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. 2017 Aug 18;6:e29397. doi: 10.7554/eLife.29397

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© 2017, Vassallo et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 3. — (A) SitA3-CTD is a toxic tRNase. Expression of the indicated CTDs was induced with arabinose in E. coli, and cell growth monitored by measuring the optical density of the cultures at 600 nm (OD₆₀₀). OD₆₀₀ values are reported as the average ± standard error for three independent experiments (left). RNA was isolated after 90 min of toxin expression and analyzed by Northern blot hybridization using probes to the indicated tRNAs (right). The arrow indicates cleaved tRNA_UGC^Ala. (B) SitA1-CTD has DNase activity. E. coli cells were stained with DAPI at 0 hr and after 6 hr of toxin expression. Cells expressing sitA1-CTD became filamentous and lost DAPI staining at 6 hr (left). In contrast, sitA3-CTD expressing cells retained DAPI staining (right), though their nucleoids became compacted (yellow arrow). Bar, 5 μm.

DOI: http://dx.doi.org/10.7554/eLife.29397.008

Figure 3—figure supplement 1. — (A) SitA3-CTD is a toxic tRNase. Expression of the indicated CTDs was induced with arabinose in E. coli, and cell growth monitored by measuring the optical density of the cultures at 600 nm (OD₆₀₀). OD₆₀₀ values are reported as the average ± standard error for three independent experiments (left). RNA was isolated after 90 min of toxin expression and analyzed by Northern blot hybridization using probes to the indicated tRNAs (right). The arrow indicates cleaved tRNA_UGC^Ala. (B) SitA1-CTD has DNase activity. E. coli cells were stained with DAPI at 0 hr and after 6 hr of toxin expression. Cells expressing sitA1-CTD became filamentous and lost DAPI staining at 6 hr (left). In contrast, sitA3-CTD expressing cells retained DAPI staining (right), though their nucleoids became compacted (yellow arrow). Bar, 5 μm.

DOI: http://dx.doi.org/10.7554/eLife.29397.008