Identification of Target Sites of the α2–Mcm1 Repressor Complex in the Yeast Genome

The DNA-binding activity for the ASG7 site in EMSAs. (A) Purified α2 protein was used in an EMSA with labeled fragments containing the indicated α2-Mcml-binding sites. The protein concentration was 4.0 × 10⁻⁷ m(lanes 1,5,9), 8.0 × 10⁻⁸ m (lanes2,6,10), 1.6 × 10⁻⁸ m (lanes3,7,11), 3.2 × 10⁻⁹ m (lanes 4,8,12). (B) Purified Mcm1 protein was used in an EMSA with labeled fragments at a concentration of 1.7 × 10⁻⁸ m (lanes1,5,9), 1.7 × 10⁻⁹ m (lanes2,6,10), 1.7 × 10⁻¹⁰ m (lanes3,7,11), and 1.7 × 10⁻¹¹ m (lanes4,8,12). (C) Purified α2 protein at a concentration of 8.0 × 10⁻⁸ m (lanes 1,6,11), 1.6 × 10⁻⁸ m (lanes 2,7,12), 3.2 × 10⁻⁹ m (lanes 3,8,13), 6.4 × 10⁻¹⁰ m (lanes 4,9,14), 1.3 × 10⁻¹⁰ m (lanes 5,10,15) was mixed with Mcm1_1–96 fragment at a concentration of 1.7 × 10⁻⁹ m in the EMSAs. The α2–Mcm1 complex includes two molecules of α2 and two molecules of Mcm1 and the Mcm1 complex includes two molecules of Mcm1. the EMSAs shown are phosphorimages of the gels.