FIG 5.
Ras1 is repolarized in a mating-dependent manner. Mating mixtures were prepared from overnight cultures of MATα wild-type cells expressing either GFP-Ras1 (CBN96) or the palmitoylation-defective GFP-Ras1C203S, C204S (CBN167) with wild-type MATa cells (KN99a). Mixtures were spotted onto MS medium plates in 100-μl aliquots. At regular intervals, cells were scraped off the plate, resuspended in PBS, and examined for GFP-Ras localization. No fluorescence was detected in wild-type MATa cells (representative cells are marked by asterisks in the DIC image). Top panel, DIC images (magnification, ×100). Middle and bottom panels, fluorescent images (magnification, ×100). Arrowheads specify cells containing GFP-Ras1 polarized to emerging buds. Bar, 5 μm.