Superoxide dismutase is involved in high tolerance to copper in the deep-sea yeast, Cryptococcus sp. N6 |
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Authors: | Takeshi Miura Fumiyoshi Abe Akira Inoue Ron Usami Koki Horikoshi |
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Affiliation: | (1) Department of Applied Chemistry, Faculty of Engineering, Toyo University, 2100 Kujirai, Kawagoe, 350-0815, Japan;(2) The DEEPSTAR Group, Japan Marine Science and Technology Center, 2-15 Natsushima-cho, Yokosuka, 237-0061, Japan |
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Abstract: | The activity and expression of superoxide dismutase (SOD) was analyzed in a copper-tolerant yeast, Cryptococcus sp. N6. Using cell extracts, two distinct bands exhibiting SOD activity appeared on native PAGE: one band, with higher mobility, appeared when the cells were grown without CuSO4, and the other band appeared when the cells were grown with 10 mM CuSO4. Cells grown with 3 mM CuSO4 produced both SOD isoforms. Western blot analysis, using a monoclonal antibody against human SOD-1, showed that SOD protein was expressed in the absence of CuSO4 and that the expression level increased when the cells were grown with 3 or 10 mM CuSO4. The molecular weight of SOD from strain N6 was approx. 18 kDa. Treatment of the cells with the protein synthesis inhibitor, cycloheximide at 0.5 g ml–1, did not affect cell growth in the absence of CuSO4 but significantly inhibited growth in the presence of 10 mM CuSO4 and inhibited expression of SOD protein. This suggests that SOD may play a role in cell growth in the presence of high concentrations of CuSO4. |
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Keywords: | catalase copper tolerance Cryptococcus sp. N6 deep-sea yeast superoxide dismutase |
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