Cameron, Todd A.Vega, Daniel E.Yu, ChenfeiXiao, HanMargolin, William2022-01-212022-01-212021Cameron, Todd A., Vega, Daniel E., Yu, Chenfei, et al.. "ZipA Uses a Two-Pronged FtsZ-Binding Mechanism Necessary for Cell Division." <i>mBio,</i> 12, no. 6 (2021) American Society for Microbiology: https://doi.org/10.1128/mbio.02529-21.https://hdl.handle.net/1911/111938In most bacteria, cell division is centrally organized by the FtsZ protein, which assembles into dynamic filaments at the division site along the cell membrane that interact with other key cell division proteins. In gammaproteobacteria such as Escherichia coli, FtsZ filaments are anchored to the cell membrane by two essential proteins, FtsA and ZipA. Canonically, this interaction was believed to be mediated solely by the FtsZ C-terminal peptide (CTP) domain that interacts with these and several other regulatory proteins. However, we now provide evidence of a second interaction between FtsZ and ZipA. Using site-specific photoactivated cross-linking, we identified a noncanonical FtsZ-binding site on ZipA on the opposite side from the FtsZ CTP-binding pocket. Cross-linking at this site was unaffected by the truncation of the FtsZ linker and CTP domains, indicating that this noncanonical site must interact directly with the globular core domain of FtsZ. Mutations introduced into either the canonical or noncanonical binding sites on ZipA disrupted photo-cross-linking with FtsZ and normal ZipA function in cell division, suggesting that both binding modes are important for normal cell growth and division. One mutation at the noncanonical face was also found to suppress defects of several other canonical and noncanonical site mutations in ZipA, suggesting there is some interdependence between the two sites. Taken together, these results suggest that ZipA employs a two-pronged FtsZ-binding mechanism.engThis is is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.Journal articlembio-02529-21https://doi.org/10.1128/mbio.02529-21