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J. Biol. Chem., Vol. 282, Issue 24, 17777-17784, June 15, 2007

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Three Hydroxyproline-rich Glycopeptides Derived from a Single Petunia Polyprotein Precursor Activate defensin I, a Pathogen Defense Response Gene^*

Gregory Pearce, William F. Siems, Ramcharan Bhattacharya^¶, Yu-Chi Chen, and Clarence A. Ryan¹

From the Institute of Biological Chemistry and the Department of Chemistry, Washington State University, Pullman, Washington 99164-6340 and the ^¶National Research Center on Plant Biotechnology, Indian Agricultural Research Institute, New Delhi 110012, India

Hydroxyproline-rich glycopeptides (HypSys peptides) are recently discovered 16–20-amino acid defense signals in tobacco and tomato leaves that are derived from cell wall-associated precursors. The peptides are powerful wound signals that activate the expression of defensive genes in tobacco and tomato leaves in response to herbivore attacks. We have isolated a cDNA from petunia (Petunia hybrida) leaves encoding a putative protein of 214 amino acids that is a homolog of tobacco and tomato HypSys peptide precursors and is inducible by wounding and MeJA. The deduced protein contains a leader sequence and four predicted proline-rich peptides of 18–21 amino acids. Three of the four peptides were isolated from leaves, and each peptide contained hydroxylated prolines and glycosyl residues. Each of the peptides has a -GR- motif at its N terminus, indicating that it may be the substrate site for a processing enzyme. The peptides were active in a petunia suspension culture bioassay at nanomolar concentrations, but they did not induce the expression of defense genes that are directed against herbivores, as found in tobacco and tomato leaves. They did, however, activate expression of defensin 1, a gene associated with inducible defense responses against pathogens.

Received for publication, February 21, 2007 , and in revised form, April 19, 2007.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) EF552428 and Ef552429.

^* This research was supported in part by National Science Foundation Grant IBN 0090766 (to C. A. R.); the Charlotte Y. Martin Foundation; Washington State University College of Agriculture, Human, and Natural Resources Sciences; and a long term associateship award from the Ministry of Science and Technology, Department of Biotechnology, from the Government of India (to R. B.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed: Institute of Biological Chemistry, Washington State University, Pullman, WA 99164-6340. E-mail: cabudryan{at}hotmail.com.

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