FIG. 1.

Nonoverlapping regions of Bdf1 are required for phosphorylation and TFIID binding. (A) Recombinant GST-Bdf1 proteins carrying the indicated amino acids (numbers within parentheses) were expressed in bacteria and purified (top panel). Some proteolysis of the C-terminal regions of Bdf1 was observed, but intact fusion proteins are marked with asterisks. The proteins were immobilized on glutathione-agarose beads and incubated with yeast whole-cell extracts from a strain containing an HA epitope-tagged Taf7. After washing, the beads were tested for phosphorylation of the GST-Bdf1 proteins (middle panel). Binding to TFIID was assayed by immunoblotting for the HA tag on Taf7 (bottom panel). Similar TFIID binding results were obtained when the blots were probed for Taf1 and Taf5 (data not shown). (B) Deletion analysis results from panel A are summarized schematically. The bromodomains are shaded gray, while the acidic C-terminal region is shown in black.