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Yorodumi- EMDB-7481: The tether and tether head complex and I1 dynein complex region e... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-7481 | |||||||||
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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 (3736 axonemal repeats) of isolated Chlamydomonas wild type cilia | |||||||||
Map data | Cryo-electron tomography and subtomographic average (3736 axonemal repeats) of isolated axonemes from Chlamydomonas wide type | |||||||||
Sample |
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Biological species | Chlamydomonas reinhardtii (plant) | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 30.0 Å | |||||||||
Authors | Fu G / Wang Q / Phan N / Urbanska P / Joachimiak E / Lin J / Wloga D / Nicastro D | |||||||||
Funding support | United States, 1 items
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Citation | Journal: Mol Biol Cell / Year: 2018 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 |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_7481.map.gz | 432.7 KB | EMDB map data format | |
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Header (meta data) | emd-7481-v30.xml emd-7481.xml | 12.2 KB 12.2 KB | Display Display | EMDB header |
Images | emd_7481.png | 58.7 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-7481 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-7481 | HTTPS FTP |
-Validation report
Summary document | emd_7481_validation.pdf.gz | 78.1 KB | Display | EMDB validaton report |
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Full document | emd_7481_full_validation.pdf.gz | 77.3 KB | Display | |
Data in XML | emd_7481_validation.xml.gz | 494 B | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-7481 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-7481 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_7481.map.gz / Format: CCP4 / Size: 516.6 KB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Cryo-electron tomography and subtomographic average (3736 axonemal repeats) of isolated axonemes from Chlamydomonas wide type | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. generated in cubic-lattice coordinate | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 9.856 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : The tether and tether head complex and I1 dynein complex extracte...
Entire | Name: The tether and tether head complex and I1 dynein complex extracted from the cryo-electron tomography and subtomographic average (3736 axonemal repeats) of isolated Chlamydomonas wild type cilia |
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Components |
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-Supramolecule #1: The tether and tether head complex and I1 dynein complex extracte...
Supramolecule | Name: The tether and tether head complex and I1 dynein complex extracted from the cryo-electron tomography and subtomographic average (3736 axonemal repeats) of isolated Chlamydomonas wild type cilia type: organelle_or_cellular_component / ID: 1 / Parent: 0 |
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Source (natural) | Organism: Chlamydomonas reinhardtii (plant) / Strain: wild type |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | subtomogram averaging |
Aggregation state | tissue |
-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. |
-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: FIELD EMISSION GUN |
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 |
-Image processing
Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 30.0 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: IMOD Software - details: fiducial alignment and weighted backprojection Number subtomograms used: 3736 |
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Extraction | Number tomograms: 25 / Number images used: 3736 / Software - Name: MATLAB |
Final angle assignment | Type: ANGULAR RECONSTITUTION / Software - Name: PEET |