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- EMDB-50944: Cryo electron tomogram of Streptomyces coelicolor - ScoDelta4242 ... -

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Basic information

Entry
Database: EMDB / ID: EMD-50944
TitleCryo electron tomogram of Streptomyces coelicolor - ScoDelta4242 complemented membrane protein CisA strain - Ghost cells
Map data
Sample
  • Complex: The contractile injection system in Streptomyces coelicolor
KeywordsContractile injection system / cryoET / STRUCTURAL PROTEIN
Biological speciesStreptomyces coelicolor A3(2) (bacteria)
Methodelectron tomography / cryo EM
AuthorsCasu B / Sallmen JW / Hass PE / Afanasyev P / Xu J / Schlimpert S / Pilhofer M
Funding support Switzerland, European Union, United Kingdom, 4 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
Biotechnology and Biological Sciences Research Council (BBSRC)BB/X01097X/1 United Kingdom
CitationJournal: Elife / Year: 2025
Title: Function and firing of the contractile injection system requires the membrane protein CisA.
Authors: Bastien Casu / Joseph W Sallmen / Peter E Haas / Govind Chandra / Pavel Afanasyev / Jingwei Xu / Martin Pilhofer / Susan Schlimpert /
Abstract: Bacterial contractile injection systems (CIS) are phage tail-like macromolecular complexes that mediate cell-cell interactions by injecting effector proteins into target cells. CIS from (CIS) are ...Bacterial contractile injection systems (CIS) are phage tail-like macromolecular complexes that mediate cell-cell interactions by injecting effector proteins into target cells. CIS from (CIS) are localized in the cytoplasm. Under stress, they induce cell death and impact the life cycle. It remains unknown, however, whether CIS require accessory proteins to directly interact with the cytoplasmic membrane to function. Here, we characterize the putative membrane adaptor CisA, a conserved factor in CIS gene clusters across species. We show by cryo-electron tomography imaging and in vivo assays that CIS contraction and function depend on CisA. Using single-particle cryo-electron microscopy, we provide an atomic model of the extended CIS apparatus; however, CisA is not part of the complex. Instead, our findings show that CisA is a membrane protein with a cytoplasmic N-terminus predicted to interact with CIS components, thereby providing a possible mechanism for mediating CIS recruitment to the membrane and subsequent firing. Our work shows that CIS function in multicellular bacteria is distinct from type VI secretion systems and extracellular CIS, and possibly evolved due to the role CIS play in regulated cell death.
History
DepositionJul 8, 2024-
Header (metadata) releaseJun 18, 2025-
Map releaseJun 18, 2025-
UpdateJul 16, 2025-
Current statusJul 16, 2025Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_50944.png

Downloads & links

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Map

FileDownload / File: emd_50944.map.gz / Format: CCP4 / Size: 340.3 MB / Type: IMAGE STORED AS SIGNED BYTE
Voxel sizeX=Y=Z: 10.74 Å
Density
Minimum - Maximum-128.0 - 127.0
Average (Standard dev.)-3.6228225 (±22.868728999999998)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin209-208121
Dimensions14401024242
Spacing10241440242
CellA: 10997.76 Å / B: 15465.6 Å / C: 2599.0798 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 K3 BIOQUANTUM (6k x 4k) / 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
CTF correctionType: NONE

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