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

ABSTRACT: 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. M-NM-^TluxS and M-NM-^TlsrM-NM-^TluxS strains were grown in a chemically defined media (CDM) to two different time-points with or without the addition of 3M-BM-5M synthetic AI-2. RNA was extracted from these sampling conditions in two separate cultures (biological replicates). Each of these RNA samples was run on two different H. influenzae supragenome arrays (technical replicates). 3-13 probes represent each subcluster(gene) and two replicate probe sets are included on each custom NTHi supragenome chip thus under each sampling condition 8 separate values are obtained per strain and used for WT:Lsr-KO comparisons.

ORGANISM(S): Haemophilus influenzae

SUBMITTER: Benjamin Janto

PROVIDER: E-GEOD-57288 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Expression analysis of a full Lsr locus knockout in Haemophilus influenzae strain CZ4126/02

Project description:Investigation of whole genome gene expression differences in a full (nine-gene) Lsr locus knockout strain compared to the wild-type strain (CZ4126/02). This Lsr mutant is unable to import the autoinducer-2 (AI-2) quorum sensing molecule nor mediate any potential LsrR-based transcriptional regulation. WT and Lsr-KO strains were grown in two different media to two or three different time-points. RNA was extracted from these sampling conditions in two separate cultures (biological replicates). Each of these RNA samples was run on two different H. influenzae supragenome arrays (technical replicates). 3-13 probes represent each subcluster (gene) and two replicate probe sets are included on each custom NTHi supragenome chip. Thus, under each sampling condition, 8 separate values are obtained per strain and used for WT:Lsr-KO comparisons.

2012-10-19 | E-GEOD-41690 | biostudies-arrayexpress

Expression analysis of a full SlrV locus 1 knockout in Haemophilus influenzae strain PittII

Project description:Investigation of whole genome gene expression differences between a full (four-gene) SlrV locus 1 deletion compared to the WT PittII strain. Removal of the four SlrV genes (aka macrophage survival factors : ?msf) results in a competetive defect with the WT strain in in vitro biofilms and in the otitis media chinchilla model. It also results in an inability to persist within macrophages long-term in vitro and to escape from the middle-ear space and infect other sites in the chinchilla model. WT and ?msf strains were grown in a chemically defined media (CDM) to early stationary phase (OD ~1.0). RNA was extracted from two separate cultures (biological replicates). 3-13 probes represent each subcluster(gene) and two replicate probe sets are included on each custom NTHi supragenome chip (technical replicates) thus under each sampling condition 4 separate values are obtained per strain and used for WT:Msf-KO comparisons.

2015-06-23 | E-GEOD-70172 | biostudies-arrayexpress

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 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

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

luxS-Dependent Gene Regulation in Escherichia coli K-12 Revealed by Genomic Expression Profiling (II)

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. Keywords: Genetic modification

2007-11-14 | GSE9587 | GEO

luxS-Dependent Gene Regulation in Escherichia coli K-12 Revealed by Genomic Expression Profiling

2007-11-14 | GSE9582 | GEO

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

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

Transcription profiling of E. coli K12 luxS-dependent gene regulation

2008-06-16 | E-GEOD-9582 | biostudies-arrayexpress

3 quorum sensing signals add in microarray comparison in Yersinia pestis at 30°C

Project description:The 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. Yersinia 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 3 quorum sensing singlas in Y. pestis, microarray study was conducted comparing the 3 quorum sensing signals added in to control at 30°C to mimic the flea gut. Here, the ΔPgm (pigmentation-negative) R88 strain was used. The three signals are one AI-2 and two AHLs (N-(3-Oxooctanoyl)-L-homoserine lactone and N-Hexanoyl-DL-homoserine lactone).The control consisted of cells grown and treated under the same conditions without added signals.

2016-01-01 | GSE30109 | GEO

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