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Transcriptional response of Enterococcus faecali to the glycosylated bacteriocin glycocin F

ABSTRACT: 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.

ORGANISM(S): Enterococcus faecalis

PROVIDER: GSE138552 | GEO | 2020/06/30

REPOSITORIES: GEO

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Project description:Emerging antibiotic resistance among clinically relevant bacteria, paired with their ability to form biofilms on medical and technical devices, represents a serious problem in terms of effective and long-term decontamination in health care environments and gives rise to an urgent need for new antimicrobial materials. Here we present the first study of the impact of AGXXÂ®, a novel broad-spectrum antimicrobial surface coating consisting of micro galvanic elements formed by silver and ruthenium, on the transcriptome of the nosocomial pathogen Enterococcus faecalis. E. faecalis was subjected to metal stress by growing it for different periods of time in the presence of AGXXÂ® or silver-coated steel meshes. Subsequently, total RNA was isolated and next-generation RNA sequencing was performed to analyze variations in gene expression levels in the presence of the antimicrobial materials with focus on known stress genes. Exposure to AGXXÂ® had a large impact on the transcriptome of E. faecalis. After 24 minutes almost 1/5 of the E. faecalis genome displayed differential expression. At each time-point the cop operon was strongly up-regulated, providing indirect evidence for the presence of free Ag+-ions. Moreover, exposure to AGXXÂ® induced a broad general stress response in E. faecalis. Genes coding for the chaperones GroEL and GroES as well as the Clp proteases ClpE and ClpB were among the top up-regulated heat shock genes. Furthermore, differential expression of genes coding for thioredoxin, superoxide dismutase and glutathione synthetase indicates a high level of oxidative stress. We postulate a mechanism of action where the combination of Ag+-ions and reactive oxygen species generated by AGXXÂ® results in a synergistic antimicrobial effect, which is superior to that of conventional silver coatings. Gene expression analysis of Enterococcus faecalis 12030 either subjected to metal stress by exposure to an antimicrobial AGXXÂ®- or Ag-coated V2A steel mesh or exposed to an uncoated V2A steel mesh or left untreated performing RNA Sequencing with an Ion ProtonTM Sequencer and subsequent data analysis with a T-REx RNA-Sequencing expression analysis pipeline.

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Transcriptional response of Enterococcus faecali to the glycosylated bacteriocin glycocin F

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