Fig. 5: Subcellular distributions and co-localization of Zorya components with a proposed model.
From: Structure and mechanism of the Zorya anti-phage defence system
a, Exemplary denoised TIRF images of the subcellular distributions of WT ZorB and ZorB(D26N) fused to HT with or without Bas24. Scale bars, 1 µm. b, Comparison of the detected maxima of the ZorAB complex foci between the untreated or Bas24-exposed (MOI, 5; 30 min) conditions. n > 250 cells, 3 replicates. P = 0.022 (WT) and P = 0.027 (D26N). c, Exemplary denoised TIRF images of the subcellular distributions of mNG-tagged ZorC and ZorD, with or without Bas24; mNG was fused to either the ZorC N terminus (mNG–ZorC) or ZorD C terminus (ZorD–mNG). Scale bars, 1 µm. d, Comparison of the detected maxima of the ZorC and ZorD foci between the untreated and Bas24-exposed (MOI 5, 30 min) conditions. n > 250 cells. P = 0.004 and P = 0.04. e, Exemplary denoised TIRF images of co-localization of mNG–ZorC with ZorB(WT)–HT with or without Bas24. Scale bars, 1 µm. The white arrows highlight co-localization. f, Co-localization analysis of ZorB(WT)–HT or ZorB(D26N)–HT with ZorC–mNG with or without Bas24 (MOI, 5; 30 min). P = 0.0002 (left), P = 0.0042 (right). g, Exemplary denoised TIRF images of co-localization of ZorD–mNG with ZorB–HT, with or without Bas24. Scale bars, 1 µm. h, Co-localization analysis of ZorB(WT)–HT or mutants (ZorB(D26N)–HT and ZorA(∆483–739) (tail-tip deletion)) with ZorD–mNG, with or without Bas24 (MOI, 5; 30 min). ****P < 0.0001. i–k, The proposed Zorya defence model. OM, outer membrane. i, Inactive ZorAB diffuses laterally within the inner membrane. j, Inactive ZorAB detects cell envelope perturbation during phage infection and ZorB PGBDs anchor to PG. Ion translocation through ZorAB triggers ZorA and its tail to rotate around ZorB. k, The ZorAB motor signal is transferred through the ZorA tail, which recruits and/or activates ZorC and ZorD. ZorC and ZorD bind to and degrade phage DNA, preventing phage replication. Datapoints represent the mean focus counts for each of three replicates and the shaded bars represent the mean between replicates. For b and d, data are mean values, with the whiskers representing the minimum to maximum values. Statistical analysis was performed using unpaired t-tests (b,d,f) or two-way ANOVA (h)33. ZorB(WT)–HT and ZorB(D26N)–HT expression data are provided in Extended Data Fig. 6e.
