A membrane protein, EzrA, regulates assembly dynamics of FtsZ by interacting with the C-terminal tail of FtsZ

Biochemistry. 2007 Sep 25;46(38):11013-22. doi: 10.1021/bi700710j. Epub 2007 Aug 24.


FtsZ polymerizes to form a dynamic ring structure called the Z-ring at the midcell of bacteria. EzrA, a membrane protein, has been shown to prevent the formation of aberrant Z-rings in the low GC Gram-positive bacteria by inhibiting FtsZ assembly. In this study, we show that Bacillus subtilis (B. subtilis) EzrA inhibited the assembly and bundling of B. subtilis FtsZ. It increased the critical concentration of FtsZ assembly and depolymerized the preformed FtsZ polymers in vitro. We obtained evidence suggesting that B. subtilis EzrA forms complex with B. subtilis FtsZ in vitro. EzrA was found to bind to FtsZ at a single site with a dissociation constant of 4.3 +/- 0.6 microM. EzrA-FtsZ interaction has a significant electrostatic contribution as apparent from the effect of salt on their binding interactions. To elucidate the site of interaction between EzrA and FtsZ, we deleted 16 amino acid residues from the extreme C-terminal tail of B. subtilis FtsZ, which are conserved in FtsZ orthologues. EzrA did not inhibit the assembly of C-terminal truncated B. subtilis FtsZ. It also did not bind to the C-terminal truncated FtsZ detectably, suggesting that EzrA interacts with FtsZ through its conserved C-terminal tail residues. Further, a 17-residue synthetic peptide (365-382) of the C-terminal tail of FtsZ (CTP17) was used to probe the interaction of EzrA with the C-terminal tail of FtsZ. CTP17 bound to EzrA, inhibited the binding of EzrA to FtsZ, and surmounted the inhibitory effects of EzrA on the assembly of FtsZ in vitro. The data together showed that EzrA binds to the C-terminal tail of FtsZ. FtsA, a positive regulator of FtsZ assembly, is also known to interact with the C-terminal tail of FtsZ. The results indicated an interesting possibility that the assembly dynamics of FtsZ in the Z-ring is regulated by the competition between positive and negative regulators sharing the same binding site on FtsZ.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacillus subtilis / metabolism
  • Bacillus subtilis / ultrastructure
  • Bacterial Proteins / antagonists & inhibitors
  • Bacterial Proteins / metabolism*
  • Bacterial Proteins / ultrastructure
  • Binding Sites
  • Binding, Competitive
  • Cell Division / physiology
  • Cytokinesis / drug effects
  • Cytoskeletal Proteins / antagonists & inhibitors
  • Cytoskeletal Proteins / metabolism*
  • Cytoskeletal Proteins / ultrastructure
  • Cytoskeleton / metabolism
  • Cytoskeleton / ultrastructure
  • Gene Expression Regulation, Bacterial
  • Guanosine Triphosphate / metabolism
  • Hydrolysis
  • Kinetics
  • Membrane Proteins / genetics
  • Membrane Proteins / isolation & purification
  • Membrane Proteins / metabolism*
  • Mutation
  • Peptides / chemical synthesis
  • Peptides / metabolism
  • Protein Binding
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism*
  • Recombinant Fusion Proteins / ultrastructure
  • Sequence Deletion


  • Bacterial Proteins
  • Cytoskeletal Proteins
  • EzrA protein, Bacillus subtilis
  • FtsA protein, Bacteria
  • FtsZ protein, Bacteria
  • Membrane Proteins
  • Peptides
  • Recombinant Fusion Proteins
  • Guanosine Triphosphate