Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Anthrax toxins cooperatively inhibit endocytic recycling by the Rab11/Sec15 exocyst

Nature volume 467, pages 854–858 (2010)Cite this article

Subjects

Abstract

Bacillus anthracis is the causative agent of anthrax in humans and other mammals1,2. In lethal systemic anthrax, proliferating bacilli secrete large quantities of the toxins lethal factor (LF) and oedema factor (EF), leading to widespread vascular leakage and shock. Whereas host targets of LF (mitogen-activated protein-kinase kinases) and EF (cAMP-dependent processes)3 have been implicated in the initial phase of anthrax1,2, less is understood about toxin action during the final stage of infection. Here we use Drosophila melanogaster to identify the Rab11/Sec15 exocyst, which acts at the last step of endocytic recycling, as a novel target of both EF and LF. EF reduces levels of apically localized Rab11 and indirectly blocks vesicle formation by its binding partner and effector Sec15 (Sec15–GFP), whereas LF acts more directly to reduce Sec15–GFP vesicles. Convergent effects of EF and LF on Rab11/Sec15 inhibit expression of and signalling by the Notch ligand Delta and reduce DE-cadherin levels at adherens junctions. In human endothelial cells, the two toxins act in a conserved fashion to block formation of Sec15 vesicles, inhibit Notch signalling, and reduce cadherin expression at adherens junctions. This coordinated disruption of the Rab11/Sec15 exocyst by anthrax toxins may contribute to toxin-dependent barrier disruption and vascular dysfunction during B. anthracis infection.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Learn more

Prices may be subject to local taxes which are calculated during checkout

Figure 1: LF and EF synergistically inhibit Notch signalling.
Figure 2: LF and EF inhibit Rab11/Sec15-dependent recycling.
Figure 3: Conserved activity of anthrax toxins in mammals.

References

  1. Mourez, M. Anthrax toxins. Rev. Physiol. Biochem. Pharmacol. 152, 135–164 (2004)

    Article  CAS  Google Scholar 

  2. Tournier, J. N., Quesnel-Hellmann, A., Cleret, A. & Vidal, D. R. Contribution of toxins to the pathogenesis of inhalational anthrax. Cell. Microbiol. 9, 555–565 (2007)

    Article  CAS  Google Scholar 

  3. Lacy, D. B. & Collier, R. J. Structure and function of anthrax toxin. Curr. Top. Microbiol. Immunol. 271, 61–85 (2002)

    CAS  PubMed  Google Scholar 

  4. Duesbery, N. S. et al. Proteolytic inactivation of MAP-kinase-kinase by anthrax lethal factor. Science 280, 734–737 (1998)

    Article  ADS  CAS  Google Scholar 

  5. Vitale, G. et al. Anthrax lethal factor cleaves the N-terminus of MAPKKs and induces tyrosine/threonine phosphorylation of MAPKs in cultured macrophages. Biochem. Biophys. Res. Commun. 248, 706–711 (1998)

    Article  CAS  Google Scholar 

  6. Leppla, S. H. Anthrax toxin edema factor: a bacterial adenylate cyclase that increases cyclic AMP concentrations of eukaryotic cells. Proc. Natl Acad. Sci. USA 79, 3162–3166 (1982)

    Article  ADS  CAS  Google Scholar 

  7. Moayeri, M. & Leppla, S. H. The roles of anthrax toxin in pathogenesis. Curr. Opin. Microbiol. 7, 19–24 (2004)

    Article  CAS  Google Scholar 

  8. Guichard, A., Park, J. M., Cruz-Moreno, B., Karin, M. & Bier, E. Anthrax lethal factor and edema factor act on conserved targets in Drosophila . Proc. Natl Acad. Sci. USA 103, 3244–3249 (2006)

    Article  ADS  CAS  Google Scholar 

  9. Pezard, C., Berche, P. & Mock, M. Contribution of individual toxin components to virulence of Bacillus anthracis . Infect. Immun. 59, 3472–3477 (1991)

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Fortini, M. E. & Bilder, D. Endocytic regulation of Notch signaling. Curr. Opin. Genet. Dev. 19, 323–328 (2009)

