Fig. 4: Acb1 and Apyc1 disrupt CBASS and Pycsar host defence.
From: Phage anti-CBASS and anti-Pycsar nucleases subvert bacterial immunity
a, Agarose gel analysis of uncut plasmid DNA incubated with the CBASS effector Cap5 and 3′3′-cGAMP that was treated with wild-type (WT) Acb1, catalytically inactive Acb1-H44A/H113A or WT Apyc1. Data are representative of three independent experiments. For unprocessed gels, see Supplementary Fig. 1. ds, double-stranded DNA. b, Release of fluorescent substrate from an NAD+ analogue incubated with the Pycsar effector PycTIR and cUMP that was treated with WT Acb1, WT Apyc1 or catalytically inactive Apyc1-H64A/H66A/H69A. Data are presented as mean ± s.d. from n = 3 independent experiments. RFUs, relative fluorescence units. c, E. coli carrying plasmids encoding a type III CBASS operon from E. coli KTE188 and/or T4 Acb1 were challenged with serial dilutions of P1 phage. Data are presented as mean ± s.d. from n = 3 independent experiments. PFUs, plaque-forming units. d, E. coli carrying plasmids encoding a Pycsar operon and/or SBSphiJ Apyc1 were challenged with serial dilutions of T5 phage. Data are presented as mean ± s.d. from n = 3 independent experiments. e, Representative plaque assays of E. coli carrying a plasmid encoding an active or catalytically inactive CBASS operon from Yersinia aleksiciae and challenged with WT phage T4 or phage T4 engineered to remove Acb1 (Δacb1). f, Summary of plaque assay results of WT or Δacb1 phage T4 infection of E. coli carrying CBASS operons from Y. aleksiciae or E. coli. Data are presented as mean ± s.d. from n = 4 (Y. aleksiciae operon) or n = 3 (E. coli operon) technical replicates and are representative of at least 3 biologically independent experiments. Statistical significance in c, d and f was determined using an unpaired two-tailed t-test.
