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Cloning and functional characterization of the copper/zinc superoxide dismutase gene from the heavy-metal-tolerant yeast Cryptococcus liquefaciens strain N6

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Abstract

The deep-sea yeast Cryptococcus liquefaciens strain N6 possesses high superoxide dismutase (SOD) activity and a high tolerance toward metal ions. To clarify the relationship between metal tolerance and SOD activity in this strain, we cloned the Cu/Zn SOD gene. This gene (Cl-SOD1) consists of 471 bp encoding 157 amino acids; the associated protein had 59.9–76.7% identity with Cu/Zn SOD proteins of other yeast species. The highest identity corresponded to Cryptococcus gattii (76.7%). Cl-SOD1 expression in the sod1 mutant of Saccharomyces cerevisiae revealed that this SOD protein was functional in S. cerevisiae. The Cl-SOD1 protein possessed approximately fourfold greater activity than S. cerevisiae SOD1 (Sc-SOD1) at 30°C. The amount of Cl-SOD1 mRNA in strain N6 increased in the presence of copper ion. However, the level of this transcript was not dependent on an increase in copper ion concentration and did not correlate well with changes in the amount of Cu/Zn SOD protein. This result suggests that strain N6 possesses other Cu/Zn SOD genes induced in a manner different from Cl-SOD1 as found in Candida albicans, or that the Cl-SOD1 gene undergoes posttranscriptional regulation upon increase of copper ion.

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Acknowledgment

This study was supported by Grants-in-Aid for the Protein 3000 Project (National Project on Protein Structural and Functional Analyses) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan.

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Authors and Affiliations

  1. Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259-B5 Nagatsuta, Midori-ku, Yokohama, Kanagawa, 226-8501, Japan

    Shin Kanamasa, Koichiro Sumi, Naho Yamuki, Takashi Kumasaka & Susumu Kajiwara

  2. Extremobiosphere Research Center, Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima-Cho, Yokohama, Kanagawa, 237-0061, Japan

    Takeshi Miura & Fumiyoshi Abe

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  1. Shin Kanamasa
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  2. Koichiro Sumi
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  3. Naho Yamuki
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  4. Takashi Kumasaka
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  5. Takeshi Miura
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  6. Fumiyoshi Abe
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  7. Susumu Kajiwara
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Correspondence to Susumu Kajiwara.

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Communicated by P. Punt.

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Kanamasa, S., Sumi, K., Yamuki, N. et al. Cloning and functional characterization of the copper/zinc superoxide dismutase gene from the heavy-metal-tolerant yeast Cryptococcus liquefaciens strain N6. Mol Genet Genomics 277, 403–412 (2007). https://doi.org/10.1007/s00438-006-0197-6

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  • DOI: https://doi.org/10.1007/s00438-006-0197-6

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