    Article  CAS  Google Scholar 

  11. Jafar-Nejad, H. et al. Sec15, a component of the exocyst, promotes notch signaling during the asymmetric division of Drosophila sensory organ precursors. Dev. Cell 9, 351–363 (2005)

    Article  CAS  Google Scholar 

  12. Wu, H., Rossi, G. & Brennwald, P. The ghost in the machine: small GTPases as spatial regulators of exocytosis. Trends Cell Biol. 18, 397–404 (2008)

    Article  CAS  Google Scholar 

  13. Zhang, J. et al. Thirty-one flavors of Drosophila rab proteins. Genetics 176, 1307–1322 (2007)

    Article  CAS  Google Scholar 

  14. Emery, G. et al. Asymmetric Rab 11 endosomes regulate delta recycling and specify cell fate in the Drosophila nervous system. Cell 122, 763–773 (2005)

    Article  CAS  Google Scholar 

  15. Langevin, J. et al. Drosophila exocyst components Sec5, Sec6, and Sec15 regulate DE-Cadherin trafficking from recycling endosomes to the plasma membrane. Dev. Cell 9, 365–376 (2005)

    Article  CAS  Google Scholar 

  16. Guo, W., Roth, D., Walch-Solimena, C. & Novick, P. The exocyst is an effector for Sec4p, targeting secretory vesicles to sites of exocytosis. EMBO J. 18, 1071–1080 (1999)

    Article  CAS  Google Scholar 

  17. Salminen, A. & Novick, P. J. The Sec15 protein responds to the function of the GTP binding protein, Sec4, to control vesicular traffic in yeast. J. Cell Biol. 109, 1023–1036 (1989)

    Article  CAS  Google Scholar 

  18. Zhang, X. M., Ellis, S., Sriratana, A., Mitchell, C. A. & Rowe, T. Sec15 is an effector for the Rab11 GTPase in mammalian cells. J. Biol. Chem. 279, 43027–43034 (2004)

    Article  CAS  Google Scholar 

  19. Wu, S., Mehta, S. Q., Pichaud, F., Bellen, H. J. & Quiocho, F. A. Sec15 interacts with Rab11 via a novel domain and affects Rab11 localization in vivo . Nature Struct. Mol. Biol. 12, 879–885 (2005)

    Article  CAS  Google Scholar 

  20. Roca, C. & Adams, R. H. Regulation of vascular morphogenesis by Notch signaling. Genes Dev. 21, 2511–2524 (2007)

    Article  CAS  Google Scholar 

  21. Dejana, E., Tournier-Lasserve, E. & Weinstein, B. M. The control of vascular integrity by endothelial cell junctions: molecular basis and pathological implications. Dev. Cell 16, 209–221 (2009)

    Article  CAS  Google Scholar 

  22. Janes, B. K. & Stibitz, S. Routine markerless gene replacement in Bacillus anthracis . Infect. Immun. 74, 1949–1953 (2006)

    Article  CAS  Google Scholar 

  23. Firoved, A. M. et al. Bacillus anthracis edema toxin causes extensive tissue lesions and rapid lethality in mice. Am. J. Pathol. 167, 1309–1320 (2005)

    Article  CAS  Google Scholar 

  24. Kuo, S. R. et al. Anthrax toxin-induced shock in rats is associated with pulmonary edema and hemorrhage. Microb. Pathog. 44, 467–472 (2007)

    Article  Google Scholar 

  25. Gozes, Y., Moayeri, M., Wiggins, J. F. & Leppla, S. H. Anthrax lethal toxin induces ketotifen-sensitive intradermal vascular leakage in certain inbred mice. Infect. Immun. 74, 1266–1272 (2006)

    Article  CAS  Google Scholar 

  26. Tessier, J. et al. Contributions of histamine, prostanoids, and neurokinins to edema elicited by edema toxin from Bacillus anthracis . Infect. Immun. 75, 1895–1903 (2007)

