The Antituberculosis Drug Ethambutol Selectively Blocks Apical Growth in CMN Group Bacteria

Schubert, Karin; Sieger, Boris; Giacomelli, Giacomo; Böhm, Kati; Rieblinger, Angela; Lindenthal, Laura; Sachs, Nadja; Wanner, Gerhard; Bramkamp, Marc

doi:10.1128/mbio.02213-16

Cited by 36 publications

(50 citation statements)

References 54 publications

(92 reference statements)

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“…We found that the multilayered cell envelope of C. glutamicum and M. smegmatis is assembled sequentially at the septum from PG to MM during cytokinesis, unlike the apparently synchronous assembly observed during polar growth, thus suggesting that C. glutamicum and M. smegmatis use two different mechanisms (and machineries) for envelope assembly during polar growth versus cytokinesis. This conclusion was further supported by our observations with EMB treatment, after which polar PG synthesis and cell elongation were stalled while septal PG synthesis and septation were not affected, in agreement with the observations in a recent report 39 . The sequential assembly of septal PG and MM in the mycolata contrasts substantially with the synchronous assembly of septal PG and OM in Gram-negative bacteria.…”

Section: Discussionsupporting

confidence: 93%

Sequential assembly of the septal cell envelope prior to V snapping in Corynebacterium glutamicum

et al. 2019

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Members of the Corynebacterineae, including Corynebacterium and Mycobacterium, have an atypical cell envelope characterized by an additional mycomembrane outside of the peptidoglycan layer. How this multilayered cell envelope is assembled remains unclear. Here, we tracked the assembly dynamics of different envelope layers in Corynebacterium glutamicum and Mycobacterium smegmatis by using metabolic labeling and found that the septal cell envelope is assembled sequentially in both species. Additionally, we demonstrate that in C. glutamicum, the peripheral peptidoglycan layer at the septal junction remains contiguous throughout septation, forming a diffusion barrier for the fluid mycomembrane. This diffusion barrier is resolved through perforations in the peripheral peptidoglycan, thus leading to the confluency of the mycomembrane before daughter cell separation (V snapping). Furthermore, the same junctional peptidoglycan also serves as a mechanical link holding the daughter cells together and undergoes mechanical fracture during V snapping. Finally, we show that normal V snapping in C. glutamicum depends on complete assembly of the septal cell envelope. Reprints and permissions information is available at www.nature.com/reprints.

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Section: Discussionsupporting

confidence: 93%

Sequential assembly of the septal cell envelope prior to V snapping in Corynebacterium glutamicum

et al. 2019

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“…Mycobacteria have been shown to expand from their poles ( Aldridge et al, 2012 ; Santi et al, 2013 ; Meniche et al, 2014 ; Thanky et al, 2007 ; Kieser and Rubin, 2014 ; Singh et al, 2013 ; Joyce et al, 2012 ) but published micrographs suggest that d -amino acid probes may label both the poles and sidewall of these organisms ( Meniche et al, 2014 ; Siegrist et al, 2013 ; Boutte et al, 2016 ; Botella et al, 2017 ; Schubert et al, 2017 ; Rodriguez-Rivera et al, 2018 ). Metabolic labeling can be achieved by a one-step process, in which the fluorophore is directly appended to the probe, or a two-step process in which a small chemical tag on the d -amino acid is detected by subsequent reaction with a fluorescent reactive partner ([ Siegrist et al, 2015 ], Figure 1A ).…”

Section: Resultsmentioning

confidence: 99%

“…Because cell wall peptidoglycan synthesis is critical for bacterial replication, it is often used to localize the sites of growth and division. Intriguingly, d -amino acid probes, which in other species have been shown to incorporate into peptidoglycan ( de Pedro et al, 1997 ; Kuru et al, 2012 ; Siegrist et al, 2013 ), label both the poles and sidewall of mycobacteria ( Meniche et al, 2014 ; Siegrist et al, 2013 ; Boutte et al, 2016 ; Botella et al, 2017 ; Schubert et al, 2017 ; Rodriguez-Rivera et al, 2018 ). The localization of these molecules is supported by the detection of peptidoglycan synthetic enzymes at the mycobacterial cell tips and periphery ( Meniche et al, 2014 ; Joyce et al, 2012 ; Hett et al, 2010 ; Kieser et al, 2015a ; Plocinski et al, 2011 ).…”

