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  1. IPB Halle
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    • Trenner 0
    • Molecular Signal Processing
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Publications

Lee, J.; Klüsener, B.; Tsiamis, G.; Stevens, C.; Neyt, C.; Tampakaki, A. P.; Panopoulos, N. J.; Nöller, J.; Weiler, E. W.; Cornelis, G. R.; Mansfield, J. W.; Nürnberger, T.; HrpZPsph from the plant pathogen Pseudomonas syringae pv. phaseolicola binds to lipid bilayers and forms an ion-conducting pore in vitro Proc. Natl. Acad. Sci. U.S.A. 98 289-294 (2001) DOI: 10.1073/pnas.98.1.289
  • Abstract
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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.

Publications

Zimmermann, S.; Nürnberger, T.; Frachisse, J.-M.; Wirtz, W.; Guern, J.; Hedrich, R.; Scheel, D.; Receptor-mediated activation of a plant Ca2+-permeable ion channel involved in pathogen defense Proc. Natl. Acad. Sci. U.S.A. 94 2751-2755 (1997) DOI: 10.1073/pnas.94.6.2751
  • Abstract
  • BibText
  • RIS

Pathogen recognition at the plant cell surface typically results in the initiation of a multicomponent defense response. Transient influx of Ca2+ across the plasma membrane is postulated to be part of the signaling chain leading to pathogen resistance. Patch-clamp analysis of parsley protoplasts revealed a novel Ca2+-permeable, La3+-sensitive plasma membrane ion channel of large conductance (309 pS in 240 mM CaCl2). At an extracellular Ca2+ concentration of 1 mM, which is representative of the plant cell apoplast, unitary channel conductance was determined to be 80 pS. This ion channel (LEAC, for large conductance elicitor-activated ion channel) is reversibly activated upon treatment of parsley protoplasts with an oligopeptide elicitor derived from a cell wall protein of Phytophthora sojae. Structural features of the elicitor found previously to be essential for receptor binding, induction of defense-related gene expression, and phytoalexin formation are identical to those required for activation of LEAC. Thus, receptor-mediated stimulation of this channel appears to be causally involved in the signaling cascade triggering pathogen defense in parsley.

Publications

Nürnberger, T.; Nennstiel, D.; Hahlbrock, K.; Scheel, D.; Covalent cross-linking of the Phytophthora megasperma oligopeptide elicitor to its receptor in parsley membranes. Proc. Natl. Acad. Sci. U.S.A. 92 2338-2342 (1995) DOI: 10.1073/pnas.92.6.2338
  • Abstract
  • BibText
  • RIS

An oligopeptide elicitor from Phytophthora megasperma f.sp. glycinea (Pep-13) that induces phytoalexin accumulation in cultured parsley cells was radioiodinated and chemically cross-linked to its binding site in microsomal and plasma membrane preparations with each of three homobifunctional reagents. Analysis by SDS/PAGE and autoradiography of solubilized membrane proteins demonstrated labeling of a 91-kDa protein, regardless of which reagent was used. Cross-linking of this protein was prevented by addition of excess unlabeled Pep-13. The competitor concentration found to half-maximally reduce the intensity of the cross-linked band was 6 nM, which is in good agreement with the IC50 value of 4.7 nM, obtained from ligand binding assays. No crosslinking of 125I-labeled Pep-13 was observed by using microsomal membranes from three other plant species, indicating species-specific occurrence of the binding site. Coupling of 125I-Pep-13 to the parsley 91-kDa protein required the same structural elements within the ligand as was recently reported for binding of 125I-Pep-13 to parsley microsomes, elicitor-induced stimulation of ion fluxes across the plasma membrane, the oxidative burst, the expression of defense-related genes, and phytoalexin production. These findings suggest that the 91-kDa protein identified in parsley membranes is the oligopeptide elicitor receptor mediating activation of a multicomponent defense response.

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