FIG. 7.

Recruitment of the Srb8-11 complex to the STA1 promoter requires Nrg1 and Sfl1. (A) Cell extracts prepared from the integrated SRB10-HA strain were incubated with 5 μg of GST, GST-Nrg1, or GST-Sfl1. GST proteins and their interacting proteins were precipitated with glutathione-agarose beads. Fractions of the input (1/10) and pellet (1/2) were analyzed by Western blot analysis with an α-HA antibody. (B) Cells tagged with integrated SRB10-HA, isogenic mutants, or nontagged strains were grown to mid-log phase in synthetic medium containing 2% glucose and were then fixed with formaldehyde. An α-HA ChIP assay was performed, and the core promoter region of the STA1 promoter was amplified by PCR using purified DNA to detect Srb10 binding to the STA1 promoter.