Investigation of protein expression of Saccharomyces cerevisiae cells in quiescent and proliferating state before and after toxic stress

The focus of the present study is to determine proteins responsible for the oxidative and toxic stress response in proliferating and stationary phase (G0) cultures. Therefore, the yeast Saccharomyces cerevisiae was treated with oxidative and drug compounds (H2O2, menadione, zeocin, and ibuprofen) in...

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Main Authors: Asya Daskalova, Ventsislava Petrova, Lyudmila Velkova, Anna Kujumdzieva, Anna Tomova, Wolfgang Voelter, Pavlina Dolashka
Format: Article
Language:English
Published: Taylor & Francis Group 2021-01-01
Series:Biotechnology & Biotechnological Equipment
Subjects:
Online Access:http://dx.doi.org/10.1080/13102818.2021.1879677
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author Asya Daskalova
Ventsislava Petrova
Lyudmila Velkova
Anna Kujumdzieva
Anna Tomova
Wolfgang Voelter
Pavlina Dolashka
author_facet Asya Daskalova
Ventsislava Petrova
Lyudmila Velkova
Anna Kujumdzieva
Anna Tomova
Wolfgang Voelter
Pavlina Dolashka
author_sort Asya Daskalova
collection DOAJ
description The focus of the present study is to determine proteins responsible for the oxidative and toxic stress response in proliferating and stationary phase (G0) cultures. Therefore, the yeast Saccharomyces cerevisiae was treated with oxidative and drug compounds (H2O2, menadione, zeocin, and ibuprofen) in both phases. These substances were chosen to determine the redox status of the yeast. S. cerevisiae appeared to employ different strategies to ensure their antioxidant defence metabolism. Analysis, including sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) coupled with mass spectrometry, was used in the search. The proteins were identified by SDS-PAGE, matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) mass spectrometry analysis, and Mascot database-fingerprint. The final step was determination of protein profiles of yeast S. cerevisiae in proliferating (M) and stationary phase (G0). Seven bands were determined and the corresponding proteins were proposed: cytochrome c peroxidase, glutathione S-transferase omega-like, NAPDH-dependent diflavin reductase, DNA replication fork-blocking protein, putative aryl alcohol dehydrogenase, AP-1-like transcription factor YAP5, GTP-binding protein. All putative proteins coincide with the literature database. A typical example of such an adaptation mechanism in the defence against oxidative damage is the synthesis of several glutathione and thioredoxin peroxidases in the yeast cell. A deeper investigation of the conserved mechanisms responsible for entry into, survival, and exit from quiescence in higher eukaryotes will help the development of new anticancer therapies, the study in the process of ageing and neurodegenerative diseases.
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spelling doaj.art-f7b06f652d6548f99054b86e60499d082022-12-21T18:35:24ZengTaylor & Francis GroupBiotechnology & Biotechnological Equipment1310-28181314-35302021-01-0135136637610.1080/13102818.2021.18796771879677Investigation of protein expression of Saccharomyces cerevisiae cells in quiescent and proliferating state before and after toxic stressAsya Daskalova0Ventsislava Petrova1Lyudmila Velkova2Anna Kujumdzieva3Anna Tomova4Wolfgang Voelter5Pavlina Dolashka6Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of SciencesDepartment of General and Industrial Microbiology, Faculty of Biology, Sofia University ‘St. Kliment Ohridski’Institute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of SciencesDepartment of General and Industrial Microbiology, Faculty of Biology, Sofia University ‘St. Kliment Ohridski’Department of General and Industrial Microbiology, Faculty of Biology, Sofia University ‘St. Kliment Ohridski’Interfaculty Institute of Biochemistry, University of TübingenInstitute of Organic Chemistry with Centre of Phytochemistry, Bulgarian Academy of SciencesThe focus of the present study is to determine proteins responsible for the oxidative and toxic stress response in proliferating and stationary phase (G0) cultures. Therefore, the yeast Saccharomyces cerevisiae was treated with oxidative and drug compounds (H2O2, menadione, zeocin, and ibuprofen) in both phases. These substances were chosen to determine the redox status of the yeast. S. cerevisiae appeared to employ different strategies to ensure their antioxidant defence metabolism. Analysis, including sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) coupled with mass spectrometry, was used in the search. The proteins were identified by SDS-PAGE, matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) mass spectrometry analysis, and Mascot database-fingerprint. The final step was determination of protein profiles of yeast S. cerevisiae in proliferating (M) and stationary phase (G0). Seven bands were determined and the corresponding proteins were proposed: cytochrome c peroxidase, glutathione S-transferase omega-like, NAPDH-dependent diflavin reductase, DNA replication fork-blocking protein, putative aryl alcohol dehydrogenase, AP-1-like transcription factor YAP5, GTP-binding protein. All putative proteins coincide with the literature database. A typical example of such an adaptation mechanism in the defence against oxidative damage is the synthesis of several glutathione and thioredoxin peroxidases in the yeast cell. A deeper investigation of the conserved mechanisms responsible for entry into, survival, and exit from quiescence in higher eukaryotes will help the development of new anticancer therapies, the study in the process of ageing and neurodegenerative diseases.http://dx.doi.org/10.1080/13102818.2021.1879677saccharomyces cerevisiaemaldi-tof/tof spectraoxidative and toxic stressproliferating (m) and stationary phase (g0)
spellingShingle Asya Daskalova
Ventsislava Petrova
Lyudmila Velkova
Anna Kujumdzieva
Anna Tomova
Wolfgang Voelter
Pavlina Dolashka
Investigation of protein expression of Saccharomyces cerevisiae cells in quiescent and proliferating state before and after toxic stress
Biotechnology & Biotechnological Equipment
saccharomyces cerevisiae
maldi-tof/tof spectra
oxidative and toxic stress
proliferating (m) and stationary phase (g0)
title Investigation of protein expression of Saccharomyces cerevisiae cells in quiescent and proliferating state before and after toxic stress
title_full Investigation of protein expression of Saccharomyces cerevisiae cells in quiescent and proliferating state before and after toxic stress
title_fullStr Investigation of protein expression of Saccharomyces cerevisiae cells in quiescent and proliferating state before and after toxic stress
title_full_unstemmed Investigation of protein expression of Saccharomyces cerevisiae cells in quiescent and proliferating state before and after toxic stress
title_short Investigation of protein expression of Saccharomyces cerevisiae cells in quiescent and proliferating state before and after toxic stress
title_sort investigation of protein expression of saccharomyces cerevisiae cells in quiescent and proliferating state before and after toxic stress
topic saccharomyces cerevisiae
maldi-tof/tof spectra
oxidative and toxic stress
proliferating (m) and stationary phase (g0)
url http://dx.doi.org/10.1080/13102818.2021.1879677
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