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Journal of Bacteriology, October 1998, p. 5203-5210, Vol. 180, No. 19
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
HrpW of Erwinia amylovora, a New Harpin
That Contains a Domain Homologous to Pectate Lyases of a Distinct
Class
Jihyun F.
Kim and
Steven V.
Beer*
Department of Plant Pathology, Cornell
University, Ithaca, New York 14853
Received 14 April 1998/Accepted 21 July 1998
Harpins, such as HrpN of Erwinia amylovora, are
extracellular glycine-rich proteins that elicit the hypersensitive
reaction (HR). We identified hrpW of E. amylovora, which encodes a protein similar to known harpins in
that it is acidic, rich in glycine and serine, and lacks cysteine. A
putative HrpL-dependent promoter was identified upstream of
hrpW, and Western blot analysis of hrpL mutants
indicated that the production of HrpW is regulated by hrpL.
HrpW is secreted via the Hrp (type III) pathway based on analysis of
wild-type strains and hrp secretion mutants. When infiltrated into plants, HrpW induced rapid tissue collapse, which required active plant metabolism. The HR-eliciting activity was heat
stable and protease sensitive. Thus, we concluded that HrpW is a new
harpin. HrpW of E. amylovora consists of two domains connected by a Pro and Ser-rich sequence. A fragment containing the
N-terminal domain was sufficient to elicit the HR. Although no pectate
lyase activity was detected, the C-terminal region of HrpW is
homologous to pectate lyases of a unique class, suggesting that HrpW
may be targeted to the plant cell wall. Southern analysis indicated
that hrpW is conserved among several Erwinia
species, and hrpW, provided in trans, enhanced
the HR-inducing ability of a hrpN mutant. However, HrpW did
not increase the virulence of a hrpN mutant in host tissue,
and hrpW mutants retained the wild-type ability to elicit
the HR in nonhosts and to cause disease in hosts.
*
Corresponding author. Mailing address: 410 Plant
Science Bldg., Department of Plant Pathology, Cornell University,
Ithaca, NY 14853. Phone: (607) 255-7870. Fax: (607) 255-4471. E-mail: svb1{at}cornell.edu.
Journal of Bacteriology, October 1998, p. 5203-5210, Vol. 180, No. 19
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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