Extended Data Fig. 2: T5 phage tail assembly protein triggers pSlfn5-mediated phage defense.
a, T5 phage titer in supernatants of cells with no defense, RorSlfn5, or catalytic mutant of RorSlfn5 (RorSlfn5dead) was quantified at 50 min post infection with T5 phage at MOI = 10. Phage titers were normalized to the mean of the no-defense control. Data is shown as the mean of three biological replicates ± S.D. One-way ANOVA with post hoc Tukey HSD test was used to compare the experimental groups. The resulting p values are shown in the plot. b, Antiviral activities of SfoSlfn5 ortholog against a panel of phages. EOP – efficiency of plating. Data is shown as the mean of three biological replicates ± S.D. White dots show the average of three technical replicates for each biological replicate. c-g, Cellular toxicity assay in E. coli K-12 MG1655 cells co-transformed with a plasmid expressing RorSlfn5 (c), PagSlfn5 (d), or SfoSlfn5 (e) and a plasmid for arabinose-inducible expression of T5.142 homologs (T5.142 – T5 phage, S114 – Salmonella phage S114, DU_PP_V - Pectobacterium phage DU_PP_V, My1 - Pectobacterium phage My1, KKP_3711 - Enterobacter phage KKP_3711, vB_PagS_AAS21 - Pantoea phage vB_PagS_AAS21). Assay was performed in three biological replicates, with one representative replicate shown in the figure. Asterisk (*) indicates toxicity phenotypes. pBAD-GFP was used as a positive control for inducible protein expression. f-g, Cellular toxicity assay in MG1655 transformed only with T5.142 homologs (f) or co-transformed with a plasmid expressing RorSlfn5E15A,D20A and T5.142 homologs (g).
