OpaR controls a network of downstream transcription factors in Vibrio parahaemolyticus BB22OP
Vibrio 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 re...
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| description | Vibrio 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. |
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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.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0121863</identifier><identifier>PMID: 25901572</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Analysis ; Bacterial Proteins - genetics ; Biofilms ; Blotting, Western ; Cell density ; DNA binding proteins ; DNA microarrays ; Electrophoretic mobility ; Electrophoretic Mobility Shift Assay ; Gastroenteritis ; Gene expression ; Gene Expression Regulation, Bacterial ; Gene regulation ; Gene sequencing ; Genomes ; Genomics ; High-Throughput Nucleotide Sequencing - methods ; Humans ; Identification methods ; Nucleotide sequence ; Promoter Regions, Genetic - genetics ; Quorum Sensing ; Real-Time Polymerase Chain Reaction ; Regulatory Sequences, Nucleic Acid - genetics ; Regulon - genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Reverse transcription ; Ribonucleic acid ; RNA ; RNA, Messenger - genetics ; Seafood ; Strains (organisms) ; Transcription factors ; Transcription Factors - genetics ; Transcription Factors - metabolism ; Vibrio parahaemolyticus ; Vibrio parahaemolyticus - genetics ; Vibrio parahaemolyticus - growth & development ; Vibrio parahaemolyticus - metabolism ; Virulence ; Virulence (Microbiology) ; Virulence factors ; Virulence Factors - genetics ; Water-borne diseases ; Waterborne diseases</subject><ispartof>PloS one, 2015-04, Vol.10 (4), p.e0121863-e0121863</ispartof><rights>COPYRIGHT 2015 Public Library of Science</rights><rights>2015 Kernell Burke et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2015 Kernell Burke et al 2015 Kernell Burke et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c758t-e5c0ac73409d645189b7f7c57f62509b4b8af99b4d7a36556f1e9a864c4efcdd3</citedby><cites>FETCH-LOGICAL-c758t-e5c0ac73409d645189b7f7c57f62509b4b8af99b4d7a36556f1e9a864c4efcdd3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4406679/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4406679/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79569,79570</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25901572$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kernell Burke, Alison</creatorcontrib><creatorcontrib>Guthrie, Leah T C</creatorcontrib><creatorcontrib>Modise, Thero</creatorcontrib><creatorcontrib>Cormier, Guy</creatorcontrib><creatorcontrib>Jensen, Roderick V</creatorcontrib><creatorcontrib>McCarter, Linda L</creatorcontrib><creatorcontrib>Stevens, Ann M</creatorcontrib><title>OpaR controls a network of downstream transcription factors in Vibrio parahaemolyticus BB22OP</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Vibrio 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. 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genetics</subject><subject>Quorum Sensing</subject><subject>Real-Time Polymerase Chain Reaction</subject><subject>Regulatory Sequences, Nucleic Acid - genetics</subject><subject>Regulon - genetics</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>Reverse transcription</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>RNA, Messenger - genetics</subject><subject>Seafood</subject><subject>Strains (organisms)</subject><subject>Transcription factors</subject><subject>Transcription Factors - genetics</subject><subject>Transcription Factors - metabolism</subject><subject>Vibrio parahaemolyticus</subject><subject>Vibrio parahaemolyticus - genetics</subject><subject>Vibrio parahaemolyticus - growth & development</subject><subject>Vibrio parahaemolyticus - metabolism</subject><subject>Virulence</subject><subject>Virulence (Microbiology)</subject><subject>Virulence factors</subject><subject>Virulence