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- EMDB-5845: Cryo-electron tomography of Chlamydomonas axoneme labeled with st... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-5845 | |||||||||
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Title | Cryo-electron tomography of Chlamydomonas axoneme labeled with streptavidin (Wild type) | |||||||||
![]() | Averaged tomogram of axonemal doublet microtubules (Wild type) | |||||||||
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![]() | cilia and flagella / radial spokes | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 68.0 Å | |||||||||
![]() | Oda T / Yanagisawa H / Yagi T / Kikkawa M | |||||||||
![]() | ![]() Title: Mechanosignaling between central apparatus and radial spokes controls axonemal dynein activity. Authors: Toshiyuki Oda / Haruaki Yanagisawa / Toshiki Yagi / Masahide Kikkawa / ![]() Abstract: Cilia/flagella are conserved organelles that generate fluid flow in eukaryotes. The bending motion of flagella requires concerted activity of dynein motors. Although it has been reported that the ...Cilia/flagella are conserved organelles that generate fluid flow in eukaryotes. The bending motion of flagella requires concerted activity of dynein motors. Although it has been reported that the central pair apparatus (CP) and radial spokes (RSs) are important for flagellar motility, the molecular mechanism underlying CP- and RS-mediated dynein regulation has not been identified. In this paper, we identified nonspecific intermolecular collision between CP and RS as one of the regulatory mechanisms for flagellar motility. By combining cryoelectron tomography and motility analyses of Chlamydomonas reinhardtii flagella, we show that binding of streptavidin to RS heads paralyzed flagella. Moreover, the motility defect in a CP projection mutant could be rescued by the addition of exogenous protein tags on RS heads. Genetic experiments demonstrated that outer dynein arms are the major downstream effectors of CP- and RS-mediated regulation of flagellar motility. These results suggest that mechanosignaling between CP and RS regulates dynein activity in eukaryotic flagella. | |||||||||
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 | ![]() | 41.3 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 8.8 KB 8.8 KB | Display Display | ![]() |
Images | ![]() | 203.9 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 5846C ![]() 5847C ![]() 5848C ![]() 5849C ![]() 5850C ![]() 5851C ![]() 5852C ![]() 5853C ![]() 5854C ![]() 5855C C: citing same article ( |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Averaged tomogram of axonemal doublet microtubules (Wild type) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 6.07 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 : Chlamydomonas axoneme, wild type
Entire | Name: Chlamydomonas axoneme, wild type |
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Components |
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-Supramolecule #1000: Chlamydomonas axoneme, wild type
Supramolecule | Name: Chlamydomonas axoneme, wild type / type: sample / ID: 1000 / Number unique components: 1 |
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-Supramolecule #1: Axoneme
Supramolecule | Name: Axoneme / type: organelle_or_cellular_component / ID: 1 / Recombinant expression: No / Database: NCBI |
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Source (natural) | Organism: ![]() ![]() |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | subtomogram averaging |
Aggregation state | filament |
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Sample preparation
Concentration | 0.01 mg/mL |
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Buffer | pH: 7.2 Details: 30 mM Hepes-NaOH pH 7.2, 5 mM MgCl2, 1 mM dithiothreitol, 1 mM EGTA, and 50 mM NaCl |
Grid | Details: 300 mesh copper grid with home-made holey carbon support |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 93 K / Instrument: LEICA EM GP / Method: Blot for 5 seconds before plunging |
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Electron microscopy
Microscope | JEOL 3100FFC |
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Temperature | Min: 90 K / Max: 95 K |
Specialist optics | Energy filter - Name: JEOL Omega / Energy filter - Lower energy threshold: 0.0 eV / Energy filter - Upper energy threshold: 23.0 eV |
Date | Aug 15, 2013 |
Image recording | Category: CCD / Film or detector model: TVIPS TEMCAM-F416 (4k x 4k) / Average electron dose: 90 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Calibrated magnification: 25700 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.2 mm / Nominal magnification: 15000 |
Sample stage | Specimen holder model: GATAN LIQUID NITROGEN / Tilt series - Axis1 - Min angle: -60.5 ° / Tilt series - Axis1 - Max angle: 60.5 ° |
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Image processing
Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 68.0 Å / Resolution method: OTHER / Software - Name: PEET, RUBY-HELIX / Number subtomograms used: 644 |
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