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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 | |||||||||
Map data | The cryo-EM map for the U2 part of human pre-A complex | |||||||||
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Keywords | human spliceosome / pre-A complex / PRP5 / A complex / U1 snRNP / U2 snRNP / SPLICING | |||||||||
| Function / homology | Function and homology informationnegative regulation of protein refolding / U11/U12 snRNP / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / U7 snRNP / histone pre-mRNA 3'end processing complex / nuclear histone mRNA catabolic process / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs ...negative regulation of protein refolding / U11/U12 snRNP / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / U7 snRNP / histone pre-mRNA 3'end processing complex / nuclear histone mRNA catabolic process / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / 7-methylguanosine cap hypermethylation / U12-type spliceosomal complex / U1 snRNP binding / methylosome / pICln-Sm protein complex / RNA splicing, via transesterification reactions / small nuclear ribonucleoprotein complex / splicing factor binding / SMN-Sm protein complex / spliceosomal tri-snRNP complex / commitment complex / P granule / U4 snRNP / snRNP binding / U2-type precatalytic spliceosome / mRNA cis splicing, via spliceosome / U2-type prespliceosome assembly / U2-type spliceosomal complex / telomerase holoenzyme complex / U2-type catalytic step 2 spliceosome / telomerase RNA binding / pre-mRNA binding / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / U1 snRNP / U2 snRNP / RNA Polymerase II Transcription Termination / protein methylation / pre-mRNA 5'-splice site binding / U2-type prespliceosome / negative regulation of chaperone-mediated autophagy / precatalytic spliceosome / mRNA 5'-splice site recognition / mRNA 3'-splice site recognition / regulation of RNA splicing / positive regulation of transcription by RNA polymerase III / mRNA Splicing - Minor Pathway / positive regulation of transcription by RNA polymerase I / spliceosomal complex assembly / negative regulation of mRNA splicing, via spliceosome / spliceosomal tri-snRNP complex assembly / U5 snRNP / cellular response to transforming growth factor beta stimulus / U2 snRNA binding / U4/U6 x U5 tri-snRNP complex / Cajal body / U1 snRNA binding / cellular response to retinoic acid / spliceosomal snRNP assembly / stem cell differentiation / catalytic step 2 spliceosome / spliceosomal complex / mRNA Splicing - Major Pathway / lysosomal lumen / Gene and protein expression by JAK-STAT signaling after Interleukin-12 stimulation / RNA splicing / regulation of DNA repair / Hsp70 protein binding / positive regulation of neuron projection development / cellular response to tumor necrosis factor / negative regulation of protein catabolic process / mRNA splicing, via spliceosome / B-WICH complex positively regulates rRNA expression / mRNA processing / mRNA Polyadenylation / nuclear matrix / transcription corepressor activity / Dengue Virus-Host Interactions / snRNP Assembly / spermatogenesis / SARS-CoV-2 modulates host translation machinery / nuclear body / RNA helicase activity / nuclear speck / single-stranded RNA binding / RNA helicase / chromatin remodeling / ribosome / mRNA binding / nucleolus / positive regulation of DNA-templated transcription / protein-containing complex binding / chromatin / enzyme binding / positive regulation of transcription by RNA polymerase II / protein homodimerization activity / ATP hydrolysis activity / DNA binding / DNA-templated transcription / RNA binding / extracellular exosome Similarity search - Function | |||||||||
| Biological species | Homo sapiens (human) / unidentified adenovirus | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.0 Å | |||||||||
Authors | Zhang X / Zhan X / Shi Y | |||||||||
| Funding support | China, 1 items
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Citation | Journal: Nat Struct Mol Biol / Year: 2024Title: 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 | emd_32074.map.gz | 398.8 MB | EMDB map data format | |
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| Header (meta data) | emd-32074-v30.xml emd-32074.xml | 47.3 KB 47.3 KB | Display Display | EMDB header |
| Images | emd_32074.png | 64.2 KB | ||
| Filedesc metadata | emd-32074.cif.gz | 13.1 KB | ||
| Archive directory | https://data.pdbj.org/pub/emdb/structures/EMD-32074 ftp://data.pdbj.org/pub/emdb/structures/EMD-32074 | 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 F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_32074.map.gz / Format: CCP4 / Size: 421.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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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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Processing | 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: FIELD EMISSION GUN |
| 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 |
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About Yorodumi




Keywords
Homo sapiens (human)
unidentified adenovirus
Authors
China, 1 items
Citation
























Z (Sec.)
Y (Row.)
X (Col.)





















FIELD EMISSION GUN
