FIG. 7.

Δsin4, but not Δpgd1, nuclear extracts (NEs) can form stable Scaffold complexes. PICs (without NTPs) were formed for 40 min in the presence of the activator Gal4-VP16 and washed. To form Scaffold complexes, washed PICs were resuspended in transcription mix and initiated with NTPs (+) for 3.0 min. Washed promoter-bound Scaffold complexes were either immediately digested from the beads (0 min) or resuspended in transcription mix plus competitor DNA for 40 min before being washed and digested (40 min). For more accurate quantitation, 2× Δpgd1 and 4× Δsin4 reactions were performed. The graph compares the retention of Scaffold components in mutants to their retention in wild-type extracts at time 0. The calculations are based on at least five experiments.