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Regulation of PTSBio by transcriptional repressor NigR

ABSTRACT: In this study we analyzed the transcriptional regulation of the operon (OBio) encoding the PTSBio and showed that it was repressed by NigR, a LacI-like transcriptional regulator. To elucidate the role of NigR in regulation of S. mutans carbohydrate transporters we performed full genome expression microarrays using custom Affymetrix chip. Expression microarray analysis was performed with ΔnigR grown in CDM supplemented with glucose and compared to that of WT grown under identical conditions. Deletion of the nigR gene did not exert broad transcription changes in S. mutans under tested conditions. OBio was the only operon that was significantly upregulated in the ΔnigR strain (Supplemental Table 1). This data confirmed that NigR is the repressor of OBio and that OBio is its only PTS target under tested condition.

ORGANISM(S): Streptococcus mutans UA159 Streptococcus mutans

PROVIDER: GSE63213 | GEO | 2015/06/22

SECONDARY ACCESSION(S): PRJNA266999

REPOSITORIES: GEO

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Project description:Salmonella is an important enteric pathogen that causes a spectrum of diseases varying from mild gastroenteritis to life threatening typhoid fever. Salmonella does not have lac operon. However, E. Coli, Salmonellaâs close relative, has lac operon. Being an enteric pathogen like E. coli, Salmonella will also benefit from lac operon. Then, why Salmonella has lost lac operon?. To address this question, lacI, an important component of lac operon was expressed in Salmonella via pTrc99A plasmid. As a control, pTrc99A without lacI was also expressed in Salmonella. The effect of LacI on the transcription profile of Salmonella was analyzed using microarray technique. Overall design The total RNA was isolated from Salmonella using âRNeasy Mini Kitâ (QIAGEN). Organism used: Salmonella enterica serovar Typhimurium LT2 * Slides: Agilentâs arrays (8x15k) AMADID: NO: 17809 * Starting material: Salmonella RNA in nuclease-free water * RNA Samples used: Wild type (WT), WT + pTrc99A+LacI (PTRC), WT + pTrc99A-LacI(Delta lac) * Labeling kit: MessageAmpâ¢ II-Bacteria RNA Amplification Kit from Ambion Cat # AM1790 * Labeling Method: T7 promoter based-linear amplification to generate labeled complementary RNA * Total RNA and cRNA Purification Kit: Qiagenâs RNeasy minikit Cat#74104 * Hybridization Kit: Agilentâs In situ Hybridzation kit 5184-3568 * RNA quality was checked using Bioanalyzer. The array slides were scanned immediately by PerkinElmer Scan array Gx Microarray scanner. The Scan array software (PerkinElmer) was used for grid wise normalization of array images. Two arrays were used with mutant and rescue experiments and dye swap experiments were included in the final analysis. The data was analysed by GeneSpring GX and Biointerpreter software from Genotypic Technology, Bangalore. The differential expression was considered if the Log 2 mean of at least -1 for the down regulated genes and +1 for the upregulated genes. We considered only the genes that were reproducible from both replicates.

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Regulation of PTSBio by transcriptional repressor NigR

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