Bacterial actin MreB forms antiparallel double filaments

Ent, F. van den; Izoré, Thierry; Bharat, Tanmay A. M.; Johnson, Christopher M.; Löwe, Jan

doi:10.7554/elife.02634

Cited by 156 publications

(360 citation statements)

References 93 publications

(152 reference statements)

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“…As the concentration of A22 is increased, MreB becomes localized to the membrane and more fluorescence is found in the cytoplasm. These results are consistent with previous work showing that MreB bound to A22 weakens interprotofilament contacts, thus decreasing the stability of MreB polymers (27,28).…”

Section: Mreb Polymers In A22-treated Cells and The Effect Of A22 On supporting

confidence: 93%

“…In cells treated with A22, the drug interacts with nucleotide-bound MreB to prevent nucleotide hydrolysis and destabilize filaments (28,32). A22 treatment and mutations perturbing the nucleotide-binding pocket of MreB have the potential to alter the geometry of the polymer as it has been shown that the angle between adjacent MreB monomers depends on the state of the bound nucleotide (34).…”

Section: Discussionmentioning

confidence: 99%

“…A22 has been shown to inhibit rod shape through a specific interaction with MreB (27,28). Growing E. coli in the presence of different sublethal concentrations of A22 resulted in the growth of cells with varying steady-state cell diameters (Fig.…”

Section: Generating Different Cell Shapes By Perturbing Mrebmentioning

confidence: 98%

“…Although high concentrations of A22 abolish MreB polymers (28,31), the effect of sublethal concentrations on MreB structure is not known. We measured MreB polymer length and the amount of MreB that appears to be on the surface, and we found that these quantities appeared to be inversely correlated with the concentration of A22 in wild-type cells when moderate amounts of the drug are used (Fig.…”

Section: Mreb Polymers In A22-treated Cells and The Effect Of A22 On mentioning

confidence: 99%

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MreB Orientation Correlates with Cell Diameter in Escherichia coli

Ouzounov

Nguyen

Bratton

et al. 2016

Biophysical Journal

127

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Bacteria have remarkably robust cell shape control mechanisms. For example, cell diameter only varies by a few percent across a given population. The bacterial actin homolog, MreB, is necessary for establishment and maintenance of rod shape although the detailed properties of MreB that are important for shape control remained unknown. In this study, we perturb MreB in two ways: by treating cells with the polymerization-inhibiting drug A22 and by creating point mutants in mreB. These perturbations modify the steady-state diameter of cells over a wide range, from 790 5 30 nm to 1700 5 20 nm. To determine which properties of MreB are important for diameter control, we correlated structural characteristics of fluorescently tagged MreB polymers with cell diameter by simultaneously analyzing three-dimensional images of MreB and cell shape. Our results indicate that the helical pitch angle of MreB inversely correlates with the cell diameter of Escherichia coli. Other correlations between MreB and cell diameter are not found to be significant. These results demonstrate that the physical properties of MreB filaments are important for shape control and support a model in which MreB organizes the cell wall growth machinery to produce a chiral cell wall structure and dictate cell diameter.

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Section: Mreb Polymers In A22-treated Cells and The Effect Of A22 On supporting

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Section: Generating Different Cell Shapes By Perturbing Mrebmentioning

confidence: 98%

Section: Mreb Polymers In A22-treated Cells and The Effect Of A22 On mentioning

confidence: 99%

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MreB Orientation Correlates with Cell Diameter in Escherichia coli

Ouzounov

Nguyen

Bratton

et al. 2016

Biophysical Journal

127

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“…S4A), which we suggest represents a previously unidentified MreB-binding site. Because RodZ 83 does lose rod shape, we propose that the helix-turn-helix interaction site may explain the rotation-independent morphogenesis roles of RodZ, whereas the juxta-membrane RodZ-MreB interaction site may enhance MreB rotation and explain the recent report that MreB is intimately membrane associated (21). The two MreB-binding sites may also partially functionally overlap as RodZ 83 does not fully phenocopy the ΔrodZ cell shape phenotype (SI Appendix, Fig.…”

Section: Rodz Mediates Mreb Rotation By Linking Cytoplasmic Mreb Tomentioning

confidence: 92%

RodZ links MreB to cell wall synthesis to mediate MreB rotation and robust morphogenesis

