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- EMDB-16479: Cryo-electron tomogram of a Roseburia intestinalis DSM 14610 cell. -

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Basic information

Entry
Database: EMDB / ID: EMD-16479
TitleCryo-electron tomogram of a Roseburia intestinalis DSM 14610 cell.
Map dataA cryo-electron tomogram of an intact Roseburia intestinalis DSM 14610 cell.
Sample
  • Cell: Roseburia intestinalis DSM 14610
KeywordsBacteria / Roseburia / UNKNOWN FUNCTION
Biological speciesRoseburia intestinalis L1-82 (bacteria)
Methodelectron tomography / cryo EM
AuthorsWimmer BH / Medalia O
Funding support1 items
OrganizationGrant numberCountry
Not funded
CitationJournal: Microlife / Year: 2023
Title: Phylogenetic diversity of core rumen microbiota as described by cryo-ET.
Authors: Benedikt H Wimmer / Sarah Moraïs / Ran Zalk / Itzhak Mizrahi / Ohad Medalia /
Abstract: Microbial taxonomy is critical for describing ecosystem composition, yet the link between taxonomy and properties of microbes, such as their cellular architecture, remains poorly defined. We ...Microbial taxonomy is critical for describing ecosystem composition, yet the link between taxonomy and properties of microbes, such as their cellular architecture, remains poorly defined. We hypothesized that the cellular architecture represents microbial niche adaptation. We used cryo-electron microscopy and tomography to analyze microbial morphology in order to associate cellular architecture with phylogeny and genomic contents. As a model system, we chose the core rumen microbiome and imaged a large isolate collection covering 90% of its richness at the order level. Based on quantifications of several morphological features, we found that the visual similarity of microbiota is significantly related to their phylogenetic distance. Up to the level, closely related microbes have similar cellular architectures, which are highly correlated with genome similarity. However, in more distantly related bacteria, the correlation both with taxonomy and genome similarity is lost. This is the first comprehensive study of microbial cellular architecture and our results highlight that structure remains an important parameter in classification of microorganisms, along with functional parameters such as metabolomics. Furthermore, the high-quality images presented in this study represent a reference database for the identification of bacteria in anaerobic ecosystems.
History
DepositionJan 20, 2023-
Header (metadata) releaseApr 5, 2023-
Map releaseApr 5, 2023-
UpdateDec 13, 2023-
Current statusDec 13, 2023Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_16479.png

Downloads & links

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Map

FileDownload / File: emd_16479.map.gz / Format: CCP4 / Size: 401 MB / Type: IMAGE STORED AS SIGNED BYTE
AnnotationA cryo-electron tomogram of an intact Roseburia intestinalis DSM 14610 cell.
Voxel sizeX=Y=Z: 8.839 Å
Density
Minimum - Maximum-128.0 - 127.0
Average (Standard dev.)-1.4908445 (±27.767761)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin16-16-236
Dimensions928960472
Spacing960928472
CellA: 8485.439 Å / B: 8202.592 Å / C: 4172.008 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : Roseburia intestinalis DSM 14610

EntireName: Roseburia intestinalis DSM 14610
Components
  • Cell: Roseburia intestinalis DSM 14610

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Supramolecule #1: Roseburia intestinalis DSM 14610

SupramoleculeName: Roseburia intestinalis DSM 14610 / type: cell / ID: 1 / Parent: 0
Source (natural)Organism: Roseburia intestinalis L1-82 (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: 7.4 / Details: TBS
GridModel: Quantifoil R2/1 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: PLASMA CLEANING / Pretreatment - Time: 25 sec.
VitrificationCryogen name: ETHANE / Instrument: HOMEMADE PLUNGER / Details: blotting 3 - 4s.
Detailsdiluted to OD = 0.15, grown in YCFA + 1% Glucose
SectioningOther: NO SECTIONING
Fiducial markerManufacturer: Aurion / 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: 41 / Average electron dose: 3.4 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 6.0 µm / Nominal defocus min: 6.0 µm / Nominal magnification: 64000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

DetailsIMOD reconstruction
Final reconstructionAlgorithm: BACK PROJECTION / Software - Name: IMOD / Number images used: 41

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