|Entry||Database: EMDB / ID: EMD-20676|
|Title||Average of the basal body from bovine respiratory cilia|
|Sample||Nascent axoneme from bovine respiratory epithelial cells|
|Biological species||Bos taurus (cattle)|
|Method||subtomogram averaging / cryo EM / Resolution: 31.5 Å|
|Authors||Greenan GA / Vale RD / Agard DA|
|Funding support|| United States, 2 items |
|Citation||Journal: J Cell Biol / Year: 2020|
Title: Electron cryotomography of intact motile cilia defines the basal body to axoneme transition.
Authors: Garrett A Greenan / Ronald D Vale / David A Agard /
Abstract: Cells use motile cilia to generate force in the extracellular space. The structure of a cilium can be classified into three subdomains: the intracellular basal body (BB) that templates cilium ...Cells use motile cilia to generate force in the extracellular space. The structure of a cilium can be classified into three subdomains: the intracellular basal body (BB) that templates cilium formation, the extracellular axoneme that generates force, and the transition zone (TZ) that bridges them. While the BB is composed of triplet microtubules (TMTs), the axoneme is composed of doublet microtubules (DMTs), meaning the cilium must convert between different microtubule geometries. Here, we performed electron cryotomography to define this conversion, and our reconstructions reveal identifying structural features of the BB, TZ, and axoneme. Each region is distinct in terms of microtubule number and geometry, microtubule inner proteins, and microtubule linkers. TMT to DMT conversion occurs within the BB, and microtubule geometry changes to axonemal by the end of the TZ, followed by the addition of axoneme-specific components essential for cilium motility. Our results provide the highest-resolution images of the motile cilium to date and reveal how BBs template axonemes.
|Structure viewer||EM map: |
Downloads & links
|File||Download / File: emd_20676.map.gz / Format: CCP4 / Size: 12.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 8.18 Å|
|Symmetry||Space group: 1|
CCP4 map header:
-Entire Nascent axoneme from bovine respiratory epithelial cells
|Entire||Name: Nascent axoneme from bovine respiratory epithelial cells|
Number of components: 1
-Component #1: cellular-component, Nascent axoneme from bovine respiratory epith...
|Cellular-component||Name: Nascent axoneme from bovine respiratory epithelial cells|
Recombinant expression: No
|Source||Species: Bos taurus (cattle)|
|Source (natural)||Organelle: cilium|
|Specimen||Specimen state: Filament / Method: cryo EM|
|Sample solution||pH: 7.4|
|Vitrification||Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Temperature: 295 K / Humidity: 100 % / Details: 15 second blot.|
-Electron microscopy imaging
Model: Tecnai Polara / Image courtesy: FEI Company
|Imaging||Microscope: FEI POLARA 300|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 0.8 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: OTHER|
|Camera||Detector: GATAN K2 SUMMIT (4k x 4k)|
|Processing||Method: subtomogram averaging / Applied symmetry: C1 (asymmetric) / Number of subtomograms: 5768|
|3D reconstruction||Resolution: 31.5 Å / Resolution method: FSC 0.143 CUT-OFF|
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