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First published online August 17, 2007; 10.1104/pp.107.103432 Plant Physiology 145:426-436 (2007) © 2007 American Society of Plant Biologists OPEN ACCESS ARTICLE
AtHIPM, an Ortholog of the Apple HrpN-Interacting Protein, Is a Negative Regulator of Plant Growth and Mediates the Growth-Enhancing Effect of HrpN in Arabidopsis1,[C],[OA]Department of Plant Pathology, Cornell University, Ithaca, New York 14853
HrpN (harpin) protein is critical to the virulence of the fire blight pathogen Erwinia amylovora in host plants like apple (Malus x domestica). Moreover, exogenous treatment of Arabidopsis (Arabidopsis thaliana), a nonhost plant, with partially purified HrpN enhances growth. To address the bases of the effects of HrpN in disease, we sought a HrpN-interacting protein(s) in apple, using a yeast two-hybrid assay. A single positive clone, designated HIPM (HrpN-interacting protein from Malus), was found. HIPM, a 6.5-kD protein, interacted with HrpN in yeast and in vitro. Deletion analysis showed that the N-terminal 198 of 403 amino acids of HrpN are required for interaction with HIPM. HIPM orthologs were found in Arabidopsis (AtHIPM) and rice (Oryza sativa; OsHIPM). HrpN also interacted with AtHIPM in yeast and in vitro. In silico analyses revealed that the three plant proteins contain putative signal peptides and putative transmembrane domains. We showed that both HIPM and AtHIPM have functional signal peptides, and green fluorescent protein-tagged HIPM and AtHIPM associated, in clusters, with plasma membranes. Both HIPM and AtHIPM are expressed constitutively; however, they are expressed more strongly in apple and Arabidopsis flowers than in leaves and stems. The size of AtHIPM knockout mutant plants of Arabidopsis was slightly larger than the wild-type plants. Interestingly, the knockout mutant did not exhibit enhanced plant growth in response to treatment with HrpN. Overexpression of AtHIPM conversely resulted in smaller plants. These results indicate that AtHIPM functions as a negative regulator of plant growth and mediates enhanced growth that results from treatment with HrpN.
1 This work was supported in part by the U.S. Department of Agriculture Cooperative State Research, Education, and Extension Service (special grant no. 2003–34367–13158) and by the Eden Bioscience. 2 Present address: Boyce Thompson Institute for Plant Research, Tower Road, Ithaca, NY 14853. The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Steven V. Beer (svb1{at}cornell.edu). [C] Some figures in this article are displayed in color online but in black and white in the print edition. [OA] Open Access articles can be viewed online without a subscription. www.plantphysiol.org/cgi/doi/10.1104/pp.107.103432 * Corresponding author; e-mail svb1{at}cornell.edu. Received June 8, 2007; accepted July 30, 2007; published August 17, 2007.
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