Figure 1.
Isolation of constitutively degrading Ypq1 mutants. (A) Left: Western blot showing degradation of Ypq1-GFP before (0 h) or after (6 h) Lys starvation. Vec, Empty vector. Pgk1, loading control. Right: Protein levels were quantified as FL Ypq1-GFP/(FL Ypq1-GFP + free GFP). The error bars represent SD (n = 3). FL, full-length. (B) Subcellular localization of Ypq1-GFP before and after Lys starvation. Scale bar, 2 µm. (C) Predicted structure of Ypq1 showing PQ motifs and charged residues within the translocation tunnel. In top and bottom views, loops have been removed for clarity. (D) General architecture of eukaryotic PQ-loop family members. (E) Top: Degradation of constitutive Ypq1 mutants under different expression levels of Ssh4: endogenous (WT SSH4), overexpression (SSH4 OE), and deletion (ssh4Δ). Bottom: Quantification (±SD, n = 3). (F) Subcellular localization of Ypq1-GFP mutants. Scale bar, 2 µm.

Isolation of constitutively degrading Ypq1 mutants. (A) Left: Western blot showing degradation of Ypq1-GFP before (0 h) or after (6 h) Lys starvation. Vec, Empty vector. Pgk1, loading control. Right: Protein levels were quantified as FL Ypq1-GFP/(FL Ypq1-GFP + free GFP). The error bars represent SD (n = 3). FL, full-length. (B) Subcellular localization of Ypq1-GFP before and after Lys starvation. Scale bar, 2 µm. (C) Predicted structure of Ypq1 showing PQ motifs and charged residues within the translocation tunnel. In top and bottom views, loops have been removed for clarity. (D) General architecture of eukaryotic PQ-loop family members. (E) Top: Degradation of constitutive Ypq1 mutants under different expression levels of Ssh4: endogenous (WT SSH4), overexpression (SSH4 OE), and deletion (ssh4Δ). Bottom: Quantification (±SD, n = 3). (F) Subcellular localization of Ypq1-GFP mutants. Scale bar, 2 µm.

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