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Toxin-induced activation of the G protein p21 Rho by deamidation of glutamine

Nature volume 387, pages 729–733 (1997)Cite this article

Abstract

Pathogenic Escherichia coli are responsible for a variety of diseases, including diarrhoea, haemolytic uraemic syndrome, kidney infection, septicaemia, pneumonia and meningitis. Toxins called cytotoxic necrotizing factors (CNFs) are among the virulence factors produced by uropathogenic (CNF1)1 or enteropathogenic (CNF2)2 E. coli strains that cause diseases in humans and animals, respectively. CNFs induce an increase in the content of actin stress fibres and focal contacts in cultured cells3,4. Effects of CNFs on the actin cytoskeleton correlated with a decrease in the electrophoretic mobility of the GTP-binding protein Rho4,5 and indirect evidence indicates that CNF1 might constitutively activate Rho6. Here we show that CNF1 catalyses the deamidation of a glutamine residue at position 63 of Rho, turning it into glutamic acid, which inhibits both intrinsic GTP hydrolysis and that stimulated by its GTPase-activating protein (GAP). Thus, this deamidation of glutamine 63 by CNF1 leads to the constitutive activation of Rho, and induces the reorganization of actin stress fibres. To our knowledge, CNF1 is the first example of a bacterial toxin acting by deamidation of a specific target protein.

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Figure 1: Induction of actin stress fibres in Vero cells by CNF1 and modification of the electrophoretic mobility of the Rho GTP-binding protein by CNF1 in vivo.
Figure 2: CNF1-induced mobility-shifting of RhoA in vitro.
Figure 3: Deamidation of Rho glutamine 63 by CNF1.
Figure 4: Electrophoretic mobility shift, time course of [35S]GTP-γS dissociation, and determination of the intrinsic and rhoGAP-stimulated GTP hydrolysis of RhoA wild type, Q63E RhoA, and CNF1-treated RhoA.
Figure 5: Microinjection into Vero cells of CNF1-treated RhoA, Q63E RhoA and RhoA.

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Acknowledgements

We thank J. d'Alayer and M. Davi (Institut Pasteur, Paris, France) for trypsin digestion and amino-acid sequencing of Rho proteins; A. Hall for the gift of Rho and RhoGAP expression vectors; J. R. Murphy, E. Van Obberghen-Schilling. P. Cossart and A. Galmiche for discussion.

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Authors and Affiliations

  1. *INSERM U452, Faculté de Médecine, avenue de Valombrose, Nice, 06107 Cedex 2, France

    Gilles Flatau, Emmanuel Lemichez, Michel Gauthier & Patrice Boquet

  2. †Institut de Pharmacologie Moléculaire et Cellulaire du CNRS, route des Lucioles, Sophia Antipolis, 06560, Valbonne, France

    Pierre Chardin & Sonia Paris

  3. ‡Department of Ultrastructures, Istituto Superiore di Sanità, viale Regina Elena, 00161, Roma, Italy

    Carla Fiorentini

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  1. Gilles Flatau
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Correspondence to Patrice Boquet.

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Flatau, G., Lemichez, E., Gauthier, M. et al. Toxin-induced activation of the G protein p21 Rho by deamidation of glutamine. Nature 387, 729–733 (1997). https://doi.org/10.1038/42743

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