HrpZPsph from the plant pathogen Pseudomonas syringae pv. phaseolicola binds to lipid bilayers and forms an ion-conducting pore in vitro

  1. Justin Lee*,†,
  2. Birgit Klüsener†,‡,
  3. George Tsiamis†,§,
  4. Conrad Stevens§,
  5. Cécile Neyt,
  6. Anastasia P. Tampakaki‖,
  7. Nickolas J. Panopoulos‖,
  8. Joachim Nöller**,
  9. Elmar W. Weiler‡,
  10. Guy R. Cornelis,
  11. John W. Mansfield§, and
  12. Thorsten Nürnberger*,‡‡
  1. *Leibniz-Institut für Pflanzenbiochemie, Weinberg 3, D-06120 Halle, Germany; ‡Lehrstuhl für Pflanzenphysiologie, Ruhr-Universität Bochum, Universitätsstrasse 150, D-44780 Bochum, Germany; §Department of Biological Sciences, Wye College, Wye, Ashford, Kent TN25 5AH, United Kingdom; Microbial Pathogenesis Unit, Christian de Duve Institute of Cellular Pathology and Faculty of Medicine, Université Catholique de Louvain, UCL 74-49, B1200 Brussels 5, Belgium; ‖Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-HELLAS, University of Crete, POB 1527, 71110 Heraklion, Greece; and **NIMBUS Biotechnologie GmbH, Karl-Heine-Strasse 99, D-04229 Leipzig, Germany

  1. Edited by Eugene W. Nester, University of Washington, Seattle, WA, and approved November 9, 2000 (received for review June 8, 2000)

Abstract

The hrp gene clusters of plant pathogenic bacteria control pathogenicity on their host plants and ability to elicit the hypersensitive reaction in resistant plants. Some hrp gene products constitute elements of the type III secretion system, by which effector proteins are exported and delivered into plant cells. Here, we show that the hrpZ gene product from the bean halo-blight pathogen, Pseudomonas syringae pv. phaseolicola (HrpZPsph), is secreted in an hrp-dependent manner in P. syringae pv. phaseolicola and exported by the type III secretion system in the mammalian pathogen Yersinia enterocolitica. HrpZPsph was found to associate stably with liposomes and synthetic bilayer membranes. Under symmetric ionic conditions, addition of 2 nM of purified recombinant HrpZPsph to the cis compartment of planar lipid bilayers provoked an ion current with a large unitary conductivity of 207 pS. HrpZPsph-related proteins from P. syringae pv. tomato or syringae triggered ion currents similar to those stimulated by HrpZPsph. The HrpZPsph-mediated ion-conducting pore was permeable for cations but did not mediate fluxes of Cl−. Such pore-forming activity may allow nutrient release and/or delivery of virulence factors during bacterial colonization of host plants.

Footnotes

  • ↵ † J.L., B.K., and G.T. contributed equally to this work.

  • ↵ ‡‡ To whom reprint requests should be addressed at: Institut für Pflanzenbiochemie, Weinberg 3, D-06120 Halle/Saale, Germany. Email: tnuernbe{at}ipb-halle.de.

  • This paper was submitted directly (Track II) to the PNAS office.

  • Data deposition: The sequence reported in this paper has been deposited in the GenBank database (accession no. AF 268940).

  • Article published online before print: Proc. Natl. Acad. Sci. USA, 10.1073/pnas.011265298.

  • Article and publication date are at www.pnas.org/cgi/doi/10.1073/pnas.011265298

  • Abbreviations:
    POPC,
    1-hexadecanoyl-2-(cis-9-octadecenoyl)-sn-glycero-3-phosphocholine;
    POPE,
    1-hexadecanoyl-2-(cis-9-octadecenoyl)-sn-glycero-3-phosphoethanolamine
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  1. Published online before print December 26, 2000, PNAS January 2, 2001 vol. 98 no. 1 289-294
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