Studying Z-ring formation following nucleoid partitioning in escherichia coli by microscopy flow-cytometry and machine learning

ABSTRACT The min system spatially regulates division through the topological regulation of MinCD, an inhibitor of cell division. MinCD was previously shown to inhibit division by preventing assembly of the Z ring (E. Bi and J. Lutkenhaus, J. Bacteriol. 175:1118–1125, 1993); however, this was questioned in a recent report (S. S. Justice, J. Garcia-Lara, and L. I. Rothfield, Mol. Microbiol. 37:410–423, 2000) which indicated that MinCD acted after Z-ring formation and prevented the recruitment of FtsA to the Z ring. This discrepancy was due in part to alternative fixation conditions. We have therefore reinvestigated the action of MinCD and avoided fixation by using green fluorescent protein (GFP) fusions to division proteins. MinCD prevented the localization of both FtsZ-GFP and ZipA-GFP, consistent with it preventing Z-ring assembly. Consistent with a direct interaction between FtsZ and the MinCD inhibitor, we find that increased FtsZ, but not FtsA, suppresses MinCD-induced lethality. Furthermore, strains carrying various alleles offtsZ, selected on the basis of resistance to the inhibitor SulA, displayed variable resistance to MinCD. These results are consistent with FtsZ as the target of MinCD and confirm that this inhibitor prevents Z-ring assembly.

Download Full-text

Transient Membrane-Linked FtsZ Assemblies Precede Z-Ring Formation in Escherichia coli

Current Biology ◽

10.1016/j.cub.2019.12.023 ◽

2020 ◽

Vol 30 (3) ◽

pp. 499-508.e6 ◽

Cited By ~ 7

Author(s):

Bryant E. Walker ◽

Jaana Männik ◽

Jaan Männik

Keyword(s):

Escherichia Coli ◽

Ring Formation ◽

Z Ring

Download Full-text

Resveratrol antibacterial activity against Escherichia coli is mediated by Z-ring formation inhibition via suppression of FtsZ expression

Scientific Reports ◽

10.1038/srep10029 ◽

2015 ◽

Vol 5 (1) ◽

Cited By ~ 69

Author(s):

Dahyun Hwang ◽

Young-Hee Lim

Keyword(s):

Escherichia Coli ◽

Antibacterial Activity ◽

Ring Formation ◽

Z Ring

Download Full-text

New Temperature-Sensitive Alleles of ftsZ in Escherichia coli

Journal of Bacteriology ◽

10.1128/jb.187.1.358-365.2005 ◽

2005 ◽

Vol 187 (1) ◽

pp. 358-365 ◽

Cited By ~ 18

Author(s):

Stephen G. Addinall ◽

Elaine Small ◽

Duncan Whitaker ◽

Shane Sturrock ◽

William D. Donachie ◽

...

Keyword(s):

Escherichia Coli ◽

Cell Division ◽

Point Mutations ◽

Ring Formation ◽

Single Amino Acid ◽

Temperature Sensitive ◽

Permissive Temperature ◽

Nonpermissive Temperature ◽

Z Ring

ABSTRACT We isolated five new temperature-sensitive alleles of the essential cell division gene ftsZ in Escherichia coli, using P1-mediated, localized mutagenesis. The five resulting single amino acid changes (Gly109→Ser109 for ftsZ6460, Ala129→Thr129 for ftsZ972, Val157→Met157 for ftsZ2066, Pro203→Leu203 for ftsZ9124, and Ala239→Val239 for ftsZ2863) are distributed throughout the FtsZ core region, and all confer a lethal cell division block at the nonpermissive temperature of 42°C. In each case the division block is associated with loss of Z-ring formation such that fewer than 2% of cells show Z rings at 42°C. The ftsZ9124 and ftsZ6460 mutations are of particular interest since both result in abnormal Z-ring formation at 30°C and therefore cause significant defects in FtsZ polymerization, even at the permissive temperature. Neither purified FtsZ9124 nor purified FtsZ6460 exhibited polymerization when it was assayed by light scattering or electron microscopy, even in the presence of calcium or DEAE-dextran. Hence, both mutations also cause defects in FtsZ polymerization in vitro. Interestingly, FtsZ9124 has detectable GTPase activity, although the activity is significantly reduced compared to that of the wild-type FtsZ protein. We demonstrate here that unlike expression of ftsZ84, multicopy expression of the ftsZ6460, ftsZ972, and ftsZ9124 alleles does not complement the respective lethalities at the nonpermissive temperature. In addition, all five new mutant FtsZ proteins are stable at 42°C. Therefore, the novel isolates carrying single ftsZ(Ts) point mutations, which are the only such strains obtained since isolation of the classical ftsZ84 mutation, offer significant opportunities for further genetic characterization of FtsZ and its role in cell division.

