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Figure 4 | Genome Biology

Figure 4

From: Application of the comprehensive set of heterozygous yeast deletion mutants to elucidate the molecular basis of cellular chromium toxicity

Figure 4

Chromium causes errors in mRNA translation. (a) Cells transformed with the dual-luciferase plasmid [50] were exposed or not to 200 μg ml-1 paromomycin ('Paro'), 0.6 mM Cu(NO3)2 or 0.1 mM CrO3, in YNB medium for 90 minutes. The activities of the firefly and renilla luciferases in derived protein extracts were determined luminometrically. The ratio of luminescence from the firefly versus renilla luciferase indicates the short-term level of translational read-through of the UAA stop codon that separates the two open reading frames. All values are means ± standard error of the mean from at least three independent determinations. RLU, relative light units. (b, c) Exponential-phase S. cerevisiae L1494 (ade1-14) cells (OD600 ~1.0, plus a 10-fold dilution) were spotted in 6 μl aliquots on to YEPD agar supplemented or not with 150 μg ml-1 paromomycin, 8 mM Cu(NO3)2, 0.15 mM CrO3 or 3.6 mM H2O2. Plates were incubated for 3 days at 30°C either aerobically (b) or anaerobically (c). In the latter case, plates were incubated aerobically at 4°C after the 3 days incubation to allow development of the red pigment before images were captured. The stressors were supplied at doses that produced similar degrees of mild inhibition of aerobic growth (versus controls) within each experiment on the different media.

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