In vitro reconstitution of Escherichia coli divisome activation

Radler, Philipp; Baranova, Natalia S.; Caldas, Paulo; Sommer, Christoph; López-Pelegrín, Mar; Michalík, David; Loose, Martin

doi:10.1038/s41467-022-30301-y

Cited by 13 publications

(14 citation statements)

References 49 publications

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“…However, the MTH of FtsA serves only as a generic membrane anchor for FtsZ, independently of any specific sequence identity. Replacing FtsA's MTH with another membrane attachment still allows FtsZ tethering in vitro and in vivo (Radler et al, 2022; Shiomi & Margolin, 2008).…”

Section: Membrane Tethers In the Gram‐negative Model Escherichia Colimentioning

confidence: 99%

“…However, the higher‐order structure of tethered FtsZ is distinct among the different FtsA structures. Whereas FtsZ protofilaments are aligned, but kept apart, when tethered to mini‐rings of wild‐type FtsA, nonring FtsA structures (including the curved oligomers and double‐stranded filaments) pack the FtsZ protofilaments close together in bundles, probably because of the higher packing density on the membrane of the FtsA tethers in this form (Krupka et al, 2017; Radler et al, 2022). The cytoplasmic domain of FtsN, an essential divisome protein that interacts with FtsA's unique 1C domain (Busiek et al, 2012; Rico et al, 2004), can convert the mini‐rings to the double‐stranded oligomers (Nierhaus et al, 2022).…”

Section: Membrane Tethers In the Gram‐negative Model Escherichia Colimentioning

confidence: 99%

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Anchors: A way for FtsZ filaments to stay membrane bound

Naha

Haeusser

Margolin

2023

Molecular Microbiology

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Most bacteria use the tubulin homolog FtsZ to organize their cell division. FtsZ polymers initially assemble into mobile complexes that circle around a ring‐like structure at the cell midpoint, followed by the recruitment of other proteins that will constrict the cytoplasmic membrane and synthesize septal peptidoglycan to divide the cell. Despite the need for FtsZ polymers to associate with the membrane, FtsZ lacks intrinsic membrane binding ability. Consequently, FtsZ polymers have evolved to interact with the membrane through adaptor proteins that both bind FtsZ and the membrane. Here, we discuss recent progress in understanding the functions of these FtsZ membrane tethers. Some, such as FtsA and SepF, are widely conserved and assemble into varied oligomeric structures bound to the membrane through an amphipathic helix. Other less‐conserved proteins, such as EzrA and ZipA, have transmembrane domains, make extended structures, and seem to bind to FtsZ through two separate interactions. This review emphasizes that most FtsZs use multiple membrane tethers with overlapping functions, which not only attach FtsZ polymers to the membrane but also organize them in specific higher‐order structures that can optimize cell division activity. We discuss gaps in our knowledge of these concepts and how future studies can address them.

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Section: Membrane Tethers In the Gram‐negative Model Escherichia Colimentioning

confidence: 99%

Section: Membrane Tethers In the Gram‐negative Model Escherichia Colimentioning

confidence: 99%

Anchors: A way for FtsZ filaments to stay membrane bound

Naha

Haeusser

Margolin

2023

Molecular Microbiology

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“…Figure S3 explores in depth the role of hydrolysis and dissociation reactions in depolymerisation kinetics. Given that monomers detach into solution upon depolymerisation, p off is not dependent on the environment of the filament on the surface [31]. To reach steady state p on = p off must be satisfied, which defines an intrinsic size N c = −r on τ det log(1 − p on ) around which filaments fluctuate while treadmilling at constant velocity v c = r on σ .…”

Section: Model Descriptionmentioning

confidence: 99%

Self-organisation of mortal filaments: the role of FtsZ treadmilling in bacterial division ring formation

Campos

Whitley

Radler

et al. 2023

Preprint

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Functional protein filaments in the cell commonly treadmill - grow and shrink on opposite ends, driven by energy consumption. Treadmilling causes filaments to seem as if they were moving, even though the individual proteins remain static. Here we investigate the role of treadmilling, implemented as dynamic turnover, in the collective filament self-organisation. On the example of the bacterial FtsZ protein, a highly evolutionary conserved tubulin homologue, we show, in computer simulations and in vitro experiment, that treadmilling drives filament nematic ordering by means of dissolving misaligned filaments. We demonstrate that this ordering via local dissolution allows the system to quickly respond to chemical and geometrical biases in the cell, and is necessary for the formation of the bacterial division ring in live Bacillus subtilis. We finally use simulations to quantitatively explain the characteristic dynamics of FtsZ division ring formation measured in vivo. Beyond FtsZ and cytoskeletal filaments, our study identifies a novel mechanism for nematic ordering via constant birth and death of energy-consuming filaments.

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“…Proteins used in this study, wildtype FtsZ, FtsZ L169R and FtsA, were purified as previously described 53 . FtsZ, L169R was obtained by site-directed mutagenesis (SDM).…”

Section: Supplementary Figure Captionsmentioning

confidence: 99%

Chiral and nematic phases of flexible active filaments

Dunajova

Mateu

Radler

et al. 2022

Preprint

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The emergence of large-scale order in self-organized systems relies on local interactions between individual components. During bacterial cell division, the tubulin-homolog FtsZ polymerizes into treadmilling filaments that further assemble into a cytoskeletal ring. Although minimal in vitro assays have shown the striking self-organization capacity of FtsZ filaments, such as dynamic chiral assemblies, how these large-scale structures emerge and relate to individual filament properties remains poorly understood. To understand this quantitatively, we combined minimal chiral active matter simulations with biochemical reconstitution experiments. Using STED and TIRF microscopy as well as high-speed AFM, we imaged the behavior of FtsZ filaments on different spatial scales. Simulations and experiments revealed that filament density and flexibility define the local and global order of the system: At intermediate densities, flexible filaments organize into chiral rings and polar bands, while an effectively nematic organization dominates for high filament densities and for mutant filaments with increased rigidity. Our predicted phase diagram captured these features quantitatively, demonstrating how filament flexibility, density and chirality cooperate with activity to give rise to a large repertoire of collective behaviors. These properties are likely important for the dynamic organization of soft chiral matter, including that of treadmilling FtsZ filaments during bacterial cell division.

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In vitro reconstitution of Escherichia coli divisome activation

Cited by 13 publications

References 49 publications

Anchors: A way for FtsZ filaments to stay membrane bound

Anchors: A way for FtsZ filaments to stay membrane bound

Self-organisation of mortal filaments: the role of FtsZ treadmilling in bacterial division ring formation

Chiral and nematic phases of flexible active filaments

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