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Gln 63 of Rho is deamidated by Escherichia coli cytotoxic necrotizing factor-1

Nature volume 387, pages 725–729 (1997)Cite this article

Abstract

The actin cytoskeleton is regulated by GTP-hydrolysing proteins, the Rho GTPases1,2, which act as molecular switches in diverse signal-transduction processes3. Various bacterial toxins can inactivate Rho GTPases by ADP-ribosylation1 or glucosylation4. Previous research has identified Rho proteins as putative targets for Escherichia coli cytotoxic necrotizing factors 1 and 2 (CNF1 and 2)5,6. These toxins induce actin assembly and multinucleation in culture cells. Here we show that treatment of RhoA with CNF1 inhibits the intrinsic GTPase activity of RhoA and completely blocks GTPase activity stimulated by the Rho-GTPase-activating protein (rhoGAP). Analysis by mass spectrometry and amino-acid sequencing of proteolytic peptides derived from CNF1-treated RhoA indicate that CNF1 induces deamidation of a glutamine residue at position 63 (Gln 63) to give constitutively active Rho protein.

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Figure 1: Induction of multinucleation and increase in F-actin by CNF1.
Figure 2: Effects of CNF1 on the migration behaviour of Rho.
Figure 3: a, Nucleotide-binding of CNF1-treated RhoA.
Figure 4: Tandem MS spectrum of the doubly charged tryptic peptide Gln 52–Arg 68 of the CNF I-exposed RhoA.
Figure 5: GTPase activity and nucleotide-binding of Gln63Glu-RhoA.
Figure 6: Induction of stress fibres by microinjection of Gln63Glu-RhoA.

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Acknowledgements

We thank I. Just for critical reading of the manuscript.

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Authors and Affiliations

  1. *Institut für Pharmakologie und Toxikologie der Albert-Ludwigs-Universität Freiburg, Hermann-Herder-Strasse 5, Freiburg, 79104, Germany

    Gudula Schmidt, Peter Sehr, Jörg Selzer & Klaus Aktories

  2. †EMBL, 69012, Heidelberg, Germany

    Matthias Wilm & Matthias Mann

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  1. Gudula Schmidt
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Correspondence to Klaus Aktories.

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Schmidt, G., Sehr, P., Wilm, M. et al. Gln 63 of Rho is deamidated by Escherichia coli cytotoxic necrotizing factor-1. Nature 387, 725–729 (1997). https://doi.org/10.1038/42735

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