OTULIN antagonizes LUBAC signaling by specifically hydrolyzing Met1-linked polyubiquitin

Kirstin Keusekotten, Paul Ronald Elliott, Laura Glockner, Berthe Katrine Fiil, Rune Busk Damgaard, Yogesh Kulathu, Tobias Wauer, Manuela Kathrin Hospenthal, Mads Gyrd-Hansen, Daniel Krappmann, Kay Hofmann, David Komander

    Research output: Contribution to journalArticle

    185 Citations (Scopus)

    Abstract

    The linear ubiquitin (Ub) chain assembly complex (LUBAC) is an E3 ligase that specifically assembles Met1-linked (also known as linear) Ub chains that regulate nuclear factor ?B (NF-?B) signaling. Deubiquitinases (DUBs) are key regulators of Ub signaling, but a dedicated DUB for Met1 linkages has not been identified. Here, we reveal a previously unannotated human DUB, OTULIN (also known as FAM105B), which is exquisitely specific for Met1 linkages. Crystal structures of the OTULIN catalytic domain in complex with diubiquitin reveal Met1-specific Ub-binding sites and a mechanism of substrate-assisted catalysis in which the proximal Ub activates the catalytic triad of the protease. Mutation of Ub Glu16 inhibits OTULIN activity by reducing kcat 240-fold. OTULIN overexpression or knockdown affects NF-?B responses to LUBAC, TNFa, and poly(I:C) and sensitizes cells to TNFa-induced cell death. We show that OTULIN binds LUBAC and that overexpression of OTULIN prevents TNFa-induced NEMO association with ubiquitinated RIPK1. Our data suggest that OTULIN regulates Met1-polyUb signaling.
    Original languageEnglish
    Pages (from-to)1312-1326
    Number of pages15
    JournalCell
    Volume153
    Issue number6
    DOIs
    Publication statusPublished - 6 Jun 2013

    Fingerprint

    Polyubiquitin
    Ubiquitin
    Poly I-C
    Ubiquitin-Protein Ligases
    Cell death
    Catalysis
    Catalytic Domain
    Peptide Hydrolases
    Cell Death
    Crystal structure
    Binding Sites
    Association reactions
    Mutation
    Substrates
    Deubiquitinating Enzymes

    Keywords

    • Animals
    • Models, Molecular
    • Humans
    • Polyubiquitin
    • Amino Acid Sequence
    • Endopeptidases
    • Sequence Alignment
    • Molecular Sequence Data
    • Cytokines
    • Crystallography, X-Ray
    • Protein Structure, Tertiary
    • Signal Transduction
    • Catalysis

    Cite this

    Keusekotten, K., Elliott, P. R., Glockner, L., Fiil, B. K., Damgaard, R. B., Kulathu, Y., ... Komander, D. (2013). OTULIN antagonizes LUBAC signaling by specifically hydrolyzing Met1-linked polyubiquitin. Cell, 153(6), 1312-1326. https://doi.org/10.1016/j.cell.2013.05.014
    Keusekotten, Kirstin ; Elliott, Paul Ronald ; Glockner, Laura ; Fiil, Berthe Katrine ; Damgaard, Rune Busk ; Kulathu, Yogesh ; Wauer, Tobias ; Hospenthal, Manuela Kathrin ; Gyrd-Hansen, Mads ; Krappmann, Daniel ; Hofmann, Kay ; Komander, David. / OTULIN antagonizes LUBAC signaling by specifically hydrolyzing Met1-linked polyubiquitin. In: Cell. 2013 ; Vol. 153, No. 6. pp. 1312-1326.
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    abstract = "The linear ubiquitin (Ub) chain assembly complex (LUBAC) is an E3 ligase that specifically assembles Met1-linked (also known as linear) Ub chains that regulate nuclear factor ?B (NF-?B) signaling. Deubiquitinases (DUBs) are key regulators of Ub signaling, but a dedicated DUB for Met1 linkages has not been identified. Here, we reveal a previously unannotated human DUB, OTULIN (also known as FAM105B), which is exquisitely specific for Met1 linkages. Crystal structures of the OTULIN catalytic domain in complex with diubiquitin reveal Met1-specific Ub-binding sites and a mechanism of substrate-assisted catalysis in which the proximal Ub activates the catalytic triad of the protease. Mutation of Ub Glu16 inhibits OTULIN activity by reducing kcat 240-fold. OTULIN overexpression or knockdown affects NF-?B responses to LUBAC, TNFa, and poly(I:C) and sensitizes cells to TNFa-induced cell death. We show that OTULIN binds LUBAC and that overexpression of OTULIN prevents TNFa-induced NEMO association with ubiquitinated RIPK1. Our data suggest that OTULIN regulates Met1-polyUb signaling.",
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    Keusekotten, K, Elliott, PR, Glockner, L, Fiil, BK, Damgaard, RB, Kulathu, Y, Wauer, T, Hospenthal, MK, Gyrd-Hansen, M, Krappmann, D, Hofmann, K & Komander, D 2013, 'OTULIN antagonizes LUBAC signaling by specifically hydrolyzing Met1-linked polyubiquitin', Cell, vol. 153, no. 6, pp. 1312-1326. https://doi.org/10.1016/j.cell.2013.05.014

