+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-9767 | |||||||||
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Title | Doublet microtubule of fap52 null mutant | |||||||||
Map data | fap52_DMT | |||||||||
Sample |
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Biological species | Chlamydomonas reinhardtii (plant) | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 40.0 Å | |||||||||
Authors | Owa M / Uchihashi T / Yanagisawa H / Yamano T / Iguchi H / Fukuzawa H / Wakabayashi K / Ando T / Kikkawa M | |||||||||
Citation | Journal: Nat Commun / Year: 2019 Title: Inner lumen proteins stabilize doublet microtubules in cilia and flagella. Authors: Mikito Owa / Takayuki Uchihashi / Haru-Aki Yanagisawa / Takashi Yamano / Hiro Iguchi / Hideya Fukuzawa / Ken-Ichi Wakabayashi / Toshio Ando / Masahide Kikkawa / Abstract: Motile cilia are microtubule-based organelles that play important roles in most eukaryotes. Although axonemal microtubules are sufficiently stable to withstand their beating motion, it remains ...Motile cilia are microtubule-based organelles that play important roles in most eukaryotes. Although axonemal microtubules are sufficiently stable to withstand their beating motion, it remains unknown how they are stabilized while serving as tracks for axonemal dyneins. To address this question, we have identified two uncharacterized proteins, FAP45 and FAP52, as microtubule inner proteins (MIPs) in Chlamydomonas. These proteins are conserved among eukaryotes with motile cilia. Using cryo-electron tomography (cryo-ET) and high-speed atomic force microscopy (HS-AFM), we show that lack of these proteins leads to a loss of inner protrusions in B-tubules and less stable microtubules. These protrusions are located near the inner junctions of doublet microtubules and lack of both FAP52 and a known inner junction protein FAP20 results in detachment of the B-tubule from the A-tubule, as well as flagellar shortening. These results demonstrate that FAP45 and FAP52 bind to the inside of microtubules and stabilize ciliary axonemes. | |||||||||
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_9767.map.gz | 7.6 MB | EMDB map data format | |
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Header (meta data) | emd-9767-v30.xml emd-9767.xml | 8.2 KB 8.2 KB | Display Display | EMDB header |
Images | emd_9767.png | 40 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-9767 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-9767 | HTTPS FTP |
-Validation report
Summary document | emd_9767_validation.pdf.gz | 79.2 KB | Display | EMDB validaton report |
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Full document | emd_9767_full_validation.pdf.gz | 78.3 KB | Display | |
Data in XML | emd_9767_validation.xml.gz | 495 B | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9767 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9767 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_9767.map.gz / Format: CCP4 / Size: 30.5 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | fap52_DMT | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 7.1 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 : axoneme of Chlamydomonas reinhardtii
Entire | Name: axoneme of Chlamydomonas reinhardtii |
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Components |
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-Supramolecule #1: axoneme of Chlamydomonas reinhardtii
Supramolecule | Name: axoneme of Chlamydomonas reinhardtii / type: organelle_or_cellular_component / ID: 1 / Parent: 0 |
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Source (natural) | Organism: Chlamydomonas reinhardtii (plant) / Strain: fap52 |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | subtomogram averaging |
Aggregation state | filament |
-Sample preparation
Concentration | 0.02 mg/mL |
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Buffer | pH: 7.4 |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | JEOL 3100FFC |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 100.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: OTHER / Imaging mode: DIFFRACTION |
-Image processing
Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 40.0 Å / Resolution method: FSC 0.5 CUT-OFF / Number subtomograms used: 2300 |
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Extraction | Number tomograms: 10 / Number images used: 2500 |
Final angle assignment | Type: NOT APPLICABLE |