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Open data
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Basic information
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Title | The cryo-EM structure of the human pre-A complex | |||||||||
![]() | The cryo-EM map for the U2 part of human pre-A complex | |||||||||
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![]() | human spliceosome / pre-A complex / PRP5 / A complex / U1 snRNP / U2 snRNP / SPLICING | |||||||||
Function / homology | ![]() negative regulation of protein refolding / U2AF complex / U11/U12 snRNP / regulation of ATP-dependent activity / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / U7 snRNP / histone pre-mRNA 3'end processing complex / SLBP independent Processing of Histone Pre-mRNAs ...negative regulation of protein refolding / U2AF complex / U11/U12 snRNP / regulation of ATP-dependent activity / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / U7 snRNP / histone pre-mRNA 3'end processing complex / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / B-WICH complex / protein methylation / U12-type spliceosomal complex / 7-methylguanosine cap hypermethylation / RNA splicing, via transesterification reactions / U1 snRNP binding / methylosome / regulation of mRNA splicing, via spliceosome / pICln-Sm protein complex / positive regulation of mRNA splicing, via spliceosome / snRNP binding / small nuclear ribonucleoprotein complex / splicing factor binding / SMN-Sm protein complex / P granule / spliceosomal tri-snRNP complex / U2-type precatalytic spliceosome / U2-type spliceosomal complex / telomerase RNA binding / telomerase holoenzyme complex / commitment complex / mRNA cis splicing, via spliceosome / U2-type prespliceosome assembly / negative regulation of chaperone-mediated autophagy / U2-type catalytic step 2 spliceosome / SAGA complex / U4 snRNP / U2 snRNP / RNA Polymerase II Transcription Termination / U1 snRNP / U2-type prespliceosome / positive regulation of transcription by RNA polymerase III / pre-mRNA 5'-splice site binding / precatalytic spliceosome / spliceosomal complex assembly / positive regulation of transcription by RNA polymerase I / regulation of RNA splicing / mRNA 5'-splice site recognition / mRNA Splicing - Minor Pathway / mRNA 3'-splice site recognition / U5 snRNP / U2 snRNA binding / spliceosomal snRNP assembly / cellular response to transforming growth factor beta stimulus / Cajal body / U1 snRNA binding / regulation of DNA repair / intercellular bridge / U4/U6 x U5 tri-snRNP complex / cellular response to retinoic acid / catalytic step 2 spliceosome / Hsp70 protein binding / Gene and protein expression by JAK-STAT signaling after Interleukin-12 stimulation / mRNA Splicing - Major Pathway / RNA splicing / stem cell differentiation / spliceosomal complex / negative regulation of smooth muscle cell proliferation / negative regulation of protein catabolic process / mRNA splicing, via spliceosome / positive regulation of neuron projection development / B-WICH complex positively regulates rRNA expression / nuclear matrix / cytoplasmic ribonucleoprotein granule / fibrillar center / mRNA processing / transcription corepressor activity / cellular response to tumor necrosis factor / snRNP Assembly / spermatogenesis / SARS-CoV-2 modulates host translation machinery / single-stranded RNA binding / RNA helicase activity / nuclear speck / nuclear body / RNA helicase / ribosome / chromatin remodeling / mRNA binding / protein-containing complex binding / positive regulation of DNA-templated transcription / nucleolus / enzyme binding / protein homodimerization activity / positive regulation of transcription by RNA polymerase II / ATP hydrolysis activity / DNA binding / RNA binding / extracellular exosome Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.0 Å | |||||||||
