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- EMDB-16208: Cryo electron tomogram of Streptomyces coelicolor - Example 3 -

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Basic information

Entry
Database: EMDB / ID: EMD-16208
TitleCryo electron tomogram of Streptomyces coelicolor - Example 3
Map data
Sample
  • Complex: The contractile injection system in Streptomyces coelicolor
KeywordsSTRUCTURAL PROTEIN / Contractile injection system / cryoET
Biological speciesStreptomyces coelicolor A3(2) (bacteria)
Methodelectron tomography / cryo EM
AuthorsCasu B / Sallmen JW / Schlimpert S / Pilhofer M
Funding support Switzerland, European Union, United Kingdom, 3 items
OrganizationGrant numberCountry
Swiss National Science Foundation31003A_179255 Switzerland
European Research Council (ERC)679209European Union
Biotechnology and Biological Sciences Research Council (BBSRC)BB/T015349/1 United Kingdom
CitationJournal: Nat Microbiol / Year: 2023
Title: Cytoplasmic contractile injection systems mediate cell death in Streptomyces.
Authors: Bastien Casu / Joseph W Sallmen / Susan Schlimpert / Martin Pilhofer /
Abstract: Contractile injection systems (CIS) are bacteriophage tail-like structures that mediate bacterial cell-cell interactions. While CIS are highly abundant across diverse bacterial phyla, representative ...Contractile injection systems (CIS) are bacteriophage tail-like structures that mediate bacterial cell-cell interactions. While CIS are highly abundant across diverse bacterial phyla, representative gene clusters in Gram-positive organisms remain poorly studied. Here we characterize a CIS in the Gram-positive multicellular model organism Streptomyces coelicolor and show that, in contrast to most other CIS, S. coelicolor CIS (CIS) mediate cell death in response to stress and impact cellular development. CIS are expressed in the cytoplasm of vegetative hyphae and are not released into the medium. Our cryo-electron microscopy structure enabled the engineering of non-contractile and fluorescently tagged CIS assemblies. Cryo-electron tomography showed that CIS contraction is linked to reduced cellular integrity. Fluorescence light microscopy furthermore revealed that functional CIS mediate cell death upon encountering different types of stress. The absence of functional CIS had an impact on hyphal differentiation and secondary metabolite production. Finally, we identified three putative effector proteins, which when absent, phenocopied other CIS mutants. Our results provide new functional insights into CIS in Gram-positive organisms and a framework for studying novel intracellular roles, including regulated cell death and life-cycle progression in multicellular bacteria.
History
DepositionNov 22, 2022-
Header (metadata) releaseMar 15, 2023-
Map releaseMar 15, 2023-
UpdateDec 13, 2023-
Current statusDec 13, 2023Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_16208.png

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Map

FileDownload / File: emd_16208.map.gz / Format: CCP4 / Size: 510.5 MB / Type: IMAGE STORED AS SIGNED BYTE
Voxel sizeX=Y=Z: 10.74 Å
Density
Minimum - Maximum-128.0 - 127.0
Average (Standard dev.)-5.491258 (±21.400089999999999)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin-181208-209
Dimensions14401024363
Spacing10241440363
CellA: 10997.76 Å / B: 15465.6 Å / C: 3898.6199 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : The contractile injection system in Streptomyces coelicolor

EntireName: The contractile injection system in Streptomyces coelicolor
Components
  • Complex: The contractile injection system in Streptomyces coelicolor

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Supramolecule #1: The contractile injection system in Streptomyces coelicolor

SupramoleculeName: The contractile injection system in Streptomyces coelicolor
type: complex / ID: 1 / Parent: 0
Source (natural)Organism: Streptomyces coelicolor A3(2) (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: 7.5
VitrificationCryogen name: ETHANE-PROPANE / Instrument: FEI VITROBOT MARK IV
SectioningOther: NO SECTIONING
Fiducial markerManufacturer: cytodiagnostics / Diameter: 10 nm

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Number real images: 61 / Average electron dose: 2.1 e/Å2 / Details: 61 tilt series with around 130 accumulated dose.
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 8.0 µm / Nominal defocus min: 8.0 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Final reconstructionSoftware - Name: IMOD / Number images used: 61

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