FIG. 5.

Dia2 degradation requires an NLS-containing domain. (A) Diagram of N-terminal Dia2 deletion mutants used in this study. Expression of the mutant proteins is shown in the lower panel. Samples were prepared and blotted as described for Fig. 3B. (B) Dia2 mutants lacking the NLS region are mislocalized. Immunofluorescence was performed as for Fig. 4. Scale bar, 5 μm. (C) Growth phenotypes of Dia2 N-terminal deletions. The indicated strains were spotted to YPD with the indicated amounts of hydroxyurea or MMS in 10-fold dilution series and grown at room temperature. (D) Nucleus-localized Dia2 mutants are susceptible to proteolysis. Dia2 mutants were subjected to stability assays as described for Fig. 4. Quantitation of results from three independent experiments is shown on the graph. Error bars indicate standard deviations.