|Entry||Database: EMDB / ID: EMD-30878|
|Title||Focused refinement map of SF3B region of the activated human minor spliceosome|
|Sample||Structure of the Activated Human Minor Spliceosome (Bact Complex)|
|Biological species||Homo sapiens (human)|
|Method||single particle reconstruction / cryo EM / Resolution: 2.9 Å|
|Authors||Bai R / Wan R|
|Funding support|| China, 1 items |
|Citation||Journal: Science / Year: 2021|
Title: Structure of the activated human minor spliceosome.
Authors: Rui Bai / Ruixue Wan / Lin Wang / Kui Xu / Qiangfeng Zhang / Jianlin Lei / Yigong Shi /
Abstract: The minor spliceosome mediates splicing of the rare but essential U12-type precursor messenger RNA. Here, we report the atomic features of the activated human minor spliceosome determined by cryo- ...The minor spliceosome mediates splicing of the rare but essential U12-type precursor messenger RNA. Here, we report the atomic features of the activated human minor spliceosome determined by cryo-electron microscopy at 2.9-angstrom resolution. The 5' splice site and branch point sequence of the U12-type intron are recognized by the U6atac and U12 small nuclear RNAs (snRNAs), respectively. Five newly identified proteins stabilize the conformation of the catalytic center: The zinc finger protein SCNM1 functionally mimics the SF3a complex of the major spliceosome, the RBM48-ARMC7 complex binds the γ-monomethyl phosphate cap at the 5' end of U6atac snRNA, the U-box protein PPIL2 coordinates loop I of U5 snRNA and stabilizes U5 small nuclear ribonucleoprotein (snRNP), and CRIPT stabilizes U12 snRNP. Our study provides a framework for the mechanistic understanding of the function of the human minor spliceosome.
|Validation Report||Summary, Full report, XML, About validation report|
|Structure viewer||EM map: |
Downloads & links
|File||Download / File: emd_30878.map.gz / Format: CCP4 / Size: 824 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 1.0742 Å|
|Symmetry||Space group: 1|
CCP4 map header:
-Additional map: focused refinement of SF3b130
-Entire Structure of the Activated Human Minor Spliceosome (Bact Complex)
|Entire||Name: Structure of the Activated Human Minor Spliceosome (Bact Complex)|
Number of components: 1
-Component #1: protein, Structure of the Activated Human Minor Spliceosome (Bact...
|Protein||Name: Structure of the Activated Human Minor Spliceosome (Bact Complex)|
Recombinant expression: No
|Mass||Experimental: 1.7 MDa|
|Source||Species: Homo sapiens (human) / Strain: HeLa S3|
|Source (natural)||Location in cell: Nucleus|
|Specimen||Specimen state: Particle / Method: cryo EM|
|Sample solution||Specimen conc.: 0.2 mg/mL / pH: 7.9|
|Vitrification||Cryogen name: ETHANE|
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Imaging||Microscope: FEI TITAN KRIOS|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 19.531 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Imaging mode: BRIGHT FIELD / Energy filter: GIF Quantum LS|
|Specimen Holder||Model: OTHER|
|Processing||Method: single particle reconstruction / Number of projections: 101443|
|3D reconstruction||Resolution: 2.9 Å / Resolution method: FSC 0.143 CUT-OFF|
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