title: Posttranslational Modification of HOIP Blocks Toll-Like Receptor 4-Mediated Linear-Ubiquitin-Chain Formation.
abstract: Linear ubiquitination is an atypical posttranslational modification catalyzed by the linear-ubiquitin-chain assembly complex (LUBAC), containing HOIP, HOIL-1L, and Sharpin. LUBAC facilitates NF-κB activation and inflammation upon receptor stimulation by ligating linear ubiquitin chains to critical signaling molecules. Indeed, linear-ubiquitination-dependent signaling is essential to prevent pyogenic bacterial infections that can lead to death. While linear ubiquitination is essential for intracellular receptor signaling upon microbial infection, this response must be measured and stopped to avoid tissue damage and autoimmunity. While LUBAC is activated upon bacterial stimulation, the mechanisms regulating LUBAC activity in response to bacterial stimuli have remained elusive. We demonstrate that LUBAC activity itself is downregulated through ubiquitination, specifically, ubiquitination of the catalytic subunit HOIP at the carboxyl-terminal lysine 1056. Ubiquitination of Lys1056 dynamically altered HOIP conformation, resulting in the suppression of its catalytic activity. Consequently, HOIP Lys1056-to-Arg mutation led not only to persistent LUBAC activity but also to prolonged NF-κB activation induced by bacterial lipopolysaccharide-mediated Toll-like receptor 4 (TLR4) stimulation, whereas it showed no effect on NF-κB activation induced by CD40 stimulation. This study describes a novel posttranslational regulation of LUBAC-mediated linear ubiquitination that is critical for specifically directing TLR4-mediated NF-κB activation.
  
  
  OBJECTIVE
  Posttranslational modification of proteins enables cells to respond quickly to infections and immune stimuli in a tightly controlled manner. Specifically, covalent modification of proteins with the small protein ubiquitin is essential for cells to initiate and terminate immune signaling in response to bacterial and viral infection. This process is controlled by ubiquitin ligase enzymes, which themselves must be regulated to prevent persistent and deleterious immune signaling. However, how this regulation is achieved is poorly understood. This paper reports a novel ubiquitination event of the atypical ubiquitin ligase HOIP that is required to terminate bacterial lipopolysaccharide (LPS)-induced TLR4 immune signaling. Ubiquitination causes the HOIP ligase to undergo a conformational change, which blocks its enzymatic activity and ultimately terminates LPS-induced TLR4 signaling. These findings provide a new mechanism for controlling HOIP ligase activity that is vital to properly regulate a proinflammatory immune response.
authors.affiliation: Department of Molecular Microbiology and Immunology, Keck School of Medicine, Los Angeles, California, USA.
authors.forename: James
authors.initials: J
authors.lastname: Bowman
authors.affiliation: Department of Molecular Microbiology and Immunology, Keck School of Medicine, Los Angeles, California, USA Abbott Diagnostics, Abbott Park, Illinois, USA.
authors.forename: Mary A
authors.initials: MA
authors.lastname: Rodgers
authors.affiliation: Department of Molecular Microbiology and Immunology, Keck School of Medicine, Los Angeles, California, USA.
authors.forename: Mude
authors.initials: M
authors.lastname: Shi
authors.affiliation: Department of Molecular Microbiology and Immunology, Keck School of Medicine, Los Angeles, California, USA.
authors.forename: Rina
authors.initials: R
authors.lastname: Amatya
authors.affiliation: Department of Pediatrics, University of Iowa, Iowa City, Iowa, USA.
authors.forename: Bruce
authors.initials: B
authors.lastname: Hostager
authors.affiliation: Department of Molecular and Cellular Physiology, Graduate School of Medicine, Kyoto University, Yoshida, Kyoto, Japan.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Iwai
authors.affiliation: Department of Molecular Microbiology and Immunology, Keck School of Medicine, Los Angeles, California, USA.
authors.forename: Shou-Jiang
authors.initials: SJ
authors.lastname: Gao
authors.affiliation: Department of Molecular Microbiology and Immunology, Keck School of Medicine, Los Angeles, California, USA Department of Pharmacology and Pharmaceutical Sciences, School of Pharmacy, University of Southern California, Los Angeles, California, USA jaeujung@med.usc.edu.
authors.forename: Jae U
authors.initials: JU
authors.lastname: Jung
mesh_terms: D007113:Immunity, Innate; D008070:Lipopolysaccharides / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D011487:Protein Conformation; D055503:Protein Multimerization; D011499:Protein Processing, Post-Translational; D051197:Toll-Like Receptor 4 / Q000378:metabolism*; D025801:Ubiquitin / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D008070:Lipopolysaccharides; D016328:NF-kappa B; D051197:Toll-Like Receptor 4; D025801:Ubiquitin; C558338:RNF31 protein, human; D044767:Ubiquitin-Protein Ligases