title: Inhibition of Canonical NF-κB Signaling by a Small Molecule Targeting NEMO-Ubiquitin Interaction.
abstract: The IκB kinase (IKK) complex acts as the gatekeeper of canonical NF-κB signaling, thereby regulating immunity, inflammation and cancer. It consists of the catalytic subunits IKKα and IKKβ and the regulatory subunit NEMO/IKKγ. Here, we show that the ubiquitin binding domain (UBAN) in NEMO is essential for IKK/NF-κB activation in response to TNFα, but not IL-1β stimulation. By screening a natural compound library we identified an anthraquinone derivative that acts as an inhibitor of NEMO-ubiquitin binding (iNUB). Using biochemical and NMR experiments we demonstrate that iNUB binds to NEMOUBAN and competes for interaction with methionine-1-linked linear ubiquitin chains. iNUB inhibited NF-κB activation upon UBAN-dependent TNFα and TCR/CD28, but not UBAN-independent IL-1β stimulation. Moreover, iNUB was selectively killing lymphoma cells that are addicted to chronic B-cell receptor triggered IKK/NF-κB activation. Thus, iNUB disrupts the NEMO-ubiquitin protein-protein interaction interface and thereby inhibits physiological and pathological NF-κB signaling.
authors.affiliation: Research Unit Cellular Signal Integration, Helmholtz Zentrum München für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.
authors.forename: Michelle
authors.initials: M
authors.lastname: Vincendeau
authors.affiliation: Research Unit Cellular Signal Integration, Helmholtz Zentrum München für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.|Assay Development and Screening Platform, Institute of Molecular Toxicology and Pharmacology, Helmholtz Zentrum München für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.
authors.forename: Kamyar
authors.initials: K
authors.lastname: Hadian
authors.affiliation: Institute of Structural Biology, Helmholtz Zentrum München für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.|Center for Integrated Protein Science Munich at Biomolecular NMR Spectroscopy, Department Chemie, Technische Universität München, Lichtenbergstr. 4, 85747 Garching, Germany.
authors.forename: Ana C
authors.initials: AC
authors.lastname: Messias
authors.affiliation: Assay Development and Screening Platform, Institute of Molecular Toxicology and Pharmacology, Helmholtz Zentrum München für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.
authors.forename: Jara K
authors.initials: JK
authors.lastname: Brenke
authors.affiliation: Institute of Structural Biology, Helmholtz Zentrum München für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.
authors.forename: Jenny
authors.initials: J
authors.lastname: Halander
authors.affiliation: Research Unit Cellular Signal Integration, Helmholtz Zentrum München für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.
authors.forename: Richard
authors.initials: R
authors.lastname: Griesbach
authors.affiliation: Research Unit Cellular Signal Integration, Helmholtz Zentrum München für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.
authors.forename: Ute
authors.initials: U
authors.lastname: Greczmiel
authors.affiliation: Research Unit Cellular Signal Integration, Helmholtz Zentrum München für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.
authors.forename: Arianna
authors.initials: A
authors.lastname: Bertossi
authors.affiliation: Institute of Structural Biology, Helmholtz Zentrum München für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.|Center for Integrated Protein Science Munich at Biomolecular NMR Spectroscopy, Department Chemie, Technische Universität München, Lichtenbergstr. 4, 85747 Garching, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Stehle
authors.affiliation: Research Unit Cellular Signal Integration, Helmholtz Zentrum München für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Nagel
authors.affiliation: Research Unit Cellular Signal Integration, Helmholtz Zentrum München für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Demski
authors.affiliation: Institute of Structural Biology, Helmholtz Zentrum München für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.
authors.forename: Hana
authors.initials: H
authors.lastname: Velvarska
authors.affiliation: Institute of Structural Biology, Helmholtz Zentrum München für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.|Biomedical Center of the Ludwig-Maximilians University München, Department of Cell Biology, Großhadener Str. 9, 82152 Planegg-Martinsried, Germany.
authors.forename: Dierk
authors.initials: D
authors.lastname: Niessing
authors.affiliation: Protein Expression and Purification Facility, Institute of Structural Biology, Helmholtz Zentrum München für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.
authors.forename: Arie
authors.initials: A
authors.lastname: Geerlof
authors.affiliation: Institute of Structural Biology, Helmholtz Zentrum München für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.|Center for Integrated Protein Science Munich at Biomolecular NMR Spectroscopy, Department Chemie, Technische Universität München, Lichtenbergstr. 4, 85747 Garching, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Sattler
authors.affiliation: Research Unit Cellular Signal Integration, Helmholtz Zentrum München für Gesundheit und Umwelt, Ingolstädter Landstr. 1, 85764 Neuherberg, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Krappmann
mesh_terms: D000818:Animals; D000880:Anthraquinones / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical; D006367:HeLa Cells; D006801:Humans; D053583:Interleukin-1beta / Q000502:physiology; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D015398:Signal Transduction / Q000187:drug effects*; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology; D025801:Ubiquitin / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: D000880:Anthraquinones; D053583:Interleukin-1beta; D047908:Intracellular Signaling Peptides and Proteins; C514981:NEMO protein, mouse; D016328:NF-kappa B; C439524:TNF protein, human; D014409:Tumor Necrosis Factor-alpha; D025801:Ubiquitin