The distinctive cell division interactome of Neisseria gonorrhoeae.

BACKGROUND: Bacterial cell division is an essential process driven by the formation of a Z-ring structure, as a cytoskeletal scaffold at the mid-cell, followed by the recruitment of various proteins which form the divisome. The cell division interactome reflects the complement of different interactions between all divisome proteins. To date, only two cell division interactomes have been characterized, in Escherichia coli and in Streptococcus pneumoniae. The cell divison proteins encoded by Neisseria gonorrhoeae include FtsZ, FtsA, ZipA, FtsK, FtsQ, FtsI, FtsW, and FtsN. The purpose of the present study was to characterize the cell division interactome of N. gonorrhoeae using several different methods to identify protein-protein interactions. We also characterized the specific subdomains of FtsA implicated in interactions with FtsZ, FtsQ, FtsN and FtsW. RESULTS: Using a combination of bacterial two-hybrid (B2H), glutathione S-transferase (GST) pull-down assays, and surface plasmon resonance (SPR), nine interactions were observed among the eight gonococcal cell division proteins tested. ZipA did not interact with any other cell division proteins. Comparisons of the N. gonorrhoeae cell division interactome with the published interactomes from E. coli and S. pneumoniae indicated that FtsA-FtsZ and FtsZ-FtsK interactions were common to all three species. FtsA-FtsW and FtsK-FtsN interactions were only present in N. gonorrhoeae. The 2A and 2B subdomains of FtsANg were involved in interactions with FtsQ, FtsZ, and FtsN, and the 2A subdomain was involved in interaction with FtsW. CONCLUSIONS: Results from this research indicate that N. gonorrhoeae has a distinctive cell division interactome as compared with other microorganisms.

An Escherichia coli expression model reveals the species-specific function of FtsA from Neisseria gonorrhoeae in cell division.

Escherichia coli (Ec) has been used to study the function of cell division proteins from different microorganisms, especially when genetic tools are limited for studying these proteins in their native hosts. The expression of ftsA from Neisseria gonorrhoeae (Ng) disrupted cell division in E. coli resulting in a significant increase in cell length. In some cells, FtsANg localised to the division site and the poles of E. coli cells, but the majority of cells showed no specifical localisation. FtsANg did not complement an E. coli ftsA mutant strain. Bacterial two-hybrid and GST pull-down assays indicated that FtsANg interacted with FtsNEc, but no other cell division proteins from E. coli. This interaction was mediated through the 2A and 2B subdomains of FtsANg. This evidence suggests that the function of FtsANg is species specific.