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

Figure 1.

Figure 1.

TFIIB-RFX supports accurate transcription initiation from a MET17 promoter containing an upstream RFX binding site. (A) Experimental strategy. See text for details. (B) Strain Y892 (see Supplementary Table S1) containing YCp33-xMET17-GFP, and either pRS314-TFIIB (IIB) or pRS314-TFIIB-RFX (IIB-RFX), were grown in glucose-containing CSM medium supplemented with 0.5 mM methionine (+methionine), or in glucose-containing YNB medium with no methionine (−methionine). RNA levels for xMET17-GFP, MET17 and ACT1 were quantified by RT-qPCR and normalized to 25S rRNA levels. Values are expressed as a percentage of the maximum value. Error bars represent standard deviations from three independent experiments. (C) Same RNA preparations as in (B) were subjected to primer extension analysis with primers for MET17 and xMET17-GFP starting in both cases at exactly the same distance from MET17 TATA box (175 bp). U3 snoRNA was used as a control.