Determining the rate-limiting processes for cell division in Escherichia coli

• Published in: Nature communications 2024-11, Vol.15 (1), p.9948-15, Article 9948
• Main Authors: Männik, Jaana, Kar, Prathitha, Amarasinghe, Chathuddasie, Amir, Ariel, Männik, Jaan
• Format: Article
• Language: English
• Subjects: 13/62; 14/35; 14/63; 631/326/41/1969; 631/326/88; 631/80/641/2090; 639/766/747; Accumulation; Actin; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Cell cycle; Cell Division; Cell membranes; Coliforms; Constrictions; Cytoskeletal Proteins - genetics; Cytoskeletal Proteins - metabolism; E coli; Escherichia coli; Escherichia coli - cytology; Escherichia coli - growth & development; Escherichia coli - metabolism; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; Growth rate; Humanities and Social Sciences; In vivo methods and tests; Limiting factors; Membrane Proteins - genetics; Membrane Proteins - metabolism; Modelling; multidisciplinary; Peptidoglycans; Proteins; Science; Science (multidisciplinary); Stochasticity; Tubulin
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-024-54242-w

A critical cell cycle checkpoint for most bacteria is the onset of constriction when the septal peptidoglycan synthesis starts. According to the current understanding, the arrival of FtsN to midcell triggers this checkpoint in Escherichia coli . Recent structural and in vitro data suggests that recruitment of FtsN to the Z-ring leads to a conformational switch in actin-like FtsA, which links FtsZ protofilaments to the cell membrane and acts as a hub for the late divisome proteins. Here, we investigate this putative pathway using in vivo measurements and stochastic cell cycle modeling at moderately fast growth rates. Quantitatively upregulating protein concentrations and determining the resulting division timings shows that FtsN and FtsA numbers are not rate-limiting for the division in E. coli . However, at higher overexpression levels, they affect divisions: FtsN by accelerating and FtsA by inhibiting them. At the same time, we find that the FtsZ numbers in the cell are one of the rate-limiting factors for cell divisions in E. coli . Altogether, these findings suggest that instead of FtsN, accumulation of FtsZ in the Z-ring is one of the main drivers of the onset of constriction in E. coli at faster growth rates. The recruitment of protein FtsN to midcell is thought to be the main rate-liming process for cell division in the bacterial model E. coli . Here, Männik et al. use experiments with live bacteria as well as stochastic cell-cycle modeling to show that instead of FtsN, accumulation of the tubulin-like protein FtsZ in the Z-ring is one of the main drivers of the onset of cell division at fast growth rates in E. coli .