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- EMDB-6871: A microtubule-dynein tethering complex regulates the axonemal inn... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-6871 | |||||||||
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Title | A microtubule-dynein tethering complex regulates the axonemal inner dynein f (I1) | |||||||||
![]() | Chlamydomonas fap44 mutant, IDA f in ATP plus vanadate state (state III) | |||||||||
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Biological species | ![]() ![]() | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 43.0 Å | |||||||||
![]() | Kubo T / Hou Y / Cochran DA / Witman GB / Oda T | |||||||||
![]() | ![]() Title: A microtubule-dynein tethering complex regulates the axonemal inner dynein f (I1). Authors: Tomohiro Kubo / Yuqing Hou / Deborah A Cochran / George B Witman / Toshiyuki Oda / ![]() ![]() Abstract: Motility of cilia/flagella is generated by a coordinated activity of thousands of dyneins. Inner dynein arms (IDAs) are particularly important for the formation of ciliary/flagellar waveforms, but ...Motility of cilia/flagella is generated by a coordinated activity of thousands of dyneins. Inner dynein arms (IDAs) are particularly important for the formation of ciliary/flagellar waveforms, but the molecular mechanism of IDA regulation is poorly understood. Here we show using cryoelectron tomography and biochemical analyses of Chlamydomonas flagella that a conserved protein FAP44 forms a complex that tethers IDA f (I1 dynein) head domains to the A-tubule of the axonemal outer doublet microtubule. In wild-type flagella, IDA f showed little nucleotide-dependent movement except for a tilt in the f β head perpendicular to the microtubule-sliding direction. In the absence of the tether complex, however, addition of ATP and vanadate caused a large conformational change in the IDA f head domains, suggesting that the movement of IDA f is mechanically restricted by the tether complex. Motility defects in flagella missing the tether demonstrates the importance of the IDA f-tether interaction in the regulation of ciliary/flagellar beating. | |||||||||
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 | ![]() | 46.9 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 10.3 KB 10.3 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 3 KB | Display | ![]() |
Images | ![]() | 76.1 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 78.2 KB | Display | ![]() |
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Full document | ![]() | 77.3 KB | Display | |
Data in XML | ![]() | 495 B | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 6866C ![]() 6867C ![]() 6868C ![]() 6869C ![]() 6870C ![]() 6872C ![]() 6873C ![]() 6874C C: citing same article ( |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Map
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Annotation | Chlamydomonas fap44 mutant, IDA f in ATP plus vanadate state (state III) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 7.025 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 : Inner dynein f
Entire | Name: Inner dynein f |
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Components |
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-Supramolecule #1: Inner dynein f
Supramolecule | Name: Inner dynein f / type: organelle_or_cellular_component / ID: 1 / Parent: 0 Details: fap44 mutant Chlamydomonas axoneme in ATP plus vanadate state (state III) |
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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 | 0.02 mg/mL |
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Buffer | pH: 7.2 |
Grid | Model: Homemade / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Atmosphere: AIR |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV / Details: blot 5.5 sec before plunging. |
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Electron microscopy
Microscope | JEOL 3100FFC |
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Specialist optics | Energy filter - Name: In-column Omega Filter |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average exposure time: 6.0 sec. / Average electron dose: 1.6 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Sample stage | Specimen holder model: GATAN 914 HIGH TILT LIQUID NITROGEN CRYO TRANSFER TOMOGRAPHY HOLDER Cooling holder cryogen: HELIUM |