FIG. 4.

Promoter binding experiments. (A) Whole cell extracts containing (HA)₃-tagged wild-type TAF130 or TAF130-Δ16 were incubated with paramagnetic bears containing a 500-bp fragment containing a derivative of the HIS4 promoter with one Gal4 binding site (P), the same HIS4 promoter fragment with several mutations at the TATA element (−T), a 280-bp promoterless fragment from the pBluescript vector present in the two previous constructs that contains the Gal4 binding site (L), and the beads without DNA (lane B). The left panel shows the indicated proteins bound to the DNA, whereas the right panel shows the analysis of 10% of the first supernatant after incubation. (B) Effect of the Gal4-VP16 activator on TFIID binding. The promoterless template containing the Gal4 binding site was incubated with extracts containing (HA)₃-tagged wild-type TAF130 or TAF130-Δ16 in the absence (−) or presence (+) of Gal4-VP16.