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Regulation of glutamine and glutamate metabolism by GlnR and GlnA in Streptococcus pneumoniae

ABSTRACT: 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

ORGANISM(S): Streptococcus pneumoniae

PROVIDER: GSE5088 | GEO | 2006/06/23

SECONDARY ACCESSION(S): PRJNA95391

REPOSITORIES: GEO

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Regulation of glutamine and glutamate metabolism by GlnR and GlnA in Streptococcus pneumoniae

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.

2010-07-01 | E-GEOD-5088 | biostudies-arrayexpress

Streptococcus pneumoniae

Project description:Regulation of glutamine and glutamate metabolism by GlnR and GlnA in Streptococcus pneumoniae

| PRJNA95391 | ENA

Dual regulation of nitrogen metabolism by CodY and GlnR in Streptococcus mutans

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.

2010-07-01 | GSE13540 | GEO

Mycobacterium smegmatis - Comparison of glnR deletion strain MH1 under nitrogen surplus and starvation

Project description:In this approach, changes in total transcript of a glnR deletion strain of Mycobacterium smegmatis due to nitrogen starvation were monitored. This is a control experiment for the comparison of wild type and glnR deletion strain resulting in putative nitrogen-related genes which are not controlled by GlnR.

2011-07-01 | GSE30033 | GEO

The global transcriptomic profile in the manganese-stressed wild type, DglnA and DmntPDglnA E. coli strains

Project description:We have grown the E. coli strains in the presence or absence of 1mM MnCl2 to observe if GlnA has some role in mitigating maganese stress. We identified the pathways that are altered under manganese stress. From the analyses, we noticed that nitrogen metabolism genes, glutamate glutamine cycle enzymes coding genes, pH homeostasis genes play crucial roles in mitigating manganese stress. GlnA plays central role in such manganes-shock managnement.

2022-03-05 | GSE186657 | GEO

Analyses of the GlnR- and AmtR- dependent nitrogen regulatory network of Mycobacterium smegmatis.

Project description:Total transcriptome analyses were carried out to investigate changes in transcript patterns in the Mycobacterium smegmatis wild type strain SMR5 due to nitrogen starvation and to expand the knowledge about the role of the two transcriptional regulators of nitrogen metabolism, namely GlnR and AmtR, in these processes. A first experiment revealed enhanced transcript levels of 284 genes and reduced transcripts of 231 genes in the wild type under nitrogen starvation compared to nitrogen surplus. When glnR deletion strain MH1 was compared to the wild type under nitrogen starvation, decreased transcript levels of 125 genes were detected, indicating that these are activated by GlnR due to nitrogen limitation. Comparing amtR deletion strain YL1 to the wild type under nitrogen starvation, enhanced transcript levels of 2 genes were found, indicating that they are repressed by AmtR under nitrogen surplus. A comparison of YL1 and wild type under surplus, as well as a comparison of YL1 under nitrogen surplus and starvation and of MH1 under nitrogen surplus and starvation were carried out as additional control experiments. It can be concluded that GlnR is the master regulator of nitrogen control in M. smegmatis and that AmtR fulfills only a small, subordinate role in the regulation of an operon. This SuperSeries is composed of the SubSeries listed below.

2011-07-01 | GSE30236 | GEO

Mycobacterium smegmatis - Comparison of wild type SMR5 and glnR deletion strain MH1 under nitrogen starvation

Project description:In this approach, total transcript of the Mycobacterium smegmatis wild type SMR5 and a glnR deletion strain was compared under nitrogen starvation. Transcript levels of 6 genes were enhanced, 125 genes reduced in the mutant strain. This experiment confirmed GlnR as the global regulator of nitrogen metabolism in M. smegmatis. Two biological replicates were analyzed.

2011-07-01 | GSE30234 | GEO

L-Glutamine metabolism and its regulation in C. glutamicum

Project description:Corynebacterium glutamicum, a gram-positive soil bacterium used for the industrial production of amino acids such as L-glutamate and L-lysine, is able to use a number of different nitrogen sources, such as ammonium, urea, or creatinine. In this communication, we show that L-glutamine serves as an excellent nitrogen source for C. glutamicum and allows similar growth rates in glucose minimal medium as ammonium. A transcriptome comparison revealed a strong induction of the nitrogen starvation response when glutamine was used as nitrogen source. Subsequent growth experiments with a variety of mutants defective in nitrogen metabolism showed that glutamate synthase is crucial for glutamine utilization, while a putative glutaminase is dispensable under the experimental conditions used. The fact that the glutamate synthase encoding gltBD operon is under strict nitrogen control explains the necessity for induction of the nitrogen starvation response. The paradox situation that the nitrogen starvation response is induced although intracellular L-glutamine levels are high has implications on nitrogen sensing. In contrast to other gram-positive and gram-negative bacteria such as Bacillus subtilis, Salmonella typhimurium, and Klebsiella pneumoniae, a drop in glutamine concentration obviously does not serve as a nitrogen starvation signal in C. glutamicum.

2010-10-05 | GSE19779 | GEO

Transcriptional analysis of glnA deletion mutant in Haloferax mediterranei

Project description:Transcriptional profiling of Haloferax mediterranei DglnA and HM26 strains in two different culture media: a) cells were grown exponentially on complex media + 40 mM Gln, b) cells were shifted to nitrogen starvation conditions (72h). In the Nsta(∆glnA)-Cx and Nsta-Cx contrasts, glnA2 was up-regulated, and glnA3 did not show any difference. Therefore, glnA2 and glnA3 were unable to compensate lack of glnA because either they do not properly encode GS or are not expressed under the same conditions.

2020-04-30 | GSE135303 | GEO

Transcription profiling by array of the complete GlnR nitrogen-response regulon in Mycobacterium smegmatis

Project description:BACKGROUND: Nitrogen is an essential element for bacterial growth and an important component of biological macromolecules. Consequently, responding to nitrogen limitation is critical for bacterial survival and involves the interplay of signalling pathways and transcriptional regulation of nitrogen assimilation and scavenging genes. In the soil dwelling saprophyte Mycobacterium smegmatis the OmpR-type response regulator GlnR is thought to mediate the transcriptomic response to nitrogen limitation. However, to date only ten genes have been shown to be in the GlnR regulon, a vastly reduced number compared to other organisms. RESULTS: We investigated the role of GlnR in the nitrogen limitation response and determined the entire GlnR regulon, by combining expression profiling of M. smegmatis wild type and glnR deletion mutant, with GlnR-specific chromatin immunoprecipitation and high throughput sequencing. We identify 53 GlnR binding sites during nitrogen limitation that control the expression of over 100 genes, demonstrating that GlnR is the regulator controlling the assimilation and utilisation of nitrogen. We also determine a consensus GlnR binding motif and identify key residues within the motif that are required for specific GlnR binding. CONCLUSIONS: We have demonstrated that GlnR is the global nitrogen response regulator in M. smegmatis, directly regulating the expression of more than 100 genes. GlnR controls key nitrogen stress survival processes including primary nitrogen metabolism pathways, the ability to utilise nitrate and urea as alternative nitrogen sources, and the potential to use cellular components to provide a source of ammonium. These studies further our understanding of how mycobacteria survive nutrient limiting conditions. [Data is also available from http://bugs.sgul.ac.uk/E-BUGS-143]

2013-04-24 | E-BUGS-143 | biostudies-arrayexpress

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