TY - JOUR
T1 - Overexpression of L-glutamine
T2 - D-fructose-6-phosphate amidotransferase provides resistance to methylmercury in Saccharomyces cerevisiae
AU - Miura, Nobuhiko
AU - Kaneko, Satoshi
AU - Hosoya, Shinji
AU - Furuchi, Takemitsu
AU - Miura, Kyoko
AU - Kuge, Shusuke
AU - Naganuma, Akira
PY - 1999/9/17
Y1 - 1999/9/17
N2 - 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. Copyright (C) 1999 Federation of European Biochemical Societies.
AB - 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. Copyright (C) 1999 Federation of European Biochemical Societies.
KW - Cytotoxicity
KW - L-Glutamine:D-fructose-6-phosphate amidotransferase
KW - Methylmercury
KW - Resistance gene
KW - Screening
KW - Yeast
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U2 - 10.1016/S0014-5793(99)01158-8
DO - 10.1016/S0014-5793(99)01158-8
M3 - Article
C2 - 10481068
AN - SCOPUS:0032869418
VL - 458
SP - 215
EP - 218
JO - FEBS Letters
JF - FEBS Letters
SN - 0014-5793
IS - 2
ER -