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Pathogen blocks host death receptor signalling by arginine GlcNAcylation of death domains

Nature volume 501, pages 242–246 (2013)Cite this article

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Abstract

The tumour necrosis factor (TNF) family is crucial for immune homeostasis, cell death and inflammation. These cytokines are recognized by members of the TNF receptor (TNFR) family of death receptors, including TNFR1 and TNFR2, and FAS and TNF-related apoptosis-inducing ligand (TRAIL) receptors1. Death receptor signalling requires death-domain-mediated homotypic/heterotypic interactions between the receptor and its downstream adaptors, including TNFR1-associated death domain protein (TRADD) and FAS-associated death domain protein (FADD)2. Here we discover that death domains in several proteins, including TRADD, FADD, RIPK1 and TNFR1, were directly inactivated by NleB, an enteropathogenic Escherichia coli (EPEC) type III secretion system effector known to inhibit host nuclear factor-κB (NF-κB) signalling3,4. NleB contained an unprecedented N-acetylglucosamine (GlcNAc) transferase activity that specifically modified a conserved arginine in these death domains (Arg 235 in the TRADD death domain). NleB GlcNAcylation (the addition of GlcNAc onto a protein side chain) of death domains blocked homotypic/heterotypic death domain interactions and assembly of the oligomeric TNFR1 complex, thereby disrupting TNF signalling in EPEC-infected cells, including NF-κB signalling, apoptosis and necroptosis. Type-III-delivered NleB also blocked FAS ligand and TRAIL-induced cell death by preventing formation of a FADD-mediated death-inducing signalling complex (DISC). The arginine GlcNAc transferase activity of NleB was required for bacterial colonization in the mouse model of EPEC infection. The mechanism of action of NleB represents a new model by which bacteria counteract host defences, and also a previously unappreciated post-translational modification.

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Figure 1: NleB blocks TNF-α signalling by directly targeting the TRADD death domain.
Figure 2: NleB GlcNAcylates the TRADD death domain and disrupts its oligomerization.
Figure 3: NleB GlcNAcylates Arg 235 that is required for TRADD function.
Figure 4: NleB GlcNAcylates FADD and other death domain proteins, and blocks FAS- and TRAIL-induced apoptosis.
Figure 5: Disrupting several death receptor pathways by NleB GlcNAc transferase activity that is required for bacterial colonization in vivo.

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Acknowledgements

We thank I. Rosenshine for providing nleBE deletion EPEC strains, H. Wu for the TNFR1/2 plasmid, K. Iwai for LUBAC antibodies, H. Sakurai for the p-TAK1 antibody, Z. Yang for yeast two-hybrid screen, L. Feng for synthesizing Ac4GlcNAc-d3, M. Dong for accessing the ETD instrument and X. Liu for assistance with mass spectrometry data analyses. We also thank members of the Shao laboratory for discussions and technical assistance. The research was supported in part by an International Early Career Scientist grant from the Howard Hughes Medical Institute to F.S. This work was also supported by the National Basic Research Program of China 973 Programs 2010CB835400 and 2012CB518700 to F.S. and 2010CB835204 to S.C.

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  1. Shan Li and Li Zhang: These authors contributed equally to this work.

Authors and Affiliations

  1. College of Biological Sciences, China Agricultural University, Beijing, 100094, China

    Shan Li & Qing Yao

  2. National Institute of Biological Sciences, Beijing, 102206, China

    Shan Li, Li Zhang, Qing Yao, Lin Li, Na Dong, Wenqing Gao, Xiaojun Ding, Liming Sun, She Chen & Feng Shao

  3. Graduate School of Chinese Academy of Medical Sciences and Beijing Union Medical College, Beijing, 100730, China

    Li Zhang

  4. Department of Chemical Biology, Beijing National Laboratory for Molecular Sciences, College of Chemistry and Molecular Engineering, Peking University, Beijing, 100871, China

    Jie Rong & Xing Chen

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Contributions

S.L. and L.Z. performed most experiments, assisted by Q.Y., N.D. and W.G. L.L., X.D. and S.C. performed mass spectrometry analyses and S.C. analysed the data. L.S., J.R. and X.C. contributed reagents and analytic tools. S.L., L.Z. and F.S. analysed the data and wrote the manuscript. All authors discussed the results and commented on the manuscript.

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Correspondence to She Chen or Feng Shao.

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Li, S., Zhang, L., Yao, Q. et al. Pathogen blocks host death receptor signalling by arginine GlcNAcylation of death domains. Nature 501, 242–246 (2013). https://doi.org/10.1038/nature12436

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