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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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Detection of ROS accumulation in LPS-treated S. cerevisiae BY4742 cells, using flow cytometry.H2DCFH-DA was used to stain the untreated (Control) and LPS-treated cells (LPS+). The result was represented by density banding coloring. Gray dots represent the lowest density, while red dots represent the highest density. The gate “E” was set by corresponding dye-unloading samples. FL1-DCF represents 2′,7′-dichlorofluorescein while FL2-H assists for data display. The majority populations (red dots) from both samples shew similar ROS levels.
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pone-0104428-g009: Detection of ROS accumulation in LPS-treated S. cerevisiae BY4742 cells, using flow cytometry.H2DCFH-DA was used to stain the untreated (Control) and LPS-treated cells (LPS+). The result was represented by density banding coloring. Gray dots represent the lowest density, while red dots represent the highest density. The gate “E” was set by corresponding dye-unloading samples. FL1-DCF represents 2′,7′-dichlorofluorescein while FL2-H assists for data display. The majority populations (red dots) from both samples shew similar ROS levels.

Mentions: ROS level in LPS-treated S. cerevisiae BY4742 cells was analyzed by staining with H2DCFH-DA [34]. H2DCFH-DA can be hydrolyzed by esterase to produce 2′,7′-dichlorofluorescin, which is then oxidized by ROS to form the fluorescent 2′,7′-dichlorofluorescein. The ROS levels in the majority of LPS-treated S. cerevisiae BY4742 cells were similar to the untreated cells (Fig. 9), indicating the ROS stability in LPS-treated S. cerevisiae BY4742 cells.


Response of Saccharomyces cerevisiae to the stimulation of lipopolysaccharide.

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

Detection of ROS accumulation in LPS-treated S. cerevisiae BY4742 cells, using flow cytometry.H2DCFH-DA was used to stain the untreated (Control) and LPS-treated cells (LPS+). The result was represented by density banding coloring. Gray dots represent the lowest density, while red dots represent the highest density. The gate “E” was set by corresponding dye-unloading samples. FL1-DCF represents 2′,7′-dichlorofluorescein while FL2-H assists for data display. The majority populations (red dots) from both samples shew similar ROS levels.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0104428-g009: Detection of ROS accumulation in LPS-treated S. cerevisiae BY4742 cells, using flow cytometry.H2DCFH-DA was used to stain the untreated (Control) and LPS-treated cells (LPS+). The result was represented by density banding coloring. Gray dots represent the lowest density, while red dots represent the highest density. The gate “E” was set by corresponding dye-unloading samples. FL1-DCF represents 2′,7′-dichlorofluorescein while FL2-H assists for data display. The majority populations (red dots) from both samples shew similar ROS levels.
Mentions: ROS level in LPS-treated S. cerevisiae BY4742 cells was analyzed by staining with H2DCFH-DA [34]. H2DCFH-DA can be hydrolyzed by esterase to produce 2′,7′-dichlorofluorescin, which is then oxidized by ROS to form the fluorescent 2′,7′-dichlorofluorescein. The ROS levels in the majority of LPS-treated S. cerevisiae BY4742 cells were similar to the untreated cells (Fig. 9), indicating the ROS stability in LPS-treated S. cerevisiae BY4742 cells.

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