Journal: Structure (London, England : 1993)

Article Title: Convergent evolution of the barnase/EndoU/colicin/RelE (BECR) fold in antibacterial tRNase toxins

doi: 10.1016/j.str.2019.08.010

Figure Lengend Snippet: Structural homologs of CdiA-CT•CdiI Kp342 and CdiA-CT•CdiI EC3006 , See also Figure S5 .

Article Snippet: E. coli K-12 sub-strain EPI100 , Epicentre/Lucigen , Cat# EC10010.

Techniques:

Journal: Structure (London, England : 1993)

Article Title: Convergent evolution of the barnase/EndoU/colicin/RelE (BECR) fold in antibacterial tRNase toxins

doi: 10.1016/j.str.2019.08.010

Figure Lengend Snippet: A) Toxin activation inside E. coli cells leads to tRNAGAUIle cleavage. Toxin expression was induced L-arabinose where indicated and RNA isolated for Northern blot hybridization using probes to the indicated tRNAs. Aminoacyl acceptor stem sequences and toxin cleavage sites are indicated on the right. B) tRNAGAUIle sequence showing the hybridized S1 probe and oligonucleotide standards used to map toxin cleavage sites. C) S1 protection analysis of tRNAGAUIle cleaved by CdiA-CTKp342. RNA samples were isolated from i) cells intoxicated by intracellular CdiA-CTKp342 expression, ii) competition co-cultures and iii) in vitro nuclease reactions. Where indicated, the neutralizing effect of CdiIKp342 was examined. Samples were hybridized with 3′-radiolabeled S1 probe and treated with S1 nuclease as described in Methods. A portion of the S1 probe-tRNAGAUIle heteroduplex sequence is shown to the right of the autoradiogram. D) S1 protection analysis of tRNAGAUIle cleaved by CdiA-CTEC3006. Samples were analyzed as described for panel C.

Article Snippet: E. coli K-12 sub-strain EPI100 , Epicentre/Lucigen , Cat# EC10010.

Techniques: Activation Assay, Expressing, Isolation, Northern Blot, Hybridization, Sequencing, In Vitro

Journal: Structure (London, England : 1993)

Article Title: Convergent evolution of the barnase/EndoU/colicin/RelE (BECR) fold in antibacterial tRNase toxins

doi: 10.1016/j.str.2019.08.010

Figure Lengend Snippet: 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.

Article Snippet: E. coli K-12 sub-strain EPI100 , Epicentre/Lucigen , Cat# EC10010.

Techniques: In Vitro, Isolation, Incubation, Purification, Northern Blot, Activity Assay

Journal: Structure (London, England : 1993)

Article Title: Convergent evolution of the barnase/EndoU/colicin/RelE (BECR) fold in antibacterial tRNase toxins

doi: 10.1016/j.str.2019.08.010

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: E. coli K-12 sub-strain EPI100 , Epicentre/Lucigen , Cat# EC10010.

Techniques: Virus, Recombinant, Software