FIG 4.

Gcn5p- and Rpd3p-dependent regulation of lysine triplet 3 acetylation controls Ume6p DNA binding and repression activity. (A) Ume6p occupancy at the SPO13 promoter was monitored by ChIP in mid-log-phase cultures expressing Ume6p, Ume6p^K3R, or Ume6p^K3Q. (B) Same as panel A except in host strains with the indicated genotypes. The anti-acetylated Lys (Ac-Lys) antibodies detect acetylated proteins associated with the SPO13 promoter but not acetylated Ume6p (15). Mean values from three independent biological replicates are presented (±standard errors of the means) relative to the values for the NUP85 internal control. (C and D) SPO13 mRNA levels from mid-log-phase cultures expressing the indicated UME6 allele in either the rpd3Δ (C) or the gcn5Δ (D) background were monitored by RT-qPCR. The different UME6 alleles examined are given below the graph. (E) SPO13 mRNA abundance measured as described above for panel C except that the cultures expressing the indicated UME6 alleles were grown to mid-log phase in acetate medium. Mean values from three independent biological replicates are presented (±standard errors of the means) relative to the values for the NUP85 internal control. In all panels, the asterisks indicate a P value of <0.05 (Student's t test) for the difference from the wild type. In panel B, # indicates that the two values indicated by the line are significantly different (P < 0.05).