Figure 9.

Cdc12–GFP is localized abnormally in Yck-deficient cells. (A) Wild-type cells (strain LRB758) carrying pTD150-CDC12 (pRS316:CDC12: GFP) were grown to log phase in synthetic medium at 37°C. (B) yck^ts cells (strain LRB756) carrying pTD150-CDC12 were grown to log phase in synthetic medium at 24°C, and then a portion of the culture was shifted to 30°C for 3 h. Note the presence of thick neck rings in approximately half of the cells and the lack of clear separation of bright rings in large-budded cells. Cells with abnormal septin fluorescence (most morphologically normal) are marked by arrowheads. (C) The cells shown are from the same original culture as those shown in B, but the cells in C were shifted from 24° to 37°C for 3 h before observation. Note the lack of normal neck structures in both severely misshapen and relatively normal-appearing cells. Arrowheads mark abnormal septin structures. In all cases, cells were placed on agar pads for observation and photography by DIC and fluorescence optics. Bar, 2 μm.