Figure 1.

Uso1 and Sed5 colocalize with Grh1 in CUPS. (A) Cells coexpressing Uso1-2×GFP and Grh1-2×mCherry were grown in complete medium and imaged by fluorescence microscopy (Growth), or centrifuged and resuspended in 2% potassium acetate to induce starvation and visualized by fluorescence microscopy at 0.5, 2, and 4 h after incubation in starvation medium. (B and C) Wild-type (B) and uso1-1 (C) cultures expressing Grh1-2×GFP were grown at 25°C, then either held at 25°C or shifted for 45 min to 37°C, followed by incubation in starvation medium at 25°C or 37°C, respectively, for 2 h. Cells were observed by fluorescence microscopy just before starvation (Growth) and after 0.5 and 2 h of starvation. (D) Wild-type or grh1Δ cells expressing Uso1-GFP were cultured in growth medium or starved for 2 h, and the distribution of Uso1-GFP was monitored by fluorescence microscopy. (E) Cells expressing GFP-Sed5 and Grh1-2×mCherry were processed and analyzed as described in A. Broken lines indicate cell boundaries. Bars, 2 µm.