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Transcriptional and metabolomic consequences of luxS inactivation reveal a metabolic rather than quorum sensing role for LuxS in Lactobacillus reuteri 100-23

ABSTRACT: Autoinducer-2 (AI-2)-mediated quorum sensing has been extensively studied in relation to the regulation of microbial behaviour. There are however two potential roles for the AI-2 synthase (LuxS). The first is in the production of AI-2 and the second is as an enzyme in the activated methyl cycle where it catalyses the conversion of S-ribosylhomocysteine to homocysteine. The by-product of the reaction catalysed by LuxS is (S)-4,5-dihydroxy-2,3-pentanedione (DPD), which spontaneously forms the furanones known collectively as AI-2. The mammalian gastrointestinal tract contains a complex collection of bacterial species so a method of interspecies communication might influence community structure and function. Lactobacillus reuteri 100-23 is an autochthonous inhabitant of the rodent forestomach where it adheres to the non-secretory epithelium of the forestomach forming a biofilm. Microarray comparisons of gene expression profiles of the L. reuteri 100-23 wild type and a luxS mutant under different culture conditions revealed altered transcription of genes encoding proteins involved in cysteine biosynthesis, the oxidative stress response and cell wall proteins. Metabolomic analysis revealed that the luxS mutation affected cellular levels of fermentation products, fatty acids and amino acids. Cell density-dependent changes (log phase versus stationary phase growth) in gene transcription were not detected. Overall, the results indicated that AI-2 was unlikely to be involved in gene regulation in L. reuteri 100-23 in a classical quorum-sensing type manner.

ORGANISM(S): Limosilactobacillus reuteri subsp. rodentium

PROVIDER: GSE32520 | GEO | 2011/10/01

SECONDARY ACCESSION(S): PRJNA147845

REPOSITORIES: GEO

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Transcriptional and metabolomic consequences of luxS inactivation reveal a metabolic rather than quorum sensing role for LuxS in Lactobacillus reuteri 100-23

Project description:Autoinducer-2 (AI-2)-mediated quorum sensing has been extensively studied in relation to the regulation of microbial behaviour. There are however two potential roles for the AI-2 synthase (LuxS). The first is in the production of AI-2 and the second is as an enzyme in the activated methyl cycle where it catalyses the conversion of S-ribosylhomocysteine to homocysteine. The by-product of the reaction catalysed by LuxS is (S)-4,5-dihydroxy-2,3-pentanedione (DPD), which spontaneously forms the furanones known collectively as AI-2. The mammalian gastrointestinal tract contains a complex collection of bacterial species so a method of interspecies communication might influence community structure and function. Lactobacillus reuteri 100-23 is an autochthonous inhabitant of the rodent forestomach where it adheres to the non-secretory epithelium of the forestomach forming a biofilm. Microarray comparisons of gene expression profiles of the L. reuteri 100-23 wild type and a luxS mutant under different culture conditions revealed altered transcription of genes encoding proteins involved in cysteine biosynthesis, the oxidative stress response and cell wall proteins. Metabolomic analysis revealed that the luxS mutation affected cellular levels of fermentation products, fatty acids and amino acids. Cell density-dependent changes (log phase versus stationary phase growth) in gene transcription were not detected. Overall, the results indicated that AI-2 was unlikely to be involved in gene regulation in L. reuteri 100-23 in a classical quorum-sensing type manner. Analysis of the microarray data was obtained from two or more independent biological replicates.

2011-09-30 | E-GEOD-32520 | biostudies-arrayexpress

Transcriptional analysis of luxS-null strain in Yersinia pestis CO92 at 37°C

2013-09-05 | GSE22849 | GEO

LuxS/AI-2 quorum sensing microarray comparison in Yersinia pestis at 30°C

Project description:The AI-2 quorum-sensing system has been linked to diverse phenotypes and regulatory changes in pathogenic bacteria. In strain CO92, the AI-2 signal is produced in a luxS-dependent manner, reaching maximal levels of 2.5 μM in late logarithmic growth, and both wild type and pigmentation mutant strains made equivalent levels of AI-2. Y. pestis CO92 possesses a chromosomal lsr locus encoding factors involved in the binding and import of AI-2, and confirming this assignment, an lsr deletion increased extracellular pools of AI-2. To assess the functional role of AI-2 sensing in Y. pestis, microarray study was conducted comparing the ∆Pgm strain R88 to a ∆Pgm ∆luxS mutant at 30°C to mimic the flea gut.

2016-01-01 | GSE30108 | GEO

Transcriptional analysis of luxS-null strain in Yersinia pestis CO92 at 37M-BM-0C

Project description:Yersinia pestis, the etiological agent of plague, is able to sense cell density by quorum sensing. The function of quorum sensing in Y. pestis is not clear. Here, the process of autoinducer 2 (AI-2) quorum sensing was investigated by comparing transcript profiles when luxS gene was knocked out. The luxS gene encodes S-ribosylhomocysteinase which can produce DPD, a precursor of AI-2. The strain M-bM-^HM-^Fpgm (pigmentation-negative) mutant R88 was called wild type. The M-bM-^HM-^Fpgm M-bM-^HM-^FluxS mutant was called control. Six independent RNA samples from R88 were paired with six independent RNA samples from M-bM-^HM-^Fpgm M-bM-^HM-^FluxS mutant cultures for hybridization to six two-color microarrays. A dye-swap design was used to remove the Cy5 and Cy3 dye bias.

