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How the Pseudomonas aeruginosa ExoS toxin downregulates Rac

Nature Structural Biology volume 8, pages 23–26 (2001)Cite this article

Abstract

Pseudomonas aeruginosa is an opportunistic bacterial pathogen. One of its major toxins, ExoS, is translocated into eukaryotic cells by a type III secretion pathway. ExoS is a dual function enzyme that affects two different Ras-related GTP binding proteins. The C-terminus inactivates Ras through ADP ribosylation, while the N-terminus inactivates Rho proteins through its GTPase activating protein (GAP) activity. Here we have determined the three-dimensional structure of a complex between Rac and the GAP domain of ExoS in the presence of GDP and AlF3. Composed of ∼130 residues, this ExoS domain is the smallest GAP hitherto described. The GAP domain of ExoS is an all-helical protein with no obvious structural homology, and thus no recognizable evolutionary relationship, with the eukaryotic RhoGAP or RasGAP fold. Similar to other GAPs, ExoS downregulates Rac using an arginine finger to stabilize the transition state of the GTPase reaction, but the details of the ExoS–Rac interaction are unique. Considering the intrinsic resistance of P. aeruginosa to antibiotics, this might open up a new avenue towards blocking its pathogenicity.

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Figure 1: Secondary and tertiary structure of ExoS-N.
Figure 2: Comparison of the ExoS-N and RhoGAP folds.
Figure 3: The interaction between Rac and ExoS-N.
Figure 4: The GAP activity of ExoS-N.

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Acknowledgements

We thank S. McSweeny, A. Perrakis and ESRF for beam-time allocation and help during data collection. We additionally thank E. Carrier and A. Gerhards for technical assistance, M. Hess for help with the figures, K. Scheffzek for carefully reading the manuscript, and R. Schebaum for secretarial assistance. J.T.B. was supported by the NIH, A.W. by the DFG.

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Authors and Affiliations

  1. Abteilung Strukturelle Biologie, Max-Planck-Institut für molekulare Physiologie, Otto-Hahn-Str. 11, Dortmund, 44227, Germany

    Martin Würtele, Eva Wolf, Gretel Buchwald, Mohammad Reza Ahmadian & Alfred Wittinghofer

  2. Microbiology and Molecular Genetics, Medical College of Wisconsin, Wisconsin, 53226, USA

    Kristin J. Pederson & Joseph T. Barbieri

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  1. Martin Würtele
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Correspondence to Alfred Wittinghofer.

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Würtele, M., Wolf, E., Pederson, K. et al. How the Pseudomonas aeruginosa ExoS toxin downregulates Rac. Nat Struct Mol Biol 8, 23–26 (2001). https://doi.org/10.1038/83007

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