Figure 3.

A subset of c-di-AMP–binding proteins overexpressed in E. coli whole cell lysates determined by DRaCALA. A and B, fraction bound of radiolabeled [³²P]c-di-AMP is shown for lysates from E. coli induced for the expression of the indicated genes in presence of nonspecific ATP competitor at 100 μm (A) or 200 μm (B). C and D, induced lysates overexpressing the indicated gene are tested for specificity of c-di-AMP binding to [³²P]c-di-AMP by competition assays. C, competitors are 100 μm ATP (open bars) or 100 μm ATP and 100 μm c-di-AMP (closed bars). D, competitors are 200 μm ATP (open bars) or 200 μm ATP and 100 μm c-di-AMP (closed bars). Significance of binding was determined using unpaired t test between vector-only control and indicated gene (A and B) or between ATP and c-di-AMP competitors (C and D). p values of <0.05, <0.01, and <0.001 are indicated by *, **, and ***, respectively.