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. 2014 Nov 6;5(1):21–33. doi: 10.1534/g3.114.015180

Figure 2.

Figure 2

Genetic interactions between GTF and Pol II mutants. (A) Plasmids containing tfg2 mutants were integrated into yeast strains constructed to allow shuffling of a WT RPO21/RPB1 URA3 plasmid in favor of WT or mutant rpo21/rpb1 LEU2 plasmids through use of 5FOA poisoning of URA3+ cells. On the left, SC-Leu media allows coexistence of both URA3 and LEU2 plasmids. On the right, supplementation of SC-Leu with 5FOA uncovers rpb1 phenotypes by selecting against the RPB1 URA3 plasmid. Pol II mutants that have slower elongation rate than WT in vitro (LOFs) and mutants that genetically cluster with them are annotated with a blue bar; mutants that have faster elongation rate than WT in vitro (GOFs) and mutants genetically cluster with them are labeled with a green bar. Mutants are arranged by their measured elongation rates or elongation rates inferred by strength of genetic phenotypes compared with those mutants tested biochemically (Kaplan et al. 2012). (B) sua7 allele-Pol II mutant interactions examined as for tfg2 alleles in (A). (C and D) Phenotypes of viable GTF-Pol II double mutants are shown as a heatmap with qualitative determinations of growth defects on various media. Inviable double mutants are colored in gray. In YPD, YPD 37°C, YPRaf, and SC-Leu media, single and double mutant growth levels are normalized to WT. Blue indicates decreased growth relative to WT; red indicates increased growth compared with WT. In SC-Leu+MPA, growth difference is normalized to that on SC-Leu to quantify MPA sensitivity (shown as blue) or resistance (shown as red). In YPRafGal and SC-Lys, growth on the plate is divided by ratio of WT growth to mutant growth on corresponding general media (YPD and SC-Leu) to account for GalR and Spt- phenotypes (Gal+ and Lys+, shown in red) in contrast to their underlying growth defects. (C) tfg2 mutants. (D) sua7 mutants. See Figure S2 and Figure S3 for representative spot growth assay figures used for quantification in heatmaps and see Materials and Methods for further explanation of heatmaps.