    Article  CAS  Google Scholar 

  27. Balzac, F. et al. E-cadherin endocytosis regulates the activity of Rap1: a traffic light GTPase at the crossroads between cadherin and integrin function. J. Cell Sci. 118, 4765–4783 (2005)

    Article  CAS  Google Scholar 

  28. Silvis, M. R. et al. Rab11b regulates the apical recycling of the cystic fibrosis transmembrane conductance regulator in polarized intestinal epithelial cells. Mol. Biol. Cell 20, 2337–2350 (2009)

    Article  CAS  Google Scholar 

  29. Kosman, D. et al. Multiplex detection of RNA expression in Drosophila embryos. Science 305, 846 (2004)

    Article  CAS  Google Scholar 

  30. van Sorge, N. M. et al. Anthrax toxins inhibit neutrophil signaling pathways in brain endothelium and contribute to the pathogenesis of meningitis. PLoS ONE 3, e2964 (2008)

    Article  ADS  Google Scholar 

  31. Cook, O., Biehs, B. & Bier, E. brinker and optomotor-blind act coordinately to initiate development of the L5 wing vein primordium in Drosophila . Development 131, 2113–2124 (2004)

    Article  CAS  Google Scholar 

  32. Lunde, K. et al. Activation of the knirps locus links patterning to morphogenesis of the second wing vein in Drosophila . Development 130, 235–248 (2003)

    Article  CAS  Google Scholar 

  33. O’Neill, J. W. & Bier, E. Double-label in situ hybridization using biotin and digoxigenin-tagged RNA probes. Biotechniques 17, 874–875 (1994)

    Google Scholar 

Download references

Acknowledgements

We thank A. Kurciyan for help in analysing the effects of expressing wild-type and dominant-negative forms of Rab in Drosophila, A. Cooper and members of the E.B. and V.N. laboratories and H. Bellen for comments on the manuscript and suggestions. We thank S. Leppla for providing purified preparations of LF, EF and PA, S. Stibitz for B. anthracis mutants, and the following investigators for providing antibodies: R. Cohen (anti-Rab11), A. Parks (anti-Dl), K. Irvine (anti-Serrate) and J. Collier (anti-LF). Support for these studies was provided by National Institutes of Health (NIH) R01 grants AI070654 and NS29870 (E.B.), AI077780 (V.N.), an IRACDA NIH postdoctoral fellowship GM068524 (S.M.M.) and a Biomedical Research Fellowship from The Hartwell Foundation (S.M.M.).

Author information

Author notes

  1. Shauna M. McGillivray, Beatriz Cruz-Moreno and Nina M. van Sorge: These authors contributed equally to this work.

Authors and Affiliations

  1. Section of Cell and Developmental Biology, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093-0349, USA ,

    Annabel Guichard, Beatriz Cruz-Moreno & Ethan Bier

  2. Department of Pediatrics, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093-0687, USA,

    Shauna M. McGillivray, Nina M. van Sorge & Victor Nizet

  3. Department of Biology, Texas Christian University, 2800 South University Drive, Fort Worth, 76129, Texas, USA

    Shauna M. McGillivray

  4. Skaggs School of Pharmacy & Pharmaceutical Sciences, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093-0687, USA ,

    Victor Nizet

Authors
  1. Annabel Guichard
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shauna M. McGillivray
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Beatriz Cruz-Moreno
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nina M. van Sorge
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Victor Nizet
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ethan Bier
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

All authors participated in designing the experiments. A.G. and B.C.-M. carried out the Drosophila experiments. S.M.M. and N.M.v.S. carried out the experiments with vertebrate cells and mice. E.B. wrote the manuscript with all other authors providing significant input.

Corresponding author

Correspondence to Ethan Bier.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-12 with legends, Supplementary Tables 1-2, Supplementary Results, an additional reference and a list of Supplementary Microscopy Settings. (PDF 23277 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Guichard, A., McGillivray, S., Cruz-Moreno, B. et al. Anthrax toxins cooperatively inhibit endocytic recycling by the Rab11/Sec15 exocyst. Nature 467, 854–858 (2010). https://doi.org/10.1038/nature09446

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature09446

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing