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. 2013 Aug 20;41(21):9651–9662. doi: 10.1093/nar/gkt701

Figure 7.

Figure 7.

Transcriptional activation by LexA-TBP from a MET17 derivative containing the LexA operator requires Mediator. (A) Untagged (No TAP) and Med14 TAP-tagged cells (Y14 and Y84) containing YCp33-opMET17-GFP (see schematic representation) and either pRS313-LexA (Lex) or pRS313-LexA-TBP (LexTBP) were grown as shown in Figure 2. Pol II and Med14-TAP occupancy at opMET17-GFP and MET2 was measured by ChIP. DNA was analysed by qPCR using primers for the ORF (Pol II ChIP) or the promoter (Mediator ChIP). Occupancy levels were normalized using IME2 ORF. Error bars indicate standard deviations from three (Pol II ChIP) or four (Mediator ChIP) independent experiments. Asterisks indicate P < 0.005 in a Student’s t test. (B) The med17(srb4)-138 mutant and an isogenic wild-type strain (Y400 and Y402) containing YCp33-opMET17-GFP and either YCp91-LexA (Lex) or YCp91-LexA-TBP (LexTBP) were grown at 28°C to early log phase in CSM medium supplemented with 0.5 mM methionine, and were shifted to 37°C. Pol II occupancy was measured by ChIP before and 60 min after the shift. DNA was analysed by qPCR using primers for the 5′-end of GFP ORF, ACT1 ORF and IME2 ORF. Error bars indicate standard deviations from two independent experiments.