Abstract
We have previously shown that overexpression of the human tumor suppressor protein P53 causes cell death of the yeast Saccharomyces cerevisiae. P53 overproduction led to transcriptional downregulation of some yeast genes, such as the TRX1/2 thioredoxin system, which plays a key role in cell protection against various oxidative stresses induced by reactive oxygen species (ROS). In the present work, the impact of TRX2 overexpression on apoptosis mediated by p53 overexpression in yeast is investigated. In yeast cells expressing P53 under an inducible promoter together with TRX2 under a strong constitutive promoter, we showed that Tr2p overproduction reduced the apoptotic effect exerted by P53 and increased the viability of the P53-overproducing cells. Furthermore, measurements of ROS amounts by flow cytometry and fluorescence microscopy indicated that the TRX2 protein acted probably through its increased detoxifying activity on the P53-generated ROS. The steady-state level and activity of P53 were not affected by TRX2 overexpression, as shown by western blotting and functional analysis of separated alleles in yeast (FASAY), respectively. The growth inhibitory effect of P53 was partially reversed by the antioxidant N-acetylcysteine. Our data strengthen the idea that overexpression of a single gene (trx2) decreases the p53-mediated cell death by decreasing ROS accumulation.
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Acknowledgments
This work is dedicated to Pr Daniel Thomas, UTC Compiègne, France, and to our colleague Dr. Noomen Hadj Taieb. We deeply thank Pr Richard Iggo for the gift of strain yIG397 and for his encouragement. Pr Gerard Rosset, Pr Denis Pompon, and Pr François Lacroute are thanked for the gift of BFG1, pDP, and pFL39 vectors. Mrs. Rim Besbes is thanked for her help in fluorescent microscopy. Dr. Wajdi Ayadi is thanked for the antibody anti-tubulin. This work is a part of the contract program concluded with the «Ministère de l’Enseignement Supérieur et de la Recherche Scientifique» of Tunisia.
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This article does not contain any studies with human participants or animals performed by any of the authors.
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This study was funded by the Ministry of Higher Education and Scientific Research, Tunisian Government.
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Kamoun, Y., Mabrouk, I., Delahodde, A. et al. Overexpression of yeast thioredoxin TRX2 reduces p53-mediated cell death in yeast. Appl Microbiol Biotechnol 99, 8619–8628 (2015). https://doi.org/10.1007/s00253-015-6886-5
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DOI: https://doi.org/10.1007/s00253-015-6886-5