MreB Orientation Correlates with Cell Diameter in Escherichia coli

Ouzounov, Nikolay; Nguyen, Jeffrey P.; Bratton, Benjamin P.; Jacobowitz, David M.; Gitai, Zemer; Shaevitz, Joshua W.

doi:10.1016/j.bpj.2016.07.017

Cited by 93 publications

(138 citation statements)

References 39 publications

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“…In all cases, the cells producing mCherry‐tagged MreB were slightly thicker than the corresponding untagged MreB producing cells. This is consistent with the previous reports that cells producing fluorescent protein‐tagged MreB are slightly thicker than cells producing untagged MreB (Billaudeau et al, ; Ouzounov et al, ). However, the general tendency was that cells producing MreB A174T were thicker and cells producing MreB A125V were thinner than WT cells.…”

Section: Resultssupporting

confidence: 93%

“…Growth and morphology of cells producing MreB‐mCherry SW were comparable with those of WT cells (Bendezú et al, ). During this project, it was reported that msfGFP does not affect the function of MreB and thus msfGFP is a better choice than mCherry for fusion with MreB (Ouzounov et al, ). Thus, we re‐evaluated the effect of fusion of MreB with mCherry by analyzing the length, width and sensitivity to A22 of cells producing MreB mutants tagged with mCherry or untagged (Supporting Information Figures S1 and S2, Table S1).…”

Section: Resultsmentioning

confidence: 99%

“…It is noteworthy that the width of cells producing MreB A125V or MreB A174T was different from that of cells producing MreB. It has been shown that the orientation of MreB polymers correlates with the cell width (Ouzounov et al, ; Tropini et al, ), but not with polymer number and polymer size (Ouzounov et al, ). Although cells become thicker by treatment with mecillinam or A22, MreB speed decreased if cells were treated with mecillinam whereas it was not affected by treatment with A22 (Tropini et al, ).…”

Section: Introductionmentioning

confidence: 99%

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Relation between rotation of MreB actin and cell width of Escherichia coli

et al. 2019

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Bacterial cells, including Escherichia coli and Bacillus subtilis, continuously elongate and divide. Although the cell width is maintained during cell cycle, the molecular mechanisms involved in its regulation remain unknown. MreB has been implicated to play a role in maintaining cell width. Several point mutations in mreB that affect cell width have been identified. The MreB protein forms clusters or polymers in the cell and moves along annular tracks perpendicular to the long axis. This rotation is coupled with peptidoglycan synthesis. Here, we focused on two MreB mutants, MreBA125V and MreBA174T. Cells producing MreBA125V and MreBA174T were thinner and thicker than WT cells, and MreBA125V and MreBA174T rotated faster and slower than WT MreB, respectively. We observed that the rotation rate correlated with the cell wall synthesis rate. Thus, we conclude that the velocity of MreB rotation also affects cell width, that is, the faster the MreB rotates, the thinner the cell width is.

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Section: Resultssupporting

confidence: 93%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Relation between rotation of MreB actin and cell width of Escherichia coli

et al. 2019

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“…100 MreB monomers polymerize into filaments that rotate circumferentially around the long axis of cells. 101–104 E. coli MreB is positioned in contact with the cytoplasmic membrane through an N-terminal amphipathic helix 105 and has been hypothesized to localize a protein complex containing proteins that perform peptidoglycan synthesis and degradation to regions of the cell wall with negative curvature. 106,107 The directed motion of MreB in these regions of the cell is correlated with peptidoglycan assembly and enables cells to maintain a rod shape, 107 as inhibiting MreB function causes cells to gradually change shape from a rod to a sphere.…”

Section: Bacterial Proteins That Hypothetically Contribute To Cell Stmentioning

confidence: 99%

Bacterial Cell Mechanics

2017

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Cellular mechanical properties play an integral role in bacterial survival and adaptation. Historically, the bacterial cell wall and, in particular, the layer of polymeric material called the peptidoglycan were the elements to which cell mechanics could be primarily attributed. Disrupting the biochemical machinery that assembles the peptidoglycan (e.g., using the β-lactam family of antibiotics) alters the structure of this material, leads to mechanical defects, and results in cell lysis. Decades after the discovery of peptidoglycan-synthesizing enzymes, the mechanisms that underlie their positioning and regulation are still not entirely understood. In addition, recent evidence suggests a diverse group of other biochemical elements influence bacterial cell mechanics, may be regulated by new cellular mechanisms, and may be triggered in different environmental contexts to enable cell adaptation and survival. This review summarizes the contributions that different biomolecular components of the cell wall (e.g., lipopolysaccharides, wall and lipoteichoic acids, lipid bilayers, peptidoglycan, and proteins) make to Gram-negative and Gram-positive bacterial cell mechanics. We discuss the contribution of individual proteins and macromolecular complexes in cell mechanics and the tools that make it possible to quantitatively decipher the biochemical machinery that contributes to bacterial cell mechanics. Advances in this area may provide insight into new biology and influence the development of antibacterial chemotherapies.

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“…However, Γ values are only directly comparable if two particles have the same α. Therefore, to directly compare the speed over short distances of particles with different α, we report τ* as the time it takes a particle to explore a set area (0.5 μm, the typical size of an MreB focus) (16).…”

Section: Significancementioning

confidence: 99%

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.

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111

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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MreB Orientation Correlates with Cell Diameter in Escherichia coli

Cited by 93 publications

References 39 publications

Relation between rotation of MreB actin and cell width of Escherichia coli

Relation between rotation of MreB actin and cell width of Escherichia coli

Bacterial Cell Mechanics

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

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