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Shigella flexneri suppresses NF-κB activation by inhibiting linear ubiquitin chain ligation

Nature Microbiology volume 1, Article number: 16084 (2016) Cite this article

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Abstract

The linear ubiquitin chain assembly complex (LUBAC) is a multimeric E3 ligase that catalyses M1 or linear ubiquitination of activated immune receptor signalling complexes (RSCs). Mutations that disrupt linear ubiquitin assembly lead to complex disease pathologies including immunodeficiency and autoinflammation in both humans and mice, but microbial toxins that target LUBAC function have not yet been discovered. Here, we report the identification of two homologous Shigella flexneri type III secretion system effector E3 ligases IpaH1.4 and IpaH2.5, which directly interact with LUBAC subunit Heme-oxidized IRP2 ubiquitin ligase-1 (HOIL-1L) and conjugate K48-linked ubiquitin chains to the catalytic RING-between-RING domain of HOIL-1-interacting protein (HOIP). Proteasomal degradation of HOIP leads to irreversible inactivation of linear ubiquitination and blunting of NF-κB nuclear translocation in response to tumour-necrosis factor (TNF), IL-1β and pathogen-associated molecular patterns. Loss of function studies in mammallian cells in combination with bacterial genetics explains how Shigella evades a broad spectrum of immune surveillance systems by cooperative inhibition of receptor ubiquitination and reveals the critical importance of LUBAC in host defence against pathogens.

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Figure 1: Screening S. flexneri T3SS effector proteins for disrupting the TNF-R signalling pathway.
Figure 2: IpaH1.4/2.5 mediates proteasomal degradation of HOIP.
Figure 3: IpaH1.4/2.5 post-translationally modifies Lys residues 735, 783 and 875 in HOIP.
Figure 4: IpaH1.4 is the major E3 ligase regulator of cytokine RSCs.
Figure 5: Innate immune pathways are disrupted by IpaH1.4 during infection.
Figure 6: IpaH1.4 and OspI cooperate to limit IL-1R signalling during Shigella infection.

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Acknowledgements

The authors thank K. Iwai (Kyoto University) and T. Sixma (The Netherlands Cancer Institute) for providing plasmids, R. Potts (UTSW) for reagents and advice, and the Alto laboratory for discussions. The authors acknowledge the services of the University of Texas Southwestern Medical Center proteomics core. This work was supported by grants from the National Institutes of Health (AI083359 and GM100486), the Welch Foundation (#I-1704) and the Burroughs Wellcome Fund to N.M.A.

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  1. Department of Microbiology, University of Texas Southwestern Medical Center, Dallas, 75390, Texas, USA

    Maarten F. de Jong, Zixu Liu, Didi Chen & Neal M. Alto

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  1. Maarten F. de Jong
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  2. Zixu Liu
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Contributions

M.F.d.J. and N.M.A. were responsible for study design, the analysis and interpretation of data and for writing the manuscript. M.F.d.J. performed and analysed the experiments. Z.L. and D.C. developed essential assays systems.

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Correspondence to Neal M. Alto.

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de Jong, M., Liu, Z., Chen, D. et al. Shigella flexneri suppresses NF-κB activation by inhibiting linear ubiquitin chain ligation. Nat Microbiol 1, 16084 (2016). https://doi.org/10.1038/nmicrobiol.2016.84

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