Successful infection by enteric bacterial pathogens depends on the ability of the bacteria to colonize the gut, replicate in host tissues and disseminate to other hosts. Pathogens such as Salmonella, Shigella and enteropathogenic and enterohaemorrhagic (EPEC and EHEC, respectively) Escherichia coli use a type III secretion system (T3SS) to deliver virulence effector proteins into host cells during infection that promote colonization and interfere with antimicrobial host responses1,2,3. Here we report that the T3SS effector NleB1 from EPEC binds to host cell death-domain-containing proteins and thereby inhibits death receptor signalling. Protein interaction studies identified FADD, TRADD and RIPK1 as binding partners of NleB1. NleB1 expressed ectopically or injected by the bacterial T3SS prevented Fas ligand or TNF-induced formation of the canonical death-inducing signalling complex (DISC) and proteolytic activation of caspase-8, an essential step in death-receptor-induced apoptosis. This inhibition depended on the N-acetylglucosamine transferase activity of NleB1, which specifically modified Arg 117 in the death domain of FADD. The importance of the death receptor apoptotic pathway to host defence was demonstrated using mice deficient in the FAS signalling pathway, which showed delayed clearance of the EPEC-like mouse pathogen Citrobacter rodentium and reversion to virulence of an nleB mutant. The activity of NleB suggests that EPEC and other attaching and effacing pathogens antagonize death-receptor-induced apoptosis of infected cells, thereby blocking a major antimicrobial host response.

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  1. 1.

    et al. Enteropathogenic and enterohaemorrhagic Escherichia coli: even more subversive elements. Mol. Microbiol. 80, 1420–1438 (2011)

  2. 2.

    , , & Salmonella effector proteins and host-cell responses. Cell. Mol. Life Sci. 68, 3687–3697 (2011)

  3. 3.

    et al. Shigella deploy multiple countermeasures against host innate immune responses. Curr. Opin. Microbiol. 14, 16–23 (2011)

  4. 4.

    et al. The death domain superfamily in intracellular signaling of apoptosis and inflammation. Annu. Rev. Immunol. 25, 561–586 (2007)

  5. 5.

    , & The many roles of FAS receptor signaling in the immune system. Immunity 30, 180–192 (2009)

  6. 6.

    et al. Compartmentalization of TNF receptor 1 signaling: internalized TNF receptosomes as death signaling vesicles. Immunity 21, 415–428 (2004)

  7. 7.

    & Induction of TNF receptor I-mediated apoptosis via two sequential signaling complexes. Cell 114, 181–190 (2003)

  8. 8.

    et al. The type III effectors NleE and NleB from enteropathogenic E. coli and OspZ from Shigella block nuclear translocation of NF-κB p65. PLoS Pathog. 6, e1000898 (2010)

  9. 9.

    et al. Cysteine methylation disrupts ubiquitin-chain sensing in NF-κB activation. Nature 481, 204–208 (2012)

  10. 10.

    et al. NleB, a bacterial effector with glycosyltransferase activity, targets GAPDH function to inhibit NF-κB activation. Cell Host Microbe 13, 87–99 (2013)

  11. 11.

    , & Structural requirements for signal-induced target binding of FADD determined by functional reconstitution of FADD deficiency. J. Biol. Chem. 280, 31360–31367 (2005)

  12. 12.

    et al. The Fas–FADD death domain complex structure reveals the basis of DISC assembly and disease mutations. Nature Struct. Mol. Biol. 17, 1324–1329 (2010)

  13. 13.

    , , , & Bcl-2 and Fas/APO-1 regulate distinct pathways to lymphocyte apoptosis. EMBO J. 14, 6136–6147 (1995)

  14. 14.

    et al. Essential role of the type III secretion system effector NleB in colonization of mice by Citrobacter rodentium. Infect. Immun. 74, 2328–2337 (2006)

  15. 15.

    et al. XIAP discriminates between type I and type II FAS-induced apoptosis. Nature 460, 1035–1039 (2009)

  16. 16.

    et al. The BH3-only protein bid is dispensable for DNA damage- and replicative stress-induced apoptosis or cell-cycle arrest. Cell 129, 423–433 (2007)

  17. 17.

    et al. Identification of a novel Citrobacter rodentium type III secreted protein, EspI, and roles of this and other secreted proteins in infection. Infect. Immun. 72, 2288–2302 (2004)

  18. 18.

    et al. Dissecting virulence: systematic and functional analyses of a pathogenicity island. Proc. Natl Acad. Sci. USA 101, 3597–3602 (2004)

  19. 19.

