Global Transcriptional Response of Clostridium difficile Carrying the ϕCD38-2 Prophage
Clostridium difficile is one of the most dangerous pathogens in hospital settings. Most strains of C. difficile carry one or more prophages, and some of them, like ϕCD38-2 and ϕCD119, can influence the expression of toxin genes. However, little is known about the global host response in the presence...
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Veröffentlicht in: | Applied and Environmental Microbiology 2015-02, Vol.81 (4), p.1364-1374 |
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description | Clostridium difficile
is one of the most dangerous pathogens in hospital settings. Most strains of
C. difficile
carry one or more prophages, and some of them, like ϕCD38-2 and ϕCD119, can influence the expression of toxin genes. However, little is known about the global host response in the presence of a given prophage. In order to fill this knowledge gap, we used high-throughput RNA sequencing (RNA-seq) to conduct a genome-wide transcriptomic analysis of the epidemic
C. difficile
strain R20291 carrying the ϕCD38-2 prophage. A total of 39 bacterial genes were differentially expressed in the R20291 lysogen, 26 of them being downregulated. Several of the regulated genes encode transcriptional regulators and phosphotransferase system (PTS) subunits involved in glucose, fructose, and glucitol/sorbitol uptake and metabolism. ϕCD38-2 also upregulated the expression of a group of regulatory genes located in phi-027, a resident prophage common to most ribotype 027 isolates. The most differentially expressed gene was that encoding the conserved phase-variable cell wall protein CwpV, which was upregulated ∼20-fold in the lysogen. Quantitative PCR and immunofluorescence showed that the increased
cwpV
expression results from a greater proportion of cells actively transcribing the gene. Indeed, ∼95% of lysogenic cells express
cwpV
, as opposed to only ∼5% of wild-type cells. Furthermore, the higher proportion of cells expressing
cwpV
results from a higher frequency of recombination of the genetic switch controlling phase variation, which we confirmed to be dependent on the host-encoded recombinase RecV. In summary, ϕCD38-2 interferes with phase variation of the surface protein CwpV and the expression of metabolic genes. |
doi_str_mv | 10.1128/AEM.03656-14 |
format | Article |
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is one of the most dangerous pathogens in hospital settings. Most strains of
C. difficile
carry one or more prophages, and some of them, like ϕCD38-2 and ϕCD119, can influence the expression of toxin genes. However, little is known about the global host response in the presence of a given prophage. In order to fill this knowledge gap, we used high-throughput RNA sequencing (RNA-seq) to conduct a genome-wide transcriptomic analysis of the epidemic
C. difficile
strain R20291 carrying the ϕCD38-2 prophage. A total of 39 bacterial genes were differentially expressed in the R20291 lysogen, 26 of them being downregulated. Several of the regulated genes encode transcriptional regulators and phosphotransferase system (PTS) subunits involved in glucose, fructose, and glucitol/sorbitol uptake and metabolism. ϕCD38-2 also upregulated the expression of a group of regulatory genes located in phi-027, a resident prophage common to most ribotype 027 isolates. The most differentially expressed gene was that encoding the conserved phase-variable cell wall protein CwpV, which was upregulated ∼20-fold in the lysogen. Quantitative PCR and immunofluorescence showed that the increased
cwpV
expression results from a greater proportion of cells actively transcribing the gene. Indeed, ∼95% of lysogenic cells express
cwpV
, as opposed to only ∼5% of wild-type cells. Furthermore, the higher proportion of cells expressing
cwpV
results from a higher frequency of recombination of the genetic switch controlling phase variation, which we confirmed to be dependent on the host-encoded recombinase RecV. In summary, ϕCD38-2 interferes with phase variation of the surface protein CwpV and the expression of metabolic genes.</description><identifier>ISSN: 0099-2240</identifier><identifier>EISSN: 1098-5336</identifier><identifier>EISSN: 1098-6596</identifier><identifier>DOI: 10.1128/AEM.03656-14</identifier><identifier>PMID: 25501487</identifier><language>eng</language><publisher>1752 N St., N.W., Washington, DC: American Society for Microbiology</publisher><subject>Clostridium difficile ; Genetics and Molecular Biology</subject><ispartof>Applied and Environmental Microbiology, 2015-02, Vol.81 (4), p.1364-1374</ispartof><rights>Copyright © 2015, American Society for Microbiology. All Rights Reserved. 2015 American Society for Microbiology</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c361t-90d03eedbb918aa01671c387b41535d29eaeee75d90c77fc1204fe1032ab15db3</citedby><cites>FETCH-LOGICAL-c361t-90d03eedbb918aa01671c387b41535d29eaeee75d90c77fc1204fe1032ab15db3</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/PMC4309704/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4309704/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,3188,27924,27925,53791,53793</link.rule.ids></links><search><contributor>Schaffner, D. W.</contributor><creatorcontrib>Sekulovic, Ognjen</creatorcontrib><creatorcontrib>Fortier, Louis-Charles</creatorcontrib><title>Global Transcriptional Response of Clostridium difficile Carrying the ϕCD38-2 Prophage</title><title>Applied and Environmental Microbiology</title><description>Clostridium difficile
is one of the most dangerous pathogens in hospital settings. Most strains of
C. difficile
carry one or more prophages, and some of them, like ϕCD38-2 and ϕCD119, can influence the expression of toxin genes. However, little is known about the global host response in the presence of a given prophage. In order to fill this knowledge gap, we used high-throughput RNA sequencing (RNA-seq) to conduct a genome-wide transcriptomic analysis of the epidemic
