Fig. 1. Mitotic Lte1 localisation is actin independent but requires activation of Cdc28/Cln kinase. (A) cln1cln2cln3 GAL1-CLN3 strain (SJ123) was arrested in G1 and released into YEPGalactose to induce expression of Cln3 at 30°C. Aliquots were removed at indicated times for analysis by real time microscopy, determination of budding index and cell cycle progression by DAPI stain. () Budding index; () cortical Lte1; () cells with pre-anaphase nuclear morphology; () cells with anaphase nuclear morphology; (•) cells with completely separated DNA masses (telophase). (B) Cells described in (A) were arrested in G1 (a) and released into the cell cycle in the presence of LatB (b) or DMSO as control (c). Localisation of Lte1GFP monitored after 2 hours. The percentage of cells with cortical Lte1 is indicated below. (C) Wild-type cells (SY125) were arrested in G1 with -factor (a). Shmoos were treated with LatB for 15 minutes and cells examined by microscopy (b-c). Numbers indicate the percentage of cells that localised Lte1GFP to the tip of the mating projection. (D) Localisation of Lte1GFP in cdc15-1 mutant cells expressing CFP-tubulin (SY126) at the restrictive temperature (a), following 1 hour nocodazole treatment at 37°C (b) and 1 hour after release at permissive temperature (c). Numbers indicate the percentage of cells with cortical Lte1. Bar, 10 µm.

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