    , , , & Virulence is positively selected by transmission success between mammalian hosts. Curr. Biol. 17, 783–788 (2007)

  20. 20.

    et al. Bacterial genetic determinants of non-O157 STEC outbreaks and hemolytic-uremic syndrome after infection. J. Infect. Dis. 194, 819–827 (2006)

  21. 21.

    et al. Association of genomic O island 122 of Escherichia coli EDL 933 with verocytotoxin-producing Escherichia coli seropathotypes that are linked to epidemic and/or serious disease. J. Clin. Microbiol. 41, 4930–4940 (2003)

  22. 22.

    et al. Host–microbe interactions have shaped the genetic architecture of inflammatory bowel disease. Nature 491, 119–124 (2012)

  23. 23.

    & One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc. Natl Acad. Sci. USA 97, 6640–6645 (2000)

  24. 24.

    O-GlcNAc-specific antibody CTD110.6 cross-reacts with N-GlcNAc2-modified proteins induced under glucose deprivation. PLoS ONE 6, e18959 (2011)

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We gratefully acknowledge P. Bouillet for the gift of Bid−/− mice and T. Cumming for assistance with animal work. This work was supported by the Australian National Health and Medical Research Council (Program Grant no.606788 to E.L.H., Project Grants no.637332, no.1009145 to A.S., no.1009145 to L.O.R., Australia Fellowship to A.S.), the Wellcome Trust to G.F., the Juvenile Diabetes Foundation; the Leukaemia and the Lymphoma Society (New York; SCOR grant no.7413) to A.S. E.L.H was supported by an Australian Research Council Future Fellowship. J.S.P., M.K., T.W., C.G. and P.R. were supported by Australian Postgraduate Awards. This work was made possible through Victorian State Government Operational Infrastructure Support and Australian Government NHMRC IRIISS.

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  1. Department of Microbiology and Immunology, University of Melbourne, Victoria 3010, Australia

    • Jaclyn S. Pearson
    • , Cristina Giogha
    • , Sze Ying Ong
    • , Catherine L. Kennedy
    • , Michelle Kelly
    • , Tania Wong Fok Lung
    • , Patrice Riedmaier
    • , Clare V. L. Oates
    • , Ali Zaid
    • , Sabrina Mühlen
    •  & Elizabeth L. Hartland
  2. MRC Centre for Molecular Bacteriology and Infection, Department of Life Sciences, Imperial College, London SW7 2AZ, UK

    • Keith S. Robinson
    • , Valerie F. Crepin
    •  & Gad Frankel
  3. Centre for Innate Immunity and Infectious Diseases, Monash Institute of Medical Research, Victoria 3010, Australia

    • Ashley Mansell
  4. Centre for Immunology and Infectious Disease, Blizard Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London E1 2AT, UK

    • Olivier Marches
  5. Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Victoria 3010, Australia

    • Ching-Seng Ang
    •  & Nicholas A. Williamson
  6. The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia

    • Lorraine A. O’Reilly
    • , Aleksandra Bankovacki
    • , Ueli Nachbur
    • , Giuseppe Infusini
    • , Andrew I. Webb
    • , John Silke
    •  & Andreas Strasser
  7. Department of Medical Biology, University of Melbourne, Victoria 3010, Australia

    • Lorraine A. O’Reilly
    • , Andrew I. Webb
    • , John Silke
    •  & Andreas Strasser
  8. Murdoch Children’s Research Institute, Royal Children’s Hospital, Parkville, Victoria 3052, Australia

    • Elizabeth L. Hartland


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J.S.P., C.G., S.Y.O., C.L.K., M.K., K.S.R., T.W.F.L., P.R., C.V.O. and A.B. designed and performed the experiments. V.F.C. and O.M. generated reagents. C.S.A., N.A.W., G.I. and A.I.W. performed mass spectrometry analyses. L.A.O., A.Z., S.M., U.N., A.M., A.S., J.S., G.F., J.S.P. and E.L.H. contributed to experimental design. J.S.P., A.S., J.S., G.F. and E.L.H. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Elizabeth L. Hartland.

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