Nε−Lysine Acetylation of a Bacterial Transcription Factor Inhibits Its DNA-Binding Activity
Figure 4
Acetylation of Lys180 inhibits RcsB DNA-binding activity, mimicking the effect of substitutions that block interactions at this position.
Reactions of RcsB proteins with the flhDC operator sequence contained equimolar RcsA, and were analyzed by EMSAs. A. The DNA-binding activities of wild-type RcsB (RcsBWT, top panel) and acetylated RcsB (RcsBAc, bottom panel) were assessed. RcsB protein concentrations from left to right: 31.25, 62.5, 125, 250, 500 nM. B. Wild-type and variant RcsB proteins were incubated with Pat and [14C, C-1]-Ac-CoA, resolved on a denaturing polyacrylamide gel (top), and exposed to a phosphor screen (bottom). C. Assessment of the DNA-binding activities of wild-type and variant RcsB proteins. RcsB protein concentrations from left to right: 125, 250, 500 nM.