title: Mechanism and Regulation of DNA-Protein Crosslink Repair by the DNA-Dependent Metalloprotease SPRTN.
abstract: Covalent DNA-protein crosslinks (DPCs) are toxic DNA lesions that interfere with essential chromatin transactions, such as replication and transcription. Little was known about DPC-specific repair mechanisms until the recent identification of a DPC-processing protease in yeast. The existence of a DPC protease in higher eukaryotes is inferred from data in Xenopus laevis egg extracts, but its identity remains elusive. Here we identify the metalloprotease SPRTN as the DPC protease acting in metazoans. Loss of SPRTN results in failure to repair DPCs and hypersensitivity to DPC-inducing agents. SPRTN accomplishes DPC processing through a unique DNA-induced protease activity, which is controlled by several sophisticated regulatory mechanisms. Cellular, biochemical, and structural studies define a DNA switch triggering its protease activity, a ubiquitin switch controlling SPRTN chromatin accessibility, and regulatory autocatalytic cleavage. Our data also provide a molecular explanation on how SPRTN deficiency causes the premature aging and cancer predisposition disorder Ruijs-Aalfs syndrome.
authors.affiliation: The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK.
authors.forename: Julian
authors.initials: J
authors.lastname: Stingele
authors.affiliation: The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK.
authors.forename: Roberto
authors.initials: R
authors.lastname: Bellelli
authors.affiliation: Center for Integrated Protein Science at the Department Chemie, Lehrstuhl für Biochemie, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching, Germany.
authors.forename: Ferdinand
authors.initials: F
authors.lastname: Alte
authors.affiliation: The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK.
authors.forename: Graeme
authors.initials: G
authors.lastname: Hewitt
authors.affiliation: The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK.
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Sarek
authors.affiliation: MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.
authors.forename: Sarah L
authors.initials: SL
authors.lastname: Maslen
authors.affiliation: Molecular Biophysics and Integrated Bioimaging, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
authors.forename: Susan E
authors.initials: SE
authors.lastname: Tsutakawa
authors.affiliation: The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK.
authors.forename: Annabel
authors.initials: A
authors.lastname: Borg
authors.affiliation: The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK.
authors.forename: Svend
authors.initials: S
authors.lastname: Kjær
authors.affiliation: Molecular Biophysics and Integrated Bioimaging, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA; Department of Molecular and Cellular Oncology, The University of Texas M.D. Anderson Cancer Center, Houston, TX 77030, USA.
authors.forename: John A
authors.initials: JA
authors.lastname: Tainer
authors.affiliation: MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK.
authors.forename: J Mark
authors.initials: JM
authors.lastname: Skehel
authors.affiliation: Center for Integrated Protein Science at the Department Chemie, Lehrstuhl für Biochemie, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Groll
authors.affiliation: The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK. Electronic address: simon.boulton@crick.ac.uk.
authors.forename: Simon J
authors.initials: SJ
authors.lastname: Boulton
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D017173:Caenorhabditis elegans / Q000187:drug effects / Q000201:enzymology / Q000235:genetics / Q000528:radiation effects; D029742:Caenorhabditis elegans Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D002945:Cisplatin / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000737:chemistry; D018360:Crystallography, X-Ray; D004247:DNA / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000528:radiation effects; D005557:Formaldehyde / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D051379:Mice; D008958:Models, Molecular; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D017433:Protein Structure, Secondary; D012568:Schizosaccharomyces / Q000201:enzymology / Q000235:genetics; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D014466:Ultraviolet Rays; D051760:Xeroderma Pigmentosum Group A Protein / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D029742:Caenorhabditis elegans Proteins; D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; C574636:SPRTN protein, human; D029702:Schizosaccharomyces pombe Proteins; C553374:XPA-1 protein, C elegans; D051760:Xeroderma Pigmentosum Group A Protein; D005557:Formaldehyde; D004247:DNA; D002945:Cisplatin
keywords: DNA repair; DNA-protein crosslinks; DVC1; Ruijs-Aalfs syndrome; SPRTN; Spartan; Wss1; formaldehyde; hepatocellular carcinoma; progeria; protease; topoisomerase