de la Cruz-Herrera, Carlos FCampagna, MichelaGarcía, Maria AMarcos-Villar, LauraLang, ValerieBaz-Martínez, MaiteGutiérrez, SylviaVidal, AnxoRodríguez, Manuel SEsteban, MarianoRivas, Carmen2016-07-012016-07-012014-09-19de la Cruz-Herrera CF, Campagna M, García MA, Marcos-Villar L, Lang V, Baz-Martínez M, et al. Activation of the double-stranded RNA-dependent protein kinase PKR by small ubiquitin-like modifier (SUMO). J. Biol. Chem.. 2014 ; 289(38):26357-670021-9258http://hdl.handle.net/10668/2245Journal Article; Research Support, Non-U.S. Gov't;The dsRNA-dependent kinase PKR is an interferon-inducible protein with ability to phosphorylate the α subunit of the eukaryotic initiation factor (eIF)-2 complex, resulting in a shut-off of general translation, induction of apoptosis, and inhibition of virus replication. Here we analyzed the modification of PKR by the small ubiquitin-like modifiers SUMO1 and SUMO2 and evaluated the consequences of PKR SUMOylation. Our results indicate that PKR is modified by both SUMO1 and SUMO2, in vitro and in vivo. We identified lysine residues Lys-60, Lys-150, and Lys-440 as SUMOylation sites in PKR. We show that SUMO is required for efficient PKR-dsRNA binding, PKR dimerization, and eIF2α phosphorylation. Furthermore, we demonstrate that SUMO potentiates the inhibition of protein synthesis induced by PKR in response to dsRNA, whereas a PKR SUMOylation mutant is impaired in its ability to inhibit protein synthesis and shows reduced capability to control vesicular stomatitis virus replication and to induce apoptosis in response to vesicular stomatitis virus infection. In summary, our data demonstrate the important role of SUMO in processes mediated by the activation of PKR.enDouble-stranded RNA (dsRNA)Protein Kinase RNA-activated (PKR)SumoylationTranslation ControlVirusActivación enzimáticaInteracciones huésped-patógenoInmunidad innataMapeo peptídicoUnión proteicaARN bicatenarioARN viralProteína SUMO-1Análisis de secuencia de proteínaSumoilaciónReplicación viraleIF-2 quinasaCélulas 3T3AnimalesMultimerización de proteínasRatonesMedical Subject Headings::Organisms::Eukaryota::AnimalsMedical Subject Headings::Phenomena and Processes::Chemical Phenomena::Biochemical Phenomena::Biochemical Processes::Enzyme ActivationMedical Subject Headings::Phenomena and Processes::Microbiological Phenomena::Microbiological Processes::Host-Pathogen InteractionsMedical Subject Headings::Phenomena and Processes::Immune System Phenomena::Immunity::Immunity, InnateMedical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Chemistry Techniques, Analytical::Peptide MappingMedical Subject Headings::Phenomena and Processes::Chemical Phenomena::Biochemical Phenomena::Biochemical Processes::Protein BindingMedical Subject Headings::Chemicals and Drugs::Nucleic Acids, Nucleotides, and Nucleosides::Nucleic Acids::RNA::RNA, Double-StrandedMedical Subject Headings::Chemicals and Drugs::Nucleic Acids, Nucleotides, and Nucleosides::Nucleic Acids::RNA::RNA, ViralMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Ubiquitins::Small Ubiquitin-Related Modifier Proteins::SUMO-1 ProteinMedical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Genetic Techniques::Sequence Analysis::Sequence Analysis, ProteinMedical Subject Headings::Phenomena and Processes::Chemical Phenomena::Biochemical Phenomena::Biochemical Processes::Peptide Biosynthesis::Protein Biosynthesis::Protein Modification, Translational::Protein Processing, Post-Translational::Ubiquitination::SumoylationMedical Subject Headings::Organisms::Viruses::RNA Viruses::Mononegavirales::Rhabdoviridae::VesiculovirusMedical Subject Headings::Phenomena and Processes::Microbiological Phenomena::Microbiological Processes::Virus Physiological Processes::Virus ReplicationMedical Subject Headings::Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Transferases::Phosphotransferases::Phosphotransferases (Alcohol Group Acceptor)::Protein Kinases::Protein-Serine-Threonine Kinases::eIF-2 KinaseMedical Subject Headings::Anatomy::Cells::Cells, Cultured::Cell Line::3T3 CellsMedical Subject Headings::Phenomena and Processes::Chemical Phenomena::Biochemical Phenomena::Biochemical Processes::Protein MultimerizationMedical Subject Headings::Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Rodentia::Muridae::Murinae::Miceresearch article25074923open access10.1074/jbc.M114.5609611083-351XPMC4176227