Extended Data Fig. 1 |. Analysis of the bactericidal activity of DddA and its activity against dsDNA and RNA substrates.

a, Genomic context of dddA (purple) and dddI_A (blue) in B. cenocepacia H111. b, Viability of B. cenocepacia ΔdddA ΔdddI_A (recipient) over time during competition with B. cenocepacia donor strains carrying wild-type dddA_tox or dddA_tox^E1347A. Values and error bars represent the mean ± s.d. of three technical replicates. The experiment was repeated three times with similar results. c, α-VSV-g western blot analysis of total cell lysates of E. coli expressing the indicated deaminases tagged with VSV-G epitope. RNAP-β was used as a loading control. Results are representative of n = 2 independent biological replicates. d, In vitro DNA cytidine deamination assays using double-stranded 36-nt DNA substrates containing AC, TC, CC, and GC with a FAM fluorophore on the forward (A) or reverse (B) strand. Deamination activity results in a cleaved product (P). Images are representative of n = 2 independent biological replicates. e, f, Poisoned primer extension assay to detect deamination of cytidine in single-stranded (e) or double-stranded (f) RNA substrates. Images are representative of n = 2 independent biological replicates. A mix of RNA substrates containing the sequences GUCG or GUUG at the indicated ratios were incubated with purified DddA_tox and reverse transcriptase. Primer extension was performed in reactions with ddGTP to terminate primer extension at cytidines. Cytidine deamination yields the 31-mer product.