Figure 5

The Mediator is required for Swi/Snf recruitment to the GAL1 UAS_G. (A) ChIP analysis of the binding of Swi1-Myc to the GAL1 UAS_G in the absence of the Mediator. Temperature-sensitive srb4-138 mutant (KLY042) containing Swi1-Myc was grown at 28°C and either shifted to 37°C for 45 min to inactivate srb4 (37°C) or kept at the permissive temperature (28°C). Galactose was added to media as in previous experiments GAL gene expression. (B) Quantification of experiments as shown in (A). (C, D) Similar experiment as in (A, B) except that binding of Gal11-Myc was analyzed (KLY090). PCRs contain chromosome II telomere primers used here as an internal control to normalize signals for each lane. (E) Persistence of the Swi/Snf complex in the absence of the Mediator. KLY042 cells were grown at the permissive temperature and galactose was added to the media for 30 min to induced GAL gene expression. Cells were then shifted to inducing media at permissive (28°C) or nonpermissive temperature (37°C). Aliquots were taken 30, 60, 120, and 180 min after a shift to the nonpermissive temperature. PCRs contain chromosome II telomere primers as an internal control. (F) Quantification of experiments as illustrated in (E).