A High-Content Microscopy Screening Identifies New Genes Involved in Cell Width Control in Bacillus subtilis

Juillot, Dimitri; Cornilleau, Charlène; Deboosère, Nathalie; Billaudeau, Cyrille; Evouna-Mengue, Parfait; Lejard, Véronique; Brodin, Priscille; Carballido-López, Rut; Chastanet, Arnaud

doi:10.1128/msystems.01017-21

Cited by 10 publications

(5 citation statements)

References 78 publications

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“…subtilis across the cell cycle and growth conditions (63) facilitates correction for membrane curvature. Second, Gram-positive bacteria lack an outer membrane.…”

Section: Discussionmentioning

confidence: 99%

Quantification of membrane fluidity in bacteria using TIR-FCS

Barbotin,

Billaudeau,

Sezgin

et al. 2024

Biophysical Journal

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“…subtilis across the cell cycle and growth conditions (63) facilitates correction for membrane curvature. Second, Gram-positive bacteria lack an outer membrane.…”

Section: Discussionmentioning

confidence: 99%

Quantification of membrane fluidity in bacteria using TIR-FCS

Barbotin,

Billaudeau,

Sezgin

et al. 2024

Biophysical Journal

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“…Since antibiotic dilution predominantly depends on cell width ( equation 5 ), a central question is how width is determined ( 36 – 38 ) and how fast width is remodeled in response to antibiotic treatments. During steady-state growth, cell width is one of the most tightly controlled cellular parameters in both E. coli and B. subtilis ( 39 , 40 ) such that fluctuations in width are restored within 4 to 5 generations ( 17 ), commensurate with the typical time necessary for bacterial width remodeling ( 41 ). Under non-steady-state conditions, the cell width reaches a new equilibrium only within a few generations after nutrient shifts ( 42 ) or antibiotic exposure ( 15 , 18 ).…”

Section: Discussionmentioning

confidence: 99%

Antibiotic Resistance via Bacterial Cell Shape-Shifting

Ojkic

Serbanescu

Banerjee

2022

mBio

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Bacteria have evolved to develop multiple strategies for antibiotic resistance by effectively reducing intracellular antibiotic concentrations or antibiotic binding affinities, but the role of cell morphology in antibiotic resistance remains poorly understood. By analyzing cell morphological data for different bacterial species under antibiotic stress, we find that bacteria increase or decrease the cell surface-to-volume ratio depending on the antibiotic target.

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“…Although the mRNA counts achieved for S. aureus biofilm are less than those reported for another widely studied gram-positive pathogen, Bacillus subtilis, where 200-300 mRNA reads per cell were captured during planktonic growth, 20,22 this discrepancy can be explained by its larger cell volume (~4-6X) compared to S. aureus. [41][42][43] Studies from eukaryotic scRNA-seq have established that low-coverage sequencing is sufficient to fully capture sample heterogeneity within a large number of cells. [44][45][46][47] Quality control measures throughout the BaSSSh-seq process are presented in the Methods and Figure S1B-E.…”

Section: Basssh-seq Enables Bacterial Scrna-seq Of Biofilm and Incorp...mentioning

confidence: 99%

Bacterial single-cell RNA sequencing captures biofilm transcriptional heterogeneity and differential responses to immune pressure

Korshoj,

Kielian

2024

Preprint

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Biofilm formation is an important mechanism of survival and persistence for many bacterial pathogens. These multicellular communities contain subpopulations of cells that display vast metabolic and transcriptional diversity along with high recalcitrance to antibiotics and host immune defenses. Investigating the complex heterogeneity within biofilm has been hindered by the lack of a sensitive and high-throughput method to assess stochastic transcriptional activity and regulation between bacterial subpopulations, which requires single-cell resolution. We have developed an optimized bacterial single-cell RNA sequencing method, BaSSSh-seq, to studyStaphylococcus aureusdiversity during biofilm growth and transcriptional adaptations following immune cell exposure. We validated the ability of BaSSSh-seq to capture extensive transcriptional heterogeneity during biofilm compared to planktonic growth. Application of new computational tools revealed transcriptional regulatory networks across the heterogeneous biofilm subpopulations and identification of gene sets that were associated with a trajectory from planktonic to biofilm growth. BaSSSh-seq also detected alterations in biofilm metabolism, stress response, and virulence that were tailored to distinct immune cell populations. This work provides an innovative platform to explore biofilm dynamics at single-cell resolution, unlocking the potential for identifying biofilm adaptations to environmental signals and immune pressure.

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A High-Content Microscopy Screening Identifies New Genes Involved in Cell Width Control in Bacillus subtilis

Abstract: Bacterial shape is primarily dictated by the external cell wall, a vital structure that, as such, is the target of countless antibiotics. Our understanding of how bacteria synthesize and maintain this structure is therefore a cardinal question for both basic and applied research.

Cited by 10 publications

References 78 publications

Quantification of membrane fluidity in bacteria using TIR-FCS

Quantification of membrane fluidity in bacteria using TIR-FCS

Antibiotic Resistance via Bacterial Cell Shape-Shifting

Bacterial single-cell RNA sequencing captures biofilm transcriptional heterogeneity and differential responses to immune pressure

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