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- EMDB-7487: The tether and tether head complex and I1 dynein complex region e... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-7487 | |||||||||
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Title | The tether and tether head complex and I1 dynein complex region extracted from the cryo-electron tomography and subtomographic average (1711 axonemal repeats) of isolated Chlamydomonas fap43 mutant cilia (axonemal repeats missing the tether and tether head complex were not included) | |||||||||
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Biological species | ![]() ![]() | |||||||||
Method | electron tomography / cryo EM / Resolution: 37.0 Å | |||||||||
![]() | Fu G / Wang Q / Phan N / Urbanska P / Joachimiak E / Lin J / Wloga D / Nicastro D | |||||||||
![]() | ![]() Title: The I1 dynein-associated tether and tether head complex is a conserved regulator of ciliary motility. Authors: Gang Fu / Qian Wang / Nhan Phan / Paulina Urbanska / Ewa Joachimiak / Jianfeng Lin / Dorota Wloga / Daniela Nicastro / ![]() ![]() Abstract: Motile cilia are essential for propelling cells and moving fluids across tissues. The activity of axonemal dynein motors must be precisely coordinated to generate ciliary motility, but their ...Motile cilia are essential for propelling cells and moving fluids across tissues. The activity of axonemal dynein motors must be precisely coordinated to generate ciliary motility, but their regulatory mechanisms are not well understood. The tether and tether head (T/TH) complex was hypothesized to provide mechanical feedback during ciliary beating because it links the motor domains of the regulatory I1 dynein to the ciliary doublet microtubule. Combining genetic and biochemical approaches with cryoelectron tomography, we identified FAP44 and FAP43 (plus the algae-specific, FAP43-redundant FAP244) as T/TH components. WT-mutant comparisons revealed that the heterodimeric T/TH complex is required for the positional stability of the I1 dynein motor domains, stable anchoring of CK1 kinase, and proper phosphorylation of the regulatory IC138-subunit. T/TH also interacts with inner dynein arm d and radial spoke 3, another important motility regulator. The T/TH complex is a conserved regulator of I1 dynein and plays an important role in the signaling pathway that is critical for normal ciliary motility. | |||||||||
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 | ![]() | 435 KB | ![]() | |
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Header (meta data) | ![]() ![]() | 12.2 KB 12.2 KB | Display Display | ![]() |
Images | ![]() | 63.8 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 | ![]() | 494 B | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Voxel size | X=Y=Z: 10.77 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 : The tether and tether head complex and I1 dynein complex region e...
Entire | Name: The tether and tether head complex and I1 dynein complex region extracted from the cryo-electron tomography and subtomographic average (1711 axonemal repeats) of isolated Chlamydomonas fap43 ...Name: The tether and tether head complex and I1 dynein complex region extracted from the cryo-electron tomography and subtomographic average (1711 axonemal repeats) of isolated Chlamydomonas fap43 mutant cilia (axonemal repeats missing the tether and tether head complex were not included) |
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Components |
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-Supramolecule #1: The tether and tether head complex and I1 dynein complex region e...
Supramolecule | Name: The tether and tether head complex and I1 dynein complex region extracted from the cryo-electron tomography and subtomographic average (1711 axonemal repeats) of isolated Chlamydomonas fap43 ...Name: The tether and tether head complex and I1 dynein complex region extracted from the cryo-electron tomography and subtomographic average (1711 axonemal repeats) of isolated Chlamydomonas fap43 mutant cilia (axonemal repeats missing the tether and tether head complex were not included) type: organelle_or_cellular_component / ID: 1 / Parent: 0 |
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Source (natural) | Organism: ![]() ![]() |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | electron tomography |
Aggregation state | tissue |
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Sample preparation
Buffer | pH: 7.4 Component:
Details: Solutions were freshly made before use. | ||||||||||||
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Grid | Model: Quantifoil R2/2 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Atmosphere: AIR / Pretreatment - Pressure: 101.325 kPa | ||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber temperature: 100 K / Instrument: HOMEMADE PLUNGER Details: back-side blotting for 1.5-2.5 seconds before plunging. | ||||||||||||
Sectioning | Other: NO SECTIONING | ||||||||||||
Fiducial marker | Manufacturer: Sigma-Aldrich / Diameter: 10 nm |
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Electron microscopy
Microscope | FEI TECNAI F30 |
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Specialist optics | Energy filter - Name: GIF 2000 / Energy filter - Lower energy threshold: 0 eV / Energy filter - Upper energy threshold: 20 eV |
Image recording | Film or detector model: GATAN ULTRASCAN 1000 (2k x 2k) / Average electron dose: 1.5 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 8.0 µm / Nominal defocus min: 6.0 µm / Nominal magnification: 13500 |
Sample stage | Specimen holder model: GATAN LIQUID NITROGEN / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Tecnai F30 / Image courtesy: FEI Company |
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Image processing
Final reconstruction | Algorithm: BACK PROJECTION / Resolution.type: BY AUTHOR / Resolution: 37.0 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: ![]() Software - details: fiducial alignment and weighted backprojection Number images used: 780 |
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