Fig. 1: Rpd3L complex deacetylates Ada3 and its deacetylase activity is repressed by sucrose treatment.
a Analysis of Ada3 acetylation in the deletion mutants of Rpd3L, Rpd3S, Rpd3μ by immunoblots with indicated antibodies. b Analysis of Ada3 deacetylase activity of Rpd3 complex from WT (Wt Rpd3 comp.) and ash1Δ mutant (Rpd3 ash1Δ comp.) using purified SAGA as the substrate. P-values refer to the statistical difference of Ada3-K14ac/Ada3 and Ada3-K182ac/Ada3 between Wt Rpd3 and Rpd3 ash1Δ mutant. c Rpd3L deacetylases Ada3 at a faster rate than Rpd3S using purified SAGA as the substrate. d Loss of Ash1 enhanced the interaction between Ada3-FLAG-containing SAGA and Ada3-Myc-containing SAGA as determined by Co-IP assay. e Analysis of Ada3 deacetylase activity of Rpd3L purified from cells grown in YP + 2% glucose (Rpd3L-Glu) and YP + 2% sucrose (Rpd3L-Suc), respectively. f WT and deletion mutants of Rpd3 complex were grown in YP + 2% sucrose and treated with 0.1% H2O2 or 5 M NaCl for 2 h. The survival percentage was calculated by counting the number of colonies formed on YPD agar. For (a–c, e, f), data represent means ± SE; n = 3 biological independent experiments; one-way ANOVA with Dunnett’s multiple comparison tests (a), two-way ANOVA with Å Ãdák’s multiple comparisons tests (b, c, e), and two tailed unpaired t-tests (f) were used for statistical analysis. For (d), shown is the typical example of two biological independent experiments.
