FIG. 5.

Mutants have a general defect in single-round and multiround transcription on an immobilized template. PICs were formed on an immobilized template for 40 min. As described for Fig. 2, NTPs were added to initiate transcription and the reactions were stopped after 2.5 (A) or 40 (B) min. Transcripts were assayed by primer extension (gels). Results of typical transcription reactions with wild-type, Δpgd1, and Δsin4 extracts with no activator (−), Gal4-AH (A), Gal4-VP16 (V), or Gal4-Gcn4 (G) are shown. Levels of activation (n-fold) are indicated below the gels. Data shown in the graphs are the averages from at least two experiments, with the level of mutant transcription calculated as a percentage of wild-type transcription. (C) Rounds of transcription were calculated by dividing multiround transcription by single-round transcription. The results shown are an average of two experiments.