FIG. 3.
Kap95 is important for efficient RPA import, whereas Msn5 is not essential. (A) RFA1-GFP was expressed in wild-type, msn5 deletion (msn5Δ), and kap95 temperature-sensitive cells (rsl1-1 and rsl1-4 alleles of kap95ts) and grown at 24°C. Cells were retained at 24°C or shifted to 37°C for 2 h and observed for localization of Rfa1-GFP. Wild-type and msn5Δ cells contain Rfa1-GFP predominantly in the nucleus at both temperatures. kap95ts cells exhibit an increased level of cytoplasmic fluorescence at both temperatures and nearly equivalent nuclear and cytoplasmic fluorescence in the rsl1-4/kap95ts allele at 37°C. (B) The same experiment was performed with Rfa3-GFP expressed in wild-type, msn5Δ, and kap95ts cells. Rfa3-GFP remained apparent in the nucleus in all three strains, with increased cytoplasmic fluorescence in the kap95ts mutant.