Coupled, Circumferential Motions of the Cell Wall Synthesis Machinery and MreB Filaments in
            <i>B. subtilis</i>

Garner, Ethan C.; Bernard, Rémi; Wang, Wen-Qin; Zhuang, Xiaowei; Rudner, David Z.; Mitchison, Tim

doi:10.1126/science.1203285

Cited by 497 publications

(602 citation statements)

References 19 publications

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“…S4). Although not explored here, the MreB polymers we observe are dynamic and move circumferentially over long-time scales as reported previously (7,25,26).…”

Section: Mreb Msfgfp Is Minimally Perturbativesupporting

confidence: 73%

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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“…S4). Although not explored here, the MreB polymers we observe are dynamic and move circumferentially over long-time scales as reported previously (7,25,26).…”

Section: Mreb Msfgfp Is Minimally Perturbativesupporting

confidence: 73%

MreB Orientation Correlates with Cell Diameter in Escherichia coli

Ouzounov

Nguyen

Bratton

et al. 2016

Biophysical Journal

127

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“…It is thus plausible that MreB filaments are physically linked to the enzymes responsible for cell-wall insertion. In fact, some of the cellwall-synthesis enzymes have been seen to move in a similar manner as MreB filaments in the Gram-positive B. subtilis (Garner et al 2011;Domínguez-Escobar et al 2011), supporting the hypothesis of physical interaction. In Gram-negative E. coli, at least one important synthesising enzyme, the transpeptidase PBP2, moves rapidly and diffusively, showing no processivity on the sub-second time scale (Lee et al 2014), thus suggesting a more transient interaction of the cell-wall synthesis proteins.…”

Section: Necessity For Regulationmentioning

confidence: 57%

“…The length of the MreB filaments is currently under debate, in particular because native-expression-level MreB filaments have not been detected in whole cells by electron microscopy (Swulius and Jensen 2012). Irrespective of their exact length, it has been shown by fluorescence microscopy that MreB filaments rotate around the long cell axis in a processive manner in Gram-negative (Teeffelen et al 2011) and Gram-positive bacteria (Garner et al 2011;Domín-guez-Escobar et al 2011;Olshausen et al 2013;Reimold et al 2013). This rotation depends on PG synthesis and proceeds at a speed compatible with processive insertion of single glycan strands into the PG meshwork (Teeffelen et al 2011), as already suggested by Burmann and Park in the 1980s (Burman and Park 1984).…”

Section: Necessity For Regulationmentioning

confidence: 99%

Getting into shape: How do rod-like bacteria control their geometry?

Amir¹,

Teeffelen²

2014

Syst Synth Biol

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Rod-like bacteria maintain their cylindrical shapes with remarkable precision during growth. However, they are also capable to adapt their shapes to external forces and constraints, for example by growing into narrow or curved confinements. Despite being one of the simplest morphologies, we are still far from a full understanding of how shape is robustly regulated, and how bacteria obtain their near-perfect cylindrical shapes with excellent precision. However, recent experimental and theoretical findings suggest that cell-wall geometry and mechanical stress play important roles in regulating cell shape in rod-like bacteria. We review our current understanding of the cell wall architecture and the growth dynamics, and discuss possible candidates for regulatory cues of shape regulation in the absence or presence of external constraints. Finally, we suggest further future experimental and theoretical directions which may help to shed light on this fundamental problem.

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“…5B and C). Peaks of fluorescence, corresponding to the sites of PG insertion, were evenly distributed with an average spacing between approximately 0.5 and 1 μm, consistent with the reported spacing between the tracks followed by MreB‐associated PG synthetic machineries along the sidewalls of B. subtilis (Dominguez‐Escobar et al ., 2011; Garner et al ., 2011). …”

Section: Resultsmentioning

confidence: 99%

Hydrolysis of peptidoglycan is modulated by amidation of meso‐diaminopimelic acid and Mg²⁺ in Bacillus subtilis

Dajkovic

Tesson

Chauhan

et al. 2017

Molecular Microbiology

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SummaryThe ability of excess Mg2+ to compensate the absence of cell wall related genes in Bacillus subtilis has been known for a long time, but the mechanism has remained obscure. Here, we show that the rigidity of wild‐type cells remains unaffected with excess Mg2+, but the proportion of amidated meso‐diaminopimelic (mDAP) acid in their peptidoglycan (PG) is significantly reduced. We identify the amidotransferase AsnB as responsible for mDAP amidation and show that the gene encoding it is essential without added Mg2+. Growth without excess Mg2+ causes ΔasnB mutant cells to deform and ultimately lyse. In cell regions with deformations, PG insertion is orderly and indistinguishable from the wild‐type. However, PG degradation is unevenly distributed along the sidewalls. Furthermore, ΔasnB mutant cells exhibit increased sensitivity to antibiotics targeting the cell wall. These results suggest that absence of amidated mDAP causes a lethal deregulation of PG hydrolysis that can be inhibited by increased levels of Mg2+. Consistently, we find that Mg2+ inhibits autolysis of wild‐type cells. We suggest that Mg2+ helps to maintain the balance between PG synthesis and hydrolysis in cell wall mutants where this balance is perturbed in favor of increased degradation.

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Coupled, Circumferential Motions of the Cell Wall Synthesis Machinery and MreB Filaments in B. subtilis

Cited by 497 publications

References 19 publications

MreB Orientation Correlates with Cell Diameter in Escherichia coli

MreB Orientation Correlates with Cell Diameter in Escherichia coli

Getting into shape: How do rod-like bacteria control their geometry?

Hydrolysis of peptidoglycan is modulated by amidation of meso‐diaminopimelic acid and Mg²⁺ in Bacillus subtilis

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Coupled, Circumferential Motions of the Cell Wall Synthesis Machinery and MreB Filaments in B. subtilis

Cited by 497 publications

References 19 publications

MreB Orientation Correlates with Cell Diameter in Escherichia coli

MreB Orientation Correlates with Cell Diameter in Escherichia coli

Getting into shape: How do rod-like bacteria control their geometry?

Hydrolysis of peptidoglycan is modulated by amidation of meso‐diaminopimelic acid and Mg2+ in Bacillus subtilis

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Hydrolysis of peptidoglycan is modulated by amidation of meso‐diaminopimelic acid and Mg²⁺ in Bacillus subtilis