formation of RAD51 foci, but without affecting the upstream HR steps of DNA end resection and RPA loading. Furthermore, TOPBP1 BRCT domains 7/8 are essential for RAD51 foci formation. Mechanistically, TOPBP1 physically binds PLK1 and promotes PLK1 kinase-mediated phosphorylation of RAD51 at serine 14, a modification required for RAD51 recruitment to chromatin. Overall, our results provide mechanistic insights into TOPBP1's role in HR, with potential clinical implications for cancer treatment.
authors.affiliation: Danish Cancer Society Research Center, DK-2100 Copenhagen, Denmark Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, 779 00 Olomouc, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Moudry
authors.affiliation: Danish Cancer Society Research Center, DK-2100 Copenhagen, Denmark.
authors.forename: Kenji
authors.initials: K
authors.lastname: Watanabe
authors.affiliation: Danish Cancer Society Research Center, DK-2100 Copenhagen, Denmark.
authors.forename: Kamila M
authors.initials: KM
authors.lastname: Wolanin
authors.affiliation: Danish Cancer Society Research Center, DK-2100 Copenhagen, Denmark Department of Medical Biochemistry and Biophysics, Science For Life Laboratory, Division of Translational Medicine and Chemical Biology, Karolinska Institute, 17121 Solna, Sweden.
authors.forename: Jirina
authors.initials: J
authors.lastname: Bartkova
authors.affiliation: The Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, England, UK.
authors.forename: Isabel E
authors.initials: IE
authors.lastname: Wassing
authors.affiliation: Danish Cancer Society Research Center, DK-2100 Copenhagen, Denmark.
authors.forename: Sugiko
authors.initials: S
authors.lastname: Watanabe
authors.affiliation: Danish Cancer Society Research Center, DK-2100 Copenhagen, Denmark.
authors.forename: Robert
authors.initials: R
authors.lastname: Strauss
authors.affiliation: Department of Biology, University of Copenhagen, DK-2200 Copenhagen, Denmark.
authors.forename: Rune
authors.initials: R
authors.lastname: Troelsgaard Pedersen
authors.affiliation: Department of Biology, University of Copenhagen, DK-2200 Copenhagen, Denmark.
authors.forename: Vibe H
authors.initials: VH
authors.lastname: Oestergaard
authors.affiliation: Department of Biology, University of Copenhagen, DK-2200 Copenhagen, Denmark.
authors.forename: Michael
authors.initials: M
authors.lastname: Lisby
authors.affiliation: Department of Pathology, Familial and Hereditary Cancer Unit, University Hospital, 35010 Las Palmas de Gran Canaria, Spain.
authors.forename: Miguel
authors.initials: M
authors.lastname: Andújar-Sánchez
authors.affiliation: Danish Cancer Society Research Center, DK-2100 Copenhagen, Denmark.
authors.forename: Apolinar
authors.initials: A
authors.lastname: Maya-Mendoza
authors.affiliation: The Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, England, UK.
authors.forename: Fumiko
authors.initials: F
authors.lastname: Esashi
authors.affiliation: Danish Cancer Society Research Center, DK-2100 Copenhagen, Denmark Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, DK-2200 Copenhagen, Denmark.
authors.forename: Jiri
authors.initials: J
authors.lastname: Lukas
authors.affiliation: Danish Cancer Society Research Center, DK-2100 Copenhagen, Denmark Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, 779 00 Olomouc, Czech Republic Department of Medical Biochemistry and Biophysics, Science For Life Laboratory, Division of Translational Medicine and Chemical Biology, Karolinska Institute, 17121 Solna, Sweden jb@cancer.dk.
authors.forename: Jiri
authors.initials: J
authors.lastname: Bartek
mesh_terms: D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000378:metabolism; D002843:Chromatin / Q000378:metabolism*; D042002:Chromatin Assembly and Disassembly; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005260:Female; D057809:HEK293 Cells; D006367:HeLa Cells; D059765:Homologous Recombination; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D010766:Phosphorylation; D010793:Phthalazines / Q000494:pharmacology*; D010879:Piperazines / Q000494:pharmacology*; D000067856:Poly(ADP-ribose) Polymerase Inhibitors / Q000494:pharmacology*; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D034622:RNA Interference; D051135:Rad51 Recombinase / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013997:Time Factors; D014162:Transfection; D000097869:Polo-Like Kinase 1
chemical_list: D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D002843:Chromatin; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D010793:Phthalazines; D010879:Piperazines; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D051135:Rad51 Recombinase; D000097869:Polo-Like Kinase 1; C110513:TOPBP1 protein, human; C495153:RAD51 protein, human; C531550:olaparib
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