Figure 1. Proteomic analysis of eIF5A... | Rockefeller University Press

Figure 1.

Proteomic analysis of eIF5A depletion shows downregulation of mitochondrial proteins. (A) Volcano plot showing the log2 fold change values (tif51Ats relative to wild-type) of 1,358 detected proteins plotted against their associated −log10 P values. tif51Ats stands for the average values obtained for the two eIF5A temperature-sensitive strains (tif51A-1 and tif51A-3). Dots representing individual proteins were divided in five different groups: P value >0.05 (black); P value <0.05 (grey); OXPHOS/TCA proteins (red); mitochondrial transport proteins (orange); translation proteins (blue); and drug membrane transport proteins (magenta). (B) Cumulative distributions of log2 fold changes of all proteins detected (grey), OXPHOS/TCA (red), and ribosomal proteins (blue). (C–G) Wild-type, tif51A-1, and tif51A-3 strains were cultured in YPD at 25°C until the early exponential phase and transferred to 25°C or 41°C for 4 h. eIF5A, Por1, Hsp60, Cor1, and Idh1 protein levels were determined by western blotting (C) and quantified (D and F). G6pdh levels were used as loading control. A representative image is shown from three independent experiments. (E and F)POR1, HSP60, COR1, and IDH1 mRNA relative levels were determined by RT-qPCR. Data information: In D–G, results are presented as mean ± SD from three independent experiments. The statistical significance was measured by using a two-tailed paired Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001. The asterisks located above the columns of the graph represent a comparison between 41°C and 25°C conditions within the same strain. The asterisks found at the top of the bars additionally indicate comparisons between the indicated strains at the same treatment condition. n.s means no significant differences. Source data are available for this figure: SourceData F1.

Proteomic analysis of eIF5A depletion shows downregulation of mitochondrial proteins. (A) Volcano plot showing the log₂ fold change values (tif51A^ts relative to wild-type) of 1,358 detected proteins plotted against their associated −log₁₀ P values. tif51A^ts stands for the average values obtained for the two eIF5A temperature-sensitive strains (tif51A-1 and tif51A-3). Dots representing individual proteins were divided in five different groups: P value >0.05 (black); P value <0.05 (grey); OXPHOS/TCA proteins (red); mitochondrial transport proteins (orange); translation proteins (blue); and drug membrane transport proteins (magenta). (B) Cumulative distributions of log₂ fold changes of all proteins detected (grey), OXPHOS/TCA (red), and ribosomal proteins (blue). (C–G) Wild-type, tif51A-1, and tif51A-3 strains were cultured in YPD at 25°C until the early exponential phase and transferred to 25°C or 41°C for 4 h. eIF5A, Por1, Hsp60, Cor1, and Idh1 protein levels were determined by western blotting (C) and quantified (D and F). G6pdh levels were used as loading control. A representative image is shown from three independent experiments. (E and F)POR1, HSP60, COR1, and IDH1 mRNA relative levels were determined by RT-qPCR. Data information: In D–G, results are presented as mean ± SD from three independent experiments. The statistical significance was measured by using a two-tailed paired Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001. The asterisks located above the columns of the graph represent a comparison between 41°C and 25°C conditions within the same strain. The asterisks found at the top of the bars additionally indicate comparisons between the indicated strains at the same treatment condition. n.s means no significant differences. Source data are available for this figure: SourceData F1.