Overexpression of peptide-methionine sulfoxide reductase inSaccharomyces cerevisiaeand human T cells provides them with high resistance to oxidative stress
| Autor: | Poston Jm, Barbara S. Berlett, Jackob Moskovitz, E Flescher, Earl R. Stadtman, J Azare |
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| Rok vydání: | 1998 |
| Předmět: |
Paraquat
T-Lymphocytes Saccharomyces cerevisiae Mutant Amidines Transfection Polymerase Chain Reaction Gene Expression Regulation Enzymologic Cell Line chemistry.chemical_compound Escherichia coli Animals Humans Cloning Molecular Hydrogen peroxide Growth medium Multidisciplinary Methionine biology Methionine sulfoxide Hydrogen Peroxide Biological Sciences Oxidants biology.organism_classification Recombinant Proteins Oxidative Stress chemistry Biochemistry Methionine Sulfoxide Reductases Methionine sulfoxide reductase Cattle Oxidoreductases MSRA |
| Zdroj: | Proceedings of the National Academy of Sciences. 95:14071-14075 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.95.24.14071 |
| Popis: | The yeast peptide-methionine sulfoxide reductase (MsrA) was overexpressed in aSaccharomyces cerevisiaenull mutant ofmsrAby using a high-copy plasmid harboring themsrAgene and its promoter. The resulting strain had about 25-fold higher MsrA activity than its parent strain. When exposed to either hydrogen peroxide, paraquat, or 2,2′-azobis-(2-amidinopropane) dihydrochloride treatment, the MsrA overexpressed strain grew better, had lower free and protein-bound methionine sulfoxide and had a better survival rate under these conditions than did themsrAmutant and its parent strain. Substitution of methionine with methionine sulfoxide in a medium lacking hydrogen peroxide had little effect on the growth pattern, which suggests that the oxidation of free methionine in the growth medium was not the main cause of growth inhibition of themsrAmutant. Ultraviolet A radiation did not result in obvious differences in survival rates among the three strains. An enhanced resistance to hydrogen peroxide treatment was shown in human T lymphocyte cells (Molt-4) that were stably transfected with the bovinemsrAand exposed to hydrogen peroxide. The survival rate of the transfected strain was much better than its parent strain when grown in the presence of hydrogen peroxide. These results support the proposition that themsrAgene is involved in the resistance of yeast and mammalian cells to oxidative stress. |
| Databáze: | OpenAIRE |
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