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The pathogen protein EspFU hijacks actin polymerization using mimicry and multivalency

Nature volume 454, pages 1005–1008 (2008)Cite this article

Abstract

Enterohaemorrhagic Escherichia coli attaches to the intestine through actin pedestals that are formed when the bacterium injects its protein EspFU (also known as TccP) into host cells1. EspFU potently activates the host WASP (Wiskott–Aldrich syndrome protein) family of actin-nucleating factors, which are normally activated by the GTPase CDC42, among other signalling molecules. Apart from its amino-terminal type III secretion signal, EspFU consists of five-and-a-half 47-amino-acid repeats. Here we show that a 17-residue motif within this EspFU repeat is sufficient for interaction with N-WASP (also known as WASL). Unlike most pathogen proteins that interface with the cytoskeletal machinery, this motif does not mimic natural upstream activators: instead of mimicking an activated state of CDC42, EspFU mimics an autoinhibitory element found within N-WASP. Thus, EspFU activates N-WASP by competitively disrupting the autoinhibited state. By mimicking an internal regulatory element and not the natural activator, EspFU selectively activates only a precise subset of CDC42-activated processes. Although one repeat is able to stimulate actin polymerization, we show that multiple-repeat fragments have notably increased potency. The activities of these EspFU fragments correlate with their ability to coordinate activation of at least two N-WASP proteins. Thus, this pathogen has used a simple autoinhibitory fragment as a component to build a highly effective actin polymerization machine.

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Figure 1: EspF U is a potent activator of N-WASP.
Figure 2: EspF U mimics the N-WASP autoinhibitory C-helix.
Figure 3: Multiple repeats of EspF U are required for potent activation of actin polymerization.

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Acknowledgements

We thank O. Akin for reagents and assistance with the bead motility experiments; J. C. Anderson, A. Chau, R. Howard, M. Lohse, A. Remenyi and L. Weaver for assistance; and members of the Lim laboratory for discussion. This work was supported by grants from the NIH (NIGMS and Nanomedicine Development Centers, NIH Roadmap), the NSF and the Packard Foundation. G.M.R. was supported by the American Heart Association.

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Author notes

  1. John E. Dueber

    Present address: Present address: Department of Synthetic Biology, California Institute of Quantitative Biomedical Research (QB3), University of California, Berkeley, 327 Stanley Hall, Berkeley, California 94720, USA.,

Authors and Affiliations

  1. Graduate Program in Chemistry and Chemical Biology and,,

    Nathan A. Sallee

  2. Department of Cellular and Molecular Pharmacology and the Cell Propulsion Laboratory (an NIH Nanomedicine Development Center), University of California, San Francisco, 600 16th Street, San Francisco, California 94158, USA,

    Nathan A. Sallee, John E. Dueber, R. Dyche Mullins & Wendell A. Lim

  3. Department of Genetics and Developmental Biology and Center for Cell Analysis and Modeling (CCAM), Raymond and Beverly Sackler Laboratory of Genetics and Molecular Medicine, University of Connecticut Health Center, Farmington, Connecticut 06030, USA,

    Gonzalo M. Rivera, Dan Vasilescu & Bruce J. Mayer

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Correspondence to Wendell A. Lim.

Supplementary information

The file contains Supplementary Figures S1-S13 and Legends; Supplementary Methods and additional references.

The Supplementary Figures include: presentation of additional data from experiments shown in the main paper, circular dichroism, quantitative binding measurements, data for the synthetic linker construct, in vivo clustering experiment controls, analytical ultracentrifugation results and description of the Comet Detector algorithm. (PDF 5004 kb)

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Sallee, N., Rivera, G., Dueber, J. et al. The pathogen protein EspFU hijacks actin polymerization using mimicry and multivalency. Nature 454, 1005–1008 (2008). https://doi.org/10.1038/nature07170

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