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- EMDB-40892: Ciliary cap complex from Tetrahymena thermophila -

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Basic information

Entry
Database: EMDB / ID: EMD-40892
TitleCiliary cap complex from Tetrahymena thermophila
Map data
Sample
  • Organelle or cellular component: Ciliary cap complex from Tetrahymena thermphila
KeywordsCilia / Axoneme / central pair / microtubules / STRUCTURAL PROTEIN
Biological speciesTetrahymena thermophila (eukaryote)
Methodsubtomogram averaging / cryo EM / Resolution: 130.0 Å
AuthorsLegal T / Bui KH
Funding support Canada, 1 items
OrganizationGrant numberCountry
Canadian Institutes of Health Research (CIHR)PJT-156354 Canada
CitationJournal: J Cell Biol / Year: 2023
Title: CEP104/FAP256 and associated cap complex maintain stability of the ciliary tip.
Authors: Thibault Legal / Mireya Parra / Maxwell Tong / Corbin S Black / Ewa Joachimiak / Melissa Valente-Paterno / Karl Lechtreck / Jacek Gaertig / Khanh Huy Bui /
Abstract: Cilia are essential organelles that protrude from the cell body. Cilia are made of a microtubule-based structure called the axoneme. In most types of cilia, the ciliary tip is distinct from the rest ...Cilia are essential organelles that protrude from the cell body. Cilia are made of a microtubule-based structure called the axoneme. In most types of cilia, the ciliary tip is distinct from the rest of the cilium. Here, we used cryo-electron tomography and subtomogram averaging to obtain the structure of the ciliary tip of the ciliate Tetrahymena thermophila. We show that the microtubules at the tip are highly crosslinked with each other and stabilized by luminal proteins, plugs, and cap proteins at the plus ends. In the tip region, the central pair lacks typical projections and twists significantly. By analyzing cells lacking a ciliary tip-enriched protein CEP104/FAP256 by cryo-electron tomography and proteomics, we discovered candidates for the central pair cap complex and explained the potential functions of CEP104/FAP256. These data provide new insights into the function of the ciliary tip and the mechanisms of ciliary assembly and length regulation.
History
DepositionMay 26, 2023-
Header (metadata) releaseSep 13, 2023-
Map releaseSep 13, 2023-
UpdateOct 11, 2023-
Current statusOct 11, 2023Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_40892.png

Downloads & links

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Map

FileDownload / File: emd_40892.map.gz / Format: CCP4 / Size: 52.7 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
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AxesZ (Sec.)Y (Row.)X (Col.)
8.48 Å/pix.
x 240 pix.
= 2035.2 Å		8.48 Å/pix.
x 240 pix.
= 2035.2 Å		8.48 Å/pix.
x 240 pix.
= 2035.2 Å

Surface

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Images are generated by Spider.

Voxel sizeX=Y=Z: 8.48 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.11
Minimum - Maximum-0.40681398 - 1.0102475
Average (Standard dev.)-0.000024601397 (±0.052982975)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions240240240
Spacing240240240
CellA=B=C: 2035.2 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #1

Fileemd_40892_half_map_1.map
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Half map: #2

Fileemd_40892_half_map_2.map
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Sample components

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Entire : Ciliary cap complex from Tetrahymena thermphila

EntireName: Ciliary cap complex from Tetrahymena thermphila
Components
  • Organelle or cellular component: Ciliary cap complex from Tetrahymena thermphila

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Supramolecule #1: Ciliary cap complex from Tetrahymena thermphila

SupramoleculeName: Ciliary cap complex from Tetrahymena thermphila / type: organelle_or_cellular_component / ID: 1 / Parent: 0
Source (natural)Organism: Tetrahymena thermophila (eukaryote) / Strain: CU428 / Organelle: Cilia

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Experimental details

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Structure determination

Methodcryo EM
Processingsubtomogram averaging
Aggregation statecell

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Sample preparation

Concentration1.8 mg/mL
BufferpH: 7.4
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeTFS KRIOS
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 4.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 4.0 µm / Nominal defocus min: 2.0 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Final reconstructionResolution.type: BY AUTHOR / Resolution: 130.0 Å / Resolution method: FSC 0.143 CUT-OFF / Number subtomograms used: 46
ExtractionNumber tomograms: 48 / Number images used: 46
Final angle assignmentType: MAXIMUM LIKELIHOOD
FSC plot (resolution estimation)

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