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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 / 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 / U1 snRNP binding / methylosome / RNA splicing, via transesterification reactions / pICln-Sm protein complex / regulation of mRNA splicing, via spliceosome / snRNP binding / positive regulation of mRNA splicing, via spliceosome / splicing factor binding / SMN-Sm protein complex / small nuclear ribonucleoprotein complex / spliceosomal tri-snRNP complex / P granule / telomerase holoenzyme complex / U2-type precatalytic spliceosome / commitment complex / mRNA cis splicing, via spliceosome / telomerase RNA binding / U2-type spliceosomal complex / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / SAGA complex / RNA Polymerase II Transcription Termination / U2 snRNP / negative regulation of chaperone-mediated autophagy / U1 snRNP / U4 snRNP / U2-type prespliceosome / positive regulation of transcription by RNA polymerase III / pre-mRNA 5'-splice site binding / precatalytic spliceosome / mRNA 5'-splice site recognition / regulation of RNA splicing / spliceosomal complex assembly / mRNA Splicing - Minor Pathway / mRNA 3'-splice site recognition / positive regulation of transcription by RNA polymerase I / U5 snRNP / U2 snRNA binding / spliceosomal snRNP assembly / cellular response to transforming growth factor beta stimulus / U1 snRNA binding / Cajal body / 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 / mRNA Splicing - Major Pathway / Gene and protein expression by JAK-STAT signaling after Interleukin-12 stimulation / RNA splicing / stem cell differentiation / spliceosomal complex / negative regulation of smooth muscle cell proliferation / positive regulation of neuron projection development / mRNA splicing, via spliceosome / negative regulation of protein catabolic process / B-WICH complex positively regulates rRNA expression / nuclear matrix / fibrillar center / cytoplasmic ribonucleoprotein granule / mRNA processing / cellular response to tumor necrosis factor / transcription corepressor activity / snRNP Assembly / spermatogenesis / SARS-CoV-2 modulates host translation machinery / RNA helicase activity / single-stranded RNA binding / nuclear speck / nuclear body / RNA helicase / ribosome / chromatin remodeling / mRNA binding / positive regulation of DNA-templated transcription / protein-containing complex binding / 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 | 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 | http://ftp.pdbj.org/pub/emdb/structures/EMD-32074 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-32074 | HTTPS FTP |
-Validation report
| Summary document | emd_32074_validation.pdf.gz | 468.1 KB | Display | EMDB validaton report |
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| Full document | emd_32074_full_validation.pdf.gz | 467.7 KB | Display | |
| Data in XML | emd_32074_validation.xml.gz | 7.6 KB | Display | |
| Data in CIF | emd_32074_validation.cif.gz | 8.8 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-32074 ftp://ftp.pdbj.org/pub/emdb/validation_reports/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
