Fig. 4: Wild-type Orf65 inactivates distantly related type IV restriction enzymes.
From: An enterococcal phage protein inhibits type IV restriction enzymes involved in antiphage defense
A Multiple sequence alignment of residues 25–42 from Orf65 homologs identified in the genomes of 9 closely related enterococcal phages. The residues are colored according to their percent similarity at that site. B Growth curves of E. coli expressing GC1503 and either Orf65 or Orf65L32F, or empty vector. Arrow indicates when protein expression was induced. Data represent mean ± s.d. of three technical replicates. C Western blot analysis of the Co-immunoprecipitation of VSV-G-tagged Orf65 or Orf65L32F with FLAG-tagged GC1503. Tagged proteins were co-expressed in E. coli for 30 min upon which cells were lysed and FLAG-tagged EF_B0059 was precipitated using anti-FLAG antibody conjugated to agarose beads. Data are representative of three independent experiments. D Outcome of coliphage infection measured by PFU/mL in E. coli strain XL-1 blue co-expressing BrxU and either Orf65 or Orf65L32F. Data represent mean ± s.d. of three technical replicates. E Outcome of Co-immunoprecipitation as in (C) but with FLAG-tagged BrxU. Source data are provided as a Source Data file.
