Figure 1

ChIP assays of promoter fragments that are predicted to be targets of the a 1-α 2 complex. ChIP assays with antibody to the α 2 protein were performed on lysates from MAT a/MATα (a/α) and mat a Δ/MATα(Δ/α) cells. Total chromatin (TC) and immunoprecipitated (IP) samples were subjected to multiplex PCR with primers flanking potential a 1-α 2 sites (labeled hsg) in the indicated promoters. Primers for the promoter region of STE6, an a-specific gene that is repressed by the α 2-Mcm1 complex in both cell types were used as a positive control for the ChIPs. Primers that hybridize in YDL223C, a gene that is not regulated by α 2, was used as the negative control. The presence of a band over background levels from the test promoter from a/α lysates but not mat Δ/α indicates that the a 1-α 2 complex is specifically binding to the promoter. Assays for the GPA1, FAR1, NEJ1, RDH54, MET31, PDE1, AMN1/CST13 and KAR1 are shown. A summary of the all of the ChIPs performed is in Table 1.