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Nod1 and Nod2 direct autophagy by recruiting ATG16L1 to the plasma membrane at the site of bacterial entry

Nature Immunology volume 11, pages 55–62 (2010)Cite this article

Abstract

Autophagy is emerging as a crucial defense mechanism against bacteria, but the host intracellular sensors responsible for inducing autophagy in response to bacterial infection remain unknown. Here we demonstrated that the intracellular sensors Nod1 and Nod2 are critical for the autophagic response to invasive bacteria. By a mechanism independent of the adaptor RIP2 and transcription factor NF-κB, Nod1 and Nod2 recruited the autophagy protein ATG16L1 to the plasma membrane at the bacterial entry site. In cells homozygous for the Crohn's disease–associated NOD2 frameshift mutation, mutant Nod2 failed to recruit ATG16L1 to the plasma membrane and wrapping of invading bacteria by autophagosomes was impaired. Our results link bacterial sensing by Nod proteins to the induction of autophagy and provide a functional link between Nod2 and ATG16L1, which are encoded by two of the most important genes associated with Crohn's disease.

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Figure 1: Nod agonists induce autophagy in vivo and in vitro.
Figure 2: Nod1-dependent autophagy contributes to the control of intracellular bacteria.
Figure 3: Nod1 and Nod2 interact with ATG16L1.
Figure 4: Colocalization of Nod proteins and ATG16 does not depend on RIP2.
Figure 5: The ATG16L1 polymorphism associated with Crohn's disease impairs MDP- and peptidoglycan-mediated induction of autophagy.
Figure 6: Nod proteins drive ATG16L1 to the bacterial entry site to promote autophagy.
Figure 7: The most common Nod2 mutant associated with Crohn's disease fails to induce autophagy.

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Acknowledgements

We thank K. Croitoru (University of Toronto) for Mode-K cells; N. Mizushima (Tokyo Medical and Dental Center) for ATG5-deficient MEFs; T. Mak (University of Toronto) for NEMO-deficient MEFs; R. Flavell (Yale University) for pEasyFlox; N. Mizushima (Tokyo Medical and Dental University Graduate School) for Flag–ΔN85-ATG16L1, Flag–ΔN85-ATG16L1*300A and HA-ATG5; R. Xavier (Harvard Medical School) for FL-ATG16L1–Myc and FL-ATG16L1*300A–Myc; M. D'Amato (Karolinska Institutet) for HA-Nod1-HA, HA-Nod2, HA–Nod2 L1007fs; F. Takeshita (Yokohama City University School of Medicine) for GFP-HA and pHR-SIN-CSGWΔNotI-GFP-LC3 GFP-Flag; C. Münz (University of Zurich) for rabbit polyclonal antibody to LC3 (anti-LC3 and for pHR-SIN-CSGWΔNotI-GFP-LC3); T.A. Kufer and E. Kremmer (University of Cologne and University of Munich) for rat monoclonal anti-Nod2; P. Sansonetti (Institute Pasteur) for S. flexneri M90T; D. Portnoy (University of California) for L. monocytogenes 10403S and the listeriolysin O deletion mutant; all other donors for reagents, antibodies, plasmids and bacterial strains; and M. Silverberg and J. Brumell for discussions. Supported by the Canadian Institutes for Health Research (L.H.T. and L.A.M.C.; MOP480142 to D.J.P.; and MOP81360 to S.E.G.), the Canadian Association Gastroenterology (S.H.), the University of Michigan (Y.-G.K.), Fondation Bettencourt-Schueller (J.G.M.), Fundação para Ciência e Tecnologia de Portugal (J.G.M.), the National Institutes of Health (DK61707 to G.N.), the Crohn's & Colitis Foundation of Canada (N.L.J., D.J.P. and S.E.G.) European Research Council (202283-PGN from SHAPE to VIR to I.G.B.) and the Howard Hughes Medical Institute (D.J.P.).

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Authors and Affiliations

  1. Department of Immunology, University of Toronto, Toronto, Canada

    Leonardo H Travassos, Seamus Hussey, João G Magalhães, Linda Yuan, Evelyn Chea, Lionel Le Bourhis & Dana J Philpott

  2. Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Canada

    Leticia A M Carneiro, Mahendrasingh Ramjeet, Fraser Soares & Stephen E Girardin

  3. Hospital for Sick Children, Toronto, Canada

    Seamus Hussey & Nicola L Jones

  4. Department of Pathology, The University of Michigan, Ann Arbor, Michigan, USA

    Yun-Gi Kim & Gabriel Nuñez

  5. Biology and Genetics of the Bacterial Cell Wall Group, Institut Pasteur, Paris, France, and Avenir Group, Institut National de la Santé et de la Recherche Médicale, Paris, France

    Ivo G Boneca

  6. Laboratoire de Bactériologie Moléculaire, Université Libre de Bruxelles, Brussels, Belgium

    Abdelmounaaim Allaoui

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Contributions

L.H.T. and L.A.M.C. designed and did experiments, analyzed data and wrote the paper; M.R., S.H., J.G.M., L.Y., F.S., E.C., L.L.B., I.G.B., A.A. and N.L.J. did experiments; Y.-G.K. and G.N. contributed tools; and S.E.G. and D.J.P. designed experiment, analyzed data and wrote the paper.

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Correspondence to Dana J Philpott.

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Travassos, L., Carneiro, L., Ramjeet, M. et al. Nod1 and Nod2 direct autophagy by recruiting ATG16L1 to the plasma membrane at the site of bacterial entry. Nat Immunol 11, 55–62 (2010). https://doi.org/10.1038/ni.1823

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