Overexpression of L‐glutamine:D‐fructose‐6‐phosphate amidotransferase provides resistance to methylmercury in Saccharomyces cerevisiae

FEBS Letters

Miura, Nobuhiko³  Furuchi, Takemitsu³  Miura, Kyoko²  Kaneko, Satoshi³  Naganuma, Akira³  Hosoya, Shinji³  Kuge, Shusuke¹ 

[1] Department of Microbiology, Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo 108-8639, Japan;Department of Environmental Sciences, Wako University, Machida 195-0071, Japan;Laboratory of Molecular and Biochemical Toxicology, Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai 980-8578, Japan

关键词: Methylmercury;    Cytotoxicity;    Yeast;    Screening;    Resistance gene;    L-Glutamine:D-fructose-6-phosphate amidotransferase;   

DOI  :  10.1016/S0014-5793(99)01158-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

To identify novel genes that confer resistance to methylmercury (MeHg), a yeast genomic DNA library was transfected into Saccharomyces cerevisiae. Two functional plasmids were isolated from transfected yeast clones D1 and H5 that exhibited resistance to MeHg. The yeast transfected with plasmid isolated from clone H5 was several-fold more resistant than yeast transfected with plasmid from clone D1. Functional characterization of the genomic DNA fragment obtained from clone H5 determined that the GFA1 gene conferred resistance to MeHg. GFA1 was reported to encode L-glutamine:D-fructose-6-phosphate amidotransferase (GFAT) which catalyzes the synthesis of glucosamine-6-phosphate from glutamine and fructose-6-phosphate. Accumulation of mercury in yeast clone W303B/pGFA1, which contains the transfected GFA1 gene, did not differ from that in control yeast clone W303B/pYES2. The W303B/pGFA1 strain did not show resistance to mercuric chloride, zinc chloride, cadmium chloride or copper chloride, suggesting that the resistance acquired by GFA1 gene transfection might be specific to MeHg. This is the first report of a gene involved in MeHg resistance in eukaryotic cells identified by screening a DNA library.

【 授权许可】

Unknown