Mechanism and regulation of DNA-protein crosslink repair by the DNA-dependent metalloprotease SPRTN
journal contributionposted on 2020-10-02, 14:13 authored by Julian Stingele, Roberto Bellelli, Ferdinand Alte, Graeme Hewitt, Grzegorz Sarek, Sarah L Maslen, Susan E Tsutakawa, Annabel Borg, Svend Kjær, John A Tainer, J Mark Skehel, Michael Groll, Simon J Boulton
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.
DNA repairDNA-protein crosslinksDVC1Ruijs-Aalfs syndromeSPRTNSpartanWss1formaldehydehepatocellular carcinomaprogeriaproteasetopoisomeraseAmino Acid SequenceAnimalsBinding SitesCaenorhabditis elegansCaenorhabditis elegans ProteinsCell LineCisplatinCross-Linking ReagentsCrystallography, X-RayDNADNA RepairDNA-Binding ProteinsFibroblastsFormaldehydeHeLa CellsHumansKineticsMiceModels, MolecularProtein BindingProtein Interaction Domains and MotifsProtein Structure, SecondarySchizosaccharomycesSchizosaccharomyces pombe ProteinsSequence AlignmentSequence Homology, Amino AcidSubstrate SpecificityUltraviolet RaysXeroderma Pigmentosum Group A ProteinHela CellsBoulton FC00104806 Biological Sciences11 Medical and Health SciencesDevelopmental Biology