Correlating cell wall polysaccharide genotypes and structures of Lactococcus lactis
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PROVIDER: PRJEB18135 | ENA |
REPOSITORIES: ENA
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Project description:Analysis of Pseudomonas aeruginosa PA01 (ATCC 15692) Psl polysaccharide deficient mutant. Psl polysaccharide deficient mutant is evaluated with microarray to understand the genes affected by this polysaccharide. Our results provide new vision on the roles played by Psl polysaccharide in P. aeruginosa. Exopolysaccharide Psl is a critical biofilm matrix component in Pseudomonas aeruginosa, which forms a fiber-like matrix to enmesh bacteria communities. Iron has been shown to serve as a signal in P. aeruginosa biofilm development, yet how iron controls biofilm formation is not clear. Here we perform a transcriptomic analysis to compare Psl negative strain versus its isogenic wild-type strain PAO1. The results indicate that the expression of genes involved in iron homeostasis and oxidative stress response increased drastically at transcriptional level in Psl negative strain, suggesting Psl deficiency induces iron limitation. Subsequent studies confirm that Psl can efficiently bind iron in vitro and Psl fibers functions as an iron storage channel in P. aeruginosa biofilms. We used two wild type (WT) replicates and two mutant (MT) replicates to compare the differential gene expression.
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Project description:Lcn972 is a non-modified bacteriocin that targets exclusively Lactococcus sp. Addition of Lcn972 inhibits cell wall biosynthesis at the septum by binding to the cell wall precursor lipid II. Resistance to Lcn972 develops upon selection with subinhibitory concentrations. The transcriptome of the highly resistant L. lactis D1 derived from the susceptible strain L. lactis MG1614 was compared. Fourteen genes were significantly up-regulated and 29 were down-regulated (expression change > 2-fold, p<0.001). Down-regulation was mostly found in sugar catabolic genes. Up-regulated genes included members of the cell envelope stress (CesR) regulon, the penicillin-binding protein pbpX and llmg2447, which may encode a putative extracytoplasmic function (ECF) anti-sigma factor located downstream of a non-functional ECF-sigX. Up-regulation of llmg2447 was linked to integration of IS981 that exchanged the -35 promoter region of llmg2447. Over-expression of llmg2447 resulted in highly Lcn972-resistant L. lactis transformants. The transcriptomes of Lactococcus lactis MG1614 susceptible to Lcn972 and L. lactis D1 resistant to Lcn972, grown under laboratory conditions, were compared using four biological replicates.
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