Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptional adaptation of L. monocytogenes EGD to oxygen availability

ABSTRACT: Comparison of aerobic and anaerobic transcriptome of L. monocytogenes EGD L. monocytogenes EGD cells used in this study were grown either aerobically or anaerobically to an OD600 = 0.70 - 0.75 and the transcriptome of these two conditions was compared Six biologically independent cultures of each aerobically and anaerobically grown L. monocytogenes EGD were used for total RNA isolation (six RNA sets). 4 array experiments were performed with Cy5-labelled cDNA derived from aerobically grown Listeria and Cy3-labelled cDNA derived from anaerobcially grown cells. For the two other RNA sets a dye swap was performed, that means the two other array experiments were performed with Cy3-labelled cDNA derived form aerobically grown Listeria and Cy5-labelled cDNA from anaerobically grown cells. As all oligonuceotides are spotted twice on an array, technical duplicates were performed for each experiment. The overall design results in 12 data points for the expression of each gene.

ORGANISM(S): Listeria monocytogenes EGDe

SUBMITTER: Stefanie MM-CM-<ller-Herbst

PROVIDER: E-GEOD-52325 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Identification of genes essential for anaerobic growth of Listeria monocytogenes.

Müller-Herbst Stefanie S Wüstner Stefanie S Mühlig Anna A Eder Daniela D M Fuchs Thilo T Held Claudia C Ehrenreich Armin A Scherer Siegfried S

Microbiology (Reading, England) 20140122 Pt 4

The facultative anaerobic bacterium Listeria monocytogenes encounters microaerophilic or anaerobic conditions in various environments, e.g. in soil, in decaying plant material, in food products and in the host gut. To elucidate the adaptation of Listeria monocytogenes to variations in oxygen tension, global transcription analyses using DNA microarrays were performed. In total, 139 genes were found to be transcribed differently during aerobic and anaerobic growth; 111 genes were downregulated and ...[more]

PMID: 24451174

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Project description:PrfA activity was studied in L. monocytogenes strain EGD and in an isogenic prfA deletion mutant (EGDΔprfA) carrying multiple copies of the wild-type prfA or the mutant prfA* gene (strains EGDΔprfApPrfA and EGDΔprfApPrfA*) after growth in brain heart infusion (BHI), Luria-Bertani broth (LB) or a defined minimal medium (MM) supplemented either with one of the three PTS-carbohydrates, glucose, mannose and cellobiose, or the non-PTS carbon source glycerol. Low PrfA activity was observed in the wild-type EGD strain in BHI and LB with either of these carbon sources, while PrfA activity was high in minimal medium in presence of glycerol but significantly reduced in presence of cellobiose. The strains expressing the prfA and prfA* gene under the prfA promoters, P1 and P2, produced equally large amounts of PrfA protein and high PrfA activity was observed in strain EGDΔprfApPrfA* under all growth conditions. In contrast, high PrfA activity in strain EGDΔprfApPrfA was only observed when this strain was cultured in BHI but not in LB or MM (in presence of either carbon source). A ptsH mutant (lacking a functional HPr) was able to grow in BHI suggesting that growth of L. monocytogenes in this culture medium is supported by carbon sources whose uptake and metabolism are independent of the PTS pathway. However, this mutant was unable to grow in LB and MM regardless which of the four carbon sources was added, suggesting that uptake of the used carbohydrates and the catabolism of glycerol depend fully on the functional common PTS pathway. Furthermore, the growth rates of L. monocytogenes are strongly reduced in presence of large amounts of PrfA protein when growing MM but less in LB and only slightly in BHI. The expression profiles of the genes encoding PTS permeases were determined in the three strains under various growth conditions. The data suggest that PrfA activity correlates with the expression level and the phosphorylation state of specific PTS permeases. This SuperSeries is composed of the following subset Series: GSE12143: Listeria monocytogenes EGD after growth BHI vs. LB vs. MM GSE12145: Listeria monocytogenes EGDΔprfApPrfA and EGDΔprfApPrfA* compared to the wild type strain EGD GSE12146: Listeria monocytogenes EGD and EGD-e

Dataset Information

Transcriptional adaptation of L. monocytogenes EGD to oxygen availability

Publications

Identification of genes essential for anaerobic growth of Listeria monocytogenes.

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