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GalR Acts as a Transcriptional Activator of galKT in the Presence of Galactose in Streptococcus pneumoniae

ABSTRACT: We explored the regulatory mechanism of Leloir pathway genes in Streptococcus pneumoniae D39. Here, we demonstrate that the expression of galKT is galactose dependent. By microarray analysis and quantitative RT-PCR, we further show the role of the transcriptional regulator GalR, present upstream of galKT, as a transcriptional activator of galKT in the presence of galactose. Moreover, we predict a 19-bp regulatory site (5'-GATAGTTTAGTAAAATTTT-3' ) for the transcriptional regulator GalR in the promoter region of galK, which is also highly conserved in other streptococci. Growth comparison of Δ galK with the D39 wildtype strain grown in the presence of galactose shows that galK is required for the proper growth of S. pneumoniae on galactose. This SuperSeries is composed of the SubSeries listed below.

ORGANISM(S): Streptococcus pneumoniae D39

PROVIDER: GSE74502 | GEO | 2015/10/30

SECONDARY ACCESSION(S): PRJNA300565

REPOSITORIES: GEO

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Comparison of the Streptococcus pneumoniae D39 wild-type to the D39 galR mutant [GalM17]

Project description:We explored the regulatory mechanism of Leloir pathway genes in Streptococcus pneumoniae D39. Here, we demonstrate that the expression of galKT is galactose dependent. By microarray analysis and quantitative RT-PCR, we further show the role of the transcriptional regulator GalR, present upstream of galKT, as a transcriptional activator of galKT in the presence of galactose. Moreover, we predict a 19-bp regulatory site (5'-GATAGTTTAGTAAAATTTT-3' ) for the transcriptional regulator GalR in the promoter region of galK, which is also highly conserved in other streptococci. Growth comparison of Î galK with the D39 wildtype strain grown in the presence of galactose shows that galK is required for the proper growth of S. pneumoniae on galactose. Comparison of the Streptococcus pneumoniae D39 galR mutant compared to D39 wild type in M17 medium+ 0.5% (w/v) galactose (GalM17). One condition design comparison of two strains including a dye swap.

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GalR Acts as a Transcriptional Activator of galKT in the Presence of Galactose in Streptococcus pneumoniae

Project description:GalR Acts as a Transcriptional Activator of galKT in the Presence of Galactose in Streptococcus pneumoniae

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Project description:Transcriptome comparison of the Streptococcus pneumoniae strain D39 grown in the presence of either lactose or galactose with that of the strain grown in the presence of glucose, revealed the elevated expression of various genes and operons, including the lac gene cluster that is organized into two operons i.e. lac operon-I (lacABCD) and lac operon-II (lacTFEG). Deletion of the DeoR family transcriptional regulator lacR that is present downstream of the lac gene cluster, revealed elevated expression of lac operon-I, even in the absence of lactose. This suggests a function of LacR as a transcriptional repressor of lac operon-I that encodes enzymes involved in the Tagatose-6-P pathway in the absence of lactose or galactose. Deletion of lacR did not affect the expression of lac operon-II that encodes for a lactose-specific PTS. This finding was further confirmed by ?-galactosidase assays with PlacA-lacZ and PlacT-lacZ in the presence of either lactose or glucose as a sole carbon source in the medium. This suggests the presence of another transcriptional regulator in the regulation of lac operon-II, which could be the BglG-family transcriptional antiterminator LacT. We demonstrate the role of LacT as a transcriptional activator of lac operon-II in the presence of lactose and CcpA-independent regulation of the lac gene cluster in S. pneumoniae. This SuperSeries is composed of the SubSeries listed below.

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