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Export of a Vibrio parahaemolyticus toxin by the Sec and type III secretion machineries in tandem

Nature Microbiology volume 4, pages 781–788 (2019)Cite this article

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Abstract

Many Gram-negative pathogens utilize dedicated secretion systems to export virulence factors such as exotoxins and effectors1,2,3,4. Several exotoxins are synthesized as precursors containing amino-terminal Sec signal peptides and are exported through the inner-membrane-bound Sec machinery to the periplasm, followed by secretion across the outer membrane to the exterior using a type II secretion system (T2SS)3,5. Here, we report that thermostable direct haemolysin (TDH), an exotoxin of the food-borne pathogen Vibrio parahaemolyticus, can be exported through the type III secretion system (T3SS), which engages in one-step secretion of effectors4, despite possessing a Sec signal peptide and being mainly secreted via the T2SS. Although the precursor of TDH is targeted to the Sec pathway, a fraction of mature TDH was observed to re-enter the bacterial cytoplasm. The N terminus of mature TDH comprises a T3SS signal sequence, allowing it to be loaded into the T3SS. We also show that T3SS-delivered TDH as an effector contributes to intestinal fluid accumulation in a rabbit diarrhoeal model of V. parahaemolyticus infection. Thus, our results show that an unconventional export mechanism for a bacterial toxin via the T3SS in tandem with the Sec machinery facilitates the virulence trait of V. parahaemolyticus.

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Fig. 1: TDH is required for the VopV-independent/T3SS2-dependent induction of intestinal fluid accumulation by V. parahaemolyticus.
Fig. 2: TDH is translocated into host cells via T3SS2.
Fig. 3: cm-TDH is generated by Lep cleavage in the course of Sec translocation.
Fig. 4: NFR facilitates m-TDH secretion from the T3SS2, with the assistance of VocC.

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Data availability

All data supporting the findings of this study are available upon reasonable request from the corresponding authors.

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Acknowledgements

We thank Y. Akiyama, H. Mori, Y. Hizukuri and E. Ishii (Kyoto University, Kyoto, Japan) for providing E. coli strains AD202, secY39, IT41 and IT42, and for critical discussions and reading of the manuscript. We also thank S. Miyoshi for the kind gift of the anti-VPP1 antibody and Y. Yamaichi for critical reading of the manuscript. This study was in part supported by Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science, grant 17K08828 (to S.M.) and grant 17K08829 (to T.K.). This work was also supported by Grants-in-Aid from the Institute for Fermentation, Osaka, from the Naito Foundation, and from the Takeda Science Foundation, Japan (to T.K.).

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Author notes

  1. Ryu Okada

    Present address: The Research Foundation for Microbial Diseases of Osaka University, Kagawa, Japan

  2. Sarunporn Tandhavanant

    Present address: Department of Microbiology and Immunology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand

  3. Hirotaka Hiyoshi

    Present address: Department of Medical Microbiology and Immunology, School of Medicine, University of California, Davis, CA, USA

  4. Kazuyoshi Gotoh

    Present address: Department of Bacteriology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan

Authors and Affiliations

  1. Department of Bacterial Infections, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan

    Shigeaki Matsuda, Ryu Okada, Sarunporn Tandhavanant, Hirotaka Hiyoshi, Kazuyoshi Gotoh, Tetsuya Iida & Toshio Kodama

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Contributions

S.M., S.T., H.H., K.G. and T.K. performed the experiments. R.O. contributed to experimental design and assisted in performing the experiments. T.I. provided valuable input on the manuscript. S.M. and T.K. designed the study, constructed the bacterial strains, analysed results and wrote the paper. All authors reviewed the manuscript and discussed the results.

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Correspondence to Shigeaki Matsuda or Toshio Kodama.

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Matsuda, S., Okada, R., Tandhavanant, S. et al. Export of a Vibrio parahaemolyticus toxin by the Sec and type III secretion machineries in tandem. Nat Microbiol 4, 781–788 (2019). https://doi.org/10.1038/s41564-019-0368-y

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