FIGURE 6.

The ulp1-CSDN mutant inhibits INO1 targeting to the NE. (A) Shown are images of WT cells producing exogenously expressed, plasmid-encoded Ulp1-GFP or Ulp1^CSDN-GFP. (B) The localization of plasmid-encoded Ulp1-GFP or Ulp1^CSDN-GFP in cells producing mCherry-tagged endogenous wild-type Ulp1 (Ulp1-mCherry) was examined. Shown are cells containing relatively high or low levels of the GFP fusion, likely stemming from cell to cell variability in plasmid copy number. Competition of NE-associated binding between the plasmid-encoded Ulp1-GFP or Ulp1^CSDN-GFP and endogenous Ulp1-mCherry proteins is indicated by the relative GFP and mCherry signal intensities at the NE. Scale bars = 2 μm. (C) Recruitment of INO1 to the NPCs was measured as in Figure 2 in a WT strain containing INO1-lacO₂₅₆/GFP-lacI and the indicated plasmid. Results are the means ± SD of three or more biological replicates. At least 50 cells were counted for each sample. (D) Levels of mRNA encoded by the INO1 gene were evaluated by qRT-PCR as described in Figure 2 following induction for the specified times in the indicated strains. Asterisks indicate a significant difference from WT (pEMPTY) as indicated by a Student’s unpaired t-test, *p < 0.05, **p < 0.01.