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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">The Spatial Organization of the VirR Boxes Is Critical for VirR-Mediated Expression of the Perfringolysin O Gene, <italic>pfoA</italic>, from <italic>Clostridium perfringens</italic></title><author><name sortKey="Cheung, Jackie K" sort="Cheung, Jackie K" uniqKey="Cheung J" first="Jackie K." last="Cheung">Jackie K. Cheung</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Dupuy, Bruno" sort="Dupuy, Bruno" uniqKey="Dupuy B" first="Bruno" last="Dupuy">Bruno Dupuy</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Deveson, Deanna S" sort="Deveson, Deanna S" uniqKey="Deveson D" first="Deanna S." last="Deveson">Deanna S. Deveson</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Rood, Julian I" sort="Rood, Julian I" uniqKey="Rood J" first="Julian I." last="Rood">Julian I. Rood</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">15150217</idno><idno type="pmc">415773</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC415773</idno><idno type="RBID">PMC:415773</idno><idno type="doi">10.1128/JB.186.11.3321-3330.2004</idno><date when="2004">2004</date><idno type="wicri:Area/Pmc/Corpus">001002</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">001002</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">The Spatial Organization of the VirR Boxes Is Critical for VirR-Mediated Expression of the Perfringolysin O Gene, <italic>pfoA</italic>, from <italic>Clostridium perfringens</italic></title><author><name sortKey="Cheung, Jackie K" sort="Cheung, Jackie K" uniqKey="Cheung J" first="Jackie K." last="Cheung">Jackie K. Cheung</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Dupuy, Bruno" sort="Dupuy, Bruno" uniqKey="Dupuy B" first="Bruno" last="Dupuy">Bruno Dupuy</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Deveson, Deanna S" sort="Deveson, Deanna S" uniqKey="Deveson D" first="Deanna S." last="Deveson">Deanna S. Deveson</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Rood, Julian I" sort="Rood, Julian I" uniqKey="Rood J" first="Julian I." last="Rood">Julian I. Rood</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></analytic><series><title level="j">Journal of Bacteriology</title><idno type="ISSN">0021-9193</idno><idno type="eISSN">1098-5530</idno><imprint><date when="2004">2004</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>The transcriptional regulation of toxin production in the gram-positive anaerobe <italic>Clostridium perfringens</italic> involves a two-component signal transduction system that comprises the VirS sensor histidine kinase and its cognate response regulator, VirR. Previous studies showed that VirR binds independently to a pair of imperfect direct repeats, now designated VirR box 1 and VirR box 2, located immediately upstream of the promoter of the <italic>pfoA</italic> gene, which encodes the cholesterol-dependent cytolysin, perfringolysin O. For this study, we introduced mutated VirR boxes into a <italic>C. perfringens pfoA</italic> mutant and found that both VirR boxes are essential for transcriptional activation. Furthermore, the spacing between the VirR boxes and the distance between the VirR boxes and the −35 region are shown to be critical for perfringolysin O production. Other VirR boxes that were previously identified from the strain 13 genome sequence were also analyzed, with perfringolysin O production used as a reporter system. The results showed that placement of the different VirR boxes at the same position upstream of the <italic>pfoA</italic> promoter yields different levels of perfringolysin O activity. In all of these constructs, VirR was still capable of binding to the target DNA, indicating that DNA binding alone is not sufficient for transcriptional activation. Finally, we show that the <italic>C. perfringens</italic> RNA polymerase binds more efficiently to the <italic>pfoA</italic> promoter in the presence of VirR, indicating that interactions must occur between these proteins. We propose that these interactions are required for VirR-mediated transcriptional activation.</p></div></front></TEI><pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Bacteriol</journal-id><journal-id journal-id-type="publisher-id">jb</journal-id><journal-title>Journal of Bacteriology</journal-title><issn pub-type="ppub">0021-9193</issn><issn pub-type="epub">1098-5530</issn><publisher><publisher-name>American Society for Microbiology</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">15150217</article-id><article-id pub-id-type="pmc">415773</article-id><article-id pub-id-type="publisher-id">1546-03</article-id><article-id pub-id-type="doi">10.1128/JB.186.11.3321-3330.2004</article-id><article-categories><subj-group subj-group-type="heading"><subject>Gene Regulation</subject></subj-group></article-categories><title-group><article-title>The Spatial Organization of the VirR Boxes Is Critical for VirR-Mediated Expression of the Perfringolysin O Gene, <italic>pfoA</italic>, from <italic>Clostridium perfringens</italic></article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Cheung</surname><given-names>Jackie K.</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Dupuy</surname><given-names>Bruno</given-names></name><xref ref-type="aff" rid="aff1">2</xref></contrib><contrib contrib-type="author"><name><surname>Deveson</surname><given-names>Deanna S.</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Rood</surname><given-names>Julian I.</given-names></name><xref ref-type="aff" rid="aff1">1</xref><xref ref-type="corresp" rid="cor1">*</xref></contrib></contrib-group><aff id="aff1">Australian Bacterial Pathogenesis Program, Department of Microbiology, Monash University, Victoria 3800, Australia,<label>1</label> Unité de Génétique Moléculaire Bactérienne, Institut Pasteur, Paris, France<label>2</label></aff><author-notes><fn id="cor1"><label>*</label><p>Corresponding author. Mailing address: Australian Bacterial Pathogenesis Program, Department of Microbiology, Monash University, Victoria 3800, Australia. Phone: 61 3 9905 4825. Fax: 61 3 9905 4811. E-mail: <email>julian.rood@med.monash.edu.au</email>.</p></fn></author-notes><pub-date pub-type="ppub"><month>6</month><year>2004</year></pub-date><volume>186</volume><issue>11</issue><fpage>3321</fpage><lpage>3330</lpage><history><date date-type="received"><day>29</day><month>11</month><year>2003</year></date><date date-type="accepted"><day>23</day><month>2</month><year>2004</year></date></history><copyright-statement>Copyright © 2004, American Society for Microbiology</copyright-statement><copyright-year>2004</copyright-year><abstract><p>The transcriptional regulation of toxin production in the gram-positive anaerobe <italic>Clostridium perfringens</italic> involves a two-component signal transduction system that comprises the VirS sensor histidine kinase and its cognate response regulator, VirR. Previous studies showed that VirR binds independently to a pair of imperfect direct repeats, now designated VirR box 1 and VirR box 2, located immediately upstream of the promoter of the <italic>pfoA</italic> gene, which encodes the cholesterol-dependent cytolysin, perfringolysin O. For this study, we introduced mutated VirR boxes into a <italic>C. perfringens pfoA</italic> mutant and found that both VirR boxes are essential for transcriptional activation. Furthermore, the spacing between the VirR boxes and the distance between the VirR boxes and the −35 region are shown to be critical for perfringolysin O production. Other VirR boxes that were previously identified from the strain 13 genome sequence were also analyzed, with perfringolysin O production used as a reporter system. The results showed that placement of the different VirR boxes at the same position upstream of the <italic>pfoA</italic> promoter yields different levels of perfringolysin O activity. In all of these constructs, VirR was still capable of binding to the target DNA, indicating that DNA binding alone is not sufficient for transcriptional activation. Finally, we show that the <italic>C. perfringens</italic> RNA polymerase binds more efficiently to the <italic>pfoA</italic> promoter in the presence of VirR, indicating that interactions must occur between these proteins. We propose that these interactions are required for VirR-mediated transcriptional activation.</p></abstract></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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