{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T23:00:11Z","timestamp":1785366011540,"version":"3.55.0"},"reference-count":53,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2015,12,4]],"date-time":"2015-12-04T00:00:00Z","timestamp":1449187200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"US Department of Agriculture (USDA) National Institute of Food and Agriculture"},{"name":"USDA-Hatch Project","award":["ILLU-802-913"],"award-info":[{"award-number":["ILLU-802-913"]}]}],"content-domain":{"domain":["bsppjournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Molecular Plant Pathology"],"published-print":{"date-parts":[[2016,6]]},"abstract":"Summary<\/jats:title>\n \n The type\n III<\/jats:styled-content>\n secretion system (\n T3SS<\/jats:styled-content>\n ) is a key pathogenicity factor in\n \n E<\/jats:italic>\n <\/jats:styled-content>\n rwinia amylovora<\/jats:italic>\n . Previous studies have demonstrated that the\n T3SS<\/jats:styled-content>\n in\n \n E<\/jats:italic>\n <\/jats:styled-content>\n .\u2009amylovora<\/jats:italic>\n is transcriptionally regulated by an\n RpoN<\/jats:styled-content>\n \u2013\n HrpL<\/jats:styled-content>\n sigma factor cascade, which is activated by the bacterial alarmone (p)pp\n G<\/jats:styled-content>\n pp. In this study, the binding site of\n HrpS<\/jats:styled-content>\n , an enhancer binding protein, was identified for the first time in plant\u2010pathogenic bacteria. Complementation of the\n hrp<\/jats:italic>\n \n L<\/jats:italic>\n <\/jats:styled-content>\n mutant with promoter deletion constructs of the\n hrp<\/jats:italic>\n \n L<\/jats:italic>\n <\/jats:styled-content>\n gene and promoter activity analyses using various lengths of the\n hrp<\/jats:italic>\n \n L<\/jats:italic>\n <\/jats:styled-content>\n promoter fused to a promoter\u2010less\n green fluorescent protein<\/jats:italic>\n (\n gfp<\/jats:italic>\n ) reporter gene delineated the upstream region for\n HrpS<\/jats:styled-content>\n binding. Sequence analysis revealed a dyad symmetry sequence between \u2212138 and \u2212125\u2009nucleotides (\n TGCAA<\/jats:styled-content>\n \u2010N4\u2010\n TTGCA<\/jats:styled-content>\n ) as the potential\n HrpS<\/jats:styled-content>\n binding site, which is conserved in the promoter of the\n hrp<\/jats:italic>\n \n L<\/jats:italic>\n <\/jats:styled-content>\n gene among plant enterobacterial pathogens. Results of quantitative real\u2010time reverse transcription\u2010polymerase chain reaction (q\n RT<\/jats:styled-content>\n \u2010\n PCR<\/jats:styled-content>\n ) and electrophoresis mobility shift assay coupled with site\u2010directed mutagenesis (\n SDM<\/jats:styled-content>\n ) analysis showed that the intact dyad symmetry sequence was essential for\n HrpS<\/jats:styled-content>\n binding, full activation of\n T3SS<\/jats:styled-content>\n gene expression and virulence. In addition, the role of the\n GAYTGA<\/jats:styled-content>\n motif (\n RpoN<\/jats:styled-content>\n binding site) of\n HrpS<\/jats:styled-content>\n in the regulation of\n T3SS<\/jats:styled-content>\n gene expression in\n \n E<\/jats:italic>\n <\/jats:styled-content>\n .\u2009amylovora<\/jats:italic>\n was characterized by complementation of the\n hrp<\/jats:italic>\n \n S<\/jats:italic>\n <\/jats:styled-content>\n mutant using mutant variants generated by\n SDM<\/jats:styled-content>\n . Results showed that a\n Y100F<\/jats:styled-content>\n substitution of\n HrpS<\/jats:styled-content>\n complemented the\n hrpS<\/jats:italic>\n mutant, whereas\n Y100A<\/jats:styled-content>\n and\n Y101A<\/jats:styled-content>\n substitutions did not. These results suggest that tyrosine (\n Y<\/jats:styled-content>\n ) and phenylalanine (\n F<\/jats:styled-content>\n ) function interchangeably in the conserved\n GAYTGA<\/jats:styled-content>\n motif of\n HrpS<\/jats:styled-content>\n in\n \n E<\/jats:italic>\n <\/jats:styled-content>\n .\u2009amylovora<\/jats:italic>\n .\n <\/jats:p>","DOI":"10.1111\/mpp.12324","type":"journal-article","created":{"date-parts":[[2015,10,6]],"date-time":"2015-10-06T07:42:40Z","timestamp":1444117360000},"page":"691-702","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["Identification of the HrpS binding site in the\n hrpL<\/i>\n promoter and effect of the RpoN binding site of HrpS on the regulation of the type III secretion system in\n Erwinia amylovora<\/i>"],"prefix":"10.1111","volume":"17","author":[{"given":"Jae Hoon","family":"Lee","sequence":"first","affiliation":[{"name":"Department of Crop Sciences University of Illinois at Urbana\u2010Champaign Urbana IL 61801 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"George W.","family":"Sundin","sequence":"additional","affiliation":[{"name":"Department of Plant, Soil, and Microbial Sciences Michigan State University East Lansing MI 48824 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Youfu","family":"Zhao","sequence":"additional","affiliation":[{"name":"Department of Crop Sciences University of Illinois at Urbana\u2010Champaign Urbana IL 61801 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