FIG 6.

Cluster 3 acetylation is required for timely meiotic gene induction and meiotic progression. (A) Ume6p, Ume6p^K3R, or Ume6p^K3Q levels were determined by Western blotting of samples taken before and following the shift to SPM (hours). Tub1p served as a loading control. (B and C) URS1^SPO13 occupancy by either Ume6p (B) or Ume6p^K3R (C) was determined by ChIP. The presence of acetylated lysine proteins at the SPO13 promoter was determined by ChIP using pan-anti-acetylated Lys antibodies. Mean values from three independent biological replicates are presented (±standard errors of the means) relative to the values for the NUP85 internal control. (D to G) Meiotic SPO13 (D), SPS4 (E), SPS100 (F), and IME1 (G) mRNA levels were determined by RT-qPCR in Ume6p- or Ume6p^K3R-expressing cultures, as indicated. Mean values from three independent biological replicates are presented (±standard errors of the means) relative to the values for the NUP85 internal control. Asterisks indicate statistical differences at a given time point using the Student t test (P < 0.05). (H) The percentages of Ume6p-, Ume6p^K3R-, or Ume6p^K3Q-expressing cultures exhibiting bi- and tetranucleated cells were determined by DAPI analysis. Mean values from three independent biological replicates are given (±standard errors of the means). Asterisks indicate statistically significant differences relative to the wild type (P < 0.05).