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Yorodumi- EMDB-20567: The axonemal doublet microtubule focusing on the I1 dynein region... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-20567 | |||||||||
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Title | The axonemal doublet microtubule focusing on the I1 dynein region extracted from the cryo-electron tomography and subtomographic average of isolated Chlamydomonas ida7-1 mutant cilia | |||||||||
Map data | The I1 dynein region of the Chlamydomonas ida7-1 mutant | |||||||||
Sample |
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Biological species | Chlamydomonas reinhardtii (plant) | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 40.0 Å | |||||||||
Authors | Fu G / Nicastro D | |||||||||
Funding support | United States, 2 items
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Citation | Journal: FASEB J / Year: 2021 Title: Structural organization of the intermediate and light chain complex of Chlamydomonas ciliary I1 dynein. Authors: Gang Fu / Chasity Scarbrough / Kangkang Song / Nhan Phan / Maureen Wirschell / Daniela Nicastro / Abstract: Axonemal I1 dynein (dynein f) is the largest inner dynein arm in cilia and a key regulator of ciliary beating. It consists of two dynein heavy chains, and an intermediate chain/light chain (ICLC) ...Axonemal I1 dynein (dynein f) is the largest inner dynein arm in cilia and a key regulator of ciliary beating. It consists of two dynein heavy chains, and an intermediate chain/light chain (ICLC) complex. However, the structural organization of the nine ICLC subunits remains largely unknown. Here, we used biochemical and genetic approaches, and cryo-electron tomography imaging in Chlamydomonas to dissect the molecular architecture of the I1 dynein ICLC complex. Using a strain expressing SNAP-tagged IC140, tomography revealed the location of the IC140 N-terminus at the proximal apex of the ICLC structure. Mass spectrometry of a tctex2b mutant showed that TCTEX2B dynein light chain is required for the stable assembly of TCTEX1 and inner dynein arm interacting proteins IC97 and FAP120. The structural defects observed in tctex2b located these 4 subunits in the center and bottom regions of the ICLC structure, which overlaps with the location of the IC138 regulatory subcomplex, which contains IC138, IC97, FAP120, and LC7b. These results reveal the three-dimensional organization of the native ICLC complex and indicate potential protein-protein interactions that are involved in the pathway by which I1 regulates 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_20567.map.gz | 394 KB | EMDB map data format | |
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Header (meta data) | emd-20567-v30.xml emd-20567.xml | 10.4 KB 10.4 KB | Display Display | EMDB header |
Images | emd_20567.png | 52.6 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-20567 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-20567 | HTTPS FTP |
-Validation report
Summary document | emd_20567_validation.pdf.gz | 306.7 KB | Display | EMDB validaton report |
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Full document | emd_20567_full_validation.pdf.gz | 306.3 KB | Display | |
Data in XML | emd_20567_validation.xml.gz | 4.8 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-20567 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-20567 | HTTPS FTP |
-Related structure data
Related structure data | C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_20567.map.gz / Format: CCP4 / Size: 469.7 KB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | The I1 dynein region of the Chlamydomonas ida7-1 mutant | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. generated in cubic-lattice coordinate | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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
-Sample components
-Entire : The I1 dynein averaged from Chlamydomonas ida7-1 mutant cilia
Entire | Name: The I1 dynein averaged from Chlamydomonas ida7-1 mutant cilia |
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Components |
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-Supramolecule #1: The I1 dynein averaged from Chlamydomonas ida7-1 mutant cilia
Supramolecule | Name: The I1 dynein averaged from Chlamydomonas ida7-1 mutant cilia type: organelle_or_cellular_component / ID: 1 / Parent: 0 |
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Source (natural) | Organism: Chlamydomonas reinhardtii (plant) |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | subtomogram averaging |
Aggregation state | cell |
-Sample preparation
Buffer | pH: 7.4 |
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Grid | Support film - Material: CARBON / Support film - topology: HOLEY / Details: unspecified |
Vitrification | Cryogen name: ETHANE / Chamber temperature: 298 K / Instrument: HOMEMADE PLUNGER Details: back-side blotting with No.1 Whitman filter 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 - Slit width: 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 | Calibrated defocus max: 8.0 µm / Calibrated defocus min: 6.0 µm / Calibrated magnification: 13500 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Sample stage | Specimen holder model: OTHER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Tecnai F30 / Image courtesy: FEI Company |
-Image processing
Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Algorithm: BACK PROJECTION / Resolution.type: BY AUTHOR / Resolution: 40.0 Å / Resolution method: FSC 0.5 CUT-OFF / Software - Name: IMOD / Number subtomograms used: 1051 |
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Extraction | Number tomograms: 9 / Number images used: 1051 / Software - Name: MATLAB |
Final angle assignment | Type: OTHER |