Cloning and sequencing of the GMP synthetase-encoding gene of Saccharomyces cerevisiae.
We have localised, within a Saccharomyces cerevisiae genomic fragment, the GUA1 gene whose amplification leads to the accumulation of several polypeptides on the two-dimensional (2-D) map of yeast proteins. Comparison of the sequence of the putative GUA1 protein with a data library shows a strong similarity with Escherichia coli, Bacillus subtilis and Dictyostelium discoideum GMP synthetases (GMPS) and other glutamine amidotransferases. The fact that disruption of the chromosomal copy of the gene leads to guanine auxotrophy, that the gual::URA3 disruption does not complement an independently obtained gual-3 mutation deficient in GMPS and that GUA1 complements this latter mutation, confirms the identification of the cloned gene as GUA1 encoding the S. cerevisiae GMPS. Finally, using microsequencing, we have identified one of the polypeptides, which is overproduced in response to GUA1 amplification, as corresponding to GUA1.
Dujardin
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Centre de Génétique Moléculaire du CNRS, 91198 Gif-sur-Yvette, France.
Kermorgant
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Slonimski
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PP
Boucherie
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D000595:Amino Acid Sequence; D000818:Animals; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D019731:Carbon-Nitrogen Ligases; D003001:Cloning, Molecular / Q000379:methods; D004023:Dictyostelium / Q000201:enzymology / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005800:Genes, Fungal; D008025:Ligases / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D017386:Sequence Homology, Amino Acid
D011994:Recombinant Proteins; D008025:Ligases; D019731:Carbon-Nitrogen Ligases; C020828:GMP synthase (glutamine-hydrolyzing)