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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-1941 | |||||||||
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Title | Radial Spoke from Chlamydomonas flagella | |||||||||
![]() | This is a density map of Radial Spokes from Chlamydomonas flagella. | |||||||||
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![]() | radial spoke / flagella / cilia / Chlamydomonas | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 39.18 Å | |||||||||
![]() | Pigino G / Bui KH / Maheshwari A / Lupetti P / Diener D / Ishikawa T | |||||||||
![]() | ![]() Title: Cryoelectron tomography of radial spokes in cilia and flagella. Authors: Gaia Pigino / Khanh Huy Bui / Aditi Maheshwari / Pietro Lupetti / Dennis Diener / Takashi Ishikawa / ![]() Abstract: Radial spokes (RSs) are ubiquitous components in the 9 + 2 axoneme thought to be mechanochemical transducers involved in local control of dynein-driven microtubule sliding. They are composed of >23 ...Radial spokes (RSs) are ubiquitous components in the 9 + 2 axoneme thought to be mechanochemical transducers involved in local control of dynein-driven microtubule sliding. They are composed of >23 polypeptides, whose interactions and placement must be deciphered to understand RS function. In this paper, we show the detailed three-dimensional (3D) structure of RS in situ in Chlamydomonas reinhardtii flagella and Tetrahymena thermophila cilia that we obtained using cryoelectron tomography (cryo-ET). We clarify similarities and differences between the three spoke species, RS1, RS2, and RS3, in T. thermophila and in C. reinhardtii and show that part of RS3 is conserved in C. reinhardtii, which only has two species of complete RSs. By analyzing C. reinhardtii mutants, we identified the specific location of subsets of RS proteins (RSPs). Our 3D reconstructions show a twofold symmetry, suggesting that fully assembled RSs are produced by dimerization. Based on our cryo-ET data, we propose models of subdomain organization within the RS as well as interactions between RSPs and with other axonemal components. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 787.5 KB | ![]() | |
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Header (meta data) | ![]() ![]() | 9.6 KB 9.6 KB | Display Display | ![]() |
Images | ![]() | 270.2 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 175.4 KB | Display | ![]() |
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Full document | ![]() | 174.5 KB | Display | |
Data in XML | ![]() | 4.8 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | This is a density map of Radial Spokes from Chlamydomonas flagella. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 6.78 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : Radial spoke from flagella/cilia
Entire | Name: Radial spoke from flagella/cilia |
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Components |
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-Supramolecule #1000: Radial spoke from flagella/cilia
Supramolecule | Name: Radial spoke from flagella/cilia / type: sample / ID: 1000 / Details: Radial spokes from Axoneme / Number unique components: 1 |
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-Supramolecule #1: radial spoke
Supramolecule | Name: radial spoke / type: organelle_or_cellular_component / ID: 1 / Name.synonym: radial spoke / Recombinant expression: Yes |
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Source (natural) | Organism: ![]() ![]() |
Recombinant expression | Organism: ![]() ![]() |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | subtomogram averaging |
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Sample preparation
Concentration | 1 mg/mL |
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Buffer | pH: 7.4 / Details: HMDKP |
Grid | Details: lacey carbon grids |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 100 K / Instrument: OTHER / Details: Vitrification instrument: Vitrobot / Method: 3 second blotting |
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Electron microscopy
Microscope | FEI TECNAI 20 |
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Temperature | Min: 100 K / Max: 100 K / Average: 100 K |
Specialist optics | Energy filter - Name: GIF / Energy filter - Lower energy threshold: 0.0 eV / Energy filter - Upper energy threshold: 20.0 eV |
Image recording | Category: CCD / Film or detector model: GATAN ULTRASCAN 1000 (2k x 2k) / Average electron dose: 0.6 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Calibrated magnification: 19000 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 1.5 mm / Nominal defocus max: 6.0 µm / Nominal defocus min: 3.0 µm / Nominal magnification: 27500 |
Sample stage | Specimen holder: Gatan 626 / Specimen holder model: GATAN LIQUID NITROGEN / Tilt series - Axis1 - Min angle: -60 ° / Tilt series - Axis1 - Max angle: 60 ° |
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Image processing
Details | Subtomograms were averaged. Average number of tilts used in the 3D reconstructions: 61. Average tomographic tilt angle increment: 2. |
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Final reconstruction | Algorithm: OTHER / Resolution.type: BY AUTHOR / Resolution: 39.18 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: ![]() |