Figure 3. The Effect of the she Mutants on Particle Localization and Formation

Yeast strains disrupted for each one of the five SHE genes were transformed with the ASH1 reporter RNA and the GFP-MS2 fusion protein and the resultant particles observed by epifluorescence after fixation. Bar, 5 µm. (A) Wild-type cells (K699). Localization of the particle and its formation is inhibited in (B) she5 deletion strain (K5205): 36% of cells with signal formed bright, single particles; approximately half of the particles were localized at the bud neck and 2% were localized in the bud. (C) she3 deletion strain (K5235): 6% formed bright, single particles and 0% were localized in the bud. (D) she1 deletion strain (K5209): 16% of cells with signal formed bright, single particles and 0% were localized in the bud. (E) she2 deletion strain (K5547): 0% formed bright, single particles and 0% were localized in the bud. (F) she4 deletion strain (K5560): 32% formed bright, single particles and 16% with signal were localized in the bud. Colocalization (yellow) of a functional myc-tagged she protein (red) with the particle (green). (G) She1myc. (H) She2myc. (I) She3myc. (J-I) She1myc with nonlocalized particle in nonbudding cell. (Half of the particles showed colocalization with She1myc.)