Fig. 7: Tpk2-catalyzed Rpd3L phosphorylation promotes SAGA dimerization, histone acetylation and metabolic gene transcription.

a, b Mutation of Rpd3-2SA (a) and Ash1-2SA (b) reduced the interaction between Ada3-FLAG-containing SAGA and Ada3-Myc-containing SAGA as determined by Co-IP and reciprocal IP. c Analysis of H3K9ac and H3K14ac in WT, Ash1-2SA, Rpd3-2SA and Rpd3-2SA/Ash1-2SA when treated with YP + 2% glucose (Glu) or YP + 2% sucrose (Suc) for 0.5 h. d RT-qPCR analysis of SUC2 transcription in WT, Ash1-2SA, Rpd3-2SA and Rpd3-2SA/Ash1-2SA mutants when treated with YP + 2% glucose or YP + 2% sucrose for 0.5 h. e ChIP-qPCR analysis of H3K9ac/H3 and H3K14ac/H3 at SUC2 promoter regions (NUC1, NUC2, NUC3, NUC4) in WT and Rpd3-2SA mutant when treated with YP + 2% glucose (Glu) or YP + 2% sucrose (Suc) for 0.5 h. f Venn diagrams showing the role of Ada3 acetylation and Rpd3 phosphorylation in sucrose-induced gene transcription (Fold change ≥ 1.5, fold change ≤ 0.75; P ≤ 0.05). g GSEA analysis of TCA cycle gene transcription in WT, Ada3-3KR, and Rpd3-2SA mutants when cells were treated with YP + 2% sucrose for 0.5 h. The RNA-seq data for Ada3-3KR were retrieved from GEO database GSE161887. One-side hypergeometric test was used for computing P-values. h RT-qPCR analysis of TCA cycle gene transcription in WT, Ash1-2SA and Rpd3-2SA mutants when treated with YP + 2% glucose (Glu) or YP + 2% sucrose (Suc) for 0.5 h. i ChIP-qPCR analysis of H3K9ac/H3 and H3K14ac/H3 at TCA cycle genes in WT and Rpd3-2SA mutant when treated with YP + 2% glucose (YP+Glu) or YP + 2% sucrose (YP+Suc) for 0.5 h. j, k LC-MS analysis of TCA cycle metabolites in WT, Ash1-2SA and Rpd3-2SA mutants when treated with YP + 2% glucose (YP+Glu) or YP + 2% sucrose (YP+Suc) for 0.5 h. For (a–e, h–k), data represent means ± SE; n = 3 biological independent experiments; two tailed unpaired t-tests (a, b) and two-way ANOVA with Tukey’s multiple comparisons tests (c–e, h–i, k) were used for statistical analysis.