Project description:Several genes involved in nitrogen metabolism are known to contribute to the virulence of pathogenic bacteria. Here, we studied the function of the nitrogen regulatory protein GlnR in the Gram-positive human pathogen Streptococcus pneumoniae. We demonstrate that GlnR mediates transcriptional repression of genes involved in glutamine synthesis and uptake (glnA, glnPQ), glutamate synthesis (gdhA), and the gene encoding the pentose phosphate pathway enzyme Zwf, which forms an operon with glnPQ. Moreover, the expression of gdhA is also repressed by the pleiotropic regulator CodY. The GlnR-dependent regulation occurs through a conserved operator sequence and is responsive to the concentration of glutamate, glutamine and ammonium in the growth medium. By means of in vitro binding studies and transcriptional analyses we show that the regulatory function of GlnR is dependent on GlnA. Mutants of glnA and glnP displayed significantly reduced adhesion to Detroit 562 human pharyngeal epithelial cells, suggesting a role for these genes in the colonization of the host by S. pneumoniae. Thus, our results provide a thorough insight into the regulation of glutamine and glutamate metabolism of S. pneumoniae as mediated by both GlnR and GlnA. Keywords: genetic modification
Project description:Several genes involved in nitrogen metabolism are known to contribute to the virulence of pathogenic bacteria. Here, we studied the function of the nitrogen regulatory protein GlnR in the Gram-positive human pathogen Streptococcus pneumoniae. We demonstrate that GlnR mediates transcriptional repression of genes involved in glutamine synthesis and uptake (glnA, glnPQ), glutamate synthesis (gdhA), and the gene encoding the pentose phosphate pathway enzyme Zwf, which forms an operon with glnPQ. Moreover, the expression of gdhA is also repressed by the pleiotropic regulator CodY. The GlnR-dependent regulation occurs through a conserved operator sequence and is responsive to the concentration of glutamate, glutamine and ammonium in the growth medium. By means of in vitro binding studies and transcriptional analyses we show that the regulatory function of GlnR is dependent on GlnA. Mutants of glnA and glnP displayed significantly reduced adhesion to Detroit 562 human pharyngeal epithelial cells, suggesting a role for these genes in the colonization of the host by S. pneumoniae. Thus, our results provide a thorough insight into the regulation of glutamine and glutamate metabolism of S. pneumoniae as mediated by both GlnR and GlnA. Each amplicon was spotted twice (technical replicates) on the DNA microarray. These replicates are indicated in the platforms and DNA microarray data by the addition of _rep1, _rep2, etc. The RNA for wild-type and mutant strains was isolated in 3 independent biological replicates which were hybridized in (partly dye-swap) to the 3 slides for glnA and 3 slides for glnR.
Project description:This microarray experiment is part of a study addressing the importance of glutamine metabolism in Streptococcus pneumoniae for the virulence of this bacterium. To be able to gain more insight into the phenotype of a double mutant of glnA (TIGR 4 locus tag SP0502, encoding glutamine synthetase) and glnP (SP1241, encoding a permease component of a glutmine uptake ABC transporter), the transcriptome of this mutant, the glnPA double mutant, was determined in strain S. pneumoniae D39 grown in rich GM17 medium with 0.5 mg/ml glutamine. This revealed a big change in transcriptome in the mutant, especially a lot of amino acid and peptide metabolic genes. Keywords: transcriptome analysis of D39 wild-type compared with its isogenic glnPA double mutant
Project description:Comparing transcriptome by means of DNA microarrays of Streptococcus pneumoniae D39 versus D39SM in medium with uracil. Details of this experiment are described in a paper with the title 'Interplay between capsule expression and uracil metabolism in Streptococcus pneumoniae'
Project description:Comparing transcriptome by means of DNA microarrays of Streptococcus pneumoniae D39 versus D39SM in medium without uracil. Details of this experiment are described in a paper with the title 'Interplay between capsule expression and uracil metabolism in Streptococcus pneumoniae'
Project description:Comparing transcriptome by means of DNA microarrays of Streptococcus pneumoniae D39 versus D39PcpsT=>C in medium with uracil. Details of this experiment are described in a paper with the title 'Interplay between capsule expression and uracil metabolism in Streptococcus pneumoniae'
Project description:Comparing transcriptome by means of DNA microarrays of Streptococcus pneumoniae D39 versus D39PcpsT=>C in medium without uracil. Details of this experiment are described in a paper with the title 'Interplay between capsule expression and uracil metabolism in Streptococcus pneumoniae'
Project description:CodY and GlnR are two major transcriptional regulators in nitrogen metabolism in Gram-positive bacteria. CodY regulates genes involved in the adaptive response to poor growth conditions, especially to nutrient limitation. GlnR controls nitrogen utilization according to the availability of nitrogen source. In this study we used microarray to investigate the regulatory roles that CodY and GlnR play in nitrogen metabolism in Streptococcus mutans.
Project description:In our study, deletion of methionine synthesis gene metE (SPD_0510) impaired the growth of Streptococcus pneumoniae strain D39 significantly when bacteria were cultured in chemically defined medium with a low concentration of methionine. Bacteria undergo methionine starvation in this condition. We aimed to know how S. pneumoniae responses to methionine starvation, particularly at the transcriptional level. RNA-seq results show that 804 genes (41.2 % of whole genome) were differentially expressed under methionine starvation. Among them, 258 genes (32.1 %) are related to cell metabolism, with 75 genes involved in sugar and carbon metabolism, 88 genes for synthesis of metabolic products and 95 genes for uptake of amino acids and sugars. Combined with other experiments data, this showed the reprogrammed metabolism in S. pneumoniae under methionine starvation.
Project description:This SuperSeries is composed of the following subset Series: GSE31815: ccpA mutant compared to D39 wild-type in Streptococcus pneumoniae in CDM + Glucose at MID-log growth phase GSE31816: ccpA mutant compared to D39 wild-type in Streptococcus pneumoniae in CDM + GLucose at transition-phase of growth (TS) GSE31817: ccpA mutant compared to D39 wild-type in Streptococcus pneumoniae in CDM + Galactose at MID-log growth phase GSE31818: ccpA mutant compared to D39 wild-type in Streptococcus pneumoniae in CDM + galactose at transition-phase of growth (TS) Refer to individual Series