FIG 5.
Localization of mRNAs during meiosis. (A) Untagged (AN120), SPS4 tag (yLJ99), SSP2 tag (yJL139), LDS2 tag (yLJ112), or SPO20 tag (yLJ121) diploids containing MS2CP-GFP and the prospore membrane marker SPO2051–91-mCherry were sporulated at 30°C. Cells in mid- to late meiosis II were identified by the prospore membrane morphology and the specific mRNA localization revealed by GFP fluorescence. SPS4 and SSP2 are protected transcripts. LDS2 and SPO20 are nonprotected members of the NDT80 regulon. (B) Effect of IME2ΔC241 on SPS4 mRNA foci. Strain yJL123, carrying the SPS4-MS2 loop transcriptional fusion and homozygous for IME2ΔC241, was sporulated and examined for localization of the SPS4-MS2 mRNA. (C) Effect of rim4-F349L on SPS4 mRNA foci. Strain yJL159, carrying the SPS4-MS2 loop transcriptional fusion and homozygous for rim4-F349L, was sporulated and examined for localization of the SPS4-MS2 mRNA. For all strains, the image shown for each tagged mRNA is representative of the pattern seen in >95% of the meiosis II cells, with >20 cells scored in each strain. Scale bar, 1 μm.