title: The role of ADP-ribosylation in regulating DNA interstrand crosslink repair.
abstract: ADP-ribosylation by ADP-ribosyltransferases (ARTs) has a well-established role in DNA strand break repair by promoting enrichment of repair factors at damage sites through ADP-ribose interaction domains. Here, we exploit the simple eukaryote Dictyostelium to uncover a role for ADP-ribosylation in regulating DNA interstrand crosslink repair and redundancy of this pathway with non-homologous end-joining (NHEJ). In silico searches were used to identify a protein that contains a permutated macrodomain (which we call aprataxin/APLF-and-PNKP-like protein; APL). Structural analysis reveals that this permutated macrodomain retains features associated with ADP-ribose interactions and that APL is capable of binding poly(ADP-ribose) through this macrodomain. APL is enriched in chromatin in response to cisplatin treatment, an agent that induces DNA interstrand crosslinks (ICLs). This is dependent on the macrodomain of APL and the ART Adprt2, indicating a role for ADP-ribosylation in the cellular response to cisplatin. Although adprt2-cells are sensitive to cisplatin, ADP-ribosylation is evident in these cells owing to redundant signalling by the double-strand break (DSB)-responsive ART Adprt1a, promoting NHEJ-mediated repair. These data implicate ADP-ribosylation in DNA ICL repair and identify that NHEJ can function to resolve this form of DNA damage in the absence of Adprt2.
authors: Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
authors: Alasdair R
authors: AR
authors: Gunn
authors: Dunn School of Pathology, University of Oxford, South Parks Road, Oxford, OX1 3RE, UK.
authors: Benito
authors: B
authors: Banos-Pinero
authors: Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
authors: Peggy
authors: P
authors: Paschke
authors: MRC Human Genetics Unit, The MRC Institute of Genetics and Molecular Medicine, University of Edinburgh, Western General Hospital, Crewe Road, Edinburgh EH4 2XU, Scotland, UK.
authors: Luis
authors: L
authors: Sanchez-Pulido
authors: Dunn School of Pathology, University of Oxford, South Parks Road, Oxford, OX1 3RE, UK.
authors: Antonio
authors: A
authors: Ariza
authors: Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
authors: Joseph
authors: J
authors: Day
authors: Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
authors: Mehera
authors: M
authors: Emrich
authors: Manchester Institute of Biotechnology, University of Manchester, Princess Street 131, Manchester, M1 7DN, UK.
authors: David
authors: D
authors: Leys
authors: MRC Human Genetics Unit, The MRC Institute of Genetics and Molecular Medicine, University of Edinburgh, Western General Hospital, Crewe Road, Edinburgh EH4 2XU, Scotland, UK.
authors: Chris P
authors: CP
authors: Ponting
authors: Dunn School of Pathology, University of Oxford, South Parks Road, Oxford, OX1 3RE, UK ivan.ahel@path.ox.ac.uk nicholas.lakin@bioch.ox.ac.uk.
authors: Ivan
authors: 0000-0002-9446-3756
authors: I
authors: Ahel
authors: Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK ivan.ahel@path.ox.ac.uk nicholas.lakin@bioch.ox.ac.uk.
authors: Nicholas D
authors: 0000-0003-4999-5814
authors: ND
authors: Lakin
mesh_terms: D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D002843:Chromatin / Q000378:metabolism; D002945:Cisplatin / Q000494:pharmacology; D003432:Cross-Linking Reagents / Q000378:metabolism*; D004247:DNA / Q000378:metabolism*; D004249:DNA Damage; D059766:DNA End-Joining Repair / Q000187:drug effects; D004260:DNA Repair / Q000187:drug effects; D004023:Dictyostelium / Q000187:drug effects / Q000378:metabolism*; D008958:Models, Molecular; D011064:Poly Adenosine Diphosphate Ribose / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D000072417:Protein Domains; D015800:Protozoan Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D002843:Chromatin; D003432:Cross-Linking Reagents; D015800:Protozoan Proteins; D011064:Poly Adenosine Diphosphate Ribose; D004247:DNA; D002945:Cisplatin
keywords: ADP-ribosyltransferases; Dictyostelium; Interstrand crosslink; PARPs