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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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Assessment of mitochondrial membrane potential in LPS-treated S. cerevisiae BY4742 cells, using flow cytometry.Rh123 was used to stain the untreated (Control), LPS-treated (LPS+) and 100% heat-treated cells (Heated). The result was represented by density banding coloring. Gray dots represent the lowest density, while red dots represent the highest density. The gate set “G” was used to show the differences among the three experiments.
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pone-0104428-g008: Assessment of mitochondrial membrane potential in LPS-treated S. cerevisiae BY4742 cells, using flow cytometry.Rh123 was used to stain the untreated (Control), LPS-treated (LPS+) and 100% heat-treated cells (Heated). The result was represented by density banding coloring. Gray dots represent the lowest density, while red dots represent the highest density. The gate set “G” was used to show the differences among the three experiments.

Mentions: Since mitochondria are preferential targets for various stresses in S. cerevisiae[3], mitochondrial function or integrity in S. cerevisiae BY4742 cells treated with LPS was assessed by staining with Rh123 [33]. Rh123 can distribute into the mitochondrial matrix in response to ΔΨm. The untreated cells and heat-treated S. cerevisiae cells were used as the negative and positive controls, respectively. As shown in Fig. 8, about half of the LPS-treated S. cerevisiae BY4742 cells had similar ΔΨm to the heat-treated cells. This increase of ΔΨm in LPS-treated S. cerevisiae cells might be due to many factors such as the inhibition of ATP synthase [36], [39].


Response of Saccharomyces cerevisiae to the stimulation of lipopolysaccharide.

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

Assessment of mitochondrial membrane potential in LPS-treated S. cerevisiae BY4742 cells, using flow cytometry.Rh123 was used to stain the untreated (Control), LPS-treated (LPS+) and 100% heat-treated cells (Heated). The result was represented by density banding coloring. Gray dots represent the lowest density, while red dots represent the highest density. The gate set “G” was used to show the differences among the three experiments.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0104428-g008: Assessment of mitochondrial membrane potential in LPS-treated S. cerevisiae BY4742 cells, using flow cytometry.Rh123 was used to stain the untreated (Control), LPS-treated (LPS+) and 100% heat-treated cells (Heated). The result was represented by density banding coloring. Gray dots represent the lowest density, while red dots represent the highest density. The gate set “G” was used to show the differences among the three experiments.
Mentions: Since mitochondria are preferential targets for various stresses in S. cerevisiae[3], mitochondrial function or integrity in S. cerevisiae BY4742 cells treated with LPS was assessed by staining with Rh123 [33]. Rh123 can distribute into the mitochondrial matrix in response to ΔΨm. The untreated cells and heat-treated S. cerevisiae cells were used as the negative and positive controls, respectively. As shown in Fig. 8, about half of the LPS-treated S. cerevisiae BY4742 cells had similar ΔΨm to the heat-treated cells. This increase of ΔΨm in LPS-treated S. cerevisiae cells might be due to many factors such as the inhibition of ATP synthase [36], [39].

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