Figure 4.

Ama1p is required for Clb1p degradation during meiosis. Wild-type (WT), ama1Δ, or cdc16-1 mutants harboring plasmids expressing Ume3p-myc (pKC337), Pds1p-HA (pOC40), or chromosomally integrated CLB1-3HA allele were induced to enter meiosis and timepoints taken as indicated. Western blot analyses of protein extracts (see Experimental Procedures) were conducted to detect the proteins indicated (arrows, A–C). MI and MII indicate the approximate times of meiosis I (MI) and meiosis II (MII) completion in wild-type strains as determined by DAPI analysis. Tub1p serves as protein loading control. (D) Northern blot analysis from the timecourse described in C. Northern blots were probed for the genes indicated. IME2 represents the early expression class, CLB1 and SPS2 middle genes and SPS100, CTT1, YMC322C are late genes. ENO1 served as a loading control.