The N-terminal domain of EzrA binds to the C terminus of FtsZ to inhibit Staphylococcus aureus FtsZ polymerization.

Bacterial cytokinesis is accompanied by a macro-molecular complex called the "divisome." The divisome consists of two major components involving positive regulators and negative regulators that regulate the polymerization of an essential cytoskeleton protein FtsZ, which plays a key role in bacterial cell division by assembling the Z-ring, and therefore has been identified as a target for antibiotics. The negative regulators prevent the Z-ring assembly by inhibiting FtsZ polymerization. In Staphylococcus aureus, a pandemic human pathogen, one of the negative regulators, EzrA, contains a trans-membrane anchor region at the N-terminus and has five predicted coiled-coils. Recent reports indicate that the polymerization of FtsZ can be inhibited by forming a complex with EzrA. In this study, we attempted to locate the binding site for the interaction between EzrA and FtsZ in S. aureus (SaEzrA and SaFtsZ, respectively), by generating various constructs of SaEzrA and SaFtsZ proteins based on limited proteolysis. Various constructs of SaEzrA and SaFtsZ proteins were expressed and homogeneously purified. A GST pull-down assay indicated that the N-terminal domain of SaEzrA interacts with the C-terminal tail of SaFtsZ, and the elongated shape of EzrA was predicted based on the Stokes radius of each construct.