{"data":{"id":"10.25384/sage.c.4161353","type":"dois","attributes":{"doi":"10.25384/sage.c.4161353","prefix":"10.25384","suffix":"sage.c.4161353","identifiers":[],"alternateIdentifiers":[],"creators":[{"name":"L. Lei","affiliation":[],"nameIdentifiers":[]},{"name":"R.N. Stipp","affiliation":[],"nameIdentifiers":[]},{"name":"T. Chen","affiliation":[],"nameIdentifiers":[]},{"name":"S.Z. Wu","affiliation":[],"nameIdentifiers":[]},{"name":"T. Hu","affiliation":[],"nameIdentifiers":[]},{"name":"M.J. Duncan","affiliation":[],"nameIdentifiers":[]}],"titles":[{"title":"Activity of \u003ci\u003eStreptococcus mutans\u003c/i\u003e VicR Is Modulated by Antisense RNA"}],"publisher":"Figshare","container":{},"publicationYear":2018,"subjects":[{"subject":"110599 Dentistry not elsewhere classified","subjectScheme":"FOR"},{"subject":"FOS: Clinical medicine","schemeUri":"http://www.oecd.org/science/inno/38235147.pdf","subjectScheme":"Fields of Science and Technology (FOS)"},{"subject":"91299 Materials Engineering not elsewhere classified","subjectScheme":"FOR"},{"subject":"FOS: Materials engineering","schemeUri":"http://www.oecd.org/science/inno/38235147.pdf","subjectScheme":"Fields of Science and Technology (FOS)"}],"contributors":[],"dates":[{"date":"2018-07-11","dateType":"Created"},{"date":"2018-07-11","dateType":"Updated"},{"date":"2018","dateType":"Issued"}],"language":null,"types":{"ris":"GEN","bibtex":"misc","citeproc":"article","schemaOrg":"Collection","resourceType":"Collection","resourceTypeGeneral":"Collection"},"relatedIdentifiers":[{"relationType":"IsSupplementTo","relatedIdentifier":"10.1177/0022034518781765","relatedIdentifierType":"DOI"}],"relatedItems":[],"sizes":[],"formats":[],"version":null,"rightsList":[{"rights":"CC BY 4.0","rightsUri":"https://creativecommons.org/licenses/by/4.0"}],"descriptions":[{"description":"The VicRK 2-component system of \u003ci\u003eStreptococcus mutans\u003c/i\u003e regulates genes associated with cell wall biogenesis and biofilm formation. A putative RNase III–encoding gene (\u003ci\u003ernc\u003c/i\u003e) is located downstream from the \u003ci\u003evicRKX\u003c/i\u003e operon. The goals of this study were to investigate the potential role of VicR in the regulation of adjacent downstream genes and evaluate transcription levels of \u003ci\u003evicR\u003c/i\u003e during planktonic and biofilm growth. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to investigate whether \u003ci\u003evicRKX\u003c/i\u003e and adjacent downstream genes were cotranscribed. Binding of purified recombinant VicR protein to promoter regions of \u003ci\u003evicR, rnc\u003c/i\u003e, and \u003ci\u003esyfA\u003c/i\u003e genes was confirmed by electrophoretic mobility shift assay and by chromatin immunoprecipitation analyses. \u003ci\u003eVicR\u003c/i\u003e antisense (AS\u003ci\u003evicR\u003c/i\u003e) RNA was detected by Northern blotting and qRT-PCR assays. AS\u003ci\u003evicR\u003c/i\u003e overexpression mutants were constructed, and the biofilm biomass was determined by crystal violet microtiter assay. Adjacent downstream genes \u003ci\u003ernc, smc, syfA, smu.1511\u003c/i\u003e, and \u003ci\u003esyfB\u003c/i\u003e were cotranscribed with \u003ci\u003evicRKX\u003c/i\u003e. The predicted promoter regions of \u003ci\u003evicR, rnc\u003c/i\u003e, and \u003ci\u003esyfA\u003c/i\u003e genes were directly regulated by VicR. An AS\u003ci\u003evicR\u003c/i\u003e RNA transcript was detected upstream of the \u003ci\u003ernc\u003c/i\u003e gene. Expression of the AS\u003ci\u003evicR\u003c/i\u003e RNA transcript was elevated in planktonic cultures and repressed during biofilm growth. In addition, Western blot data showed that expression of the VicR protein decreased by 35% in planktonic as compared with biofilm cultures. Furthermore, we show that overexpression of AS\u003ci\u003evicR\u003c/i\u003e led to a reduction in biofilm formation. The downstream genes \u003ci\u003ernc, smc, syfA, smu.1511\u003c/i\u003e, and \u003ci\u003esyfB\u003c/i\u003e are cotranscribed with \u003ci\u003evicRKX. VicR\u003c/i\u003e is autophosphorylated, and \u003ci\u003ernc\u003c/i\u003e and \u003ci\u003esyfA\u003c/i\u003e are directly regulated by VicR. Expression of VicR protein correlated inversely with different levels of AS\u003ci\u003evicR\u003c/i\u003e RNA transcript and growth conditions. The biofilm biomass decreased in the AS\u003ci\u003evicR\u003c/i\u003e overexpression mutant. These data suggest a role for the AS\u003ci\u003evicR\u003c/i\u003e RNA transcript in posttranscriptional regulation of VicR protein production in \u003ci\u003eS. mutans\u003c/i\u003e.","descriptionType":"Abstract"}],"geoLocations":[],"fundingReferences":[],"xml":"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