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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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A. Comparison of transcriptional levels of some key genes in LPS-treated S. cerevisiae BY4742 cells analyzed by RT-PCR and transcriptome, respectively. B. Methylene blue staining of some single gene deletion mutants of S. cerevisiae BY4743 after LPS treatment. According to the transcriptome analysis, genes whose transcriptional level did not significantly change in S. cerevisiae BY4742 after LPS treatment were labeled with star symbol (*); transcriptional levels for the other genes could be found in Table S4.
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pone-0104428-g006: A. Comparison of transcriptional levels of some key genes in LPS-treated S. cerevisiae BY4742 cells analyzed by RT-PCR and transcriptome, respectively. B. Methylene blue staining of some single gene deletion mutants of S. cerevisiae BY4743 after LPS treatment. According to the transcriptome analysis, genes whose transcriptional level did not significantly change in S. cerevisiae BY4742 after LPS treatment were labeled with star symbol (*); transcriptional levels for the other genes could be found in Table S4.

Mentions: To confirm the regulation information obtained by the transcriptome analysis, a few key genes were chosen to be analyzed by RT-PCR. The overall results from RT-PCR are consistent with those obtained by transcriptome analysis. In both RT-PCR and transcriptome analysis, genes SMP1, YPR145C HSP26, GND2, HSP32 and GPD1 were found up-regulated, while genes MA(α)2 and CWP1 were down-regulated (Fig. 6A). The transcriptional levels of a gene might depend on the time of cell growth and LPS treatment. For example, the transcriptional levels of HOG1 and GPD1 in S. cerevisiae BY4742 cells treated with LPS for 90 min were larger than those treated with LPS for 180 min (Fig. 6A).


Response of Saccharomyces cerevisiae to the stimulation of lipopolysaccharide.

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

A. Comparison of transcriptional levels of some key genes in LPS-treated S. cerevisiae BY4742 cells analyzed by RT-PCR and transcriptome, respectively. B. Methylene blue staining of some single gene deletion mutants of S. cerevisiae BY4743 after LPS treatment. According to the transcriptome analysis, genes whose transcriptional level did not significantly change in S. cerevisiae BY4742 after LPS treatment were labeled with star symbol (*); transcriptional levels for the other genes could be found in Table S4.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0104428-g006: A. Comparison of transcriptional levels of some key genes in LPS-treated S. cerevisiae BY4742 cells analyzed by RT-PCR and transcriptome, respectively. B. Methylene blue staining of some single gene deletion mutants of S. cerevisiae BY4743 after LPS treatment. According to the transcriptome analysis, genes whose transcriptional level did not significantly change in S. cerevisiae BY4742 after LPS treatment were labeled with star symbol (*); transcriptional levels for the other genes could be found in Table S4.
Mentions: To confirm the regulation information obtained by the transcriptome analysis, a few key genes were chosen to be analyzed by RT-PCR. The overall results from RT-PCR are consistent with those obtained by transcriptome analysis. In both RT-PCR and transcriptome analysis, genes SMP1, YPR145C HSP26, GND2, HSP32 and GPD1 were found up-regulated, while genes MA(α)2 and CWP1 were down-regulated (Fig. 6A). The transcriptional levels of a gene might depend on the time of cell growth and LPS treatment. For example, the transcriptional levels of HOG1 and GPD1 in S. cerevisiae BY4742 cells treated with LPS for 90 min were larger than those treated with LPS for 180 min (Fig. 6A).

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