Figure 2

Genome-wide expression analysis shows that loss of function of the Mediator tail module affects specific gene classes. Microarray expression profiling was done in WT, gal11–myc med3Δ, and gal11Δmed3Δ strains by isolating total RNA from cells grown in YPD media. (A) Scatterplot of expression changes in gal11–myc med3Δ and gal11Δmed3Δ strains as compared with WT strain for those genes with altered expression in at least one mutant by at least 1.5-fold. (B) A heat map was generated by carrying out clustering of expression changes ⩾1.5-fold in gal11-myc med3Δ and gal11Δ med3Δ strains by utilizing a k-means algorithm. (C) Northern analysis was performed for the expression of CHA1, MET2, MET32, and SPS19 genes in WT, gal11/med15Δ med3Δ, gal11–myc med3Δ, srb8/med12Δ, and srb10/cdk8Δ strains using gene-specific probes. The lower band seen in the MET32 panel corresponds to the transcript that is seen upon induction. Ethidium bromide stained 25S rRNA is shown as loading control. (D) Expression of MET2, MET32, SPS19, and SPO20 was analysed by qRT–PCR. The fold differences between WT and mutant strains are shown by setting the values in the WT strain as 1 after normalizing to SNR6 expression.