Morgenstein

Bratton

Nguyen

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

113

166

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The rod shape of most bacteria requires the actin homolog, MreB. Whereas MreB was initially thought to statically define rod shape, recent studies found that MreB dynamically rotates around the cell circumference dependent on cell wall synthesis. However, the mechanism by which cytoplasmic MreB is linked to extracytoplasmic cell wall synthesis and the function of this linkage for morphogenesis has remained unclear. Here we demonstrate that the transmembrane protein RodZ mediates MreB rotation by directly or indirectly coupling MreB to cell wall synthesis enzymes. Furthermore, we map the RodZ domains that link MreB to cell wall synthesis and identify mreB mutants that suppress the shape defect of ΔrodZ without restoring rotation, uncoupling rotation from rod-like growth. Surprisingly, MreB rotation is dispensable for rodlike shape determination under standard laboratory conditions but is required for the robustness of rod shape and growth under conditions of cell wall stress.bacterial cytoskeleton | bacterial cell shape | cell growth | cytoskeleton dynamics | robust rod shape B acterial cell shape is structurally determined by a rigid peptidoglycan (PG) cell wall built outside of the cytoplasmic membrane by a series of cell wall assembly enzymes (1). In many rod-shaped species these enzymes are coordinated by the actinlike protein, MreB, though the mechanism coupling this cytoplasmic protein to the extracellular cell wall enzymes and the specific functions executed by MreB have remained largely mysterious. Polymeric MreB is necessary to maintain rod-shaped cells, as inhibition of MreB polymerization or deletion of mreB cause cells to lose their rod shape. Initially, MreB was thought to form long helical structures that statically define rod shape (2, 3). Later, improved fluorescent fusion proteins and imaging methods revealed that MreB forms short polymers that dynamically rotate around the cell circumference (4-7).This circumferential rotation requires cell wall synthesis and is conserved across both Gram-negative and Gram-positive species (5-7), leading multiple groups to conclude that rotation promotes rod-shape formation. However, experimentally testing this hypothesis has proven difficult because all previous attempts to disrupt rotation have either led to cell death or massive cell shape changes, making it impossible to isolate the specific function of MreB rotation (5, 6). Furthermore, it remained difficult to explain the mechanistic link between cell wall growth and MreB rotation because of their separation in space by the cytoplasmic membrane. Here, we address both the coupling of MreB to cell wall synthesis and the function of MreB rotation. Results and DiscussionRodZ Rotates Similarly to MreB. We initially set out to identify proteins necessary for MreB rotation. In Escherichia coli, multiple proteins have been suggested to interact with MreB, including the penicillin binding protein (PBP) cell wall synthesis enzymes and RodZ, an integral membrane protein that directly binds MreB (8-11). PBP2 inhibitor...

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Cryo‐EM structure of the MinCD copolymeric filament from Pseudomonas aeruginosa at 3.1 Å resolution

2019

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Positioning of the division site in many bacterial species relies on the MinCDE system, which prevents the cytokinetic Z‐ring from assembling anywhere but the mid‐cell, through an oscillatory diffusion‐reaction mechanism. MinD dimers bind to membranes and, via their partner MinC, inhibit the polymerization of cell division protein FtsZ into the Z‐ring. MinC and MinD form polymeric assemblies in solution and on cell membranes. Here, we report the high‐resolution cryo‐EM structure of the copolymeric filaments of Pseudomonas aeruginosa MinCD. The filaments consist of three protofilaments made of alternating MinC and MinD dimers. The MinCD protofilaments are almost completely straight and assemble as single protofilaments on lipid membranes, which we also visualized by cryo‐EM.

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Bacterial actin MreB forms antiparallel double filaments

Cited by 156 publications

References 93 publications

MreB Orientation Correlates with Cell Diameter in Escherichia coli

MreB Orientation Correlates with Cell Diameter in Escherichia coli

RodZ links MreB to cell wall synthesis to mediate MreB rotation and robust morphogenesis

Cryo‐EM structure of the MinCD copolymeric filament from Pseudomonas aeruginosa at 3.1 Å resolution

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