Extended Data Fig. 1: Gp54Bas11 activates CapRelSJ46.
From: A bacterial immunity protein directly senses two disparate phage proteins
(a) Summary of 3 independent replicates of phage spotting assay in Fig. 1a. Asterisks indicate p = 10−10 (SECΦ27) or 10−18 (Bas11) (unpaired two-tailed t-test). (b) Summary of 3 independent replicates of phage spotting assay in Fig. 1b. Asterisks indicate p = 10−18 (WT), 10−6 (clone 1) or 10−10 (clone 3 and clone 4) (unpaired two-tailed t-test). (c) Multiple sequence alignment of the major capsid proteins from phages SECΦ27, Bas5, Bas8, Bas10 and Bas11. Residues shown to be important for interaction with CapRelSJ46 are labeled by red arrows. (d) Immunoblot of FLAG-tagged Gp54Bas11 expressed from its native promoter (wild-type or mutated versions found in escape phage clones). RpoA is included as a loading control. Image shown is a representative of 2 biological replicates. (e) Cell viability assessed by serial dilutions of cells expressing an empty vector (EV) and the indicated variant of Gp54Bas11 from an arabinose-inducible promoter on media containing glucose or arabinose. (f) Circular dichroism spectra for purified His6-Gp54Bas11 and His6-Gp54Bas11(G24D).
