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An ATG16L1-dependent pathway promotes plasma membrane repair and limits Listeria monocytogenes cell-to-cell spread

Nature Microbiology volume 3, pages 1472–1485 (2018)Cite this article

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Abstract

Plasma membrane integrity is essential for the viability of eukaryotic cells. In response to bacterial pore-forming toxins, disrupted regions of the membrane are rapidly repaired. However, the pathways that mediate plasma membrane repair are unclear. Here we show that autophagy-related (ATG) protein ATG16L1 and its binding partners ATG5 and ATG12 are required for plasma membrane repair through a pathway independent of macroautophagy. ATG16L1 is required for lysosome fusion with the plasma membrane and blebbing responses that promote membrane repair. ATG16L1 deficiency causes accumulation of cholesterol in lysosomes that contributes to defective membrane repair. Cell-to-cell spread by Listeria monocytogenes requires membrane damage by the bacterial toxin listeriolysin O, which is restricted by ATG16L1-dependent membrane repair. Cells harbouring the ATG16L1 T300A allele associated with inflammatory bowel disease were also found to accumulate cholesterol and be defective in repair, linking a common inflammatory disease to plasma membrane integrity. Thus, plasma membrane repair could be an important therapeutic target for the treatment of bacterial infections and inflammatory disorders.

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Fig. 1: ATG16L1 contributes to toxin resistance by a pathway distinct from canonical autophagy.
Fig. 2: ATG16L1 contributes to toxin resistance by promoting plasma membrane repair.
Fig. 3: ATG16L1 promotes lysosome exocytosis and plasma membrane bleb formation.
Fig. 4: ATG16L1 deficiency causes intracellular cholesterol accumulation.
Fig. 5: Cholesterol accumulation in ATG16L1-deficient cells impairs membrane repair and lysosome exocytosis.
Fig. 6: ATG16L1 limits cell-to-cell spread of Lm by promoting lysosome exocytosis and plasma membrane repair.

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Data availability

The data that support the findings of this study are available from the corresponding author upon request.

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Acknowledgements

We are grateful to E.A. Creasey, A. Draeger, V. Gerke, S. Grinstein, A. Hostettles, N. Mizushima, D. Portnoy, A. Ratner, D. Rubinsztein and R. Truant for providing reagents and advice, and to the SickKids Hospital Flow and Mass Cytometry Facility for help with flow cytometry. J.H.B. holds the Pitblado Chair in Cell Biology. Infrastructure for the Brumell Laboratory was provided by a John Evans Leadership Fund grant from the Canadian Foundation for Innovation and the Ontario Innovation Trust. S.E.O. and N.M. were supported by a fellowship from the Research Training Centre at the Hospital for Sick Children. S.E.O. and J.H. were supported by CIHR fellowships. D.B. was supported by the Dutch Digestive Foundation. D.A.A. and M.A.C. were supported by a studentship from the Research Training Committee at the Hospital for Sick Children and an NSERC CGS-M scholarship. M.A.C. was supported by a University of Toronto Open Fellowship. M.C. was supported by an NSERC PGS-D scholarship and a CIHR Training Fellowship (no. TGF-53877). This work was supported by operating grants from The Arthritis Society of Canada (grant no. RG11/013) and the Canadian Institutes of Health Research (grant nos. MOP#97756, PJT#148668 and FDN154329).

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Author notes

  1. These authors contributed equally: Joel M. J. Tan, Nora Mellouk.

Authors and Affiliations

  1. Cell Biology Program, Hospital for Sick Children, Toronto, Ontario, Canada

    Joel M. J. Tan, Nora Mellouk, Suzanne E. Osborne, Dustin A. Ammendolia, Diana N. Dyer, Ren Li, Ju Huang, Mark A. Czuczman, Marija A. Cemma, Peter K. Kim, Aleixo M. Muise & John H. Brumell

  2. Institute of Medical Science, University of Toronto, Toronto, Ontario, Canada

    Joel M. J. Tan & John H. Brumell

  3. Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada

    Dustin A. Ammendolia, Diana N. Dyer, Mark A. Czuczman, Marija A. Cemma & John H. Brumell

  4. Department of Cell Biology and Institute of Biomembranes, University Medical Center Utrecht, Utrecht, the Netherlands

    Diede Brunen, Jorik M. van Rijn & Fulvio Reggiori

  5. Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada

    Amy M. Won & Christopher M. Yip

  6. Gastrointestinal Unit and Center for the Study of Inflammatory Bowel Disease, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

    Ramnik J. Xavier

  7. Department of Microbiology and Immunology, University of Illinois at Chicago College of Medicine, Chicago, IL, USA

    Donna A. MacDuff

  8. Department of Cell Biology, University of Groningen, University Medical Centre Groningen, Groningen, The Netherlands

    Fulvio Reggiori

  9. Department of Pathology and Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, CA, USA

    Jayanta Debnath

  10. Department of Cellular Regulation and Host Defense, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan

    Tamotsu Yoshimori

  11. Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada

    Peter K. Kim, Gregory D. Fairn & Aleixo M. Muise

  12. Keenan Research Centre for Biomedical Science, St. Michael’s Hospital, Toronto, Ontario, Canada

    Gregory D. Fairn

  13. Princess Margaret Cancer Centre, University Health Network, Toronto, Ontario, Canada

    Etienne Coyaud & Brian Raught

  14. Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada

    Brian Raught

  15. Division of Gastroenterology, Hepatology, and Nutrition, Department of Paediatrics, Hospital for Sick Children, Toronto, Ontario, Canada

    Aleixo M. Muise

  16. SickKids IBD Centre, Hospital for Sick Children, Toronto, Ontario, Canada

    Aleixo M. Muise & John H. Brumell

  17. Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA, USA

    Darren E. Higgins

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Contributions

J.H.B., J.M.J.T., N.M., S.E.O. J.H. and D.E.H. designed the experiments. J.M.J.T., N.M., S.E.O., D.A.A., D.D., R.L., D.B., J.M.v.R, M.A.C., M.C., E.C. and A.M.W. performed the experiments. C.M.Y., R.J.X., D.M., F.R., T.Y., J.D., G.D.F., B.R., P.K.K. and A.M.M. contributed reagents and consultations.

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Tan, J.M.J., Mellouk, N., Osborne, S.E. et al. An ATG16L1-dependent pathway promotes plasma membrane repair and limits Listeria monocytogenes cell-to-cell spread. Nat Microbiol 3, 1472–1485 (2018). https://doi.org/10.1038/s41564-018-0293-5

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