title: Phosphorylation and linear ubiquitin direct A20 inhibition of inflammation.
abstract: Inactivation of the TNFAIP3 gene, encoding the A20 protein, is associated with critical inflammatory diseases including multiple sclerosis, rheumatoid arthritis and Crohn's disease. However, the role of A20 in attenuating inflammatory signalling is unclear owing to paradoxical in vitro and in vivo findings. Here we utilize genetically engineered mice bearing mutations in the A20 ovarian tumour (OTU)-type deubiquitinase domain or in the zinc finger-4 (ZnF4) ubiquitin-binding motif to investigate these discrepancies. We find that phosphorylation of A20 promotes cleavage of Lys63-linked polyubiquitin chains by the OTU domain and enhances ZnF4-mediated substrate ubiquitination. Additionally, levels of linear ubiquitination dictate whether A20-deficient cells die in response to tumour necrosis factor. Mechanistically, linear ubiquitin chains preserve the architecture of the TNFR1 signalling complex by blocking A20-mediated disassembly of Lys63-linked polyubiquitin scaffolds. Collectively, our studies reveal molecular mechanisms whereby A20 deubiquitinase activity and ubiquitin binding, linear ubiquitination, and cellular kinases cooperate to regulate inflammation and cell death.
authors.affiliation: Discovery Oncology, Genentech, South San Francisco, California 94080, USA.|Early Discovery Biochemistry, Genentech, South San Francisco, California 94080, USA.
authors.forename: Ingrid E
authors.initials: IE
authors.lastname: Wertz
authors.affiliation: Physiological Chemistry, Genentech, South San Francisco, California 94080, USA.
authors.forename: Kim
authors.initials: K
authors.lastname: Newton
authors.affiliation: Immunology, Genentech, South San Francisco, California 94080, USA.
authors.forename: Dhaya
authors.initials: D
authors.lastname: Seshasayee
authors.affiliation: Early Discovery Biochemistry, Genentech, South San Francisco, California 94080, USA.
authors.forename: Saritha
authors.initials: S
authors.lastname: Kusam
authors.affiliation: Early Discovery Biochemistry, Genentech, South San Francisco, California 94080, USA.
authors.forename: Cynthia
authors.initials: C
authors.lastname: Lam
authors.affiliation: Immunology, Genentech, South San Francisco, California 94080, USA.
authors.forename: Juan
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Early Discovery Biochemistry, Genentech, South San Francisco, California 94080, USA.
authors.forename: Nataliya
authors.initials: N
authors.lastname: Popovych
authors.affiliation: Early Discovery Biochemistry, Genentech, South San Francisco, California 94080, USA.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Helgason
authors.affiliation: Early Discovery Biochemistry, Genentech, South San Francisco, California 94080, USA.
authors.forename: Allyn
authors.initials: A
authors.lastname: Schoeffler
authors.affiliation: Immunology, Genentech, South San Francisco, California 94080, USA.
authors.forename: Surinder
authors.initials: S
authors.lastname: Jeet
authors.affiliation: Immunology, Genentech, South San Francisco, California 94080, USA.
authors.forename: Nandhini
authors.initials: N
authors.lastname: Ramamoorthi
authors.affiliation: Discovery Oncology, Genentech, South San Francisco, California 94080, USA.|Early Discovery Biochemistry, Genentech, South San Francisco, California 94080, USA.
authors.forename: Lorna
authors.initials: L
authors.lastname: Kategaya
authors.affiliation: Molecular Biology, Genentech, South San Francisco, California 94080, USA.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Newman
authors.affiliation: Department of Cancer Biology and Therapeutics, The John Curtin School of Medical Research, The Australian National University, Canberra, Australian Capital Territory 2601, Australia.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Horikawa
authors.affiliation: Physiological Chemistry, Genentech, South San Francisco, California 94080, USA.
authors.forename: Debra
authors.initials: D
authors.lastname: Dugger
authors.affiliation: Protein Chemistry, Genentech, South San Francisco, California 94080, USA.
