Aubry, B. | Search bioRxiv

bioRxiv2025

Ancient Truncated FtsZ Paralogs Likely Tune Cell Division in Hyphomicrobiales

Bacterial cell division is a well conserved, tightly regulated process that allows the separation of two viable daughter cells. In most bacteria, the proteins that drive division, termed the divisome, are recruited to mid-cell by FtsZ after it polymerizes to form the Z- ring. Interestingly, FtsZ has undergone several independent duplication events across the bacterial kingdom. We identified FtsZ GTPase protein sequences across alphaproteobacterial genomes, from representative genera for each family, and observed numerous ftsZ duplications in the order Hyphomicrobiales. Hidden Markov Modeling (HMM) supported the maintenance of two distinct lineages of FtsZ GTPase duplications among three families. The Nitrobacteraceae duplication, occurring in only the genus Bradyrhizobium, exhibits a different substitution pattern from that shared by the Phyllobacteraceae and Rhizobiaceae families. Within the Rhizobiaceae lineage, Agrobacterium tumefaciens contains three paralogs of FtsZ including the essential FtsZAT, and paralogs FtsZ1, and FtsZ3. In A. tumefaciens, we show that FtsZ1, but not FtsZ3, inhibits cell division when overexpressed. A hyperactive allele of ftsW partially protected against overexpression of ftsZ1 suggesting that FtsZ1 may inhibit proper regulation of septal peptidoglycan biosynthesis during cell division. Overall, these observations suggest that maintenance of FtsZ paralogs in some bacteria which may fine tune the division process. Article SummaryGene duplication drives evolutionary innovation by providing raw genetic material and allowing microbes to acquire novel functions. Here, we explore the duplication of a gene encoding the essential cell division protein FtsZ in the Hyphomicrobiales and find that the duplication is primarily conserved in genera that interact with plant hosts. In A. tumefaciens FtsZ1 is not essential for cell division; however, overexpression of FtsZ1 inhibited cell division suggesting it may have a regulatory role during this essential process. This observation highlights the key role for gene duplication in the modulation of complex processes in bacteria.

microbiology↗

bioRxiv2024

The Cell Division Protein FzlA Performs a Conserved Function in Diverse Alphaproteobacteria

In almost all bacteria, the tubulin-like GTPase FtsZ polymerizes to form a "Z-ring" that marks the site of division. FtsZ recruits other proteins, collectively known as the divisome, that together remodel and constrict the envelope. Constriction is driven by peptidoglycan (PG) cell wall synthesis by the glycosyltransferase FtsW and the transpeptidase FtsI (FtsWI), but these enzymes require activation to function. How recruitment of FtsZ to the division site leads to FtsWI activation and constriction remains largely unknown. Previous work in our laboratory demonstrated that an FtsZ-binding protein, FzlA, is essential for activation of FtsWI in the alphaproteobacterium Caulobacter crescentus. Additionally, we found that FzlA also binds to a DNA translocase called FtsK, suggesting that it may link constriction activation to chromosome segregation. FzlA is conserved throughout alphaproteobacteria but has only been examined in detail in C. crescentus. Here, we explored whether FzlA function is conserved in diverse alphaproteobacteria. We assessed FzlA homologs from Rickettsia parkeri and Agrobacterium tumefaciens, and found that, similar to C. crescentus FzlA, they bind directly to FtsZ and localize to midcell. The FtsZ-FzlA interaction interface is conserved, as we demonstrated that FzlA from each of the three species examined can bind to FtsZ from any of the three in vitro. Additionally, we determined that A. tumefaciens FzlA can fulfill the essential function of FzlA when produced in C. crescentus, indicating conservation of function. These results suggest that FzlA serves as an important regulator that coordinates chromosome segregation with envelope constriction across diverse alphaproteobacteria. ImportanceCell division is essential for bacterial replication and must be highly regulated to ensure robust remodeling of the cell wall in coordination with segregation of the genome to daughter cells. In Caulobacter crescentus, FzlA plays a major role in regulating this process by activating cell wall synthesis in a manner that couples constriction to chromosome segregation. FzlA is broadly conserved in alphaproteobacteria suggesting it plays a similar function across this class of bacteria. Here we have shown that, indeed, FzlA biochemical interactions and function are conserved in diverse alphaproteobacteria. Because FzlA is conserved in alphaproteobacterial human pathogens, understanding this protein and its interactome could present therapeutic benefits by identifying potential antibiotic targets to treat infections.

microbiology↗