A Predicted Helix-Turn-Helix Core Is Critical for Bacteriophage Kil Peptide to Disrupt Escherichia coli Cell Division.

Background/objectives: FtsZ, a eukaryotic tubulin homolog and an essential component of the bacterial divisome, is the target of numerous antimicrobial compounds as well as proteins and peptides, most of which inhibit FtsZ polymerization dynamics. We previously showed that the Kil peptide from bacteriophage λ inhibits Escherichia coli cell division by disrupting FtsZ ring assembly, and this inhibition requires the presence of the essential FtsZ membrane anchor protein ZipA.

Methods

To investigate Kil's molecular mechanism further, we employed deletions, truncations, and molecular modeling to identify the minimal residues necessary for its activity.

Results

Modeling suggested that Kil's core segment folds into a helix-turn-helix (HTH) structure. Deleting either the C-terminal 11 residues or the N-terminal 5 residues of Kil still allowed the inhibition of E. coli cell division, but removing both termini nearly abolished this activity, indicating that a minimal region within the Kil HTH core is essential for its structure and function. Another Kil-like peptide from a closely related enterobacterial phage also disrupted FtsZ ring assembly and required ZipA for this activity. Consistent with its broader activity against FtsZ, λ Kil was able to efficiently inhibit cell division of a uropathogenic E. coli (UPEC) strain.

Conclusions

Understanding the structure and function of Kil and similar peptides can potentially reveal additional ways to target FtsZ for antimicrobial therapies and elucidate how FtsZ functions in bacterial cell division.