FIGURE 2.

Dependence on Mediator of Swi/Snf and SAGA association with active gene promoters. A, Snf2 and Snf5 association with the CHA1 promoter in wild type (WT; BY4742 for Snf5 and Research Genetics Snf2-TAP strain for Snf2) and gal11/med15Δ med3Δ (LS10 for Snf5 and RMY525 for Snf2) yeast grown in CSM without (uninduced) or with 1 mg/ml of serine (induced) was measured by ChIP. B, Snf2 association with the MET2 and MET6 promoters in WT (Snf2-TAP strain, Research Genetics) and gal11/med15Δ med3Δ (RMY525) yeast grown in YPD without (uninduced) or with (induced) 0.5 mm CdCl₂. C, Snf5 association with the indicated promoters was measured in WT (RMY521) and srb4/med17 ts (RMY522) yeast grown in YPD after 1 h at 37 °C. D, Snf5 association with promoters whose transcription is dependent or independent of the Mediator tail module, as indicated, was measured in WT (Snf2-TAP strain, Research Genetics) and gal11/med15Δ med3Δ (RMY525) yeast grown in YPD. E, effect of Mediator tail mutation on SAGA recruitment. Left panel: Spt3 association with the CHA1 promoter was measured by ChIP in wild type (Snf2-TAP strain, Research Genetics) and gal11/med15Δ med3Δ yeast (RMY525) grown in CSM without (uninduced) or with (induced) 1 mg/ml of serine, and association with the MET2 and MET6 promoters was measured in wild type (RMY521) and gal11/med15Δ med3Δ yeast (RMY415) grown in YPD without (uninduced) or with (induced) 0.5 mm CdCl₂. Right panel, Ada2 and Gcn5 association at the uninduced and induced MET2 and MET6 promoters were measured by ChIP, and the log₂ ratios of the association at the induced/uninduced promoters in wild type and gal11/med15Δ med3Δ yeast are shown, as indicated. Relative association in all cases was determined by quantitative PCR analysis of input and immunoprecipitated (IP) samples, and normalized to a non-transcribed region of ChrV (25). Error bars represent S.D. (n = 3–4).