Extended Data Fig. 4: Substrate preference of the AcrIII-1 proteins SIRV1 gp29 and YddF, and effective range of cA4 degradation.
From: An anti-CRISPR viral ring nuclease subverts type III CRISPR immunity
a–d, TLC images visualizing (under 254 nm UV light) cA4 and cA6 (450 μM) degradation by SIRV1 gp29 (a, b) and YddF (c, d) over time (in minutes). Both AcrIII-1 enzymes display a clear preference for cA4 over cA6. All TLC images are representative of three technical replicates. e, Denaturing PAGE showing activation of Csx1 (0.5 μM dimer) by the indicated amounts (500–0.5 μM) of HPLC-purified cA4, and its subsequent deactivation when either AcrIII-1 or Crn1 (2 μM dimer) was present to degrade cA4. The AcrIII-1 enzyme degraded 100-fold more cA4 than did Crn1. The control reaction (C) shows RNA incubated with Csx1 in the absence of cA4 (n = 3 technical replicates). For gel source data, see Supplementary Fig. 1.
