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Salmonella enterica Typhimurium grown with and without glucose

ABSTRACT: Cell to cell communication in bacteria to regulate various cellular processes with respect to their population density is termed quorum sensing and is achieved using signaling molecules called autoinducers. LuxS, which is involved in the synthesis of the autoinducer molecule-2 (AI-2), is conserved in several Gram-positive and Gram-negative bacteria including the enteric pathogen Salmonella Typhimurium. Genes that are regulated by luxS in S. Typhimurium were identified using microarrays and RNA samples from wild type S. Typhimurium and its isogenic luxS mutant, in two growth conditions (presence and absence of glucose), and at two different time points (mid-log and early-stationary phases). Minimal differential gene expression was observed in the presence of glucose. In the absence of both luxS and glucose, a total of 1560 genes were differentially expressed and 1361 genes were identified as luxS/AI-2-regulated at the mid-log phase and 199 genes at the early-stationary phase. Quantitative real-time PCR was performed on selected genes to validate the microarray results. These results suggest that although the expression of the luxS gene in S. Typhimurium is independent of the growth condition, its role in the production of AI-2 depends on the growth condition. It was found that luxS/AI-2 plays a vital role in a variety of processes such as metabolism, virulence gene expression, motility, transcription and translation. Keywords: Salmonella Typhimurium, quorum sensing, luxS mutant, autoinducer-2

ORGANISM(S): Salmonella enterica subsp. enterica serovar Typhi str. Ty2 Salmonella enterica subsp. enterica serovar Typhimurium str. LT2 Salmonella enterica subsp. enterica serovar Typhi str. CT18 Salmonella enterica subsp. enterica serovar Typhimurium

PROVIDER: GSE7558 | GEO | 2009/10/22

SECONDARY ACCESSION(S): PRJNA99359

REPOSITORIES: GEO

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Project description:LuxS is an enzyme involved in the activated methyl cycle. The by-product of this cycle, autoinducer 2 (AI-2), could be an important quorum sensing signal. LuxS was conserved and regulated many behaviors in different bacteria, but in most species, whether the regulations are related to AI-2 mediated quorum sensing is still unknown. In our previous study, Actinobacillus pleuropneumoniae, the etiologic agent of porcine contagious pleuropneumonia, was found to possess the functional LuxS affecting biofilm formation and virulence. In this study, microarray was used to compare the transcriptional profiles of the A. pleuropneumoniae wildtype strain 4074, luxS mutant and its AI-2 supplemented strain in four different growth phases. The results demonstrated that both LuxS and AI-2 played important roles in metabolism of this bacterium. AI-2 did not recover the gene expression changes caused by luxS deletion but caused more extensive metabolic alterations in the luxS mutant in a concentration dependent manner. Regulations of the genes involved in iron metabolism, carbonhydrate transport and host cell adhesion were validated by real-time RT-PCR and phenotypic investigations under different conditions. The results demonstrated that sugar uptake was repressed by LuxS independent of AI-2. However, iron metabolism, an important process of infection for A. pleuropneumoniae, could be controlled by LuxS through AI-2. Adhesion to host cells was also affected by LuxS and AI-2, but exogenous AI-2 displayed more obvious effects. Our results indicated that both LuxS and AI-2 are essential in the metabolism of A. pleuropneumoniae. The function of LuxS/AI-2 displayed pleiotropic roles in different conditions. AI-2 may not serve as a quorum sensing autoinducer but a metabolite or an environmental cue to affect the metabolism and virulence traits of this important pathogen. A. pleuropneumoniae strains were cultured in TSB medium supplemented with 10 M-NM-<g/ml of nicotinamide adenine dinucleotide (NAD) and 10% (v/v) filtered cattle serum at 37M-BM-0C. For samples from the mutant supplemented with AI-2, 100M-NM-<M and 25M-NM-<M AI-2 precursor (DPD) were added into the medium before early exponential phase as well as one hour before late exponential phase to cover the whole growth phase respectively. The samples were collected from early, middle, late exponential phase and stationary phase respectively and the total RNA were extracted using RNA-Solv Reagent (Omega) according to the manufacturerM-bM-^@M-^Ys instructions. For each time point, four biological replicates were combined into two samples. The intensities were normalized and transformed into log2 value. The values for each time point were averaged and the genes with log2 ratio >=1 or <=-1 were selected as differentially expressed genes.

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Salmonella enterica Typhimurium grown with and without glucose

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