2013-09-05 | E-GEOD-22849 | biostudies-arrayexpress

Effect of luxS gene mutation on Streptococcus pneumonaie biofilm gene expression

Project description:Streptococcus pneumoniae is opportunistic bacteria cause’s acute otitis media (AOM) in children. It colonizes the nasopharynx in the form of biofilms, and these biofilms act as reservoir, and are vital for pneumococcal infections. The pneumococcal biofilms are regulated by LuxS/AI-2 media quorum sensing. In this study, we confirmed the role of LuxS/AI-2 for in vitro formation of biofilms, assessed the effects of the absence of LuxS/AI-2 signaling, for pneumococcal middle ear infection and identified global genes regulated by LuxS/AI-2 during formation of pneumococcal biofilms. In the cDNA-microarray analysis, 117 genes were differentially expressed in D39 luxS mutant when compared with D39 wild type. Among the 66 genes encoding putative proteins and previously characterized proteins, 60 were significantly down-regulated and 6 were significantly up-regulated. The functional annotation revealed that genes involve in DNA replication and repair, ATP synthesis, capsule biosynthesis, cell division and cell cycle, signal transduction, transcription regulation, competence, virulence, and carbohydrate metabolism were down-regulated in the absence of LuxS/AI-2.

2018-05-17 | GSE109347 | GEO

Expression analysis of a full Lsr locus knockout on the luxS knockout background of Haemophilus influenzae strain CZ4126/02

Project description:Investigation of whole genome gene expression differences between a full (nine-gene) Lsr locus deletion and a luxS deletion strain (double knock-out) compared to a luxS isogenic mutant strain (each derived from the CZ4126/02 parent strain). Both strains are unable to produce the autoinducer-2 (AI-2) quorum sensing molecule. In addition, the Lsr mutant is unable to import AI-2 nor mediate any potential LsrR-based transcriptional regulation.

2014-05-06 | GSE57288 | GEO

LuxS/AI-2 quorum sensing microarray comparison in Yersinia pestis at 30Â°C

Project description:The AI-2 quorum-sensing system has been linked to diverse phenotypes and regulatory changes in pathogenic bacteria. In strain CO92, the AI-2 signal is produced in a luxS-dependent manner, reaching maximal levels of 2.5 Î¼M in late logarithmic growth, and both wild type and pigmentation mutant strains made equivalent levels of AI-2. Y. pestis CO92 possesses a chromosomal lsr locus encoding factors involved in the binding and import of AI-2, and confirming this assignment, an lsr deletion increased extracellular pools of AI-2. To assess the functional role of AI-2 sensing in Y. pestis, microarray study was conducted comparing the âPgm strain R88 to a âPgm âluxS mutant at 30Â°C to mimic the flea gut. Six independent RNA samples from Y. pestis CO92 ÎPgm cultures were paired with six independent RNA samples from ÎPgm ÎluxS cultures for hybridization to six two-color microarrays. A dye-swap design was used to remove the Cy5 and Cy3 dye bias.

2016-07-03 | E-GEOD-30108 | biostudies-arrayexpress

Expression data of Actinobacillus pleuropneumoniae 4074, the ΔluxS mutant and AI-2 supplemented ΔluxS mutant

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.

2011-11-15 | GSE21300 | GEO

Wild type E.coli SE15 vs. LuxS mutant E.coli SE15

Project description:Transcriptional profiling of E.coli SE15 comparing wild type E.coli SE15 with Autoindecur 2 synthesis gene LuxS mutnat E.coli SE15. E.coli SE15 is isolated from indwelling catheter of urinary tract infected patient. Examine change of quorum sensing related gene by deleting autoinducer 2 synthesis gene LuxS in E.coli

2013-08-02 | GSE49474 | GEO

Transcription profiling of E.coli luxS mutants vs. W3110 wild type

Project description:The bacterial quorum-sensing autoinducer 2 (AI-2) has received intense interest because the gene for its synthase, luxS, is common among a large number of bacterial species. We have identified luxS-controlled genes in Escherichia coli under two different growth conditions using DNA microarrays. Twenty-three genes were affected by luxS deletion in the presence of glucose, and 63 genes were influenced by luxS deletion in the absence of glucose. Minimal overlap among these gene sets suggests the role of luxS is condition dependent. Under the latter condition, the metE gene, the lsrACDBFG operon, and the flanking genes of the lsr operon (lsrR, lsrK, tam, and yneE) were among the most significantly induced genes by luxS. The E. coli lsr operon includes an additional gene, tam, encoding an S-adenosyl-l-methionine-dependent methyltransferase. Also, lsrR and lsrK belong to the same operon, lsrRK, which is positively regulated by the cyclic AMP receptor protein and negatively regulated by LsrR. lsrK is additionally transcribed by a promoter between lsrR and lsrK. Deletion of luxS was also shown to affect genes involved in methionine biosynthesis, methyl transfer reactions, iron uptake, and utilization of carbon. It was surprising, however, that so few genes were affected by luxS deletion in this E. coli K-12 strain under these conditions. Most of the highly induced genes are related to AI-2 production and transport. These data are consistent with the function of LuxS as an important metabolic enzyme but appear not to support the role of AI-2 as a true signal molecule for E. coli W3110 under the investigated conditions. Experiment Overall Design: Total RNA was extracted from E. coi W3110 wild type and luxS mutant grown in LB plus 0.8% glucose at OD 1.0. The genomic expression was compared between the two strains using Affymetrix E. Coli antisense genome array.

2008-06-16 | E-GEOD-9587 | biostudies-arrayexpress

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