Section: Introductionmentioning

confidence: 99%

Peptidoglycan precursor synthesis along the sidewall of pole-growing mycobacteria

García‐Heredia

Pohane

Melzer

et al. 2018

eLife

100

116

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Rod-shaped mycobacteria expand from their poles, yet d-amino acid probes label cell wall peptidoglycan in this genus at both the poles and sidewall. We sought to clarify the metabolic fates of these probes. Monopeptide incorporation was decreased by antibiotics that block peptidoglycan synthesis or l,d-transpeptidation and in an l,d-transpeptidase mutant. Dipeptides complemented defects in d-alanine synthesis or ligation and were present in lipid-linked peptidoglycan precursors. Characterizing probe uptake pathways allowed us to localize peptidoglycan metabolism with precision: monopeptide-marked l,d-transpeptidase remodeling and dipeptide-marked synthesis were coincident with mycomembrane metabolism at the poles, septum and sidewall. Fluorescent pencillin-marked d,d-transpeptidation around the cell perimeter further suggested that the mycobacterial sidewall is a site of cell wall assembly. While polar peptidoglycan synthesis was associated with cell elongation, sidewall synthesis responded to cell wall damage. Peptidoglycan editing along the sidewall may support cell wall robustness in pole-growing mycobacteria.

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“…A promising candidate for uncovering novel growth modes is the Gram-positive Corynebacterium glutamicum. C. glutamicum is broadly used as a production-organism for amino-acids and vitamins and also serves as model organism for the taxonomic related human pathogens C. diphteriae and Mycobacterium tuberculosis (12)(13)(14). The cell wall of Corynebacteria and Mycobacteria is a complex polymer composed of a classical bacterial peptidoglycan (PG) sacculus that is covalently bound to an arabinogalactan (AG) layer (15).…”

Section: Main Textmentioning

confidence: 99%

Single-cell growth inference ofCorynebacterium glutamicumreveals asymptotically linear growth

Messelink

Bramkamp

Broedersz

2020

Preprint

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Keywords| cell-size homeostasis | single-cell growth| apical growth | RodA| cell wall SignificanceRegulation of growth and cell size is crucial for the optimization of bacterial cellular function. So far, single bacterial cells have been found to grow exponentially, which implies the need for tight regulation mechanisms to maintain cell size throughout growth and division cycles. Here, we characterize the growth behavior of the apically growing bacterium Corynebacterium glutamicum, by developing a novel and broadly applicable inference method for single-cell growth dynamics. We find that this bacterium grows asymptotically linearly, enabling it to maintain a narrow distribution of cell sizes, despite having a large variability of single-cell growth features. Our results imply a novel interplay between mode of growth and division regulation mechanisms, which may extend to other bacteria with non-exponential growth modes. AbstractIn many bacteria, protein mass production is thought to be rate limiting for growth, implying exponential growth at the single cell level. To maintain cell-size homeostasis in proliferating populations of exponentially growing bacteria, tight growth and division mechanisms are required.However, it remains unclear whether these considerations set universal physical limits to bacterial growth. Here, we characterize the growth dynamics of the actinobacterium Corynebacterium glutamicum -a promising candidate for uncovering novel growth modes. This bacterium exhibits apical cell wall synthesis and division site selection systems appear to be absent, as reflected by a broad distribution of division asymmetries. We develop a novel growth inference method that averages out measurement noise and single-cell variability to obtain elongation rate curves as a function of birth length. Using this approach, we find that C. glutamicum exhibits asymptotically linear single-cell growth. To explain this growth mode, we model elongation as being rate-limited by the apical growth mechanism mediated by cell wall transglycosylases. This model accurately reproduces the observed elongation rate curves, and we further validate the model with growth measurements on a transglycosylase deficient ΔrodA mutant. Finally, with simulations we show that asymptotically linear growth yields a narrower distribution of cell lengths, suggesting that this growth mode can act as a substitute for tight division length and division symmetry regulation.

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The Antituberculosis Drug Ethambutol Selectively Blocks Apical Growth in CMN Group Bacteria

Cited by 36 publications

References 54 publications

Sequential assembly of the septal cell envelope prior to V snapping in Corynebacterium glutamicum

Sequential assembly of the septal cell envelope prior to V snapping in Corynebacterium glutamicum

Peptidoglycan precursor synthesis along the sidewall of pole-growing mycobacteria

Single-cell growth inference ofCorynebacterium glutamicumreveals asymptotically linear growth

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