Figure 5.

Perturbation of Basic Amino Acid Synthesis upon UBB⁺¹ Expression

(A) Protein alterations in crude mitochondria were quantified by SILAC in two independent experiments. Changes are shown that were significant in both experiments.

(B) UBB⁺¹-triggered cytotoxicity in strains deleted from genes encoding proteins accumulating in crude mitochondria upon UBB⁺¹ expression. Clonogenicity was determined 2 days after inducing expression followed by acetate treatment. The CFUs obtained using cells carrying vector controls were set to 100% in every experiment.

(C) Basic amino acids were isolated from cultures expressing UBB⁺¹ or vector controls, respectively. The mean values of amino acids from cells carrying vector controls were set to 1.0 for every amino acid.

(D) Arginine and ornithine biosynthetic pathway in S. cerevisiae. Green: deletion of genes encoding these enzymes does significantly prevent (green) and does not prevent (red) from UBB⁺¹-triggered oxidative stress, respectively (see E and Figure S5B); Black: no data.

(E) Oxidative stress in strains with disrupted arginine/ornithine biosynthesis was measured 2 days after inducing UBB⁺¹ expression followed by acetate treatment. The oxidative stress levels obtained using cells carrying vector controls were set to 100% in every experiment.

(F) Cell death in strains with disrupted arginine/ornithine biosynthesis and increased levels of arginine in the growth media was measured 2 days after inducing UBB⁺¹ expression. The proportion of dead cells carrying vector controls was set to 100% in every experiment.

(G) Cell death in strains with disrupted lysine biosynthesis and increased levels of lysine in the growth media was measured 2 days after inducing UBB⁺¹ expression. The proportion of dead cells carrying vector controls was set to 100% in every experiment.

Data: percentage change values (B and E–G) and mean values (C), respectively. Error bars: SE. p values: ^xp < 0.1, ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001. See Tables S1, S2, and S4 and Figure S5.