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A mitochondria-dependent pathway mediates the apoptosis of GSE-induced yeast.

Cao S, Xu W, Zhang N, Wang Y, Luo Y, He X, Huang K - PLoS ONE (2012)

Bottom Line: Proteins related to redox, cellular structure, membrane, energy and DNA repair were significantly increased.The relationship between the proteins and metabolites was analyzed by orthogonal projections to latent structures (OPLS).We found that the changes of the metabolites and the protein changes had relevant consistency.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of food safety and molecular biology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, People's Republic of China.

ABSTRACT
Grapefruit seed extract (GSE), which has powerful anti-fungal activity, can induce apoptosis in S. cerevisiae. The yeast cells underwent apoptosis as determined by testing for apoptotic markers of DNA cleavage and typical chromatin condensation by Terminal Deoxynucleotidyl Transferase-mediated dUTP Nick End Labeling (TUNEL) and 4,6'-diaminidino-2-phenylindole (DAPI) staining and electron microscopy. The changes of ΔΨmt (mitochondrial transmembrane potential) and ROS (reactive oxygen species) indicated that the mitochondria took part in the apoptotic process. Changes in this process detected by metabonomics and proteomics revealed that the yeast cells tenaciously resisted adversity. Proteins related to redox, cellular structure, membrane, energy and DNA repair were significantly increased. In this study, the relative changes in the levels of proteins and metabolites showed the tenacious resistance of yeast cells. However, GSE induced apoptosis in the yeast cells by destruction of the mitochondrial 60 S ribosomal protein, L14-A, and prevented the conversion of pantothenic acid to coenzyme A (CoA). The relationship between the proteins and metabolites was analyzed by orthogonal projections to latent structures (OPLS). We found that the changes of the metabolites and the protein changes had relevant consistency.

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Related in: MedlinePlus

Representative silver-stained 2-D gel of total cellular extracts from yeast cells treated with GSE.The spots that were altered upon treatment are represented by numbers (1–26).
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pone-0032943-g001: Representative silver-stained 2-D gel of total cellular extracts from yeast cells treated with GSE.The spots that were altered upon treatment are represented by numbers (1–26).

Mentions: Image digitization was carried out with an Image Scanner (GE Healthcare, USA) in transmission mode. The comparison of the protein expression in the 2D gel images was performed using Image Master 2D Elite software (GE Healthcare, USA). To account for experimental variation, triplicate gels, from protein extracts obtained from independent experiments, were analyzed for each treatment. Spot detection was carried out automatically, and those spots showing faint intensity near the detection limit of colloidal CBB were not included in the comparisons. Prior to the automatic matching of spots between gel images, one gel was selected as the reference gel for each treatment. The amount of a protein spot was calculated based on the volume of that spot. To reflect the quantitative variations in intensity of protein spots between the control and treated samples, the spot volume was normalized as a percentage of the total volume of all the spots on the corresponding gel. Statistical analysis of the data was performed using SPSS software version 11.5 (SPSS Inc., Chicago, IL). The normalized intensity of spots on three replicate 2D gels was averaged, and the standard deviation was calculated for each treatment. A two-tailed non-paired Student's t test was used to determine whether the relative change was statistically significant between control and GSE-treated samples. Only spots that changed significantly in averaged normalized spot volume were excised for protein identification (Figure 1).


A mitochondria-dependent pathway mediates the apoptosis of GSE-induced yeast.

Cao S, Xu W, Zhang N, Wang Y, Luo Y, He X, Huang K - PLoS ONE (2012)

Representative silver-stained 2-D gel of total cellular extracts from yeast cells treated with GSE.The spots that were altered upon treatment are represented by numbers (1–26).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3293924&req=5

pone-0032943-g001: Representative silver-stained 2-D gel of total cellular extracts from yeast cells treated with GSE.The spots that were altered upon treatment are represented by numbers (1–26).
Mentions: Image digitization was carried out with an Image Scanner (GE Healthcare, USA) in transmission mode. The comparison of the protein expression in the 2D gel images was performed using Image Master 2D Elite software (GE Healthcare, USA). To account for experimental variation, triplicate gels, from protein extracts obtained from independent experiments, were analyzed for each treatment. Spot detection was carried out automatically, and those spots showing faint intensity near the detection limit of colloidal CBB were not included in the comparisons. Prior to the automatic matching of spots between gel images, one gel was selected as the reference gel for each treatment. The amount of a protein spot was calculated based on the volume of that spot. To reflect the quantitative variations in intensity of protein spots between the control and treated samples, the spot volume was normalized as a percentage of the total volume of all the spots on the corresponding gel. Statistical analysis of the data was performed using SPSS software version 11.5 (SPSS Inc., Chicago, IL). The normalized intensity of spots on three replicate 2D gels was averaged, and the standard deviation was calculated for each treatment. A two-tailed non-paired Student's t test was used to determine whether the relative change was statistically significant between control and GSE-treated samples. Only spots that changed significantly in averaged normalized spot volume were excised for protein identification (Figure 1).

Bottom Line: Proteins related to redox, cellular structure, membrane, energy and DNA repair were significantly increased.The relationship between the proteins and metabolites was analyzed by orthogonal projections to latent structures (OPLS).We found that the changes of the metabolites and the protein changes had relevant consistency.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of food safety and molecular biology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, People's Republic of China.

ABSTRACT
Grapefruit seed extract (GSE), which has powerful anti-fungal activity, can induce apoptosis in S. cerevisiae. The yeast cells underwent apoptosis as determined by testing for apoptotic markers of DNA cleavage and typical chromatin condensation by Terminal Deoxynucleotidyl Transferase-mediated dUTP Nick End Labeling (TUNEL) and 4,6'-diaminidino-2-phenylindole (DAPI) staining and electron microscopy. The changes of ΔΨmt (mitochondrial transmembrane potential) and ROS (reactive oxygen species) indicated that the mitochondria took part in the apoptotic process. Changes in this process detected by metabonomics and proteomics revealed that the yeast cells tenaciously resisted adversity. Proteins related to redox, cellular structure, membrane, energy and DNA repair were significantly increased. In this study, the relative changes in the levels of proteins and metabolites showed the tenacious resistance of yeast cells. However, GSE induced apoptosis in the yeast cells by destruction of the mitochondrial 60 S ribosomal protein, L14-A, and prevented the conversion of pantothenic acid to coenzyme A (CoA). The relationship between the proteins and metabolites was analyzed by orthogonal projections to latent structures (OPLS). We found that the changes of the metabolites and the protein changes had relevant consistency.

Show MeSH
Related in: MedlinePlus