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Molecular characterization of Pantoea stewartii subsp. stewartii HrpY, a conserved response regulator of the Hrp type III secretion system, and its interaction with the hrpS promoter.

ABSTRACT: Pantoea stewartii subsp. stewartii is a bacterial pathogen of corn. Its pathogenicity depends on the translocation of effector proteins into host cells by the Hrp type III secretion system. We previously showed by genetic analysis that the HrpX sensor kinase and the HrpY response regulator are at the head of a complex cascade of regulators controlling hrp/hrc secretion and wts effector genes. This cascade also includes the HrpS response regulator and the HrpL alternative sigma factor. These regulators are shared among many important plant pathogens in the genera Pantoea, Erwinia, and Pseudomonas. In this study, we dissect the regulatory elements in the hrpS promoter region, using genetic and biochemical approaches, and show how it integrates various environmental signals, only some of which are dependent on phosphorylation of HrpY. Primer extension located the transcriptional start site of hrpS at a sigma70 promoter 601 bp upstream of the open reading frame. Electrophoretic mobility shift assays and DNase I footprinting analysis demonstrated that HrpY binds to conserved regulatory elements immediately adjacent to this promoter, and its binding affinity was increased by phosphorylation at D57. A consensus sequence for the two direct repeats bound by HrpY is proposed. Deletion analysis of the promoter region revealed that both the HrpY binding site and additional sequences farther upstream, including a putative integration host factor binding site, are required for hrpS expression. This finding suggests that other unknown regulatory proteins may act cooperatively with HrpY.

SUBMITTER: Merighi M

PROVIDER: S-EPMC1539970 | biostudies-literature | 2006 Jul

REPOSITORIES: biostudies-literature

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Molecular characterization of Pantoea stewartii subsp. stewartii HrpY, a conserved response regulator of the Hrp type III secretion system, and its interaction with the hrpS promoter.

Merighi Massimo M Majerczak Doris R DR Zianni Michael M Tessanne Kimberly K Coplin David L DL

Journal of bacteriology 20060701 14

Pantoea stewartii subsp. stewartii is a bacterial pathogen of corn. Its pathogenicity depends on the translocation of effector proteins into host cells by the Hrp type III secretion system. We previously showed by genetic analysis that the HrpX sensor kinase and the HrpY response regulator are at the head of a complex cascade of regulators controlling hrp/hrc secretion and wts effector genes. This cascade also includes the HrpS response regulator and the HrpL alternative sigma factor. These regu ...[more]

PMID: 16816181

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Project description:Following a request from the European Commission, the EFSA Panel on Plant Health performed a risk assessment of the entry of Pantoea stewartii subsp. stewartii on maize seed imported by the EU from the USA. This pest is a Gram-negative bacterium which causes Stewart's vascular wilt and leaf blight of maize (including sweet corn), a disease responsible for serious crop losses throughout the world. The following scenarios were considered: scenario A0 (current practice), scenario A1 (US request for modification of EU conditions for derogation), and scenario A2 (EU conditions for derogation). Results from the quantitative seed pathway model presented here show that, despite the low rates of plant-to-seed and seed-to-seedling transmission that have been reported in the literature for Stewart's wilt, given the amount of traded seed, and in the case of voluntary (i.e. not mandatory) inspections of seed production fields at the origin (i.e. scenario A0), the frequency of introducing the disease is in the order of magnitude of some hundred introductions per year (median number). The EU conditions for derogation would lead to a decrease in the likelihood of entry compared to scenarios A0 (about 10,000 times fewer introductions) and A1 (about 2,000 times fewer introductions). This protective effect is mainly due to the requirement that only genotypes resistant to Stewart's wilt are traded, with the additional field inspection (two instead of one per season) providing additional reassurance. The Panel also concluded that seed lot inspections, as currently carried out (e.g. with a sample of 400 seeds) are not likely to lead to a relevant reduction in the level of infected imported maize seed, given the low prevalence of Stewart's wilt at the origin. If, however, there is aggregation in infection among consignments, inspection would work towards identifying the highly infected consignments. Recently, outbreaks of Stewart's wilt have occurred in Italy (Emilia Romagna, Friuli, Lombardy and Veneto). A review is provided of the available information to assess the possible role of seed imports in these outbreaks.

Dataset Information

Molecular characterization of Pantoea stewartii subsp. stewartii HrpY, a conserved response regulator of the Hrp type III secretion system, and its interaction with the hrpS promoter.

Publications

Molecular characterization of Pantoea stewartii subsp. stewartii HrpY, a conserved response regulator of the Hrp type III secretion system, and its interaction with the hrpS promoter.

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