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Plant Cell, Vol. 13, 1079-1093, May 2001, Copyright © 2001, American Society of Plant Physiologists

A Harpin Binding Site in Tobacco Plasma Membranes Mediates Activation of the Pathogenesis-Related Gene HIN1 Independent of Extracellular Calcium but Dependent on Mitogen-Activated Protein Kinase Activity

Justin Leea, Daniel F. Klessigb, and Thorsten Nürnbergera
a Department of Stress and Developmental Biology, Leibniz Institute of Plant Biochemistry, Weinberg 3, D-06120 Halle/Saale, Germany
b Department of Molecular Biology and Biochemistry, Rutgers, The State University of New Jersey, Piscataway, New Jersey 08854-8020

Correspondence to: Thorsten Nürnberger, tnuernbe{at}ipb-halle.de (E-mail), 49-345-5582-1409 (fax)

Harpin from the bean halo-blight pathogen Pseudomonas syringae pv phaseolicola (harpinPsph) elicits the hypersensitive response and the accumulation of pathogenesis-related gene transcripts in the nonhost plant tobacco. Here, we report the characterization of a nonproteinaceous binding site for harpinPsph in tobacco plasma membranes, which is assumed to mediate the activation of plant defense responses in a receptor-like manner. Binding of 125I-harpinPsph to tobacco microsomal membranes (dissociation constant = 425 nM) and protoplasts (dissociation constant = 380 nM) was specific, reversible, and saturable. A close correlation was found between the abilities of harpinPsph fragments to elicit the transcript accumulation of the pathogenesis-related tobacco gene HIN1 and to compete for binding of 125I-harpinPsph to its binding site. Another elicitor of the hypersensitive response and HIN1 induction in tobacco, the Phytophthora megasperma–derived ß-elicitin ß-megaspermin, failed to bind to the putative harpinPsph receptor. In contrast to activation by ß-megaspermin, harpinPsph-induced activation of the 48-kD salicylic acid–responsive mitogen-activated protein kinase (MAPK) and HIN1 transcript accumulation were independent of extracellular calcium. Moreover, use of the MAPK kinase inhibitor U0126 revealed that MAPK activity was essential for pathogenesis-related gene expression in harpinPsph-treated tobacco cells. Thus, a receptor-mediated MAPK-dependent signaling pathway may mediate the activation of plant defense responses induced by harpinPsph.




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