title: Gliotoxin suppresses NF-κB activation by selectively inhibiting linear ubiquitin chain assembly complex (LUBAC).
abstract: A linear ubiquitin chain, which consists of ubiquitin molecules linked via their N- and C-termini, is formed by a linear ubiquitin chain assembly complex (LUBAC) composed of HOIP, HOIL-1L, and SHARPIN, and conjugation of a linear ubiquitin chain on the NF-κB essential modulator (NEMO) is deeply involved in NF-κB activation induced by various signals. Since abnormal activation of NF-κB is associated with inflammatory disease and malignancy, we searched for an inhibitor of LUBAC by high-throughput screening (HTS) with a Tb(3+)-fluorescein FRET system. As a result, we found that the fungal metabolite gliotoxin inhibits LUBAC selectively by binding to the RING-IBR-RING domain of HOIP, the catalytic center of LUBAC. Gliotoxin has been well-known as an inhibitor of NF-κB activation, though its action mechanism has remained elusive. Here, we show that gliotoxin inhibits signal-induced NF-κB activation by selectively inhibiting LUBAC-mediated linear ubiquitin chain formation.
authors.affiliation: †Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Sakamoto
authors.affiliation: ‡Open Innovation Center for Drug Discovery, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Shinichiro
authors.initials: S
authors.lastname: Egashira
authors.affiliation: ‡Open Innovation Center for Drug Discovery, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Nae
authors.initials: N
authors.lastname: Saito
authors.affiliation: §Department of Molecular Cell Biology, Graduate School of Medicine, Osaka City University, Osaka 545-8585, Japan.
authors.forename: Takayoshi
authors.initials: T
authors.lastname: Kirisako
authors.affiliation: ∥Structural Biology Research Center, Photon Factory, Institute of Materials Structure Science, High-Energy Accelerator Research Organization (KEK), Tsukuba, Ibaraki 305-8501, Japan.
authors.forename: Simon
authors.initials: S
authors.lastname: Miller
authors.affiliation: ⧫Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto-shi, Kyoto 606-8501, Japan.
authors.forename: Yoshiteru
authors.initials: Y
authors.lastname: Sasaki
authors.affiliation: ⧫Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto-shi, Kyoto 606-8501, Japan.
authors.forename: Tadahiko
authors.initials: T
authors.lastname: Matsumoto
authors.affiliation: ⊥Medical Innovation Center Laboratory for Malignancy Control Research/DSK, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto-shi, Kyoto 606-8501, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Shimonishi
authors.affiliation: †Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Komatsu
authors.affiliation: †Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Terai
authors.affiliation: †Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Tasuku
authors.initials: T
authors.lastname: Ueno
authors.affiliation: †Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Kenjiro
authors.initials: K
authors.lastname: Hanaoka
authors.affiliation: ‡Open Innovation Center for Drug Discovery, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Hirotatsu
authors.initials: H
authors.lastname: Kojima
authors.affiliation: ‡Open Innovation Center for Drug Discovery, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Takayoshi
authors.initials: T
authors.lastname: Okabe
authors.affiliation: ∥Structural Biology Research Center, Photon Factory, Institute of Materials Structure Science, High-Energy Accelerator Research Organization (KEK), Tsukuba, Ibaraki 305-8501, Japan.|#Department of Structural Biology, Stanford University School of Medicine, Stanford, California 94305, United States.|▽SLAC National Accelerator Laboratory, Menlo Park, California 94025, United States.
authors.forename: Soichi
authors.initials: S
authors.lastname: Wakatsuki
authors.affiliation: §Department of Molecular Cell Biology, Graduate School of Medicine, Osaka City University, Osaka 545-8585, Japan.|○Department of Biophysics and Biochemistry, Graduate School of Medicine, Osaka University, Suita, Osaka 565-0871, Japan.|⧫Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto-shi, Kyoto 606-8501, Japan.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Iwai
authors.affiliation: ‡Open Innovation Center for Drug Discovery, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Nagano
mesh_terms: D004305:Dose-Response Relationship, Drug; D019793:Fluorescein / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry; D005786:Gene Expression Regulation; D005912:Gliotoxin / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D051550:I-kappa B Kinase / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D007166:Immunosuppressive Agents / Q000737:chemistry / Q000494:pharmacology*; D019169:Jurkat Cells; D008213:Lymphocyte Activation / Q000187:drug effects; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000276:immunology; D015398:Signal Transduction; D013725:Terbium / Q000737:chemistry; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D025801:Ubiquitin / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000276:immunology; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000276:immunology; D054875:Ubiquitination / Q000187:drug effects; D014452:Ubiquitins / Q000235:genetics / Q000276:immunology
chemical_list: D005456:Fluorescent Dyes; C491731:IKBKG protein, human; D007166:Immunosuppressive Agents; D016328:NF-kappa B; C575183:SHARPIN protein, human; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha; D025801:Ubiquitin; D014452:Ubiquitins; D013725:Terbium; D005912:Gliotoxin; C504507:RBCK1 protein, human; C558338:RNF31 protein, human; D044767:Ubiquitin-Protein Ligases; D051550:I-kappa B Kinase; D019793:Fluorescein