Methods of using glucan synthase pathway reporter genes to screen for antifungal compounds

A method for determining the effect of a molecule upon the function or activity of the glucan synthase pathway comprising:

• (a) contacting a S. cerevisiae cell with, or recombinantly expressing within the cell the molecule;
• (b) detecting a change in RNA expression or protein expression in said cell of at least one target polynucleotide sequence relative to the expression of said target polynucleotide sequence in the absence of the molecule, each said target polynucleotide sequence being regulated by a promoter native to a gene selected from the group consisting of YOL113W (SKM1), YNR066C, YLR121C (YPS3), YHR209W, YKL161C, YAR050W (FLO1), and YFR030W (MET10);
• and
• (c) determining the effect of the molecule upon the function or activity of the glucan synthase pathway based upon the change in RNA expression or protein expression.

A method for monitoring the activity of the glucan synthase pathway in a S. cerevisiae cell exposed to a molecule comprising:

• (a) contacting the cell with, or recombinantly expressing within the cell, the molecule;
• (b) determining whether the RNA expression or protein expression in said cell of at least one target polynucleotide sequence is changed in step (a) relative to the expression of said target polynucleotide sequence in the absence of the molecule, each said target polynucleotide sequence being regulated by a promoter native to a gene selected from the group consisting of YOL113W (SKM1), YNR066C, YLR121C (YPS3), YHR209W, YKL161C, YAR050W (FLO1), and YFR030W (MET10);
• and
• (c) determining that the activity of the glucan synthase pathway in said cell is changed if the expression is determined to be changed in step (b), or determining that the activity of the glucan synthase pathway in said cell is unchanged if the expression is determined to be unchanged in step (b).

A method for determining whether a first S. cerevisiae cell is mutant for a glucan synthase pathway gene comprising:

• (a) in said first S. cerevisiae cell, determining the RNA or protein expression of at least one target polynucleotide sequence, each target polynucleotide sequence being regulated by a promoter native to a gene selected from the group consisting of YOL113W (SKM1 1), YNR066C, YLR121C (YPS3), YHR209W, and YKL161C, wherein said cell is not being exposed to an inhibitor of the glucan synthase pathway;
• (b) determining whether the RNA and/or protein expression of said at least one target polynucleotide sequence determined in step (a) is changed relative to the RNA and/or protein expression of said at least one target polynucleotide sequence in a second S. cerevisiae cell which is wildtype for glucan synthase genes;
• and
• (c) determining that the first S. cerevisiae cell is mutant for a glucan synthase pathway gene if the expression is determined to be changed in step (a), or determining that the first S. cerevisiae cell is not mutant for a glucan synthase pathway gene if the expression is determined to be unchanged in step (b).