Analyzing the Transcriptomes of Two Quorum-Sensing Controlled Transcription Factors, RcsA and LrhA, Important for Pantoea stewartii Virulence

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dc.contributorVirginia Tech. Department of Biological Sciencesen
dc.contributor.authorBurke, Alison K.en
dc.contributor.authorDuong, Duy Anen
dc.contributor.authorJensen, Roderick V.en
dc.contributor.authorStevens, Ann M.en
dc.contributor.departmentBiological Sciencesen
dc.contributor.editorCascales, Ericen
dc.date.accessioned2016-02-12T18:45:05Zen
dc.date.available2016-02-12T18:45:05Zen
dc.date.issued2015-12-23en
dc.description.abstractThe Gram-negative proteobacterium Pantoea stewartii subsp. stewartii causes wilt disease in corn plants. Wilting is primarily due to bacterial exopolysaccharide (EPS) production that blocks water transport in the xylem during the late stages of infection. EsaR, the master quorum-sensing (QS) regulator in P. stewartii, modulates EPS levels. At low cell densities EsaR represses or activates expression of a number of genes in the absence of its acyl homoserine lactone (AHL) ligand. At high cell densities, binding of AHL inactivates EsaR leading to derepression or deactivation of its direct targets. Two of these direct targets are the key transcription regulators RcsA and LrhA, which in turn control EPS production and surface motility/adhesion, respectively. In this study, RNA-Seq was used to further examine the physiological impact of deleting the genes encoding these two second-tier regulators. Quantitative reverse transcription PCR (qRT-PCR) was used to validate the regulation observed in the RNA-Seq data. A GFP transcriptional fusion reporter confirmed the existence of a regulatory feedback loop in the system between LrhA and RcsA. Plant virulence assays carried out with rcsA and lrhA deletion and complementation strains demonstrated that both transcription factors play roles during establishment of wilt disease in corn. These efforts further define the hierarchy of the QS-regulated network controlling plant virulence in P. stewartii.en
dc.description.notesThe read data for the pairs of duplicate samples for the P. stewartii wild-type, ΔrcsA, and ΔlrhA strains, have been deposited in the NCBI Sequence Read Archive (SRA) with accession numbers, GSM1691841, GSM1691842, GSM1691843, GSM1691844, GSM1691845 and GSM1691846, respectively. An Excel file summarizing the differential gene expression in total counts and normalized reads per million (RPM), using the P. stewartii DC283 version 8 NCBI gene annotations, has been deposited in the NCBI Gene Expression Omnibus (GEO) database (GEO Accession GSE69064).en
dc.description.sponsorshipVirginia Tech. Open Access Subvention Funden
dc.description.sponsorshipNational Science Foundation (U.S.)en
dc.description.sponsorshipMCB-0919984en
dc.description.sponsorshipVirginia Tech. Graduate Student Association Graduate Research Development Program Awarden
dc.description.sponsorshipVirginia Tech. Biological Sciences Graduate Student Development Awarden
dc.description.sponsorshipVirginia Tech. The Life Sciences I Building Funden
dc.format.extent18 p.en
dc.format.mimetypeapplication/pdfen
dc.identifier.citationKernell Burke A, Duong DA, Jensen RV, Stevens AM (2015) Analyzing the Transcriptomes of Two Quorum-Sensing Controlled Transcription Factors, RcsA and LrhA, Important for Pantoea stewartii Virulence. PLoS ONE 10(12): e0145358. doi:10.1371/journal.pone.0145358en
dc.identifier.doihttps://doi.org/10.1371/journal.pone.0145358en
dc.identifier.issn1932-6203en
dc.identifier.issue12en
dc.identifier.urihttp://hdl.handle.net/10919/64801en
dc.identifier.volume10en
dc.language.isoen_USen
dc.publisherPublic Library of Scienceen
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectGene expressionen
dc.subjectGene regulationen
dc.subjectPathogen motilityen
dc.subjectRNA foldingen
dc.subjectInfectious disease controlen
dc.subjectPlant bacterial pathogensen
dc.subjectVector constructionen
dc.subjectRegulator genesen
dc.titleAnalyzing the Transcriptomes of Two Quorum-Sensing Controlled Transcription Factors, RcsA and LrhA, Important for Pantoea stewartii Virulenceen
dc.title.serialPLOS Oneen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

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