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- EMDB-16288: Cryotomogram of endolysin Ply006 treated Listeria monocytogenes R... -

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Basic information

Entry
Database: EMDB / ID: EMD-16288
TitleCryotomogram of endolysin Ply006 treated Listeria monocytogenes Rev2 cell (stage 3)
Map dataCryo-tomogram
Sample
  • Cell: Cryotomogram of endolysin Ply006 treated Listeria monocytogenes Rev2 cell (stage 3)
Biological speciesListeria monocytogenes (bacteria)
Methodelectron tomography / cryo EM
AuthorsFeldmueller M / Wohlfarth JC / Loessner M / Pilhofer M
Funding support Switzerland, 1 items
OrganizationGrant numberCountry
Swiss National Science Foundation31003A_170042 Switzerland
CitationJournal: Nat Microbiol / Year: 2023
Title: L-form conversion in Gram-positive bacteria enables escape from phage infection.
Authors: Jan C Wohlfarth / Miki Feldmüller / Alissa Schneller / Samuel Kilcher / Marco Burkolter / Susanne Meile / Martin Pilhofer / Markus Schuppler / Martin J Loessner /
Abstract: At the end of a lytic bacteriophage replication cycle in Gram-positive bacteria, peptidoglycan-degrading endolysins that cause explosive cell lysis of the host can also attack non-infected bystander ...At the end of a lytic bacteriophage replication cycle in Gram-positive bacteria, peptidoglycan-degrading endolysins that cause explosive cell lysis of the host can also attack non-infected bystander cells. Here we show that in osmotically stabilized environments, Listeria monocytogenes can evade phage predation by transient conversion to a cell wall-deficient L-form state. This L-form escape is triggered by endolysins disintegrating the cell wall from without, leading to turgor-driven extrusion of wall-deficient, yet viable L-form cells. Remarkably, in the absence of phage predation, we show that L-forms can quickly revert to the walled state. These findings suggest that L-form conversion represents a population-level persistence mechanism to evade complete eradication by phage attack. Importantly, we also demonstrate phage-mediated L-form switching of the urinary tract pathogen Enterococcus faecalis in human urine, which underscores that this escape route may be widespread and has important implications for phage- and endolysin-based therapeutic interventions.
History
DepositionDec 6, 2022-
Header (metadata) releaseFeb 8, 2023-
Map releaseFeb 8, 2023-
UpdateMar 22, 2023-
Current statusMar 22, 2023Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_16288.png

Downloads & links

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Map

FileDownload / File: emd_16288.map.gz / Format: CCP4 / Size: 531 MB / Type: IMAGE STORED AS SIGNED BYTE
AnnotationCryo-tomogram
Voxel sizeX=Y=Z: 11 Å
Density
Minimum - Maximum-128.0 - 127.0
Average (Standard dev.)-11.116086 (±23.35065)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin01313
Dimensions928960625
Spacing960928625
CellA: 10560.0 Å / B: 10208.0 Å / C: 6875.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : Cryotomogram of endolysin Ply006 treated Listeria monocytogenes R...

EntireName: Cryotomogram of endolysin Ply006 treated Listeria monocytogenes Rev2 cell (stage 3)
Components
  • Cell: Cryotomogram of endolysin Ply006 treated Listeria monocytogenes Rev2 cell (stage 3)

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Supramolecule #1: Cryotomogram of endolysin Ply006 treated Listeria monocytogenes R...

SupramoleculeName: Cryotomogram of endolysin Ply006 treated Listeria monocytogenes Rev2 cell (stage 3)
type: cell / ID: 1 / Parent: 0
Source (natural)Organism: Listeria monocytogenes (bacteria)

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Experimental details

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Structure determination

Methodcryo EM
Processingelectron tomography
Aggregation statecell

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Sample preparation

BufferpH: 7.3
VitrificationCryogen name: ETHANE-PROPANE
SectioningOther: NO SECTIONING
Fiducial markerManufacturer: Sigma-Aldrich / Diameter: 10 nm

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 135.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 9.0 µm / Nominal defocus min: 9.0 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Final reconstructionNumber images used: 61

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