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Figure 5.
Pex30 participates in the regulation of the intracellular distribution of PA. (A) Diagram highlighting the main steps of PA metabolism in yeast. Individual and multiple reactions are indicated by solid and dashed lines, respectively. The enzymes Cds1, Pah1, and the Pah1 activator complex Nem1/Spo7 are indicated. (B) Schematic representation of Spo2051–91-GFP-ER, an ER-localized PA biosensor. The well-characterized PA sensor Spo2051–91-GFP was targeted to the ER by fusing it to the transmembrane domain of Ubc6, an ER-resident protein. (C) Localization of Spo2051–91-GFP-ER in cells with the indicated genotype. Cells were analyzed during the diauxic shift after overnight growth in SC medium. Individual Z-planes corresponding to the center and the periphery of the cell are shown. Where indicated, CDS1 was expressed from the strong constitutive GPD1 promoter. Bar, 5 µm. (D) Quantification of Spo2051–91-GFP-ER foci in cells with the indicated genotype grown as in C. The bars represent the mean and SD. Ordinary one-way ANOVA and Dunnett’s multiple comparisons were performed to compare the percentage of cells with foci with the WT condition (****, P < 0.0001; **, P < 0.01; ns, not significant). (E) Localization of Spo2051–91-GFP-ER in cells expressing the indicated Pex30 mutants. Cells were analyzed during the diauxic shift after overnight growth in SC medium. Individual Z-planes corresponding to the center and the periphery of the cell are shown. Bar, 5 µm. (F) Quantification of Spo2051–91-GFP-ER foci in cells with the indicated genotype grown as in E. The bars represent the mean and SD. Ordinary one-way ANOVA and Dunnett’s multiple comparisons were performed to compare the percentage of cells with foci with the WT condition (***, P < 0.001; ****, P < 0.0001). (G) Quantification of Spo2051–91-GFP-ER foci in cells with mutations in Pex30 family members grown as in E. The bars represent the mean and SD. Ordinary one-way ANOVA and Dunnett’s multiple comparisons were performed to compare the percentage of cells with foci with the WT condition (****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05). (H) Quantification of Spo2051-91-GFP-ER foci in cells with the indicated genotype grown as in E. The bars represent the mean and SD. Ordinary one-way ANOVA and Dunnett’s multiple comparisons were performed to compare the percentage of cells with foci with the WT condition (****, P < 0.0001; ***, P < 0.001; **, P < 0.01; ns, not significant).

Pex30 participates in the regulation of the intracellular distribution of PA. (A) Diagram highlighting the main steps of PA metabolism in yeast. Individual and multiple reactions are indicated by solid and dashed lines, respectively. The enzymes Cds1, Pah1, and the Pah1 activator complex Nem1/Spo7 are indicated. (B) Schematic representation of Spo2051–91-GFP-ER, an ER-localized PA biosensor. The well-characterized PA sensor Spo2051–91-GFP was targeted to the ER by fusing it to the transmembrane domain of Ubc6, an ER-resident protein. (C) Localization of Spo2051–91-GFP-ER in cells with the indicated genotype. Cells were analyzed during the diauxic shift after overnight growth in SC medium. Individual Z-planes corresponding to the center and the periphery of the cell are shown. Where indicated, CDS1 was expressed from the strong constitutive GPD1 promoter. Bar, 5 µm. (D) Quantification of Spo2051–91-GFP-ER foci in cells with the indicated genotype grown as in C. The bars represent the mean and SD. Ordinary one-way ANOVA and Dunnett’s multiple comparisons were performed to compare the percentage of cells with foci with the WT condition (****, P < 0.0001; **, P < 0.01; ns, not significant). (E) Localization of Spo2051–91-GFP-ER in cells expressing the indicated Pex30 mutants. Cells were analyzed during the diauxic shift after overnight growth in SC medium. Individual Z-planes corresponding to the center and the periphery of the cell are shown. Bar, 5 µm. (F) Quantification of Spo2051–91-GFP-ER foci in cells with the indicated genotype grown as in E. The bars represent the mean and SD. Ordinary one-way ANOVA and Dunnett’s multiple comparisons were performed to compare the percentage of cells with foci with the WT condition (***, P < 0.001; ****, P < 0.0001). (G) Quantification of Spo2051–91-GFP-ER foci in cells with mutations in Pex30 family members grown as in E. The bars represent the mean and SD. Ordinary one-way ANOVA and Dunnett’s multiple comparisons were performed to compare the percentage of cells with foci with the WT condition (****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05). (H) Quantification of Spo2051-91-GFP-ER foci in cells with the indicated genotype grown as in E. The bars represent the mean and SD. Ordinary one-way ANOVA and Dunnett’s multiple comparisons were performed to compare the percentage of cells with foci with the WT condition (****, P < 0.0001; ***, P < 0.001; **, P < 0.01; ns, not significant).

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