Extended Data Fig. 7: Expression of RIIB proteins activate gasdermin-mediated defense.
From: CARD domains mediate anti-phage defence in bacterial gasdermin systems
a Bacteria expressing RIIB homologs together with a negative control or the Lysobacter gasdermin system were plated in 10-fold serial dilution on LB-agar plates in conditions that repress expression (1% glucose) or induce expression (0.2% arabinose and 0.1 mM IPTG). b. Transformation efficiency assays of plasmids encoding rIIB or RFP into cells that contain the Lysobacter or Pedobacter systems on a pBAD plasmid. Bars represent the average of three biological replicates with individual data points overlaid. c. Phylogenetic tree of phage RIIB homologs (Supplementary Table 4). Names of phages from which RIIB homologs were experimentally tested are indicated. d. Bacteria co-expressing RIIB homologs together with the Lysobacter gasdermin system or a negative control were plated in 10-fold serial dilution on LB-agar plates in conditions that repress expression (1% glucose) or induce expression (0.2% arabinose and 1 mM IPTG). Numbering of RIIB homologs corresponds to the numbering on the tree in panel c. e. Western blot analyses of N-terminally HA-tagged gasdermin co-expressed with RIIB homologs, before (0 min) and 80 min after RIIB expression was induced. GroEL was used as a loading control. f. Genomic neighborhoods of two homologous phage CARD-only proteins with 57.5% sequence identity. g. DALI Z-scores of protein structures similar to the AlphaFold2 model of the phage CARD-only protein from Acinetobacter phage Acj9.
