Fig. 3: Subtelomere expression mediates glucose starvation tolerance.
a 107 cells/mL were serially diluted (1:6), spotted on YP plates with the indicated carbon sources and grown for 2d. b Mass spectrometry analysis of redox metabolites in cells of the indicated genotypes cultured in low glucose. Metabolite levels were normalized by the median of all detected metabolites. Significances were calculated by Kruskal–Wallis rank sum test (pNADH/NAD+ = 0.019, pNAC = 0.0011, pGSSG = 0.0011), followed by Mann–Whitney test (two-sided) with Benjamini–Hochberg correction (pNADH/NAD+_sml1Δrad53Δ_vs_sml1Δ = 0.041, pNADH/NAD+_sml1Δrad53Δsir2Δ_vs_sml1Δrad53Δ = 0.041, pNAC_sml1Δrad53Δ_vs_sml1Δ = 0.0021, pNAC_sml1Δrad53Δsir2Δ_vs_sml1Δrad53Δ = 0.0021, pGSSG_sml1Δrad53Δ_vs_sml1Δ = 0.0043, p GSSG_sml1Δrad53Δsir2Δ_vs_sml1Δrad53Δ = 0.026). n = 6 independent culture replicates. c Cells were inoculated in YP + low glucose with or without supply of 1 mM N-acetylcysteine, and division speed was determined by cell counting over 2 days. The right panel indicates the reduction of division time by N-acetylcysteine. Significances were calculated with one-way ANOVA (p = 2.4 × 10−6) with post hoc Tukey HSD test (psml1Δrad53Δ_vs_sml1Δ = 0.0000052, psml1Δrad53Δsir2Δ_vs_sml1Δrad53Δ = 0.0014). n = 3 independent experiments. d, e Jitter plots representing the distance of labeled subtelomeres 5 R and 14 R from the nuclear periphery. d Cells were cultured in normal glucose. Data from one parental sml1Δ control and two independent sml1Δrad53Δ clones (#1, #2) are shown. e sml1Δ controls were cultured in the indicated carbon sources. Significances were calculated by Kruskal–Wallis rank sum test (pD_5R = 8.5 × 10−15, pD_14R = 0.00039), followed by Mann–Whitney test (two-sided) with Benjamini–Hochberg correction (pD_5R_sml1Δrad53Δ#1_vs_sml1Δ = 3.1 × 10−13, pD_5R_sml1Δrad53Δ#2_vs_sml1Δ = 6.8 × 10−10, pD_14R_sml1Δrad53Δ#1_vs_sml1Δ = 0.27, pD_5R_sml1Δrad53Δ#2_vs_sml1Δ = 8.9 × 10−5). n cells from X independent experiments (n/X): subtelomere 5R D_sml1Δ = 301/3, D_sml1Δrad53Δ#1 = 181/2, D_sml1Δrad53Δ#2 = 353/3, EtOH_sml1Δ = 419/3, subtelomere 14R D_sml1Δ = 368/2, D_sml1Δrad53Δ#1 = 209/2, D_sml1Δrad53Δ#2 = 104/1, EtOH_sml1Δ = 415/2. Bars plots with error bars represent mean values and standard deviation, D glucose; DR glucose restriction (0.01% in solid media, 0.04% in liquid media), EtOH ethanol; NAC N-acetylcysteine; GSSG oxidized glutathione. *p < 0.05, **p < 0.01, ***p < 0.001.
