title: The Golgi apparatus acts as a platform for TBK1 activation after viral RNA sensing.
abstract: After viral infection and the stimulation of some pattern-recognition receptors, TANK-binding kinase I (TBK1) is activated by K63-linked polyubiquitination followed by trans-autophosphorylation. While the activated TBK1 induces type I interferon production by phosphorylating the transcription factor IRF3, the precise molecular mechanisms underlying TBK1 activation remain unclear.
  
  
  
  We report here the localization of the ubiquitinated and phosphorylated active form of TBK1 to the Golgi apparatus after the stimulation of RIG-I-like receptors (RLRs) or Toll-like receptor-3 (TLR3), due to TBK1 K63-linked ubiquitination on lysine residues 30 and 401. The ubiquitin-binding protein optineurin (OPTN) recruits ubiquitinated TBK1 to the Golgi apparatus, leading to the formation of complexes in which TBK1 is activated by trans-autophosphorylation. Indeed, OPTN deficiency in various cell lines and primary cells impairs TBK1 targeting to the Golgi apparatus and its activation following RLR or TLR3 stimulation. Interestingly, the Bluetongue virus NS3 protein binds OPTN at the Golgi apparatus, neutralizing its activity and thereby decreasing TBK1 activation and downstream signaling.
  
  
  
  Our results highlight an unexpected role of the Golgi apparatus in innate immunity as a key subcellular gateway for TBK1 activation after RNA virus infection.
authors.affiliation: INSERM, UMR_S 1197, Hôpital Paul Brousse, Villejuif, France.|Université Paris-Saclay, Paris, France.|Equipe Labellisée Ligue contre le Cancer, Villejuif, France.
authors.forename: Marie
authors.initials: M
authors.lastname: Pourcelot
authors.affiliation: INSERM, UMR_S 1197, Hôpital Paul Brousse, Villejuif, France.|Université Paris-Saclay, Paris, France.|Equipe Labellisée Ligue contre le Cancer, Villejuif, France.
authors.forename: Naima
authors.initials: N
authors.lastname: Zemirli
authors.affiliation: INSERM, UMR_S 1197, Hôpital Paul Brousse, Villejuif, France.|Université Paris-Saclay, Paris, France.|Equipe Labellisée Ligue contre le Cancer, Villejuif, France.
authors.forename: Leandro
authors.initials: L
authors.lastname: Silva Da Costa
authors.affiliation: INSERM, UMR_S 1197, Hôpital Paul Brousse, Villejuif, France.|Université Paris-Saclay, Paris, France.|Equipe Labellisée Ligue contre le Cancer, Villejuif, France.
authors.forename: Roxane
authors.initials: R
authors.lastname: Loyant
authors.affiliation: Department of Microbiology and Molecular Medicine, Faculty of Medicine, University of Geneva, Geneva, Switzerland.
authors.forename: Dominique
authors.initials: D
authors.lastname: Garcin
authors.affiliation: ANSES, INRA, ENVA, UPEC, UMR_1161 Virology, LabEx IBEID, Maisons-Alfort, France.
authors.forename: Damien
authors.initials: D
authors.lastname: Vitour
authors.affiliation: Laboratory of Molecular Immunology, Department of Biotechnology, University of Rijeka, Rijeka, Croatia.
authors.forename: Ivana
authors.initials: I
authors.lastname: Munitic
authors.affiliation: INSERM, UMR_S 1197, Hôpital Paul Brousse, Villejuif, France.|Université Paris-Saclay, Paris, France.|Equipe Labellisée Ligue contre le Cancer, Villejuif, France.
authors.forename: Aimé
authors.initials: A
authors.lastname: Vazquez
authors.affiliation: INSERM, UMR_S 1197, Hôpital Paul Brousse, Villejuif, France. damien.arnoult@inserm.fr.|Université Paris-Saclay, Paris, France. damien.arnoult@inserm.fr.|Equipe Labellisée Ligue contre le Cancer, Villejuif, France. damien.arnoult@inserm.fr.
authors.forename: Damien
authors.initials: D
authors.lastname: Arnoult
mesh_terms: D018797:Cell Cycle Proteins; D000071457:DEAD Box Protein 58 / Q000235:genetics / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism / Q000821:virology*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D007113:Immunity, Innate; D050838:Interferon Regulatory Factor-3 / Q000235:genetics / Q000378:metabolism; D026901:Membrane Transport Proteins; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D012327:RNA Virus Infections / Q000276:immunology*; D012328:RNA Viruses; D011971:Receptors, Immunologic; D015398:Signal Transduction; D051196:Toll-Like Receptor 3 / Q000235:genetics / Q000378:metabolism; D035582:Transcription Factor TFIIIA / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D054875:Ubiquitination; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D018797:Cell Cycle Proteins; D050838:Interferon Regulatory Factor-3; D026901:Membrane Transport Proteins; C409460:OPTN protein, human; D011971:Receptors, Immunologic; C075080:S10 protein, Bluetongue virus; C495344:TLR3 protein, human; D051196:Toll-Like Receptor 3; D035582:Transcription Factor TFIIIA; D017361:Viral Nonstructural Proteins; D017346:Protein Serine-Threonine Kinases; C403191:TBK1 protein, human; C452820:RIGI protein, human; D000071457:DEAD Box Protein 58