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- EMDB-2814: Electron cryotomography of the FtsZ ring in Caulobacter crescentus. -

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Basic information

Entry
Database: EMDB / ID: EMD-2814
TitleElectron cryotomography of the FtsZ ring in Caulobacter crescentus.
Map dataTomogram of dividing Caulobacter crescentus CB15N grown in M2G medium.
Sample
  • Sample: Caulobacter crescentus CB15N grown in M2G medium.
  • Organelle or cellular component: Caulobacter crescentus
KeywordsFtsZ / divisome / bacterial cell division / cytokinesis
Biological speciesCaulobacter vibrioides (bacteria)
Methodelectron tomography / cryo EM
AuthorsSzwedziak P / Wang Q / Bharat TAM / Tsim M / Lowe J
CitationJournal: Elife / Year: 2014
Title: Architecture of the ring formed by the tubulin homologue FtsZ in bacterial cell division.
Authors: Piotr Szwedziak / Qing Wang / Tanmay A M Bharat / Matthew Tsim / Jan Löwe /
Abstract: Membrane constriction is a prerequisite for cell division. The most common membrane constriction system in prokaryotes is based on the tubulin homologue FtsZ, whose filaments in E. coli are anchored ...Membrane constriction is a prerequisite for cell division. The most common membrane constriction system in prokaryotes is based on the tubulin homologue FtsZ, whose filaments in E. coli are anchored to the membrane by FtsA and enable the formation of the Z-ring and divisome. The precise architecture of the FtsZ ring has remained enigmatic. In this study, we report three-dimensional arrangements of FtsZ and FtsA filaments in C. crescentus and E. coli cells and inside constricting liposomes by means of electron cryomicroscopy and cryotomography. In vivo and in vitro, the Z-ring is composed of a small, single-layered band of filaments parallel to the membrane, creating a continuous ring through lateral filament contacts. Visualisation of the in vitro reconstituted constrictions as well as a complete tracing of the helical paths of the filaments with a molecular model favour a mechanism of FtsZ-based membrane constriction that is likely to be accompanied by filament sliding.
History
DepositionNov 7, 2014-
Header (metadata) releaseDec 24, 2014-
Map releaseDec 31, 2014-
UpdateFeb 17, 2016-
Current statusFeb 17, 2016Processing site: PDBe / Status: Released

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Structure visualization

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Supplemental images

emd-2814.png

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Map

FileDownload / File: emd_2814.map.gz / Format: CCP4 / Size: 370.4 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationTomogram of dividing Caulobacter crescentus CB15N grown in M2G medium.
Voxel sizeX=Y=Z: 17.98 Å
Density
Minimum - Maximum-4.76493883 - 2.89649415
Average (Standard dev.)0.08586969 (±0.14056909)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin-120-437407
Dimensions732462294
Spacing732462294
CellA: 8306.76 Å / B: 13161.359 Å / C: 5286.1196 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z17.9817.9799986338817.98
M x/y/z462732294
origin x/y/z0.0000.0000.000
length x/y/z8306.76013161.3595286.120
α/β/γ90.00090.00090.000
start NX/NY/NZ-40-32-96
NX/NY/NZ8165193
MAP C/R/S123
start NC/NR/NS-437-120407
NC/NR/NS462732294
D min/max/mean-4.7652.8960.086

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Supplemental data

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Sample components

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Entire : Caulobacter crescentus CB15N grown in M2G medium.

EntireName: Caulobacter crescentus CB15N grown in M2G medium.
Components
  • Sample: Caulobacter crescentus CB15N grown in M2G medium.
  • Organelle or cellular component: Caulobacter crescentus

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Supramolecule #1000: Caulobacter crescentus CB15N grown in M2G medium.

SupramoleculeName: Caulobacter crescentus CB15N grown in M2G medium. / type: sample / ID: 1000
Details: Dividing Caulobacter crescentus CB15N grown in M2G medium.
Number unique components: 1

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Supramolecule #1: Caulobacter crescentus

SupramoleculeName: Caulobacter crescentus / type: organelle_or_cellular_component / ID: 1
Details: Cells were grown to mid-log phase, mixed with 10 nm protein A gold.
Recombinant expression: No / Database: NCBI
Source (natural)Organism: Caulobacter vibrioides (bacteria) / Strain: CB15N / Cell: Bacterial

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Experimental details

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Structure determination

Methodcryo EM
Processingelectron tomography
Aggregation statecell

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Sample preparation

BufferDetails: M2G medium
GridDetails: Quantifoil R3.5/1 300 mesh Cu/Rh holey carbon grid
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Instrument: FEI VITROBOT MARK IV / Method: 2.5 s blot on Whatmann 1 filter paper.

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsCalibrated magnification: 26000 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal defocus max: -10.0 µm / Nominal defocus min: -10.0 µm / Nominal magnification: 26000
Specialist opticsEnergy filter - Name: Gatan Quantum / Energy filter - Lower energy threshold: 0.0 eV / Energy filter - Upper energy threshold: 20.0 eV
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Tilt series - Axis1 - Min angle: 0 ° / Tilt series - Axis1 - Max angle: 60 ° / Tilt series - Axis1 - Angle increment: 1 °
Alignment procedureLegacy - Astigmatism: Visualisation of power spectrum.
DateJul 31, 2014
Image recordingCategory: CCD / Film or detector model: GATAN K2 (4k x 4k) / Number real images: 121 / Average electron dose: 150 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Final reconstructionAlgorithm: OTHER / Software - Name: IMOD, tomo3D / Number images used: 121
DetailsTilt series data was aligned using IMOD and reconstruction was carried out using tomo3D.

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