title: The E3 ubiquitin ligase ARIH1 protects against genotoxic stress by initiating a 4EHP-mediated mRNA translation arrest.
abstract: DNA damage response signaling is crucial for genome maintenance in all organisms and is corrupted in cancer. In an RNA interference (RNAi) screen for (de)ubiquitinases and sumoylases modulating the apoptotic response of embryonic stem (ES) cells to DNA damage, we identified the E3 ubiquitin ligase/ISGylase, ariadne homologue 1 (ARIH1). Silencing ARIH1 sensitized ES and cancer cells to genotoxic compounds and ionizing radiation, irrespective of their p53 or caspase-3 status. Expression of wild-type but not ubiquitinase-defective ARIH1 constructs prevented sensitization caused by ARIH1 knockdown. ARIH1 protein abundance increased after DNA damage through attenuation of proteasomal degradation that required ATM signaling. Accumulated ARIH1 associated with 4EHP, and in turn, this competitive inhibitor of the eukaryotic translation initiation factor 4E (eIF4E) underwent increased nondegradative ubiquitination upon DNA damage. Genotoxic stress led to an enrichment of ARIH1 in perinuclear, ribosome-containing regions and triggered 4EHP association with the mRNA 5' cap as well as mRNA translation arrest in an ARIH1-dependent manner. Finally, restoration of DNA damage-induced translation arrest in ARIH1-depleted cells by means of an eIF2 inhibitor was sufficient to reinstate resistance to genotoxic stress. These findings identify ARIH1 as a potent mediator of DNA damage-induced translation arrest that protects stem and cancer cells against genotoxic stress.
authors.affiliation: Division of Toxicology, Leiden/Amsterdam Center for Drug Research, Leiden University, Leiden, The Netherlands.
authors.forename: Louise
authors.initials: L
authors.lastname: von Stechow
authors.affiliation: Department of Toxicogenetics, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Dimitris
authors.initials: D
authors.lastname: Typas
authors.affiliation: Division of Toxicology, Leiden/Amsterdam Center for Drug Research, Leiden University, Leiden, The Netherlands Department of Toxicogenetics, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Jordi
authors.initials: J
authors.lastname: Carreras Puigvert
authors.affiliation: Division of Toxicology, Leiden/Amsterdam Center for Drug Research, Leiden University, Leiden, The Netherlands.
authors.forename: Laurens
authors.initials: L
authors.lastname: Oort
authors.affiliation: Division of Toxicology, Leiden/Amsterdam Center for Drug Research, Leiden University, Leiden, The Netherlands.
authors.forename: Ramakrishnaiah
authors.initials: R
authors.lastname: Siddappa
authors.affiliation: Department of Toxicogenetics, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Alex
authors.initials: A
authors.lastname: Pines
authors.affiliation: Department of Toxicogenetics, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Harry
authors.initials: H
authors.lastname: Vrieling
authors.affiliation: Division of Toxicology, Leiden/Amsterdam Center for Drug Research, Leiden University, Leiden, The Netherlands.
authors.forename: Bob
authors.initials: B
authors.lastname: van de Water
authors.affiliation: Department of Toxicogenetics, Leiden University Medical Center, Leiden, The Netherlands l.mullenders@lumc.nl e.danen@lacdr.leidenuniv.nl.
authors.forename: Leon H F
authors.initials: LH
authors.lastname: Mullenders
authors.affiliation: Division of Toxicology, Leiden/Amsterdam Center for Drug Research, Leiden University, Leiden, The Netherlands l.mullenders@lumc.nl e.danen@lacdr.leidenuniv.nl.
authors.forename: Erik H J
authors.initials: EH
authors.lastname: Danen
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D053148:Caspase 3 / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D004249:DNA Damage; D053595:Embryonic Stem Cells / Q000378:metabolism; D039561:Eukaryotic Initiation Factor-4E / Q000378:metabolism*; D006801:Humans; D051379:Mice; D014176:Protein Biosynthesis; D039381:RNA Cap-Binding Proteins / Q000378:metabolism*; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: D002352:Carrier Proteins; C112609:EIF4E2 protein, human; D039561:Eukaryotic Initiation Factor-4E; D039381:RNA Cap-Binding Proteins; D012333:RNA, Messenger; D016159:Tumor Suppressor Protein p53; C000597857:eIF4E2 protein, mouse; C419193:ARIH1 protein, human; C486004:Arih1 protein, mouse; D044767:Ubiquitin-Protein Ligases; D053148:Caspase 3