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

Figure 2.

Figure 2.

Transcriptional activation of xMET17 by TFIIB-RFX leads to Mediator recruitment. (A) Untagged (No TAP) and TAP-tagged cells (Y892, Y893 and Y894) containing YCp33-xMET17-GFP and either pRS313-IIB or pRS313-IIB-RFX, or pRS313-IIB plus pRS315-Max-RFX, were grown in glucose-containing CSM medium supplemented with 0.5 mM methionine. Mediator and Pol II occupancy at xMET17-GFP and MET2 was measured by ChIP. DNA was quantitated by qPCR using primers specific for the ORF (Pol II ChIP) or for the promoter (Mediator ChIP). Occupancy levels were normalized using the ORF of the transcriptionally inactive gene IME2. Error bars indicate standard deviations from three independent experiments. (B) Left graph: A Med14 TAP-tagged rpb1-1 mutant and the isogenic wild-type strain (Y909 and Y911) containing YCp33-xMET17-GFP and pRS313-IIB or pRS313-IIB-RFX, were grown to early log phase at 25°C in CSM medium supplemented with 0.5 mM methionine, and were shifted to 37°C for 45 min before formaldehyde fixation. Mediator occupancy was measured by ChIP as in (A). Right graph: A met4-disrupted, Med14 TAP-tagged strain (met4Δ) and the isogenic wild-type strain (Y894 and Y963) containing YCp33-xMET17-GFP, and pRS314-IIB or pRS314-IIB-RFX, were grown and submitted to ChIP as above.