FIG 4.

Gcn4-mediated activation of Taf9 CRD-dependent genes. (A) Venn diagram showing statistically significant overlap between genes induced in Taf9 CRD-dependent (463 genes; this study) and Gcn4-dependent (515 genes [42]) manners. (B) qRT-PCR validation of genes from cluster I3 (Fig. 2) to determine Gcn4 dependence for activation. cDNA was prepared from total RNA isolated from WT (RPY72) and gcn4Δ (strain 249) strains (with or without SM) and analyzed as for Fig. 3B. (C) Recruitment of Gcn4 in the UAS regions of Taf9 CRD-dependent genes. Cross-linked chromatin extracts from the WT TAF9 (YMS94) or taf9-tCRD2 (YMS95) strain treated or not with SM were used to immunoprecipitate Gcn4 using 5 μl polyclonal anti-Gcn4 antibody per IP reaction. The fold enrichment was calculated relative to the POL1 ORF as a control. The error bars indicate SEM from a minimum of two chromatins and three IP reactions.