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Response of Saccharomyces cerevisiae to the stimulation of lipopolysaccharide.

Shen L, Li Y, Jiang L, Wang X - PLoS ONE (2014)

Bottom Line: In this study, we found that lipopolysaccharide-treated S. cerevisiae cells could be stained by methylene blue, but did not die.Significantly regulated genes (460 up-regulated genes and 135 down-regulated genes) in lipopolysaccharide-treated S. cerevisiae cells were analyzed on Gene Ontology, and used to establish physical protein-protein interaction network and protein phosphorylation network.Based on these analyses, most of the regulated genes in lipopolysaccharide-treated S. cerevisiae cells were related to cell wall, membrane, peroxisome and mitochondrion.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China; School of Biotechnology, Jiangnan University, Wuxi, China.

ABSTRACT
Lipopolysaccharide, known as endotoxin, can stimulate potent host immune responses through the complex of Toll-like-receptor 4 and myeloid differentiation protein 2; but its influence on Saccharomyces cerevisiae, a model organism for studying eukaryotes, is not clear. In this study, we found that lipopolysaccharide-treated S. cerevisiae cells could be stained by methylene blue, but did not die. Transcriptional profiling of the lipopolysaccharide-treated S. cerevisiae cells showed that 5745 genes were modulated: 2491 genes up-regulated and 3254 genes down-regulated. Significantly regulated genes (460 up-regulated genes and 135 down-regulated genes) in lipopolysaccharide-treated S. cerevisiae cells were analyzed on Gene Ontology, and used to establish physical protein-protein interaction network and protein phosphorylation network. Based on these analyses, most of the regulated genes in lipopolysaccharide-treated S. cerevisiae cells were related to cell wall, membrane, peroxisome and mitochondrion. Further experiments demonstrated that lipopolysaccharide stimulation caused the exposure of phosphatidylserine and the increase of mitochondrial membrane potential in S. cerevisiae cells, but levels of intracellular reactive oxygen species and metacaspase activation were not increased. This study demonstrated that lipopolysaccharide stimulation causes significant changes in S. cerevisiae cells, and the results would contribute to understand the response of eukaryotic cells to lipopolysaccharide stimulation.

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Analysis of metacaspase activity of LPS-treated S. cerevisiae BY4742 cells by flow cytometry.FITC-VAD-FMK was used to detect metacaspace activation. Untreated S. cerevisiae BY4742 cells were used as negative control; 100% heat-treated cells were chosen as positive control. The result was represented by density banding coloring. Gray dots represent the lowest density, while red dots represent the highest density. The gate “D” was set to show the differences among the three types of cells. FL1-FITC represents FITC-VAD-FMK while FS assists for data display.
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pone-0104428-g010: Analysis of metacaspase activity of LPS-treated S. cerevisiae BY4742 cells by flow cytometry.FITC-VAD-FMK was used to detect metacaspace activation. Untreated S. cerevisiae BY4742 cells were used as negative control; 100% heat-treated cells were chosen as positive control. The result was represented by density banding coloring. Gray dots represent the lowest density, while red dots represent the highest density. The gate “D” was set to show the differences among the three types of cells. FL1-FITC represents FITC-VAD-FMK while FS assists for data display.

Mentions: Metacaspase activation in LPS-treated S. cerevisiae BY4742 cells was detected by staining with FITC-VAD-FMK and analyzed by flow cytometry [31], [32]. The population distribution pattern of LPS-treated S. cerevisiae BY4742 cells was similar to the untreated cells (Fig. 10), suggesting the metacaspase activation in S. cerevisiae BY4742 cells was not affected by LPS treatment. This is consistent with the finding in transcriptome analysis that the transcriptional level of YCA1, the key gene encoding metacaspase in S. cerevisiae BY4742 cells, did not significantly change after LPS treatment [40].


Response of Saccharomyces cerevisiae to the stimulation of lipopolysaccharide.

Shen L, Li Y, Jiang L, Wang X - PLoS ONE (2014)

Analysis of metacaspase activity of LPS-treated S. cerevisiae BY4742 cells by flow cytometry.FITC-VAD-FMK was used to detect metacaspace activation. Untreated S. cerevisiae BY4742 cells were used as negative control; 100% heat-treated cells were chosen as positive control. The result was represented by density banding coloring. Gray dots represent the lowest density, while red dots represent the highest density. The gate “D” was set to show the differences among the three types of cells. FL1-FITC represents FITC-VAD-FMK while FS assists for data display.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4126697&req=5

pone-0104428-g010: Analysis of metacaspase activity of LPS-treated S. cerevisiae BY4742 cells by flow cytometry.FITC-VAD-FMK was used to detect metacaspace activation. Untreated S. cerevisiae BY4742 cells were used as negative control; 100% heat-treated cells were chosen as positive control. The result was represented by density banding coloring. Gray dots represent the lowest density, while red dots represent the highest density. The gate “D” was set to show the differences among the three types of cells. FL1-FITC represents FITC-VAD-FMK while FS assists for data display.
Mentions: Metacaspase activation in LPS-treated S. cerevisiae BY4742 cells was detected by staining with FITC-VAD-FMK and analyzed by flow cytometry [31], [32]. The population distribution pattern of LPS-treated S. cerevisiae BY4742 cells was similar to the untreated cells (Fig. 10), suggesting the metacaspase activation in S. cerevisiae BY4742 cells was not affected by LPS treatment. This is consistent with the finding in transcriptome analysis that the transcriptional level of YCA1, the key gene encoding metacaspase in S. cerevisiae BY4742 cells, did not significantly change after LPS treatment [40].

Bottom Line: In this study, we found that lipopolysaccharide-treated S. cerevisiae cells could be stained by methylene blue, but did not die.Significantly regulated genes (460 up-regulated genes and 135 down-regulated genes) in lipopolysaccharide-treated S. cerevisiae cells were analyzed on Gene Ontology, and used to establish physical protein-protein interaction network and protein phosphorylation network.Based on these analyses, most of the regulated genes in lipopolysaccharide-treated S. cerevisiae cells were related to cell wall, membrane, peroxisome and mitochondrion.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China; School of Biotechnology, Jiangnan University, Wuxi, China.

ABSTRACT
Lipopolysaccharide, known as endotoxin, can stimulate potent host immune responses through the complex of Toll-like-receptor 4 and myeloid differentiation protein 2; but its influence on Saccharomyces cerevisiae, a model organism for studying eukaryotes, is not clear. In this study, we found that lipopolysaccharide-treated S. cerevisiae cells could be stained by methylene blue, but did not die. Transcriptional profiling of the lipopolysaccharide-treated S. cerevisiae cells showed that 5745 genes were modulated: 2491 genes up-regulated and 3254 genes down-regulated. Significantly regulated genes (460 up-regulated genes and 135 down-regulated genes) in lipopolysaccharide-treated S. cerevisiae cells were analyzed on Gene Ontology, and used to establish physical protein-protein interaction network and protein phosphorylation network. Based on these analyses, most of the regulated genes in lipopolysaccharide-treated S. cerevisiae cells were related to cell wall, membrane, peroxisome and mitochondrion. Further experiments demonstrated that lipopolysaccharide stimulation caused the exposure of phosphatidylserine and the increase of mitochondrial membrane potential in S. cerevisiae cells, but levels of intracellular reactive oxygen species and metacaspase activation were not increased. This study demonstrated that lipopolysaccharide stimulation causes significant changes in S. cerevisiae cells, and the results would contribute to understand the response of eukaryotic cells to lipopolysaccharide stimulation.

Show MeSH
Related in: MedlinePlus