Fig. 2: EF_B0059 encodes a putative type IV restriction enzyme that prevents infection by phi47.

A Sequence alignments between the pheromone responsive plasmids pTEF2 and pCF10. Genes highlighted in yellow indicate a sequence identify greater than 95%. B Capacity of phage phi47 to form plaques measured by PFU/mL on the indicated E. faecalis strains. Data represent mean ± s.d. of three technical replicates. C Relative viability of E. faecalis V19 pTEF2 cells upon transcription knock-down of the indicated pTEF2 encoded genes of interest using CRISPR interference. Relative viability was determined by comparing the growth of each strain in the presence and absence of phi47. Statistical significance determined by one-way ANOVA with multiple corrections, *p = 0.0004. Data represent mean ± s.d. of four technical replicates. D, E Outcome of phi47 infections measured by PFU/mL in E. faecalis strains V583 (D) and V19 (E) in which the genes encoding EF_B0058 or EF_B0059 on pTEF2 are deleted. Data represent mean ± s.d. of two technical replicates. F Schematic of the EF_B0058 and EF_B0059 genes. G Reverse transcription-PCR analysis of the EF_B0058 and EF_B0059 mRNA transcript in E. faecalis V19 with or without pTEF2. Primer sets used to probe the structure of the EF_B0058-EF_B0059 transcriptional operon are shown in red (EF_B0058 specific), green (intergenic) and blue (EF_B0059 specific). H Growth curves of E. coli expressing EF_B0059, the indicated active site mutants, or empty vector. Arrow indicates when protein expression was induced. Data represent mean ± s.d. of three technical replicates. I Western blot of VSV-G-tagged EF_B0059 in E. coli strains from (H), 30 min post-induction. A non-specific band from the PAGE of these samples stained with Coomassie Brilliant Blue is used as a loading control (load). Data are representative of three independent experiments. Source data are provided as a Source Data file. ND Not Detected.