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Pathogenic Yersinia pp 211–229Cite as

Measurement of Yersinia Translocon Pore Formation in Erythrocytes

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2010))

Abstract

Many Gram-negative pathogens produce a type III secretion system capable of intoxicating eukaryotic cells with immune-modulating effector proteins. Fundamental to this injection process is the prior secretion of two translocator proteins destined for injectisome translocon pore assembly within the host cell plasma membrane. It is through this pore that effectors are believed to travel to gain access to the host cell interior. Yersinia species especially pathogenic to humans and animals assemble this translocon pore utilizing two hydrophobic translocator proteins—YopB and YopD. Although a full molecular understanding of the biogenesis, function and regulation of this translocon pore and subsequent effector delivery into host cells remains elusive, some of what we know about these processes can be attributed to studies of bacterial infections of erythrocytes. Herein we describe the methodology of erythrocyte infections by Yersinia, and how analysis of the resultant contact-dependent hemolysis can serve as a relative measurement of YopB- and YopD-dependent translocon pore formation.

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Acknowledgements

This work was performed within the framework of the Umeå Centre for Microbial Research at Umeå University. MSF acknowledges financial support from the Swedish Research Council grant 2014–2105, the Medical Research Foundation of Umeå University, and the Faculty of Science and Technology at Umeå University.

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Authors and Affiliations

  1. Department of Molecular Biology, Umeå University, Umeå, Sweden

    Tiago R. D. Costa, Monika K. Francis, Salah I. Farag, Tomas Edgren & Matthew S. Francis

  2. Umeå Centre for Microbial Research, Umeå University, Umeå, Sweden

    Tiago R. D. Costa, Monika K. Francis, Salah I. Farag, Tomas Edgren & Matthew S. Francis

  3. Department of Life Sciences, MRC Centre for Molecular Bacteriology and Infection, Imperial College, London, UK

    Tiago R. D. Costa

  4. Department of Medical Biochemistry and Microbiology, Uppsala Biomedical Center, Uppsala University, Uppsala, Sweden

    Tomas Edgren

Authors
  1. Tiago R. D. Costa
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  2. Monika K. Francis
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  3. Salah I. Farag
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  4. Tomas Edgren
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  5. Matthew S. Francis
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Corresponding author

Correspondence to Matthew S. Francis .

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Editors and Affiliations

  1. Paul G Allen School for Global Animal Health, Washington State University, Pullman, WA, USA

    Viveka Vadyvaloo

  2. Department of Microbiolgy and Immunology and Center for Predictive Medicine in Biodefense and Emerging Infectious Diseases, University of Louisville School of Medicine, Louisville, KY, USA

    Matthew B. Lawrenz

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Costa, T.R.D., Francis, M.K., Farag, S.I., Edgren, T., Francis, M.S. (2019). Measurement of Yersinia Translocon Pore Formation in Erythrocytes. In: Vadyvaloo, V., Lawrenz, M. (eds) Pathogenic Yersinia. Methods in Molecular Biology, vol 2010. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9541-7_15

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  • DOI: https://doi.org/10.1007/978-1-4939-9541-7_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9540-0

  • Online ISBN: 978-1-4939-9541-7

  • eBook Packages: Springer Protocols

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