Fig. 3
From: AID/APOBEC-like cytidine deaminases are ancient innate immune mediators in invertebrates
Enzymatic activities of SpAIDLs. a–c Deaminase activities of each indicated protein were measured in bacterial reversion assays using either a Kanamycin resistance reporter plasmid (a) or the endogenous rpoB gene as the reporter (b, c). Note that the SpAIDL1 dataset from b is shown again in c for clarity. The horizontal bars represent the median (numerical value below each column), and p-values (for each deaminase compared to empty pASK for a, b, and compared to wildtype SpAIDL1 for c) were calculated using a Willcoxon rank sum for unpaired data. Datasets with p < 0.05 are marked with an asterisk (a: HsAID p = 0.000001 and SpAIDL1 p = 0.02087; b: HsAID p = 6.6 × 10−9, SpAIDL1 p = 0.0005, and SpAIDL2 p = 0.0014; c: SpAIDL1 RQAA p = 0.00048). The inset represents a magnified view omitting all data points of HsAID (a, b) and one datapoint of SPAIDL2 (b) for clarity. SpAIDL1 RQ and SpAIDL1 RQAA are mutants of SpAIDL1 in which either two (H86R and E88Q) or four (H86R, E88Q, C123A, and C126A) of the conserved active site residues were altered. d, e Western blot analysis of the expression of the indicated V5-tagged deaminases in bacteria induced with AHT for 3 h (+) or not (−). Note that the enzymatic activities were tested with untagged versions of the proteins. The empty pASK_V5 plasmids served as a control. The predicted sizes for V5-tagged HsAID, SpAIDL1 (and the mutants thereof), 2, 3, 4a, and 9 are 25.4, 24.0, 23.5, 22.3, 23.6, and 24.6 kD, respectively
