FIG. 3.

Effects of HATs on GAL4 derivative-mediated transcription of the nucleosome array templates. (A) Diagram of the transcription protocol. Subsequent to the factor-binding reaction, 1 U of NuA4 or SAGA, in the presence or the absence of acetyl-CoA, was included and incubated at 30°C for 30 min. After the incubation, transcription reactions were started by adding HeLa cell nuclear extract and nucleotides. The same amounts of various GAL4-derivative proteins were used as for Fig. 2A. NTPs, nucleoside triphosphates; RT, room temperature. (B) HAT assays of the G5E4-5S nucleosome array template. Nucleosome templates were incubated in a binding reaction mixture with NuA4 or SAGA in the presence (lane 2) or the absence (lane 1) of ³H-labeled acetyl-CoA. After incubation, SDS-sample buffer and 3 μg of core histone were added, and the mixture was loaded onto a gel. Lanes 3 and 4 show Coomassie staining of the same gel, indicating the migratory positions of the four core histones. (C) Transcripts from the G5E4-5S array template, which were incubated with SAGA, in the presence or the absence of acetyl-CoA, without (−) or with various GAL4 derivative proteins. (D) Transcripts from the G5E4-5S array template which were incubated with NuA4 in the presence or the absence of acetyl-CoA without (−) or with various GAL4-derivative proteins.