Abstract
Bacterial pathogens developed several strategies to overcome defense systems of eukaryotic hosts. Within the infection process, they need to attach to and cross through epithelial layers, escape from the innate and adaptive immune response, and find a physiological niche to survive. One target to modulate the host–pathogen interaction in order to deceit pathogen resistance is the actin cytoskeleton and its regulators: the family of Rho GTPases. Some bacterial toxins catalyze a covalent modification of Rho GTPases to keep these molecular switches in a constitutive active or inactive state. This leads to rearrangement of the actin cytoskeleton. Toxin-treated cells show typical morphological changes depending on substrate specificity and action of the toxins. In this chapter, we discuss the classes of bacterial toxins based on their mode of action, their recombinant expression (specifically CNF1), intoxication and subsequent morphological changes of the actin cytoskeleton, and cell shape.
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References
Aktories, K., and Barbieri, J. T. (2005) Bacterial cytotoxins: targeting eukaryotic switches. Nat Rev Microbiol 3, 397–410.
Jank, T., and Aktories, K. (2008) Structure and mode of action of clostridial glucosylating toxins: the ABCD model. Trends Microbiol. 16, 222–229.
Sorg, I., Goehring, U. M., Aktories, K., and Schmidt, G. (2001) Recombinant Yersinia YopT leads to uncoupling of RhoA-effector interaction. Infect Immun 69, 7535–7543.
Aepfelbacher, M., Zumbihl, R., and Heesemann, J. (2005) Modulation of Rho GTPases and the actin cytoskeleton by YopT of Yersinia. Curr Top Microbiol Immunol 291, 167–175.
Genth, H., Gerhard, R., Maeda, A., Amano, M., Kaibuchi, K., Aktories, K., and Just, I. (2003) Entrapment of Rho ADP-ribosylated by Clostridium botulinum C3 exoenzyme in the Rho-guanine nucleotide dissociation inhibitor-1 complex. J Biol Chem 278, 28523–28527.
Aktories, K., Wilde, C., and Vogelsgesang, M. (2004) Rho-modifying C3-like ADP-ribosyltransferases. Rev Physiol Biochem Pharmacol 152, 1–22.
Aili, M., Telepnev, M., Hallberg, B., Wolf-Watz, H., and Rosqvist, R. (2003) In vitro GAP activity towards RhoA, Rac1 and Cdc42 is not a prerequisite for YopE induced HeLa cell cytotoxicity. Microb Pathog 34, 297–308.
Andor, A., Trulzsch, K., Essler, M., Roggenkamp, A., Wiedemann, A., Heesemann, J., and Aepfelbacher, M. (2001) YopE of Yersinia, a GAP for Rho GTPases, selectively modulates Rac-dependent actin structures in endothelial cells. Cell Microbiol 3, 301–310.
Von Pawel-Rammingen, U., Telepnev, M. V., Schmidt, G., Aktories, K., Wolf-Watz, H., and Rosqvist, R. (2000) GAP activity of the Yersinia YopE cytotoxin specifically targets the Rho pathway: a mechanism for disruption of actin microfilament structure. Mol Microbiol 36, 737–748.
Lerm, M., Selzer, J., Hoffmeyer, A., Rapp, U. R., Aktories, K., and Schmidt, G. (1999) Deamidation of Cdc42 and Rac by Escherichia coli cytotoxic necrotizing factor 1: activation of c-Jun N-terminal kinase in HeLa cells. Infect Immun 67, 496–503.
Schmidt, G., Sehr, P., Wilm, M., Selzer, J., Mann, M., and Aktories, K. (1997) Gln 63 of Rho is deamidated by Escherichia coli cytotoxic necrotizing factor-1. Nature 387, 725–729.
Hoffmann, C., Pop, M., Leemhuis, J., Schirmer, J., Aktories, K., and Schmidt, G. (2004) The Yersinia pseudotuberculosis cytotoxic necrotizing factor (CNFY) selectively activates RhoA. J Biol Chem 279, 16026–16032.
Horiguchi, Y., Inoue, N., Masuda, M., Kashimoto, T., Katahira, J., Sugimoto, N., and Matsuda, M. (1997) Bordetella bronchiseptica dermonecrotizing toxin induces reorganization of actin stress fibers through deamidation of Gln-63 of the GTP-binding protein Rho. Proc Natl Acad Sci USA 94, 11623–11626.
Huelsenbeck, S. C., May, M., Schmidt, G., and Genth, H. (2009) Inhibition of cytokinesis by Clostridium difficile toxin B and cytotoxic necrotizing factors--reinforcing the critical role of RhoA in cytokinesis. Cell Motil Cytoskeleton 66, 967–975.
Friebel, A., Ilchmann, H., Aepfelbacher, M., Ehrbar, K., Machleidt, W., and Hardt, W. D. (2001) SopE and SopE2 from Salmonella typhimurium activate different sets of Rho GTPases of the host cell. J Biol Chem 276, 34035–34040.
Knust, Z., Blumenthal, B., Aktories, K., and Schmidt, G. (2009) Cleavage of Escherichia coli cytotoxic necrotizing factor 1 is required for full biologic activity. Infect Immun 77, 1835–1841.
Barth, H., Aktories, K., Popoff, M. R., and Stiles, B. G. (2004) Binary bacterial toxins: biochemistry, biology, and applications of common Clostridium and Bacillus proteins. Microbiol Mol Biol Rev 68, 373–402.
Barth, H., Blocker, D., and Aktories, K. (2002) The uptake machinery of clostridial actin ADP-ribosylating toxins--a cell delivery system for fusion proteins and polypeptide drugs. Naunyn Schmiedebergs Arch Pharmacol 366, 501–512.
Stratmann, H., Schwan, C., Orth, J. H., Schmidt, G., and Aktories, K. (2010) Pleiotropic role of Rac in mast cell activation revealed by a cell permeable Bordetella dermonecrotic fusion toxin. Cell Signal 22, 1124–1131.
Berks, B. C., Sargent, F., and Palmer, T. (2000) The Tat protein export pathway. Mol Microbiol 35, 260–274.
DerMardirossian, C. and Bokoch, G. M. (2005) GDIs: central regulatory molecules in Rho GTPase activation. Trends Cell Biol 15, 356–363.
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Bielek, H., Schmidt, G. (2012). How to Analyze Bacterial Toxins Targeting Rho GTPases. In: Rivero, F. (eds) Rho GTPases. Methods in Molecular Biology, vol 827. Springer, New York, NY. https://doi.org/10.1007/978-1-61779-442-1_4
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DOI: https://doi.org/10.1007/978-1-61779-442-1_4
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