Extended Data Fig. 3: Identification of conserved motifs and distribution of WonAB.
From: West African–South American pandemic Vibrio cholerae encodes multiple distinct phage defence systems
(a) Sequence logos showing conservation of the identified ABC-ATPase motifs in the 198 WonA hits detected using MacSyFinder v.2.1.1, as compared to the equivalent residue number in V. cholerae A1552 WonA. Amino acids in logos are coloured according to chemical properties: polar (G, S, T, Y, C), green; neutral (Q, N), purple; basic (K, R, H), blue; acidic (D, E), red; and hydrophobic (A, V, L, I, P, W, F, M), black. (b) Location of the conserved ABC-ATPase motifs shown in (a) at the dimer interface of the WonA dimer structural prediction. (c) Sequence logo showing the conservation of the identified PD-(D/E)xK nuclease motif in the 198 WonB hits detected using MacSyFinder v.2.1.1, compared to the equivalent residue number in V. cholerae A1552 WonB. (d) Location of the identified PD-(D/E)xK nuclease residues in the WonB structural prediction. (e) Western blot showing the protein levels of WonA and WonB, natively expressed in strain A1552 (WT) and in derivatives encoding the indicated site-directed variants. Blots are representative of the results of three independent experiments. (f) Distribution of WonAB hits detected using MacSyFinder v.2.1.1. The tree shows the order-level phylogeny of genera in the RefSeq database with more than 500 genomes (see methods). For the full list of 198 WonAB hits see Supplementary Table 4. (g) Anti-phage activity of WonAB in E. coli. Fold-protection against E. coli phages of the BASEL collection conferred by the production of WonAB in E. coli MG1655ΔaraCBAD, as compared to a negative ‘no system’ control. The system was expressed from a chromosomally integrated transposon carrying the arabinose-inducible PBAD- promoter, induced by the addition of 0.2% arabinose. Bar chart shows the mean of two independent experiments.
