In Vitro Reconstitution of Escherichia Coli Divisome Activation

Overview

Journal Nat Commun

Specialty Biology

Date 2022 May 13

PMID 35550516

Authors

Philipp Radler

Natalia Baranova

Paulo Caldas

Christoph Sommer

Mar Lopez-Pelegrin

David Michalik

Martin Loose

Affiliations

Soon will be listed here.

Abstract

The actin-homologue FtsA is essential for E. coli cell division, as it links FtsZ filaments in the Z-ring to transmembrane proteins. FtsA is thought to initiate cell constriction by switching from an inactive polymeric to an active monomeric conformation, which recruits downstream proteins and stabilizes the Z-ring. However, direct biochemical evidence for this mechanism is missing. Here, we use reconstitution experiments and quantitative fluorescence microscopy to study divisome activation in vitro. By comparing wild-type FtsA with FtsA R286W, we find that this hyperactive mutant outperforms FtsA WT in replicating FtsZ treadmilling dynamics, FtsZ filament stabilization and recruitment of FtsN. We could attribute these differences to a faster exchange and denser packing of FtsA R286W below FtsZ filaments. Using FRET microscopy, we also find that FtsN binding promotes FtsA self-interaction. We propose that in the active divisome FtsA and FtsN exist as a dynamic copolymer that follows treadmilling filaments of FtsZ.

Citing Articles

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

Mannik J, Kar P, Amarasinghe C, Amir A, Mannik J Nat Commun. 2024; 15(1):9948.

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Self-organization of mortal filaments and its role in bacterial division ring formation.

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Role of the antiparallel double-stranded filament form of FtsA in activating the divisome.

Perkins A, Mounange-Badimi M, Margolin W mBio. 2024; 15(8):e0168724.

PMID: 39041810 PMC: 11323482. DOI: 10.1128/mbio.01687-24.

Role of the antiparallel double-stranded filament form of FtsA in activating the divisome.

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