FIGURE 3.

Ash1 recruits Rpd3(L) to HO URS2. A, strains DY8312 (GALp::CDC20 ASH1-Myc) and DY12247 (GALp::CDC20 SDS3-Myc) were synchronized, and ChIP experiments were performed to measure Ash1 and Rpd3(L) binding to the HO promoter. Binding to URS1 and URS2 was measured with primers that amplify from −1429 to −1139 and from −825 to −489, respectively. Sds3 is a subunit specific to Rpd3(L). B, ChIP samples were prepared from logarithmically growing strains DY8312 (ASH1-Myc) and DY12247 (SDS3-Myc), and binding was analyzed with 15 sets of PCR primers across the HO promoter. C, strain DY12247 (GALp::CDC20 SDS3-Myc) was synchronized, and ChIP samples were taken at various times following release from CDC20 arrest. Binding was analyzed with 15 sets of PCR primers across the HO promoter. URS1-, URS2-, Swi5-, and SBF-binding sites are shown for the HO promoter, where the ATG represents +1 and the transcription start site is at −20. D, strain DY13197 (GALp::CDC20 Ash1-Myc SIN3-HA) was synchronized, and protein samples were taken at various times following release from CDC20 arrest. Sin3-HA was immunoprecipitated with anti-HA antibody and analyzed on Western blots, along with controls corresponding to 10% of the input before immunoprecipitation, and the blots were probed with anti-HA and anti-Myc antibodies. E, strains DY12247 (GALp::CDC20 SDS3-Myc) and DY12251 (GALp::CDC20 SDS3-Myc ash1) were synchronized, and ChIP samples were taken at various times following release from CDC20 arrest. Binding to HO URS1 was measured with primers that amplify from −1429 to −1139.