C. difficile
strain R20291 carrying the ϕCD38-2 prophage. A total of 39 bacterial genes were differentially expressed in the R20291 lysogen, 26 of them being downregulated. Several of the regulated genes encode transcriptional regulators and phosphotransferase system (PTS) subunits involved in glucose, fructose, and glucitol/sorbitol uptake and metabolism. ϕCD38-2 also upregulated the expression of a group of regulatory genes located in phi-027, a resident prophage common to most ribotype 027 isolates. The most differentially expressed gene was that encoding the conserved phase-variable cell wall protein CwpV, which was upregulated ∼20-fold in the lysogen. Quantitative PCR and immunofluorescence showed that the increased
cwpV
expression results from a greater proportion of cells actively transcribing the gene. Indeed, ∼95% of lysogenic cells express
cwpV
, as opposed to only ∼5% of wild-type cells. Furthermore, the higher proportion of cells expressing
cwpV
results from a higher frequency of recombination of the genetic switch controlling phase variation, which we confirmed to be dependent on the host-encoded recombinase RecV. In summary, ϕCD38-2 interferes with phase variation of the surface protein CwpV and the expression of metabolic genes.</description><subject>Clostridium difficile</subject><subject>Genetics and Molecular Biology</subject><issn>0099-2240</issn><issn>1098-5336</issn><issn>1098-6596</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><recordid>eNpVkc1O3DAQgK2qqCxbbn2AHDk0dMY_SXxBQinQSiBQtYij5TiTXaNsHOxsJR6E5-KVCAVV4jQazadvDh9j3xCOEXn14_Ts6hhEoYoc5Se2QNBVroQoPrMFgNY55xL22UFK9wAgoai-sH2uFKCsygW7u-hDY_tsFe2QXPTj5MMw738ojWFIlIUuq_uQpuhbv9tmre8673xPWW1jfPTDOps2lD0_1T9FlfPsJoZxY9f0le11tk90-D6X7Pb8bFX_yi-vL37Xp5e5EwVOuYYWBFHbNBorawGLEp2oykaiEqrlmiwRlarV4Mqyc8hBdoQguG1QtY1YspM377hrttQ6GqZoezNGv7Xx0QTrzcfL4DdmHf4aKUCXIGfB0bsghocdpclsfXLU93agsEsGi2KmFK_0jH5_Q10MKUXq_r9BMK8tzNzC_GthUIoXTLd8MQ</recordid><startdate>20150201</startdate><enddate>20150201</enddate><creator>Sekulovic, Ognjen</creator><creator>Fortier, Louis-Charles</creator><general>American Society for Microbiology</general><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7T7</scope><scope>7TM</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope><scope>5PM</scope></search><sort><creationdate>20150201</creationdate><title>Global Transcriptional Response of Clostridium difficile Carrying the ϕCD38-2 Prophage</title><author>Sekulovic, Ognjen ; Fortier, Louis-Charles</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c361t-90d03eedbb918aa01671c387b41535d29eaeee75d90c77fc1204fe1032ab15db3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Clostridium difficile</topic><topic>Genetics and Molecular Biology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Sekulovic, Ognjen</creatorcontrib><creatorcontrib>Fortier, Louis-Charles</creatorcontrib><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Applied and Environmental Microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Sekulovic, Ognjen</au><au>Fortier, Louis-Charles</au><au>Schaffner, D. W.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Global Transcriptional Response of Clostridium difficile Carrying the ϕCD38-2 Prophage</atitle><jtitle>Applied and Environmental Microbiology</jtitle><date>2015-02-01</date><risdate>2015</risdate><volume>81</volume><issue>4</issue><spage>1364</spage><epage>1374</epage><pages>1364-1374</pages><issn>0099-2240</issn><eissn>1098-5336</eissn><eissn>1098-6596</eissn><abstract>Clostridium difficile
is one of the most dangerous pathogens in hospital settings. Most strains of
C. difficile
carry one or more prophages, and some of them, like ϕCD38-2 and ϕCD119, can influence the expression of toxin genes. However, little is known about the global host response in the presence of a given prophage. In order to fill this knowledge gap, we used high-throughput RNA sequencing (RNA-seq) to conduct a genome-wide transcriptomic analysis of the epidemic
C. difficile
strain R20291 carrying the ϕCD38-2 prophage. A total of 39 bacterial genes were differentially expressed in the R20291 lysogen, 26 of them being downregulated. Several of the regulated genes encode transcriptional regulators and phosphotransferase system (PTS) subunits involved in glucose, fructose, and glucitol/sorbitol uptake and metabolism. ϕCD38-2 also upregulated the expression of a group of regulatory genes located in phi-027, a resident prophage common to most ribotype 027 isolates. The most differentially expressed gene was that encoding the conserved phase-variable cell wall protein CwpV, which was upregulated ∼20-fold in the lysogen. Quantitative PCR and immunofluorescence showed that the increased
cwpV
expression results from a greater proportion of cells actively transcribing the gene. Indeed, ∼95% of lysogenic cells express
cwpV
, as opposed to only ∼5% of wild-type cells. Furthermore, the higher proportion of cells expressing
cwpV
results from a higher frequency of recombination of the genetic switch controlling phase variation, which we confirmed to be dependent on the host-encoded recombinase RecV. In summary, ϕCD38-2 interferes with phase variation of the surface protein CwpV and the expression of metabolic genes.</abstract><cop>1752 N St., N.W., Washington, DC</cop><pub>American Society for Microbiology</pub><pmid>25501487</pmid><doi>10.1128/AEM.03656-14</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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source | American Society for Microbiology; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Alma/SFX Local Collection |
subjects | Clostridium difficile Genetics and Molecular Biology |
title | Global Transcriptional Response of Clostridium difficile Carrying the ϕCD38-2 Prophage |
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