Figure 3.

Effects of C-terminal truncations on Gln3-Myc¹³ localization in wild type and sit4Δ cells. (A) Diagram depicting the C-terminus of each Gln3-Myc¹³ C-terminal truncation. (B–E) Wild type (JK9-3da) and sit4Δ (FV029) cells were transformed with plasmids encoding full length, wild type Gln3_1-730-Myc¹³ (pRR536) or Gln3_1-476-Myc¹³ (pRR610). Cells were grown to mid-log phase in YNB-glutamine (Gln), or -proline (Pro) medium. Rapamycin (200 ng/ml) was added to glutamine-grown cells where indicated (+Rap) for 15 minutes. (B and D) Illustrative images of Gln3-Myc¹³ localization under the growth conditions scored in the historgrams. Gln3-Myc¹³ localization was scored as cytoplasmic (red bars), nuclear-cytoplasmic (yellow bars; fluorescent material appearing in both the cytoplasm and colocalizing with DAPI-positive material, DNA) or nuclear (green bars; fluorescent material colocalizing only with DAPI-positive material). (C and E) Histograms depicting the percentage of Gln3 localized to each cellular compartment. Experimental format and scoring criteria were as described in the section “Materials and Methods.” Histogram values represent the averages derived from two biological replicates; error bars indicate standard deviations.