Journal
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
Volume 80, Issue 17, Pages 5386-5393Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/AEM.00755-14
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- Chang Gung Memorial Hospital of Taiwan [CMRPD 100013]
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Urease gene expression in Streptococcus salivarius 57.I, a strain of one of the major alkali producers in the mouth, is induced by acidic pH and excess amounts of carbohydrate. Expression is controlled primarily at the transcriptional level from a promoter, p(ureI). Recent sequencing analysis revealed a CodY box located 2 bases 5' to the -35 element of p(ureI). Using continuous chemostat culture, transcription from p(ureI) was shown to be repressed by CodY, and at pH 7 the repression was more pronounced than that in cells grown at pH 5.5 under both 20 and 100 mM glucose. The direct binding of CodY to p(ureI) was demonstrated by electrophoretic mobility shift assay and chromatin immunoprecipitation (ChIP)-quantitative real-time PCR (qPCR). The result of ChIP-qPCR also confirmed that the regulation of CodY is indeed modulated by pH and the binding of CodY at neutral pH is further enhanced by a limited supply of glucose (20 mM). In the absence of CodY, the C-terminal domain of the RNA polymerase (RNAP) alpha subunit interacted with the AT tracks within the CodY box, indicating that CodY and RNAP compete for the same binding region. Such regulation could ensure optimal urease expression when the enzyme is most required, i.e., at an acidic growth pH with an excess amount of carbon nutrients.
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