Project description:Listeria monocytogenes is a gram-positive, facultative anaerobe food-borne pathogen that is the causative agent of listeriosis. Upon ingestion, L. monocytogenes is subjected to a variety of non-specific host defenses such as bile. The main component of bile is bile salts which is known to be bactericidal through inducing DNA damage, oxidative damage, membrane instability, and protein misfolding. Previous studies have focused mainly on aerobic conditions; however the human GI tract is an environment ranging from microaerophilic to anaerobic. The bile salt hydrolase, an enzyme known to reduce toxicity of bile through hydrolysis, has been shown to have an increase in activity under anaerobic conditions. Therefore, the purpose of this study was to determine the effect of oxygen on bile resistance and determine the bile specific proteome that is responsible for this. To do this, we performed survival assays on virulent strains: F2365, 10403S, EGDe and avirulent strain, HCC23. Results showed an increase in viability for all virulent strains when exposed to porcine bile under anaerobic conditions. Interestingly, an increase in resistance was seen in HCC23 only under aerobic conditions. We then used a total proteomic analysis approach to compare the bile specific proteome under both aerobic and anaerobic conditions. Results showed many difference between all strains including an increase in abundance of proteins associated with stress responses, repair, cell morphology, and cell division under anaerobic conditions. Response to bile salt stress showed to be strain dependent. This study not only identified proteins responsible for L. monocytogenes bile resistance but also differerences between aerobic and anaerobic conditions thus suggesting that oxygen availability is not only a stressor but in some way providing assistance to overcoming bile salts.
2017-05-03 | PXD002243 | Pride
Project description:Transcriptomic analysis of Listeria monocytogenes in response to bile under anaerobic conditions at pH 7.5
Project description:Listeria monocytogenes is a gram-positive, food-borne pathogen responsible for invasive infections with high overall mortality. Early host defenses encountered by L. monocytogenes following ingestion include low pH of the stomach and bile present in the small intestinal lumen. We hypothesized that “epidemic” strains are better able to withstand exposure to low pH and bile encountered in the gastrointestinal tract as compared to most “environmental” strains. Furthermore, we hypothesized that epidemic and environmental strains would have distinct transcriptional programs upon exposure to these conditions. Our treatments included 1 hr exposure to acid (pH 5.5 and 3.5) and bile (0.3%) stress. Strains were pre-exposed to pH 5.5 (1 hr) before being treated with pH 3.5. We used a collection of 12 previously characterized epidemic and environmental strains and each strainXtreatment combination included 3 biological replicates for each microarray experiment. All microarray experiments were two color competitive hybridizations that paired experimental conditions with the same strain at neutral pH for acid stress and pH 5.5 for bile stress. Transcriptomes of environmental strains exposed to acid and bile stress showed remarkably greater number of genes with differences of ≥2-fold expression levels as compared to epidemic strains (5 and 7, respectively). Environmental strains were characterized by up-regulation of several stress related genes and down-regulation of several cell envelope biosynthesis and virulence related genes, suggesting that complex regulatory networks orchestrate the cellular changes in the environmental strains to overcome stressful environments. The transcriptome of epidemic strains, in contrast, showed muted responses to these stress conditions implying their pre-adaptability to acid and bile stress encountered during natural infection that may enable epidemic strains to survive and become “primed” for subsequent colonization and infection in the lower gastrointestinal tract. Keywords: stress response, comparative transcriptomics, acid-adaptation, differential virulence, acid-stress response, bile-stress response
2009-06-15 | GSE10603 | GEO
Project description:Transcriptomic analysis of Listeria monocytogenes aerobic conditions at pH 5.5
Project description:In this study, RNA-seq was used to compare the transcriptomes of Listeria monocytogenes 10403S and H7858 and corresponding ΔsigB mutants under exposure to pH 5.5 with or without bile. RNA-seq was performed on all strains and conditions in four independent biological replicates. Indexed and purified cDNA libraries (12 libraries per strains) were loaded together onto an independent flow cell without any other samples; sequencing was carried out by running Hiseq 2500 (single-end, 100-bp per read). Reads alignment was carried out using the Burrows-Wheeler Aligner (BWA). Differential expression of genes in different strains and conditions was statistically assessed using the BaySeq method.
2018-02-01 | GSE103443 | GEO
Project description:Transcriptomic analysis of Listeria monocytogenes under anaerobic conditions at pH 7.5
| PRJNA706703 | ENA
Project description:Transcriptomic analysis of Listeria monocytogenes in response to bile under aerobic and anaerobic conditions
| PRJNA706425 | ENA
Project description:Transcriptomic analysis of Listeria monocytogenes in response to bile under aerobic conditions at pH 7.5