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- EMDB-2641: Cryo-electron tomogram of Chlamydia trachomatis in contact with a... -

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Basic information

Entry
Database: EMDB / ID: EMD-2641
TitleCryo-electron tomogram of Chlamydia trachomatis in contact with a eukaryotic host cell (U2OS).
Map dataChlamydia trachomatis elementary body in contact with a eukaryotic U2OS cell
Sample
  • Sample: Chlamydia trachomatis elementary bodies in contact with eukaryotic U2OS cell
  • Organelle or cellular component: Chlamydia trachomatis elementary bodies and a U2OS cell
KeywordsChlamydia / type III secretion system / injectisome
Biological speciesChlamydia trachomatis (bacteria)
Methodelectron tomography / cryo EM
AuthorsNans A / Saibil HR / Hayward RD
CitationJournal: Cell Microbiol / Year: 2014
Title: Pathogen-host reorganization during Chlamydia invasion revealed by cryo-electron tomography.
Authors: Andrea Nans / Helen R Saibil / Richard D Hayward /
Abstract: Invasion of host cells is a key early event during bacterial infection, but the underlying pathogen-host interactions are yet to be fully visualized in three-dimensional detail. We have captured ...Invasion of host cells is a key early event during bacterial infection, but the underlying pathogen-host interactions are yet to be fully visualized in three-dimensional detail. We have captured snapshots of the early stages of bacterial-mediated endocytosis in situ by exploiting the small size of chlamydial elementary bodies (EBs) for whole-cell cryo-electron tomography. Chlamydiae are obligate intracellular bacteria that infect eukaryotic cells and cause sexually transmitted infections and trachoma, the leading cause of preventable blindness. We demonstrate that Chlamydia trachomatis LGV2 EBs are intrinsically polarized. One pole is characterized by a tubular inner membrane invagination, while the other exhibits asymmetric periplasmic expansion to accommodate an array of type III secretion systems (T3SSs). Strikingly, EBs orient with their T3SS-containing pole facing target cells, enabling the T3SSs to directly contact the cellular plasma membrane. This contact induces enveloping macropinosomes, actin-rich filopodia and phagocytic cups to zipper tightly around the internalizing bacteria. Once encapsulated into tight early vacuoles, EB polarity and the T3SSs are lost. Our findings reveal previously undescribed structural transitions in both pathogen and host during the initial steps of chlamydial invasion.
History
DepositionMay 12, 2014-
Header (metadata) releaseJun 11, 2014-
Map releaseJun 18, 2014-
UpdateOct 1, 2014-
Current statusOct 1, 2014Processing site: PDBe / Status: Released

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Structure visualization

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Structure viewerEM map:
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Supplemental images

emd_2641.tif

emd_2641_1.tif

emd_2641_1_1...

emd_2641_2.tif

Downloads & links

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Map

FileDownload / File: emd_2641.map.gz / Format: CCP4 / Size: 3.6 GB / Type: IMAGE STORED AS SIGNED INTEGER (2 BYTES)
AnnotationChlamydia trachomatis elementary body in contact with a eukaryotic U2OS cell
Voxel sizeX=Y=Z: 7.1 Å
Density
Minimum - Maximum0.0 - 32767.0
Average (Standard dev.)16495.9921875 (±1657.878784179999911)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin00215
Dimensions20482048477
Spacing20482048477
CellA: 14540.8 Å / B: 14540.8 Å / C: 3386.7 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Integer*27
Å/pix. X/Y/Z7.17.17.1
M x/y/z20482048477
origin x/y/z0.0000.0000.000
length x/y/z14540.80014540.8003386.700
α/β/γ90.00090.00090.000
start NX/NY/NZ0-51-100
NX/NY/NZ82103201
MAP C/R/S123
start NC/NR/NS00215
NC/NR/NS20482048477
D min/max/mean0.00032767.00016495.992

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Supplemental data

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Sample components

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Entire : Chlamydia trachomatis elementary bodies in contact with eukaryoti...

EntireName: Chlamydia trachomatis elementary bodies in contact with eukaryotic U2OS cell
Components
  • Sample: Chlamydia trachomatis elementary bodies in contact with eukaryotic U2OS cell
  • Organelle or cellular component: Chlamydia trachomatis elementary bodies and a U2OS cell

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Supramolecule #1000: Chlamydia trachomatis elementary bodies in contact with eukaryoti...

SupramoleculeName: Chlamydia trachomatis elementary bodies in contact with eukaryotic U2OS cell
type: sample / ID: 1000 / Number unique components: 2

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Supramolecule #1: Chlamydia trachomatis elementary bodies and a U2OS cell

SupramoleculeName: Chlamydia trachomatis elementary bodies and a U2OS cell
type: organelle_or_cellular_component / ID: 1 / Recombinant expression: No / Database: NCBI
Source (natural)Organism: Chlamydia trachomatis (bacteria) / Strain: LGV 2 / Cell: U2OS

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Experimental details

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Structure determination

Methodcryo EM
Processingelectron tomography
Aggregation statecell

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Sample preparation

GridDetails: Quantifoil 200 mesh gold (R 3.5/1) grids
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Instrument: FEI VITROBOT MARK III / Method: Blot for 10 seconds before plunging

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Electron microscopy

MicroscopeFEI POLARA 300
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 12.0 µm / Nominal magnification: 29000
Sample stageSpecimen holder model: OTHER / Tilt series - Axis1 - Min angle: -52 ° / Tilt series - Axis1 - Max angle: 42 ° / Tilt series - Axis1 - Angle increment: 2 °
DateApr 11, 2013
Image recordingCategory: CCD / Film or detector model: GATAN ULTRASCAN 4000 (4k x 4k) / Number real images: 49 / Average electron dose: 150 e/Å2
Experimental equipment
Model: Tecnai Polara / Image courtesy: FEI Company

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Image processing

Final reconstructionAlgorithm: OTHER / Software - Name: IMOD / Details: Anisotropic diffusion was done in Spider. / Number images used: 49
DetailsIMOD for alignment and reconstruction. Spider for anisotropic diffusion.

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