Structural changes of antitoxin HigA from Shigella flexneri by binding of its cognate toxin HigB

In toxin-antitoxin systems, many antitoxin proteins that neutralize their cognate toxin proteins also bind to DNA to repress transcription, and the DNA-binding affinity of the antitoxin is affected by its toxin. We solved crystal structures of the antitoxin HigA (apo-^SfHigA) and its complex with the toxin HigB (^SfHigBA) from Shigella flexneri. The apo-^SfHigA shows a distinctive V-shaped homodimeric conformation with sequestered N-domains having a novel fold. ^SfHigBA appears as a heterotetramer formed by N-terminal dimerization of ^SfHigB-bound ^SfHigA molecules. The conformational change in ^SfHigA upon ^SfHigB binding is mediated by rigid-body movements of its C-domains, which accompanied an overall conformational change from wide V-shaped to narrow V-shaped dimer. Consequently, the two putative DNA-binding helices (α7 in each subunit) are repositioned to a conformation more compatible with canonical homodimeric DNA-binding proteins containing HTH motifs. Collectively, this study demonstrates a conformational change in an antitoxin protein, which occurs upon toxin binding and is responsible for regulating antitoxin DNA binding.