title: HrpW of Erwinia amylovora, a new harpin that contains a domain homologous to pectate lyases of a distinct class.
abstract: 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.
authors.affiliation: Department of Plant Pathology, Cornell University, Ithaca, New York 14853, USA.
authors.forename: J F
authors.initials: JF
authors.lastname: Kim
authors.forename: S V
authors.initials: SV
authors.lastname: Beer
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology; D004268:DNA-Binding Proteins; D004885:Erwinia / Q000201:enzymology / Q000235:genetics* / Q000472:pathogenicity; D005798:Genes, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D010947:Plants, Toxic; D011133:Polysaccharide-Lyases / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012808:Sigma Factor; D014026:Nicotiana / Q000382:microbiology; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C085855:HrpL protein, Pseudomonas syringae; D011993:Recombinant Fusion Proteins; D012808:Sigma Factor; D011133:Polysaccharide-Lyases; C023911:pectate lyase