|Entry||Database: EMDB / ID: 9169|
|Title||Electron cryo-tomography and subtomogram averaging of microtubule triplet from procentriole|
|Map data||a subtomogram average of the B-tubule in procentriole from Chlamydomonas Reinhardtii|
|Source||Chlamydomonas reinhardtii (plant)|
|Method||subtomogram averaging / cryo EM / 22.3 Å resolution|
|Authors||Li S / Fernandez JJ / Marshall W / Agard DA|
|Citation||Journal: Elife / Year: 2019|
Title: Electron cryo-tomography provides insight into procentriole architecture and assembly mechanism.
Authors: Sam Li / Jose-Jesus Fernandez / Wallace F Marshall / David A Agard
Abstract: Centriole is an essential structure with multiple functions in cellular processes. Centriole biogenesis and homeostasis is tightly regulated. Using electron cryo-tomography (cryoET) we present the ...Centriole is an essential structure with multiple functions in cellular processes. Centriole biogenesis and homeostasis is tightly regulated. Using electron cryo-tomography (cryoET) we present the structure of procentrioles from . We identified a set of non-tubulin components attached to the triplet microtubule (MT), many are at the junctions of tubules likely to reinforce the triplet. We describe structure of the A-C linker that bridges neighboring triplets. The structure infers that POC1 is likely an integral component of A-C linker. Its conserved WD40 β-propeller domain provides attachment sites for other A-C linker components. The twist of A-C linker results in an iris diaphragm-like motion of the triplets in the longitudinal direction of procentriole. Finally, we identified two assembly intermediates at the growing ends of procentriole allowing us to propose a model for the procentriole assembly. Our results provide a comprehensive structural framework for understanding the molecular mechanisms underpinning procentriole biogenesis and assembly.
|Date||Deposition: Oct 1, 2018 / Header (metadata) release: Oct 24, 2018 / Map release: Feb 27, 2019 / Last update: Feb 27, 2019|
|Structure viewer||EM map: |
Downloads & links
|File||emd_9169.map.gz (map file in CCP4 format, 6913 KB)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 4.82 Å|
CCP4 map header:
|Entire||Name: Centriole / Details: isolated centriole and procentriole / Number of components: 1|
-Component #1: cellular-component, Centriole
|Cellular-component||Name: Centriole / Details: isolated centriole and procentriole / Recombinant expression: No|
|Source||Species: Chlamydomonas reinhardtii (plant) / Strain: CC849|
|Specimen||Specimen state: cell / Method: cryo EM|
|Crystal parameters||Space group: C1 / A: 578.4 Å / B: 578.4 Å / C: 578.4 Å / Alpha: 90 deg. / Beta: 90 deg. / Gamma: 90 deg.|
|Sample solution||pH: 7|
|Vitrification||Cryogen name: ETHANE|
-Electron microscopy imaging
Model: Tecnai Polara / Image courtesy: FEI Company
|Imaging||Microscope: FEI POLARA 300|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 0.6 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: OTHER|
|Camera||Detector: GATAN K2 SUMMIT (4k x 4k)|
|Image acquisition||Sampling size: 5 microns|
|Processing||Method: subtomogram averaging / Number of subtomograms: 12179|
|3D reconstruction||Resolution: 22.3 Å / Resolution method: FSC 0.143 CUT-OFF|
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