Fig. 1: AdfA^T4 is not the primary anti-DarTG1 counter-defense element for T4.

a Representative plaque assay of RB69 phage on wild-type MG1655 or a strain with a chromosomally inserted darTG1 system under its native promoter, in the presence or absence of adfA counter-defense elements from wild-type RB69 (adfA^RB69), an evolved resistant variant (adfA^RB69(R164H)), or T4 (adfA^T4). Strains were uninduced, relying on leaky expression only, or induced with vanillate. EV, empty vector. See Fig. S1 for an amino acid alignment of these proteins. b Efficiency of plaquing (EOP) measurements corresponding to (a) were calculated by normalizing the titer of each strain to the MG1655 control. *p = 0.03; unpaired, two tailed t test. n = three independent replicates, presented with error bars representing standard deviation. c Bacterial 2-hybrid assay demonstrating interactions between the three adfA alleles fused to the T18 fragment of adenylate cyclase and a catalytically inactive darT1(E157A) variant (darT1*) fused to the T25 fragment. Dark blue color indicates an association between the proteins; white means they do not associate. Representative image of three independent experiments is presented. d Heat map depicting EOP for strains with DarTG1 compared to empty vector controls for the indicated Tevenvirinae family phages. The average of three independent replicates is presented. Circles indicate phage genomes that carry adfA genes. See Fig. S2 for plaque assay image and bar graphs representing individual replicates with variance. Source data are provided as a Source Data file.