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. Author manuscript; available in PMC: 2020 Nov 5.
Published in final edited form as: Structure. 2019 Sep 9;27(11):1660–1674.e5. doi: 10.1016/j.str.2019.08.010

Figure 6. CdiA-CTKp342 and CdiA-CTC3006 are specific for deacylated tRNAGAUIle.

Figure 6.

A) In vitro nuclease reactions. Total RNA isolated from E. coli was incubated with purified CdiA-CTEC3006 and analyzed by Northern blotting as described in Methods. Where indicated, reactions were supplemented with CdiIEC3006 or 5 mM MgCl2. B) Acid-urea gel analysis of nuclease reactions. C) In vitro nuclease assays using deacylated tRNA. Where indicated, purified EF-Tu and EF-Ts were included in the reactions. D) CdiA-CTKp342 requires EF-Tu, EF-Ts and GTP to support tRNase activity. Deacylated tRNA was treated with the indicated proteins and tRNAGAUIle analyzed by Northern blotting. E) Acid-urea gel analysis of tRNAGAUIle isolated from mupirocin-treated cells. F) Northern blot analysis of tRNAGAUIle isolated from competition co-cultures in the presence of mupirocin.