Factors - genetics</subject><subject>Water-borne diseases</subject><subject>Waterborne diseases</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><sourceid>DOA</sourceid><recordid>eNqNk01v1DAQhiMEoqXwDxBEQkJw2MWOv-ILUlvxsVKlRQV6Q9bEcXZdkjjYCaX_HqebVhvUA_JhLPuZdzzjmSR5jtESE4HfXbrBt1AvO9eaJcIZzjl5kBxiSbIFzxB5uLc_SJ6EcIkQIznnj5ODjEmEmcgOkx_rDs5T7dreuzqkkLamv3L-Z-qqtHRXbei9gSbtPbRBe9v11rVpBbp3PqS2TS9s4a1LO_CwBdO4-rq3egjpyUmWrb88TR5VUAfzbLJHyfePH76dfl6crT-tTo_PFlqwvF8YphFoQSiSJacM57IQldBMVDxjSBa0yKGS0ZYCCGeMV9hIyDnV1FS6LMlR8nKn29UuqKkyQWEuqMwzkuNIrHZE6eBSdd424K-VA6tuDpzfKPDx6bVRWOhSY0GwloYCk5JignJikOFlWZQsar2fog1FY0ptYvGgnonOb1q7VRv3W1GKOBcyCryZBLz7NZjQq8YGbeoaWuOGm3ezXEhOR_TVP-j92U3UBmICtq1cjKtHUXVMs4xLTnAeqeU9VFylaWxsAVPZeD5zeDtzGNvE_Ok3MISgVl_P_59dX8zZ13vs1kDdb4Orh7G5whykO1B7F4I31V2RMVLjFNxWQ41ToKYpiG4v9j_ozum27clfevYCVA</recordid><startdate>20150422</startdate><enddate>20150422</enddate><creator>Kernell Burke, Alison</creator><creator>Guthrie, Leah T C</creator><creator>Modise, Thero</creator><creator>Cormier, Guy</creator><creator>Jensen, Roderick V</creator><creator>McCarter, Linda L</creator><creator>Stevens, Ann M</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20150422</creationdate><title>OpaR controls a network of downstream transcription factors in Vibrio parahaemolyticus BB22OP</title><author>Kernell Burke, Alison ; Guthrie, Leah T C ; Modise, Thero ; Cormier, Guy ; Jensen, Roderick V ; McCarter, Linda L ; Stevens, Ann M</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c758t-e5c0ac73409d645189b7f7c57f62509b4b8af99b4d7a36556f1e9a864c4efcdd3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Analysis</topic><topic>Bacterial Proteins - 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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.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>25901572</pmid><doi>10.1371/journal.pone.0121863</doi><oa>free_for_read</oa></addata></record> |
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| issn | 1932-6203 1932-6203 |
| language | eng |
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| subjects | Analysis Bacterial Proteins - genetics Biofilms Blotting, Western Cell density DNA binding proteins DNA microarrays Electrophoretic mobility Electrophoretic Mobility Shift Assay Gastroenteritis Gene expression Gene Expression Regulation, Bacterial Gene regulation Gene sequencing Genomes Genomics High-Throughput Nucleotide Sequencing - methods Humans Identification methods Nucleotide sequence Promoter Regions, Genetic - genetics Quorum Sensing Real-Time Polymerase Chain Reaction Regulatory Sequences, Nucleic Acid - genetics Regulon - genetics Reverse Transcriptase Polymerase Chain Reaction Reverse transcription Ribonucleic acid RNA RNA, Messenger - genetics Seafood Strains (organisms) Transcription factors Transcription Factors - genetics Transcription Factors - metabolism Vibrio parahaemolyticus Vibrio parahaemolyticus - genetics Vibrio parahaemolyticus - growth & development Vibrio parahaemolyticus - metabolism Virulence Virulence (Microbiology) Virulence factors Virulence Factors - genetics Water-borne diseases Waterborne diseases |
| title | OpaR controls a network of downstream transcription factors in Vibrio parahaemolyticus BB22OP |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-14T07%3A32%3A54IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=OpaR%20controls%20a%20network%20of%20downstream%20transcription%20factors%20in%20Vibrio%20parahaemolyticus%20BB22OP&rft.jtitle=PloS%20one&rft.au=Kernell%20Burke,%20Alison&rft.date=2015-04-22&rft.volume=10&rft.issue=4&rft.spage=e0121863&rft.epage=e0121863&rft.pages=e0121863-e0121863&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0121863&rft_dat=%3Cgale_plos_%3EA422696318%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1674982381&rft_id=info:pmid/25901572&rft_galeid=A422696318&rft_doaj_id=oai_doaj_org_article_17cdc1731c9e4a599413083e0e6ddbd5&rfr_iscdi=true |