ASH1 mRNA
localizes to the bud tip in Saccharomyces cerevisiae to establish
asymmetry of HO expression, important for mating type switching. To
visualize real time localization of the mRNA in living yeast cells,
green fluorescent protein (GFP) was fused to the RNA-binding protein
MS2 to follow a reporter mRNA containing MS2-binding sites. Formation
and localization of a GFP particle in the bud required ASH1
3'UTR (untranslated region) sequences. The SHE mutants disrupt
RNA and particle localization and SHE 2 and 3 mutants inhibit
particle formation as well. Both She3myc and She1myc colocalized with
the particle. Video microscopy demonstrated that She1p/Myo4p moved
particles to the bud tip at 200-440 nm/sec. Therefore, the ASH1
3'UTR-dependent particle serves as a marker for RNA transport and
localization.