Inhibition of Gyrase via a conserved α-hairpin in Vibrio cholerae ParE2 and neutralization by ParD2

The ParE family of toxins is known to target Gyrase through an as yet unknown molecular mechanism. Here we show that Vibrio cholerae ParE2 ( Vc ParE2) interacts with Gyrase in a DNA-dependent manner. The interaction site is located at the ParE-specific α-hairpin and involves Trp25 as a key residue. The latter is poorly conserved within the ParE family, although full toxicity is only retained upon substitution with Tyr in the Vc ParE2 context. In vitro , the Trp25Ala mutation reduces the rate of Gyrase-mediated supercoiling, while the so-called cleavable complex remains stabilized. The C-terminal domain of the antitoxin Vc ParD2, which wraps its intrinsically disordered domain around Vc ParE2 without covering the Trp25-containing interaction site, binds to Vc ParE2 with high affinity, but only partially prevents ParE-mediated growth inhibition. Full inhibition of Vc ParE2 requires full length Vc ParD2 that leads to a complex where Trp25 is fully shielded from solvent.