    OTULIN antagonizes LUBAC signaling by specifically hydrolyzing Met1-linked polyubiquitin. / Keusekotten, Kirstin; Elliott, Paul Ronald; Glockner, Laura; Fiil, Berthe Katrine; Damgaard, Rune Busk; Kulathu, Yogesh; Wauer, Tobias; Hospenthal, Manuela Kathrin; Gyrd-Hansen, Mads; Krappmann, Daniel; Hofmann, Kay; Komander, David.

    In: Cell, Vol. 153, No. 6, 06.06.2013, p. 1312-1326.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - OTULIN antagonizes LUBAC signaling by specifically hydrolyzing Met1-linked polyubiquitin

    AU - Keusekotten, Kirstin

    AU - Elliott, Paul Ronald

    AU - Glockner, Laura

    AU - Fiil, Berthe Katrine

    AU - Damgaard, Rune Busk

    AU - Kulathu, Yogesh

    AU - Wauer, Tobias

    AU - Hospenthal, Manuela Kathrin

    AU - Gyrd-Hansen, Mads

    AU - Krappmann, Daniel

    AU - Hofmann, Kay

    AU - Komander, David

    N1 - Copyright © 2013 Elsevier Inc. All rights reserved.

    PY - 2013/6/6

    Y1 - 2013/6/6

    N2 - The linear ubiquitin (Ub) chain assembly complex (LUBAC) is an E3 ligase that specifically assembles Met1-linked (also known as linear) Ub chains that regulate nuclear factor ?B (NF-?B) signaling. Deubiquitinases (DUBs) are key regulators of Ub signaling, but a dedicated DUB for Met1 linkages has not been identified. Here, we reveal a previously unannotated human DUB, OTULIN (also known as FAM105B), which is exquisitely specific for Met1 linkages. Crystal structures of the OTULIN catalytic domain in complex with diubiquitin reveal Met1-specific Ub-binding sites and a mechanism of substrate-assisted catalysis in which the proximal Ub activates the catalytic triad of the protease. Mutation of Ub Glu16 inhibits OTULIN activity by reducing kcat 240-fold. OTULIN overexpression or knockdown affects NF-?B responses to LUBAC, TNFa, and poly(I:C) and sensitizes cells to TNFa-induced cell death. We show that OTULIN binds LUBAC and that overexpression of OTULIN prevents TNFa-induced NEMO association with ubiquitinated RIPK1. Our data suggest that OTULIN regulates Met1-polyUb signaling.

    AB - The linear ubiquitin (Ub) chain assembly complex (LUBAC) is an E3 ligase that specifically assembles Met1-linked (also known as linear) Ub chains that regulate nuclear factor ?B (NF-?B) signaling. Deubiquitinases (DUBs) are key regulators of Ub signaling, but a dedicated DUB for Met1 linkages has not been identified. Here, we reveal a previously unannotated human DUB, OTULIN (also known as FAM105B), which is exquisitely specific for Met1 linkages. Crystal structures of the OTULIN catalytic domain in complex with diubiquitin reveal Met1-specific Ub-binding sites and a mechanism of substrate-assisted catalysis in which the proximal Ub activates the catalytic triad of the protease. Mutation of Ub Glu16 inhibits OTULIN activity by reducing kcat 240-fold. OTULIN overexpression or knockdown affects NF-?B responses to LUBAC, TNFa, and poly(I:C) and sensitizes cells to TNFa-induced cell death. We show that OTULIN binds LUBAC and that overexpression of OTULIN prevents TNFa-induced NEMO association with ubiquitinated RIPK1. Our data suggest that OTULIN regulates Met1-polyUb signaling.

    KW - Animals

    KW - Models, Molecular

    KW - Humans

    KW - Polyubiquitin

    KW - Amino Acid Sequence

    KW - Endopeptidases

    KW - Sequence Alignment

    KW - Molecular Sequence Data

    KW - Cytokines

    KW - Crystallography, X-Ray

    KW - Protein Structure, Tertiary

    KW - Signal Transduction

    KW - Catalysis

    U2 - 10.1016/j.cell.2013.05.014

    DO - 10.1016/j.cell.2013.05.014

    M3 - Article

    VL - 153

    SP - 1312

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    JF - Cell

    SN - 0092-8674

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    Keusekotten K, Elliott PR, Glockner L, Fiil BK, Damgaard RB, Kulathu Y et al. OTULIN antagonizes LUBAC signaling by specifically hydrolyzing Met1-linked polyubiquitin. Cell. 2013 Jun 6;153(6):1312-1326. https://doi.org/10.1016/j.cell.2013.05.014