OpaR Controls a Network of Downstream Transcription Factors in Vibrio parahaemolyticus BB22OP

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dc.contributorVirginia Tech. Department of Biological Sciencesen
dc.contributor.authorBurke, Alison K.en
dc.contributor.authorGuthrie, Leah T. C.en
dc.contributor.authorModise, Theroen
dc.contributor.authorCormier, Guyen
dc.contributor.authorJensen, Roderick V.en
dc.contributor.authorMcCarter, Linda L.en
dc.contributor.authorStevens, Ann M.en
dc.contributor.departmentBiological Sciencesen
dc.date.accessioned2016-02-12T22:02:29Zen
dc.date.available2016-02-12T22:02:29Zen
dc.date.issued2015-04-22en
dc.descriptionThe read data for the V. parahaemolyticus BB22OP LM5312 (opaR+) QS-proficient strain, and V. parahaemolyticus BB22TR LM5674 (∆opaR1) QS-deficient strain, have been deposited in the NCBI Sequence Read Archive (SRA) with accession numbers GSM1297676 and GSM1297677, respectively. An Excel file summarizing the differential gene expression in total counts and normalized RPM, using the BB22OP and RIMD2210633 annotations, has been deposited in the NCBI Gene Expression Omnibus (GEO) database (GEO Accession GSE53639).en
dc.description.abstractVibrio parahaemolyticus is an emerging world-wide human pathogen that is associated with food-borne gastroenteritis when raw or undercooked seafood is consumed. Expression of virulence factors in this organism is modulated by the phenomenon known as quorum sensing, which permits differential gene regulation at low versus high cell density. The master regulator of quorum sensing in V. parahaemolyticus is OpaR. OpaR not only controls virulence factor gene expression, but also the colony and cellular morphology associated with growth on a surface and biofilm formation. Whole transcriptome Next Generation sequencing (RNA-Seq) was utilized to determine the OpaR regulon by comparing strains BB22OP (opaR+, LM5312) and BB22TR (∆opaR1, LM5674). This work, using the published V. parahaemolyticus BB22OP genome sequence, confirms and expands upon a previous microarray analysis for these two strains that used an Affymetrix GeneChip designed from the closely related V. parahaemolyticus RIMD2210633 genome sequence. Overall there was excellent correlation between the microarray and RNA-Seq data. Eleven transcription factors under OpaR control were identified by both methods and further confirmed by quantitative reverse transcription PCR (qRT-PCR) analysis. Nine of these transcription factors were demonstrated to be direct OpaR targets via in vitro electrophoretic mobility shift assays with purified hexahistidine-tagged OpaR. Identification of the direct and indirect targets of OpaR, including small RNAs, will enable the construction of a network map of regulatory interactions important for the switch between the nonpathogenic and pathogenic states.en
dc.description.sponsorshipVirginia Tech. Fralin Life Sciences Instituteen
dc.description.sponsorshipVirginia Tech. Virginia Bioinformatics Instituteen
dc.description.sponsorshipVirginia Tech. Graduate Research Development Programen
dc.description.sponsorshipLife Sciences I Building Funden
dc.description.sponsorshipVirginia Tech. Open Access Subvention Funden
dc.format.extent18 p.en
dc.format.mimetypeapplication/pdfen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationKernell Burke A, Guthrie LTC, Modise T, Cormier G, Jensen RV, McCarter LL, et al. (2015) OpaR Controls a Network of Downstream Transcription Factors in Vibrio parahaemolyticus BB22OP. PLoS ONE 10(4): e0121863. doi:10.1371/ journal.pone.0121863en
dc.identifier.doihttps://doi.org/10.1371/journal.pone.0121863en
dc.identifier.issn1932-6203en
dc.identifier.issue4en
dc.identifier.urihttp://hdl.handle.net/10919/64802en
dc.identifier.volume10en
dc.language.isoen_USen
dc.publisherPublic Library of Scienceen
dc.relation.urihttp://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE53639en
dc.relation.urihttp://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE69064en
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectTranscription factorsen
dc.subjectGene regulationen
dc.subjectGene expressionen
dc.subjectRegulator genesen
dc.subjectMicroarraysen
dc.subjectRNA extractionen
dc.subjectChromosome 1en
dc.subjectRegulonsen
dc.titleOpaR Controls a Network of Downstream Transcription Factors in Vibrio parahaemolyticus BB22OPen
dc.title.serialPLOS Oneen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

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