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Intermediate filaments enable pathogen docking to trigger type 3 effector translocation

Nature Microbiology volume 1, Article number: 16025 (2016) Cite this article

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Abstract

Type 3 secretion systems (T3SSs) of bacterial pathogens translocate bacterial effector proteins that mediate disease into the eukaryotic cytosol. Effectors traverse the plasma membrane through a translocon pore formed by T3SS proteins. In a genome-wide selection, we identified the intermediate filament vimentin as required for infection by the T3SS-dependent pathogen S. flexneri. We found that vimentin is required for efficient T3SS translocation of effectors by S. flexneri and other pathogens that use T3SS, Salmonella enterica serovar Typhimurium and Yersinia pseudotuberculosis. Vimentin and the intestinal epithelial intermediate filament keratin 18 interact with the C-terminus of the Shigella translocon pore protein IpaC. Vimentin and its interaction with IpaC are dispensable for pore formation, but are required for stable docking of S. flexneri to cells; moreover, stable docking triggers effector secretion. These findings establish that stable docking of the bacterium specifically requires intermediate filaments, is a process distinct from pore formation, and is a prerequisite for effector secretion.

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Figure 1: Efficient T3SS translocation depends on intermediate filaments.
Figure 2: The S. flexneri T3SS translocon pore protein IpaC and its homologues in other pathogens interact with intermediate filaments and this interaction is required for efficient translocation.
Figure 3: Intermediate filaments are dispensable for translocon pore formation.
Figure 4: E. coli pSfT3SS reproduces S. flexneri phenotypes.
Figure 5: The interaction of intermediate filaments with the translocon pore is required for efficient docking of S. flexneri.
Figure 6: Activation of effector secretion requires S. flexneri docking.

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Acknowledgements

We thank Victor Faundez, Cammie Lesser, Wendy Picking and William Picking, Joan Mecsas, Victoria Auerbuch-Stone, Beth McCormick and Ana Maldonado-Contreras, and Claude Parsot for cell lines, bacterial strains, reagents, technical assistance and protocols, Alexandra Wiscovitch for technical assistance, and Matthew Brown for critical reading of the manuscript. This work was funded by National Institutes of Health (NIH) RO1 AI081724 (to M.B.G.), NIH T32 AI007061 (to B.C.R.), NIH F32 AI092967 (to L.A.B.), NIH F32 AI114162 (to B.C.R.), a career development award through the New England Regional Center of Excellence in Biodefense and Emerging Infectious Disease and the Institute of Chemistry and Cell Biology (ICCB)-Longwood Facility at Harvard Medical School (NIH U54 AI057159, to L.M.S.), and the Coordination for the Improvement of Higher Education Personnel (CAPES) Foundation, Ministry of Education of Brazil, Brasilia-DF, 70040-020, Brazil (to A.L.Q.).

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  1. Luisa M. Stamm, Matthijs Raaben, Bobby Brooke Herrera & Leigh A. Baxt

    Present address: †Present addresses: Liver Diseases Therapeutic Area, Gilead Sciences, Foster City, California 94404, USA (L.M.S.). Netherlands Cancer Institute, 1066 CX, Amsterdam, The Netherlands (M.R.). Harvard T.H. Chan School of Public Health, Boston, Massachusetts 02115, USA (B.B.H.). Center for Computational and Integrative Biology, Massachusetts General Hospital, Boston, Massachusetts 02114, USA (L.A.B.).,

Authors and Affiliations

  1. Division of Infectious Diseases, Department of Medicine, Massachusetts General Hospital, Boston, 02114, Massachusetts, USA

    Brian C. Russo, Luisa M. Stamm, Caleb M. Kim, Emily Kahoud, Ana L. Queiroz, Bobby Brooke Herrera, Leigh A. Baxt, Yang Fu, Gabriel Molina & Marcia B. Goldberg

  2. Department of Microbiology and Immunobiology, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Brian C. Russo, Luisa M. Stamm, Matthijs Raaben, Lindsey R. Robinson, Sayantan Bose, Leigh A. Baxt, Sean P. Whelan & Marcia B. Goldberg

  3. Division of Infectious Diseases, Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor, 48109, Michigan, USA

    Nirit Mor-Vaknin & David M. Markovitz

  4. 65 Landsdowne Street, Cambridge, 02139, Massachusetts, USA

    Marcia B. Goldberg

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Contributions

B.C.R., M.B.G., and L.M.S. wrote the manuscript. B.C.R., M.B.G., L.M.S., M.R., S.P.W., and L.A.B. designed experiments and interpreted data. B.C.R., L.M.S., M.R., L.R.R., A.L.Q., B.B.H., L.A.B., C.M.K., S.B., G.M., E.K., L.R., Y.F., and N.M-V. performed experiments. All authors discussed the results and commented on the manuscript.

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Correspondence to Marcia B. Goldberg.

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Supplementary Table 2, Figures 1-15 and References. (PDF 2842 kb)

Supplementary Table 1

Significantly enriched genes identified by selection. (XLSX 31 kb)

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Russo, B., Stamm, L., Raaben, M. et al. Intermediate filaments enable pathogen docking to trigger type 3 effector translocation. Nat Microbiol 1, 16025 (2016). https://doi.org/10.1038/nmicrobiol.2016.25

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