authors.forename: Wendy
authors.initials: W
authors.lastname: Sandoval
authors.affiliation: Structural Biology, Genentech, South San Francisco, California 94080, USA.
authors.forename: Susmith
authors.initials: S
authors.lastname: Mukund
authors.affiliation: Early Discovery Biochemistry, Genentech, South San Francisco, California 94080, USA.
authors.forename: Anuradha
authors.initials: A
authors.lastname: Zindal
authors.affiliation: Immunology, Genentech, South San Francisco, California 94080, USA.
authors.forename: Flavius
authors.initials: F
authors.lastname: Martin
authors.affiliation: Early Discovery Biochemistry, Genentech, South San Francisco, California 94080, USA.
authors.forename: Clifford
authors.initials: C
authors.lastname: Quan
authors.affiliation: Early Discovery Biochemistry, Genentech, South San Francisco, California 94080, USA.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Tom
authors.affiliation: Early Discovery Biochemistry, Genentech, South San Francisco, California 94080, USA.
authors.forename: Wayne J
authors.initials: WJ
authors.lastname: Fairbrother
authors.affiliation: Immunology, Genentech, South San Francisco, California 94080, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Townsend
authors.affiliation: Molecular Biology, Genentech, South San Francisco, California 94080, USA.
authors.forename: Søren
authors.initials: S
authors.lastname: Warming
authors.affiliation: Immunology, Genentech, South San Francisco, California 94080, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: DeVoss
authors.affiliation: Bioinformatics, Genentech, South San Francisco, California 94080, USA.
authors.forename: Jinfeng
authors.initials: J
authors.lastname: Liu
authors.affiliation: Early Discovery Biochemistry, Genentech, South San Francisco, California 94080, USA.
authors.forename: Erin
authors.initials: E
authors.lastname: Dueber
authors.affiliation: Pathology, Genentech, South San Francisco, California 94080, USA.
authors.forename: Patrick
authors.initials: P
authors.lastname: Caplazi
authors.affiliation: Immunology, Genentech, South San Francisco, California 94080, USA.
authors.forename: Wyne P
authors.initials: WP
authors.lastname: Lee
authors.affiliation: Immunogenomics Laboratory, Immunology Division, Garvan Institute of Medical Research, 384 Victoria Street, Darlinghurst, New South Wales 2010, Sydney, Australia.
authors.forename: Christopher C
authors.initials: CC
authors.lastname: Goodnow
authors.affiliation: Immunology, Genentech, South San Francisco, California 94080, USA.
authors.forename: Mercedesz
authors.initials: M
authors.lastname: Balazs
authors.affiliation: Protein Chemistry, Genentech, South San Francisco, California 94080, USA.
authors.forename: Kebing
authors.initials: K
authors.lastname: Yu
authors.affiliation: Molecular Biology, Genentech, South San Francisco, California 94080, USA.
authors.forename: Ganesh
authors.initials: G
authors.lastname: Kolumam
authors.affiliation: Physiological Chemistry, Genentech, South San Francisco, California 94080, USA.
authors.forename: Vishva M
authors.initials: VM
authors.lastname: Dixit
mesh_terms: D000818:Animals; D016923:Cell Death; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005260:Female; D007249:Inflammation / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008239:Lysine / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D010766:Phosphorylation; D025821:Polyubiquitin / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011494:Protein Kinases / Q000378:metabolism; D015398:Signal Transduction; D000072598:Tumor Necrosis Factor alpha-Induced Protein 3; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D025801:Ubiquitin / Q000737:chemistry* / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D014409:Tumor Necrosis Factor-alpha; D025801:Ubiquitin; D025821:Polyubiquitin; D011494:Protein Kinases; D000072598:Tumor Necrosis Factor alpha-Induced Protein 3; D003546:Cysteine Endopeptidases; C513261:Tnfaip3 protein, mouse; D008239:Lysine