MAT-ALPHA-1 CAN MEDIATE GENE ACTIVATION BY A-MATING FACTOR

In the yeast Saccharomyces cerevisiae, expression of alpha-specific genes is governed by the MAT-alpha-1 and MCM1 gene products. MAT-alpha-1 and MCM1 bind cooperatively to PQ elements upstream of alpha-specific genes. The PQ element not only directs alpha-specific expression but can also direct gene induction in response to treatment with a-mating pheromone. We have used gene fusions to investigate whether induction conferred by the PQ box is mediated through either MAT-alpha-1 or MCM1, or a combination of both. When MCM1 is fused to the DNA-binding domain of the bacterial repressor LexA, this fusion protein is capable of trans-activating a lacZ reporter gene driven by a LexA operator. However, the transcriptional activity of the MCM1-LexA fusion is not further enhanced by treatment of cells with a-factor. A MAT-alpha-1-LexA fusion protein is also capable of trans-activation through a LexA operator. Moreover, the activity of the MAT-alpha-1-LexA fusion protein can be further induced by treatment with a-factor. When progressive deletions are made from the amino terminus of MAT-alpha-1 in the fusion protein, the basal level of trans-activation progressively decreases, but the inducibility of the fusion protein increases. MAT-alpha-1-LexA fusion proteins, which have greater-than-or-equal-to 57 amino acids deleted from the amino terminus of MAT-alpha-1 are not capable of trans-activation. In addition, the activity of the MAT-alpha-1-LexA fusion protein is dependent on the functions of the STE7, STE11, and STE12 genes that encode components of the pheromone response pathway.