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Overexpression of L ‐glutamine: D ‐fructose‐6‐phosphate amidotransferase provides resistance to methylmercury in Saccharomyces cerevisiae
Author(s) -
Miura Nobuhiko,
Kaneko Satoshi,
Hosoya Shinji,
Furuchi Takemitsu,
Miura Kyoko,
Kuge Shusuke,
Naganuma Akira
Publication year - 1999
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(99)01158-8
Subject(s) - glutamine amidotransferase , saccharomyces cerevisiae , yeast , biochemistry , transfection , plasmid , chemistry , microbiology and biotechnology , glutamine , biology , gene , amino acid
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.