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. 2016 Jul 18;113(31):8699–8704. doi: 10.1073/pnas.1603531113

Fig. S5.

Fig. S5.

Analysis of hairpin-dependent pausing. (A) Structure of the nucleic acid scaffold used for pause analysis. The wild-type hisP hairpin RNA sequence is shown on the top. The starting 17-nt RNA transcript is shown in uppercase letters, and the 3′-terminal nucleotides added during transcription are shown in lowercase letters. Positions of the paused 19-nt and readthrough 21-nt transcripts are indicated. (B) Schematic of the pausing assay. The reactions were performed at 30 °C for Dra RNAP and 40 °C for Tth RNAP. (C) Effects of Tth Gfh1 on hairpin-dependent pausing by Tth RNAP. Analysis of hisP pausing was performed in the presence of the antisense RNA oligo with 10 mM MgCl2 (Upper) or MnCl2 (Lower). The paused transcripts are indicated with arrowheads. The pause t1/2 times are shown below the gels. (D) Comparison of hisP pausing by wild-type (WT; Upper) and ΔTL Dra (Lower) RNAPs in the presence of Mn2+ (the data for WT RNAP correspond to Fig. 3). Transcription with the ΔTL RNAP was performed in the presence of 1 mM NTPs, and the reaction times were from 1 to 300 min. The pause t1/2 times are shown below the gels.