title: Activation of the double-stranded RNA-dependent protein kinase PKR by small ubiquitin-like modifier (SUMO).
abstract: 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.
authors: Departamento de Biología Molecular y Celular, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas (CSIC), Darwin 3, Madrid 28049.
authors: Carlos F
authors: CF
authors: de la Cruz-Herrera
authors: Departamento de Biología Molecular y Celular, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas (CSIC), Darwin 3, Madrid 28049.
authors: Michela
authors: M
authors: Campagna
authors: Unidad de Investigación, Hospital Universitario Virgen de las Nieves, 18014 Granada.
authors: Maria A
authors: MA
authors: García
authors: Departamento de Biología Molecular y Celular, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas (CSIC), Darwin 3, Madrid 28049.
authors: Laura
authors: L
authors: Marcos-Villar
authors: Ubiquitylation and Cancer Molecular Biology Laboratory, Inbiomed, San Sebastian-Donostia, 20009 Gipuzkoa, Spain.
authors: Valerie
authors: V
authors: Lang
authors: Centro de Investigación en Medicina Molecular (CIMUS), Universidade de Santiago de Compostela, Instituto de Investigaciones Sanitarias (IDIS), Santiago de Compostela E15782.
authors: Maite
authors: M
authors: Baz-Martínez
authors: Confocal Service of Centro Nacional de Biotecnología-CSIC, Darwin 3, Madrid 28049, and.
authors: Sylvia
authors: S
authors: Gutiérrez
authors: Departamento de Fisioloxía and Centro de Investigación en Medicina Molecular (CIMUS), Universidade de Santiago de Compostela, Instituto de Investigaciones Sanitarias (IDIS), Santiago de Compostela E15782, Spain.
authors: Anxo
authors: A
authors: Vidal
authors: Ubiquitylation and Cancer Molecular Biology Laboratory, Inbiomed, San Sebastian-Donostia, 20009 Gipuzkoa, Spain.
authors: Manuel S
authors: MS
authors: Rodríguez
authors: Departamento de Biología Molecular y Celular, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas (CSIC), Darwin 3, Madrid 28049.
authors: Mariano
authors: M
authors: Esteban
authors: Departamento de Biología Molecular y Celular, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas (CSIC), Darwin 3, Madrid 28049,; Centro de Investigación en Medicina Molecular (CIMUS), Universidade de Santiago de Compostela, Instituto de Investigaciones Sanitarias (IDIS), Santiago de Compostela E15782,. Electronic address: crivas@cnb.csic.es.
authors: Carmen
authors: C
authors: Rivas
mesh_terms: D016475:3T3 Cells; D000818:Animals; D004789:Enzyme Activation; D054884:Host-Pathogen Interactions; D007113:Immunity, Innate; D051379:Mice; D010449:Peptide Mapping; D011485:Protein Binding; D055503:Protein Multimerization; D012330:RNA, Double-Stranded / Q000737:chemistry; D012367:RNA, Viral / Q000737:chemistry; D025842:SUMO-1 Protein / Q000378:metabolism*; D020539:Sequence Analysis, Protein; D058207:Sumoylation; D018116:Vesiculovirus / Q000502:physiology; D014779:Virus Replication; D019892:eIF-2 Kinase / Q000737:chemistry / Q000378:metabolism*
chemical_list: D012330:RNA, Double-Stranded; D012367:RNA, Viral; D025842:SUMO-1 Protein; C495501:SUMO1 protein, human; D019892:eIF-2 Kinase
keywords: Double-stranded RNA (dsRNA); Protein Kinase RNA-activated (PKR); Sumoylation; Translation Control; Virus