![]() | Zhang X / Zhan X / Shi Y | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Structural insights into branch site proofreading by human spliceosome. Authors: Xiaofeng Zhang / Xiechao Zhan / Tong Bian / Fenghua Yang / Pan Li / Yichen Lu / Zhihan Xing / Rongyan Fan / Qiangfeng Cliff Zhang / Yigong Shi / ![]() Abstract: Selection of the pre-mRNA branch site (BS) by the U2 small nuclear ribonucleoprotein (snRNP) is crucial to prespliceosome (A complex) assembly. The RNA helicase PRP5 proofreads BS selection but the ...Selection of the pre-mRNA branch site (BS) by the U2 small nuclear ribonucleoprotein (snRNP) is crucial to prespliceosome (A complex) assembly. The RNA helicase PRP5 proofreads BS selection but the underlying mechanism remains unclear. Here we report the atomic structures of two sequential complexes leading to prespliceosome assembly: human 17S U2 snRNP and a cross-exon pre-A complex. PRP5 is anchored on 17S U2 snRNP mainly through occupation of the RNA path of SF3B1 by an acidic loop of PRP5; the helicase domain of PRP5 associates with U2 snRNA; the BS-interacting stem-loop (BSL) of U2 snRNA is shielded by TAT-SF1, unable to engage the BS. In the pre-A complex, an initial U2-BS duplex is formed; the translocated helicase domain of PRP5 stays with U2 snRNA and the acidic loop still occupies the RNA path. The pre-A conformation is specifically stabilized by the splicing factors SF1, DNAJC8 and SF3A2. Cancer-derived mutations in SF3B1 damage its association with PRP5, compromising BS proofreading. Together, these findings reveal key insights into prespliceosome assembly and BS selection or proofreading by PRP5. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 398.8 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 47.3 KB 47.3 KB | Display Display | ![]() |
Images | ![]() | 64.2 KB | ||
Filedesc metadata | ![]() | 13.1 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 468.1 KB | Display | ![]() |
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Full document | ![]() | 467.7 KB | Display | |
Data in XML | ![]() | 7.6 KB | Display | |
Data in CIF | ![]() | 8.8 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7vpxMC ![]() 7evoC M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | The cryo-EM map for the U2 part of human pre-A complex | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.087 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
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Sample components
+Entire : human pre-A complex
+Supramolecule #1: human pre-A complex
+Supramolecule #2: human pre-A
+Supramolecule #3: pre-mRNA, 5SS
+Macromolecule #1: Splicing factor 3A subunit 2
+Macromolecule #2: Splicing factor 1
+Macromolecule #3: DnaJ homolog subfamily C member 8
+Macromolecule #5: Splicing factor 3B subunit 1
+Macromolecule #6: Splicing factor 3B subunit 2
+Macromolecule #7: Splicing factor 3B subunit 3
+Macromolecule #8: Splicing factor 3B subunit 4
+Macromolecule #9: Splicing factor 3B subunit 5
+Macromolecule #10: Splicing factor 3A subunit 1
+Macromolecule #11: Splicing factor 3A subunit 3
+Macromolecule #12: Probable ATP-dependent RNA helicase DDX46
+Macromolecule #13: U2 small nuclear ribonucleoprotein A'
+Macromolecule #14: U2 small nuclear ribonucleoprotein B''
+Macromolecule #15: Small nuclear ribonucleoprotein Sm D2
+Macromolecule #16: Small nuclear ribonucleoprotein F
+Macromolecule #17: Small nuclear ribonucleoprotein E
+Macromolecule #18: Small nuclear ribonucleoprotein G
+Macromolecule #19: Small nuclear ribonucleoprotein Sm D3
+Macromolecule #20: Small nuclear ribonucleoprotein-associated proteins B and B'
+Macromolecule #21: Small nuclear ribonucleoprotein Sm D1
+Macromolecule #25: U1 small nuclear ribonucleoprotein 70 kDa
+Macromolecule #26: U1 small nuclear ribonucleoprotein A
+Macromolecule #27: U1 small nuclear ribonucleoprotein C
+Macromolecule #28: PHD finger-like domain-containing protein 5A
+Macromolecule #4: U2 snRNA
+Macromolecule #22: pre-mRNA
+Macromolecule #23: 5SS
+Macromolecule #24: U1 snRNA
+Macromolecule #29: ZINC ION
+Macromolecule #30: spliceostatin A (form II)
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.9 |
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Vitrification | Cryogen name: ETHANE |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 1.5 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
Startup model | Type of model: INSILICO MODEL |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.0 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 419522 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |