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. 2011 Aug;31(16):3312–3325. doi: 10.1128/MCB.05318-11

Fig. 10.

Fig. 10.

The flap loop is not required for transcription initiation or promoter clearance in vitro. (A) Schematic representation of preinitiation complex formation and transcription initiation from the adenovirus major late promoter (ADMLp) by wild-type and ΔFL ettRNAPII. (B) Preinitiation complexes were assembled using wild-type or ΔFL ettRNAPII with a 30-min preincubation at 30°C. Transcription was initiated by the addition of NTPs for the indicated times, and RNA products were separated on a 20% denaturing polyacrylamide gel. The fractions of 4-mer and 23-mer RNAs were quantitated and plotted as a function of reaction time. (C) Preinitiation complexes were assembled using wild-type or ΔFL ettRNAPII without preincubation at 30°C. Transcription was initiated by the addition of NTPs; an aliquot was removed after the indicated time intervals; and RNA products were separated on a 20% denaturing polyacrylamide gel. The fractions of 4-mer and 23-mer RNAs were quantitated and plotted as a function of reaction time. (D and E) Ratio of the 23-mer to the 4-mer synthesized with a 30-min preincubation (D) or with no preincubation (E).