Fig. 4.

Involvement of H3K4me3 in chromatin transcription. (A) Diagram of PHO5 gene and promoter chromatin in the transcriptionally repressed and activated states (12), with labeling as in Fig. 1A. (B) Repressed (pho4Δ) or activated (pho80Δ) PHO5 chromatin was transcribed as in Fig. 1C with transcription proteins at 1× concentration, except for the omission of SAGA, SWI/SNF, or all transcription proteins as indicated. Activity is presented as in Fig. 1C (n = 7). (C) Effect of sgf29Δ and spt3Δ SAGA on chromatin transcription. Repressed (pho4Δ) or activated (pho80Δ) PHO5 chromatin was transcribed as in Fig. 1C with transcription proteins at 1× concentration, except that sgf29Δ or spt3Δ SAGA was substituted for WT SAGA where indicated. Transcription levels were normalized to the value obtained from the reaction with WT SAGA (percentage). (D) Effect of histone H3 peptides on chromatin transcription. Repressed (pho4Δ) or activated (pho80Δ) PHO5 chromatin was transcribed as in Fig. 1C with transcription proteins at 1× concentration with the addition of histone H3 peptides (1–10) with or without methylation at K4 as indicated (40 μM final concentration). Transcription levels were normalized to the value obtained from the reaction without H3 peptide (percentage).