Project description:Lactobacillus plantarum is a common inhabitant of mammalian gastrointestinal tracts and specific strains belonging to this species are marketed as probiotics intended to confer beneficial health effects. To assist in determining the physiological status and host-microbe interactions of L. plantarum in the digestive tract we assessed changes in the transcriptome of L. plantarum WCFS1 during colonization of the cecum of germ-free mice. According to the transcript profiles L. plantarum WCFS1 was metabolically active and not under severe stress in this intestinal compartment. Carbohydrate metabolism was the most strongly affected functional gene category whereby many genes encoding diverse sugar transport and degradation pathways were induced in mice even compared to L. plantarum grown in a mouse chow-derived laboratory medium. This suggests that the ability of L. plantarum WCFS1 to consume diverse energy sources including plant-associated and host-derived carbohydrates was increased during its residence in the digestive tract. Many of these genes were also induced in L. plantarum colonizing germ-free mice fed a humanized Western-style diet. Similarly a core set of genes encoding cell surface-related properties were differentially expressed in mice. This set includes genes required for the D-alanylation and glycosylation of lipoteichoic acids that were strongly down-regulated in mice. In total L. plantarum exhibits a distinct in vivo transcriptome directed towards adaptation to the mouse intestinal environment. Keywords: cell type comparison
Project description:Glycocin F (GccF) is a bacteriocin produced by Lactobacillus plantarum which causes susceptible cells to enter bacteriostasis within minutes of exposure. GccF is diglycosylated with two N-acetylglucosamine (GlcNAc) moieties and is active against strains of Lb. plantarum, as well as potential pathogens such as Enterococcus faecalis and E. faecium. Its mechanism of action, however, is unknown. To better understand how GccF inhibits growth, we carried out RNA sequencing (RNA-seq) on E. faecalis strain JH2-2 exposed to different concentrations of GccF for different lengths of time. Here we demonstrate that free GlcNAc both protects E. faecalis from the effects of GccF, as well as pre-sensitises it to GccF. Transcriptional analysis showed that exposure to free GlcNAc resulted in up-regulation of a GlcNAc-specific phosphotransferase system (PTS) transporter, which is a putative GccF receptor. Time course analysis showed over 100 genes were up- or down-regulated within 10 minutes of exposure, a number that increased to over 300 genes after 40 minutes. A large number of these genes were found to be regulated by the alternative sigma factor 54, which was shown to not be required for susceptibility to GccF. Interestingly, genes encoding the V-type ATPases and sodium:proton antiporters were found to be up-regulated at all times and all concentrations of GccF tested. This suggests an indirect response to GccF, possibly due to low-level permeabilization of the cell membrane.
Project description:Lactobacillus plantarum is a common inhabitant of mammalian gastrointestinal tracts and specific strains belonging to this species are marketed as probiotics intended to confer beneficial health effects. To assist in determining the physiological status and host-microbe interactions of L. plantarum in the digestive tract we assessed changes in the transcriptome of L. plantarum WCFS1 during colonization of the cecum of germ-free mice. According to the transcript profiles L. plantarum WCFS1 was metabolically active and not under severe stress in this intestinal compartment. Carbohydrate metabolism was the most strongly affected functional gene category whereby many genes encoding diverse sugar transport and degradation pathways were induced in mice even compared to L. plantarum grown in a mouse chow-derived laboratory medium. This suggests that the ability of L. plantarum WCFS1 to consume diverse energy sources including plant-associated and host-derived carbohydrates was increased during its residence in the digestive tract. Many of these genes were also induced in L. plantarum colonizing germ-free mice fed a humanized Western-style diet. Similarly a core set of genes encoding cell surface-related properties were differentially expressed in mice. This set includes genes required for the D-alanylation and glycosylation of lipoteichoic acids that were strongly down-regulated in mice. In total L. plantarum exhibits a distinct in vivo transcriptome directed towards adaptation to the mouse intestinal environment. Keywords: cell type comparison Six-week old germ-free C57 Black-6 male mice were inoculated with a single dose of 109 CFU of exponential-phase L. plantarum WCFS1 cells. The mice were sacrificed 15 days later, after sufficient time had passed for several turnovers of the intestinal epithelium and its overlying mucosal layer. Four mice were fed on Chow diet and two mice were fed on western style diet. RNA was isolated from the cecum of these mice. The transcriptome of L. plantarum in these mice was compared to that of L. plantarum grown on MRS broth, Chow broth, or on chemically defined media with either glucose or lactose as carbon- and energy source.