GENOMIC FOOTPRINTING OF THE PROMOTER REGIONS OF STE2 AND STE3 GENES IN THE YEAST SACCHAROMYCES-CEREVISIAE

Dimethyl sulfate, DNase I and micrococcal nuclease DNA cleavage were combined with the ligation-mediated polymerase chain reaction to obtain high resolution maps of the promoter regions for two cell-type-specific genes: the a-specific STE2 gene and the α-specific STE3 gene. We find that MCM1 binds in vivo in a-cells to a 16 bp P-box sequence located in the STE2 UAS. In α-cells, the footprint pattern is extended relative to a-cells, consistent with the additional binding of MATα2 to the sequences flanking each end of the P-box. A nucleosome was found adjacent to the P-box of the transcriptionally repressed a-specific STE2 UAS in α-cells, positioned so that the nucleosome overlaps the TATA-box. In contrast, such well-positioned nucleosomes were not found for the transcriptionally active STE2 UAS in a-cells, where instead the TATA box appears to be bound to the general transcription factor TFIID. These observations support the hypothesis that MATα2 repression of a-specific genes is mediated by nucleosomes, perhaps by exclusion of TFIID from the TATA-box. © 1993 Academic Press Limited.