Download Full-text

Coregulated assembly of actin-like FtsA polymers with FtsZ during Z-ring formation and division in Escherichia coli

10.1101/2021.01.12.426377 ◽

2021 ◽

Author(s):

Josiah J. Morrison ◽

Joseph Conti ◽

Jodi L. Camberg

Keyword(s):

Escherichia Coli ◽

Atp Hydrolysis ◽

Direct Interaction ◽

Ring Formation ◽

Phospholipid Vesicle ◽

E Coli ◽

C Terminus ◽

Z Ring

AbstractIn Escherichia coli, the actin homolog FtsA localizes the cell division machinery, beginning with the Z-ring, to the cytoplasmic membrane through direct interaction with FtsZ. FtsZ polymers are first to assemble at the Z-ring at midcell, where they direct constriction and septation. While FtsZ polymerization is critical for establishing a functional Z-ring that leads to constriction, the assembly state of FtsA and the role of FtsA ATP utilization during division in E. coli remain unclear. Here, we show that ATP hydrolysis, FtsZ interaction, and phospholipid vesicle remodeling by FtsA are impaired by a substitution mutation at the predicted active site for hydrolysis. This mutation, Glu 14 to Arg, also impairs Z-ring assembly and division in vivo. To further investigate the role of phospholipid engagement and ATP utilization in regulating FtsA function, we characterized a truncated E. coli FtsA variant, FtsA(ΔMTS), which lacks the region at the C-terminus important for engaging the membrane and is defective for ATP hydrolysis. We show that E. coli FtsA(ΔMTS) forms ATP-dependent actin-like filaments and assembly is antagonized by FtsZ. Polymerization of FtsZ with GTP, or a non-hydrolyzable analog, blocks inhibition of ATP-dependent FtsA assembly, and instead favors coassembly of stable FtsA/FtsZ polymers. In the cell, FtsA/FtsZ coassembly is favored at midcell, where FtsZ polymerizes, and inhibited at regions where FtsZ polymers are destabilized by regulators, such as MinC at the poles or SlmA at the nucleoid. We show that MinC prevents recruitment of FtsZ, via FtsA, to phospholipids, suggesting that local interactions of MinC with FtsZ block membrane tethering and uncouple the Z-ring from its major membrane contact. During Z-ring formation, the coassembly of FtsZ polymers with FtsA is coordinated and is a critical early step in division. This step also serves as a checkpoint by responding to the suite of FtsZ assembly regulators in the cell that modulate Z-ring position and dynamics prior to initiating cell wall synthesis.

Download Full-text

Cadmium toxicity in Escherichia coli: Cell morphology, Z-ring formation and intracellular oxidative balance

Ecotoxicology and Environmental Safety ◽

10.1016/j.ecoenv.2012.09.017 ◽

2012 ◽

Vol 86 ◽

pp. 54-59 ◽

Cited By ~ 17

Author(s):

Sk Tofajjen Hossain ◽

Ivy Mallick ◽

Samir Kumar Mukherjee

Keyword(s):

Escherichia Coli ◽

Cell Morphology ◽

Cadmium Toxicity ◽

Ring Formation ◽

Coli Cell ◽

Oxidative Balance ◽

Z Ring

Download Full-text

Recruitment of MinC, an Inhibitor of Z-Ring Formation, to the Membrane in Escherichia coli: Role of MinD and MinE

Journal of Bacteriology ◽

10.1128/jb.185.1.196-203.2003 ◽

2003 ◽

Vol 185 (1) ◽

pp. 196-203 ◽

Cited By ~ 85

Author(s):

Zonglin Hu ◽

Cristian Saez ◽

Joe Lutkenhaus

Keyword(s):

Escherichia Coli ◽

Atp Hydrolysis ◽

Ring Formation ◽

The Other ◽

Phospholipid Bilayer ◽

Dependent Manner ◽

Min Proteins ◽

Z Ring ◽

The Mind

ABSTRACT In Escherichia coli, the min system prevents division away from midcell through topological regulation of MinC, an inhibitor of Z-ring formation. The topological regulation involves oscillation of MinC between the poles of the cell under the direction of the MinDE oscillator. Since the mechanism of MinC involvement in the oscillation is unknown, we investigated the interaction of MinC with the other Min proteins. We observed that MinD dimerized in the presence of ATP and interacted with MinC. In the presence of a phospholipid bilayer, MinD bound to the bilayer and recruited MinC in an ATP-dependent manner. Addition of MinE to the MinCD-bilayer complex resulted in release of both MinC and MinD. The release of MinC did not require ATP hydrolysis, indicating that MinE could displace MinC from the MinD-bilayer complex. In contrast, MinC was unable to displace MinE bound to the MinD-bilayer complex. These results suggest that MinE induces a conformational change in MinD bound to the bilayer that results in the release of MinC. Also, it is argued that binding of MinD to the membrane activates MinC.

Download Full-text