Cloning of the glutamine:fructose-6-phosphate amidotransferase gene from yeast. Pheromonal regulation of its transcription

Abstract

The activity of the amino sugar-synthesizing enzyme L-glutamine:D-fructose-6-phosphate aminotransferase (EC 2.6.1.16) in haploid a cells of Saccharomyces cerevisiae increases 1.7-fold after alpha factor addition. The gene (the gene should be called GFA1 for glutamine:fructose-6-phosphate amidotransferase) for the enzyme has been cloned by complementing the gcn1 mutation (Whelan, W. L., and Ballou, C. E. (1975) J. Bacteriol. 125, 1545-1557). Its expression is increased 2-3 times within 15 min when the mating pheromone is added. The gene codes for a protein of 716 amino acids in length. It is highly homologous (64%) to the corresponding gene of Escherichia coli, except for a sequence coding for 83 amino acids (numbers 204-286), which is lacking in E. coli. The amino-terminal region of the coding sequence also shows a high degree of homology to the corresponding sequence of the E. coli and S. cerevisiae L-glutamine:phosphoribosylpyrophosphate amidotransferase. In the promotor region of the S. cerevisiae L-glutamine:D-fructose-6-phosphate amidotransferase gene the heptanucleotide "TGAAACA," shown to be required for pheromone control of transcription (Kronstad, J. W., Holly, J. A., and MacKay, V. L. (1987) Cell 50, 369-377), is present six times.