Extended Data Fig. 4: Ada3 acetylation-dependent SAGA dimerization is enhanced when grown in sucrose or phosphate starvation conditions.
a, b, Glycerol density gradient centrifugation analysis of Wt SAGA (a) and SAGA Ada3-3KR (b) purified from cells grown in YP + 2% glucose or YP + 2% sucrose. c, Glycerol density gradient centrifugation analysis of SAGA rpd3Δ purified when cells were grown in YP + 2% glucose. d, Quantification of fractions in Extended Data Fig. 4a-c. e, In vivo co-IP assay showing phosphate starvation induces Ada3 acetylation and promotes the association of Ada3-FLAG-containing SAGA and Ada3-Myc-containing SAGA. Diploid strains containing Ada3-FLAG/Ada3-Myc or Ada3-3KR-FLAG/Ada3-3KR-Myc were grown in SC media containing phosphate (SC + Pi) or phosphate free media (SC - Pi) for 3 hrs. Ada3-FLAG was immunoprecipitated with anti-FLAG beads. Both input and IP fractions were analyzed by western blots with the indicated antibodies. f, Regulation of Ada3 acetylation and SAGA dimerization by Rpd3 under normal or phosphate starvation medium. Diploid strains containing Spt7-TAP and Spt7-Myc were either grown in SC media containing phosphate (SC + Pi) or phosphate free media (SC - Pi) for 3 hrs. Spt7-TAP was immunoprecipitated with calmodulin beads. Both input and IP fractions were analyzed by western blots with the indicated antibodies. For Extended Data Fig. 4a-c, shown is the typical example of at least two biological independent experiments. For Extended Data Fig. 4e, f, shown is the typical example of three biological independent experiments.
