PMID- 28159912 OWN - NLM STAT- MEDLINE DCOM- 20170615 LR - 20220129 IS - 1470-8728 (Electronic) IS - 0264-6021 (Print) IS - 0264-6021 (Linking) VI - 474 IP - 7 DP - 2017 Mar 15 TI - Identification of TBK1 complexes required for the phosphorylation of IRF3 and the production of interferon beta. PG - 1163-1174 LID - 10.1042/BCJ20160992 [doi] AB - The double-stranded RNA mimetic poly(I:C) and lipopolysaccharide (LPS) activate Toll-like receptors 3 (TLR3) and TLR4, respectively, triggering the activation of TANK (TRAF family member-associated NF-kappaB activator)-binding kinase 1 (TBK1) complexes, the phosphorylation of interferon regulatory factor 3 (IRF3) and transcription of the interferon beta (IFNbeta) gene. Here, we demonstrate that the TANK-TBK1 and optineurin (OPTN)-TBK1 complexes control this pathway. The poly(I:C)- or LPS-stimulated phosphorylation of IRF3 at Ser396 and production of IFNbeta were greatly reduced in bone marrow-derived macrophages (BMDMs) from TANK knockout (KO) mice crossed to knockin mice expressing the ubiquitin-binding-defective OPTN[D477N] mutant. In contrast, IRF3 phosphorylation and IFNbeta production were not reduced significantly in BMDM from OPTN[D477N] knockin mice and only reduced partially in TANK KO BMDM. The TLR3/TLR4-dependent phosphorylation of IRF3 and IFNbeta gene transcription were not decreased in macrophages from OPTN[D477N] crossed to mice deficient in IkappaB kinase epsilon, a TANK-binding kinase related to TBK1. In contrast with the OPTN-TBK1 complex, TBK1 associated with OPTN[D477N] did not undergo phosphorylation at Ser172 in response to poly(I:C) or LPS, indicating that the interaction of ubiquitin chains with OPTN is required to activate OPTN-TBK1 in BMDM. The phosphorylation of IRF3 and IFNbeta production induced by Sendai virus infection were unimpaired in BMDM from TANK KO x OPTN[D477N] mice, suggesting that other/additional TBK1 complexes control the RIG-I-like receptor-dependent production of IFNbeta. Finally, we present evidence that, in human HACAT cells, the poly(I:C)-dependent phosphorylation of TBK1 at Ser172 involves a novel TBK1-activating kinase(s). CI - (c) 2017 The Author(s). FAU - Bakshi, Siddharth AU - Bakshi S AD - MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee DD1 5EH, U.K. FAU - Taylor, Jordan AU - Taylor J AD - MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee DD1 5EH, U.K. FAU - Strickson, Sam AU - Strickson S AD - MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee DD1 5EH, U.K. FAU - McCartney, Thomas AU - McCartney T AD - MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee DD1 5EH, U.K. FAU - Cohen, Philip AU - Cohen P AD - MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, Dundee DD1 5EH, U.K. p.cohen@dundee.ac.uk. LA - eng GR - MC_UU_12016/11/MRC_/Medical Research Council/United Kingdom GR - MR/K000985/1/MRC_/Medical Research Council/United Kingdom GR - WT100294/Wellcome Trust/United Kingdom GR - WT109112/Z/15/Z/Wellcome Trust/United Kingdom PT - Journal Article DEP - 20170315 PL - England TA - Biochem J JT - The Biochemical journal JID - 2984726R RN - 0 (Adaptor Proteins, Signal Transducing) RN - 0 (Adaptor Proteins, Vesicular Transport) RN - 0 (Cell Cycle Proteins) RN - 0 (Eye Proteins) RN - 0 (Interferon Regulatory Factor-3) RN - 0 (Irf3 protein, mouse) RN - 0 (Lipopolysaccharides) RN - 0 (Membrane Transport Proteins) RN - 0 (Optn protein, mouse) RN - 0 (RNA, Small Interfering) RN - 0 (TICAM1 protein, human) RN - 0 (TLR3 protein, mouse) RN - 0 (Tank protein, mouse) RN - 0 (Tlr4 protein, mouse) RN - 0 (Toll-Like Receptor 3) RN - 0 (Toll-Like Receptor 4) RN - 77238-31-4 (Interferon-beta) RN - EC 2.7.1.- (Tbk1 protein, mouse) RN - EC 2.7.11.1 (Protein Serine-Threonine Kinases) RN - EC 2.7.11.10 (I-kappa B Kinase) RN - EC 2.7.11.25 (MAP Kinase Kinase Kinases) RN - EC 2.7.11.25 (MAP kinase kinase kinase 7) RN - O84C90HH2L (Poly I-C) SB - IM MH - Adaptor Proteins, Signal Transducing/deficiency/genetics MH - Adaptor Proteins, Vesicular Transport/antagonists & inhibitors/genetics/immunology MH - Animals MH - Cell Cycle Proteins MH - Cell Line, Transformed MH - Eye Proteins/genetics/immunology MH - Gene Knock-In Techniques MH - Humans MH - I-kappa B Kinase/deficiency/genetics MH - Interferon Regulatory Factor-3/*genetics/immunology MH - Interferon-beta/*genetics/immunology MH - Keratinocytes/cytology/drug effects/immunology MH - Lipopolysaccharides/pharmacology MH - MAP Kinase Kinase Kinases/antagonists & inhibitors/genetics/immunology MH - Macrophages/cytology/drug effects/*immunology MH - Membrane Transport Proteins MH - Mice MH - Mice, Knockout MH - Phosphorylation MH - Poly I-C/pharmacology MH - Primary Cell Culture MH - Protein Serine-Threonine Kinases/*genetics/immunology MH - RNA, Small Interfering/genetics/metabolism MH - Toll-Like Receptor 3/*genetics/immunology MH - Toll-Like Receptor 4/*genetics/immunology PMC - PMC5350611 OTO - NOTNLM OT - IRF3 OT - LPS OT - TBK1 OT - TLR3 OT - interferon OT - ubiquitin COIS- The Authors declare that there are no competing interests associated with the manuscript. EDAT- 2017/02/06 06:00 MHDA- 2017/06/16 06:00 PMCR- 2017/03/15 CRDT- 2017/02/05 06:00 PHST- 2016/11/08 00:00 [received] PHST- 2017/01/23 00:00 [revised] PHST- 2017/02/03 00:00 [accepted] PHST- 2017/02/06 06:00 [pubmed] PHST- 2017/06/16 06:00 [medline] PHST- 2017/02/05 06:00 [entrez] PHST- 2017/03/15 00:00 [pmc-release] AID - BCJ20160992 [pii] AID - BCJ-2016-0992 [pii] AID - 10.1042/BCJ20160992 [doi] PST - epublish SO - Biochem J. 2017 Mar 15;474(7):1163-1174. doi: 10.1042/BCJ20160992.