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- EMDB-29753: Tomogram of an apical end of a Toxoplasma Gondii tachyzoite displ... -

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Basic information

Entry
Database: EMDB / ID: EMD-29753
TitleTomogram of an apical end of a Toxoplasma Gondii tachyzoite displaying an extruded conoid
Map dataToxoplasma Gondii tachyzoite apical end, with extruded conoid
Sample
  • Cell: Toxoplasma Gondii tachyzoite apical complex
KeywordsConoid / Parasite / preconoidal rings / CELL INVASION
Biological speciesToxoplasma gondii (eukaryote)
Methodelectron tomography / cryo EM
AuthorsMartinez M / Chang Y-W / Mageswaran SK
Funding support United States, 1 items
OrganizationGrant numberCountry
David and Lucile Packard Foundation United States
CitationJournal: Nat Commun / Year: 2023
Title: Origin and arrangement of actin filaments for gliding motility in apicomplexan parasites revealed by cryo-electron tomography.
Authors: Matthew Martinez / Shrawan Kumar Mageswaran / Amandine Guérin / William David Chen / Cameron Parker Thompson / Sabine Chavin / Dominique Soldati-Favre / Boris Striepen / Yi-Wei Chang /
Abstract: The phylum Apicomplexa comprises important eukaryotic parasites that invade host tissues and cells using a unique mechanism of gliding motility. Gliding is powered by actomyosin motors that ...The phylum Apicomplexa comprises important eukaryotic parasites that invade host tissues and cells using a unique mechanism of gliding motility. Gliding is powered by actomyosin motors that translocate host-attached surface adhesins along the parasite cell body. Actin filaments (F-actin) generated by Formin1 play a central role in this critical parasitic activity. However, their subcellular origin, path and ultrastructural arrangement are poorly understood. Here we used cryo-electron tomography to image motile Cryptosporidium parvum sporozoites and reveal the cellular architecture of F-actin at nanometer-scale resolution. We demonstrate that F-actin nucleates at the apically positioned preconoidal rings and is channeled into the pellicular space between the parasite plasma membrane and the inner membrane complex in a conoid extrusion-dependent manner. Within the pellicular space, filaments on the inner membrane complex surface appear to guide the apico-basal flux of F-actin. F-actin concordantly accumulates at the basal end of the parasite. Finally, analyzing a Formin1-depleted Toxoplasma gondii mutant pinpoints the upper preconoidal ring as the conserved nucleation hub for F-actin in Cryptosporidium and Toxoplasma. Together, we provide an ultrastructural model for the life cycle of F-actin for apicomplexan gliding motility.
History
DepositionFeb 13, 2023-
Header (metadata) releaseSep 13, 2023-
Map releaseSep 13, 2023-
UpdateSep 13, 2023-
Current statusSep 13, 2023Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_29753.png

Downloads & links

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Map

FileDownload / File: emd_29753.map.gz / Format: CCP4 / Size: 1.1 GB / Type: IMAGE STORED AS SIGNED BYTE
AnnotationToxoplasma Gondii tachyzoite apical end, with extruded conoid
Voxel sizeX=Y=Z: 10.6 Å
Density
Minimum - Maximum-120.0 - 123.0
Average (Standard dev.)32.665405 (±6.259918)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin-209209-401
Dimensions14401023800
Spacing10231440800
CellA: 10843.801 Å / B: 15264.001 Å / C: 8480.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : Toxoplasma Gondii tachyzoite apical complex

EntireName: Toxoplasma Gondii tachyzoite apical complex
Components
  • Cell: Toxoplasma Gondii tachyzoite apical complex

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Supramolecule #1: Toxoplasma Gondii tachyzoite apical complex

SupramoleculeName: Toxoplasma Gondii tachyzoite apical complex / type: cell / ID: 1 / Parent: 0
Source (natural)Organism: Toxoplasma gondii (eukaryote)

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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: DMEM supplemented with 5% fetal bovine serum and 2 mM glutamine
GridModel: Quantifoil / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY
VitrificationCryogen name: ETHANE-PROPANE / Chamber humidity: 95 % / Chamber temperature: 310 K / Instrument: LEICA EM GP / Details: Front blot for 4 seconds before plunging.
DetailsEgressed tachyzoites from a culture of confluent HFF cells.
SectioningOther: NO SECTIONING
Fiducial markerManufacturer: Ted Pella, Inc / Diameter: 10 nm

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Specialist opticsPhase plate: VOLTA PHASE PLATE / Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV
SoftwareName: SerialEM (ver. 3.8)
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average exposure time: 0.4 sec. / Average electron dose: 2.3 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 100.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 4.0 µm / Nominal defocus min: 1.5 µm / Nominal magnification: 33000
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

DetailsRaw frames were gain normalized in serialEM and motion corrected using the alignframes command in IMOD
Final reconstructionAlgorithm: BACK PROJECTION / Software - Name: IMOD (ver. 4.11.5) / Number images used: 61

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