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Transcriptomic profiling of L-rhamnose utilization in Lmo0753 deletion mutant of Listeria monocytogenes EGDe from human foodborne outbreak associated lineage LII

ABSTRACT: Among the three major genetic lineages of L. monocytogenes (i.e. LI, LII, and LIII), LI and LII are predominantly associated with foodborne listeriosis outbreaks, whereas LIII is rarely implicated in human infections. In a previous study, we identified a Crp/Fnr family transcription factor lmo0753 that was highly specific to outbreak-associated LI and LII but absent from LIII. Lmo0753 shares two conserved functional domains including a DNA-binding domain with the well-characterized master virulence regulator PrfA in L. monocytogenes. In this study, we constructed a lmo0753 deletion and complementation mutants of the fully sequenced L. monocytogenes LII strain EGDe. We found that deletion of lmo0753 led to the loss of L-rhamnose utilization in EGDe. Transcriptomic comparison of the EGDe lmo0753 deletion mutant and the wild type incubated in phenol-red medium containing L-rhamnose as the sole carbon source revealed 126 (4.5%) and 546 (19.5%) out of 2,798 genes in the EGDe genome that were up- and down-regulated for more than 2-fold, respectively. Genes involved in biotin biosynthesis, general stress response and rhamnose metabolism were shown to be differentially regulated by Lmo0753. Findings from this study may partially explain why LIII of L. monocytogenes is underrepresented in the environment and rarely associated with human listeriosis outbreaks due to the inability of rhamnose utilization. We report the transcriptomic profile of L. monocytogenes Δlmo0753 LII strain (EGDe) in broth media with L-rhamnose as the sole carbon source.

ORGANISM(S): Listeria monocytogenes

PROVIDER: GSE47721 | GEO | 2013/07/17

SECONDARY ACCESSION(S): PRJNA207398

REPOSITORIES: GEO

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Project description:The phenotypic and gene expression traits conferred by the alternative sigma factor protein M-OM-^CL in the food-borne pathogen L. monocytogenes were investigated. M-OM-^CL was shown to be important for the efficient growth of this pathogen exposed to food preservative measures such as low storage temperatures, elevated osmolarity and acidity. Based on high throughput phenotypic analysis, M-OM-^CL function was also found to be protective in L. monocytogenes EGDe cells exposed to several antimicrobial compounds including some of the antibiotics currently applied in listeriosis treatment. The expression of flagella genes and motility were significantly compromized upon loss of M-OM-^CL function. A comparative transcriptome analysis of exponential growth EGDe wild type and sigL null cells unveiled 394 genes that are positively controlled through M-OM-^CL dependent transcriptional regulation mechanisms in this bacterium during growth at low (3M-BM-0C) and optimized (37M-BM-0C) temperature conditions. Genes identified indicate that M-OM-^CL is a pleitropic transcription regulator mediating positive expression control of genes involved in diverse cellular processes including protein synthesis, molecular transport, energy metabolism, respiration, transcription regulation, metabolite biosynthesis, and cell envelope composition modification. Overall our observations have revealed that the loss of M-OM-^CL function leads to extensive gene expression defects in L. monocytogenes EGDe cells, and these are consistent with compromized nutrient assimilation, energy metabolism, protein synthesis, and metabolite biosynthesis processes as well as altered cell envelope composition and motility. Numerous gene expression changes imposed by M-OM-^CL loss in EGDe are thus also consistent with pleiotropic phenotypic defects detected in the L. monocytogenes EGDe M-bM-^HM-^FsigL strain. Gene expression DNA-microarray. Two-color hybridizations on 8*15K Agilent arrays. Wild type and three KO mutants in three temperatures, three biological replicates in each condition.

Project description:The foodborne pathogen Listeria monocytogenes has the ability to develop biofilm in food-processing environment, which becomes a major concern for the food safety. PrfA, a key transcriptional activator that regulates most of the known listerial virulence gene expression, has been shown to promote L. monocytogenes biofilm formation. In this study, the whole genome microarray was used to identify differentially expressed genes associated with the putative interaction between biofilm formation and PrfA in L. monocytogenes. Comparative transcriptome analyses indicated over 21.9% of the L. monocytogenes EGDe genes (627 out of 2857 predicted) were altered in their expression in biofilm cells relative to planktonic cell populations. These genes were classed into different functional categories which cover most of the biochemical functions encountered in bacterial cells, especially involved in ion transport, DNA repair, and cell wall biosynthesis based on significant enrichment of GO terms. Among them, 185 genes were identified to be associated with PrfA and biofilm formation by comparison of the whole gene expression profiles of L. monocytogenes EGDe and its M-NM-^TprfA mutant. The expression tendency of these PrfA-associated and biofilm-specific genes were mainly opposite in M-NM-^TprfA biofilm, and most of them are involved in phage-related function, membrane bioenergetics, and cell wall. Our results indicated that L. monocytogenes biofilm formation is probably controlled by the complex regulation network involved variable genes required for the different biological pathways. This regulatory network is modified in the prfA deletion mutant in order to maintain its stable biofilm lifestyle. Gene expression of planktonic cells and biofilm cells in Listeria monocytogenes EGDe and prfA isogenic deletion strain EGDeM-NM-^TprfA with cultivated in MEM and BHI for 48 hours, were mesasued using Agilent Listeria monocytogenes customized whole-genome microarray 8x15 array. Three replicates.

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Transcriptomic profiling of L-rhamnose utilization in Lmo0753 deletion mutant of Listeria monocytogenes EGDe from human foodborne outbreak associated lineage LII

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