Figure 5.
Electrophoretic mobility shift assays for Gcn4 binding site probes show weakened binding when the native RY site is converted to a YR site. Probes (30 bp with 3′-6-FAM labels) containing the RY site in the ARO1 promoter or the RY site in the STP2 ORF (left panels). The right panels show ARO1 and STP2 probes in which the native RY motif is converted to a YR motif (see Supplemental Table S5 for the sequences). DNA at a final concentration of 10 nM was incubated with 500 ng of unlabeled poly(dA/dT) as competitor and increasing Gcn4 concentrations (shown in nM above each lane).
