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- EMDB-14735: Tomogram of T. brucei flagella singlet microtubules -

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Basic information

Entry
Database: EMDB / ID: EMD-14735
TitleTomogram of T. brucei flagella singlet microtubules
Map data
Sample
  • Cell: Trypanosome cell
KeywordsMicrotubule Flagellum Protist / UNKNOWN FUNCTION
Biological speciesTrypanosoma brucei (eukaryote)
Methodelectron tomography / cryo EM
AuthorsHoog JL / Zabeo DZ
Funding support United Kingdom, European Union, 2 items
OrganizationGrant numberCountry
Wellcome Trust United Kingdom
European Molecular Biology Organization (EMBO)European Union
CitationJournal: Elife / Year: 2014
Title: Modes of flagellar assembly in Chlamydomonas reinhardtii and Trypanosoma brucei.
Authors: Johanna L Höög / Sylvain Lacomble / Eileen T O'Toole / Andreas Hoenger / J Richard McIntosh / Keith Gull /
Abstract: Defects in flagella growth are related to a number of human diseases. Central to flagellar growth is the organization of microtubules that polymerize from basal bodies to form the axoneme, which ...Defects in flagella growth are related to a number of human diseases. Central to flagellar growth is the organization of microtubules that polymerize from basal bodies to form the axoneme, which consists of hundreds of proteins. Flagella exist in all eukaryotic phyla, but neither the mechanism by which flagella grow nor the conservation of this process in evolution are known. Here, we study how protein complexes assemble onto the growing axoneme tip using (cryo) electron tomography. In Chlamydomonas reinhardtii microtubules and associated proteins are added simultaneously. However, in Trypanosoma brucei, disorganized arrays of microtubules are arranged into the axoneme structure by the later addition of preformed protein complexes. Post assembly, the T. brucei transition zone alters structure and its association with the central pair loosens. We conclude that there are multiple ways to form a flagellum and that species-specific structural knowledge is critical before evaluating flagellar defects. DOI: http://dx.doi.org/10.7554/eLife.01479.001.
History
DepositionApr 6, 2022-
Header (metadata) releaseOct 5, 2022-
Map releaseOct 5, 2022-
UpdateDec 13, 2023-
Current statusDec 13, 2023Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

Downloads & links

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Map

FileDownload / File: emd_14735.map.gz / Format: CCP4 / Size: 1.2 GB / Type: IMAGE STORED AS SIGNED BYTE
Voxel sizeX=Y=Z: 9.44 Å
Density
Minimum - Maximum-128.0 - 127.0
Average (Standard dev.)-10.548553 (±109.971109999999996)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin-2-190162
Dimensions19461802366
Spacing18021946366
CellA: 17010.879 Å / B: 18370.238 Å / C: 3455.0398 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : Trypanosome cell

EntireName: Trypanosome cell
Components
  • Cell: Trypanosome cell

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Supramolecule #1: Trypanosome cell

SupramoleculeName: Trypanosome cell / type: cell / ID: 1 / Parent: 0
Source (natural)Organism: Trypanosoma brucei (eukaryote) / Strain: 427

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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
VitrificationCryogen name: ETHANE
SectioningOther: NO SECTIONING
Fiducial markerDiameter: 15 nm

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Electron microscopy

MicroscopeFEI TECNAI F30
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 6.0 µm / Nominal defocus min: 4.0 µm
Image recordingFilm or detector model: GATAN ULTRASCAN 4000 (4k x 4k) / Average electron dose: 0.83 e/Å2
Experimental equipment
Model: Tecnai F30 / Image courtesy: FEI Company

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Image processing

Final reconstructionNumber images used: 121

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