|Entry||Database: EMDB / ID: EMD-11975|
|Title||Cryo-EM map of the transcribing Pol II-U1 snRNP complex with 155-nt RNA|
|Sample||The transcribing RNA polymerase II-U1 snRNP complex with 155-nt RNA:|
RNA polymerase II / Nucleic acidsNucleic acid / U1 snRNP
|Biological species||Sus scrofa domesticus (domestic pig) / synthetic construct (others) / Homo sapiens (human)|
|Method||single particle reconstruction / cryo EM / Resolution: 3.9 Å|
|Authors||Zhang S / Aibara S / Vos SM / Agafonov DE / Luehrmann R / Cramer P|
|Funding support||4 items |
|Citation||Journal: Science / Year: 2021|
Title: Structure of a transcribing RNA polymerase II-U1 snRNP complex.
Authors: Suyang Zhang / Shintaro Aibara / Seychelle M Vos / Dmitry E Agafonov / Reinhard Lührmann / Patrick Cramer /
Abstract: To initiate cotranscriptional splicing, RNA polymerase II (Pol II) recruits the U1 small nuclear ribonucleoprotein particle (U1 snRNP) to nascent precursor messenger RNA (pre-mRNA). Here, we report ...To initiate cotranscriptional splicing, RNA polymerase II (Pol II) recruits the U1 small nuclear ribonucleoprotein particle (U1 snRNP) to nascent precursor messenger RNA (pre-mRNA). Here, we report the cryo-electron microscopy structure of a mammalian transcribing Pol II-U1 snRNP complex. The structure reveals that Pol II and U1 snRNP interact directly. This interaction positions the pre-mRNA 5' splice site near the RNA exit site of Pol II. Extension of pre-mRNA retains the 5' splice site, leading to the formation of a "growing intron loop." Loop formation may facilitate scanning of nascent pre-mRNA for the 3' splice site, functional pairing of distant intron ends, and prespliceosome assembly. Our results provide a starting point for a mechanistic analysis of cotranscriptional spliceosome assembly and the biogenesis of mRNA isoforms by alternative splicing.
|Validation Report||Summary, Full report, XML, About validation report|
|Structure viewer||EM map: |
Downloads & links
|File||Download / File: emd_11975.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 1.05 Å|
|Symmetry||Space group: 1|
CCP4 map header:
|Projections & Slices|
-Additional map: Unsharpened map
|Projections & Slices|
-Half map: Half map 2
|Annotation||Half map 2|
|Projections & Slices|
-Half map: Half map 1
-Entire The transcribing RNA polymerase II-U1 snRNP complex with 155-nt RNA
|Entire||Name: The transcribing RNA polymerase II-U1 snRNP complex with 155-nt RNA|
Number of components: 4
-Component #1: protein, The transcribing RNA polymerase II-U1 snRNP complex with...
|Protein||Name: The transcribing RNA polymerase II-U1 snRNP complex with 155-nt RNA|
Recombinant expression: No
-Component #2: protein, RNA polymerase II
|Protein||Name: RNA polymerase II / Recombinant expression: No|
|Source||Species: Sus scrofa domesticus (domestic pig)|
-Component #3: protein, Nucleic acids
|Protein||Name: Nucleic acidsNucleic acid / Recombinant expression: No|
|Source||Species: synthetic construct (others)|
|Source (engineered)||Expression System: synthetic construct (others)|
-Component #4: protein, U1 snRNP
|Protein||Name: U1 snRNP / Recombinant expression: No|
|Source||Species: Homo sapiens (human)|
|Specimen||Specimen state: Particle / Method: cryo EM|
|Sample solution||pH: 7.5|
|Vitrification||Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Temperature: 277.15 K / Humidity: 100 %|
-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: 1.01 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: OTHER|
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