Figure 6.

The Sac3 PCI Domain and Med31 Are Required for Gene-NPC Targeting

GAL1-NPC targeting assay. The GAL1 locus is tagged with TetO repeats, which are labeled with TetR-GFP, the nuclear envelope is labeled with Nup188-GFP. sac3Δ cells were transformed with plasmids carrying either SAC3 wild-type, or the sac3 R288D allele, cdk8Δ cells contained an empty plasmid. For each strain, examples are shown both in glucose and galactose (scale bar, 2 μm). After z stack acquisition, the position of the labeled GAL1 locus was determined in those cells where the brightest GAL1 signal and the largest nuclear diameter were in the same z section. The bar graph shows the proportion of GAL loci found in the peripheral volume (zone I). Comparisons of GAL1 distributions for each strain and growth condition were performed using the one-tailed Fisher’s exact test. The p value is indicated for each test and N, the total number of cells analyzed, is shown at the bottom of the bars. Results were reproduced in an independent experiment: wild-type: n = 201 (glucose) and 230 (galactose), p < 0.0001; sac3 R288D: n = 190 (glucose) and 198 (galactose), p = 0,3112; cdk8Δ: n = 252 (glucose) and 214 (galactose), p < 0,0001. For analysis of a control gene not affected by TREX-2 see Figure S5C.

See also Figure S6.