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Open data
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Basic information
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| Title | The cryo-EM map of the human 17S U2 snRNP core region | |||||||||
Map data | The EM map of U2 snRNP | |||||||||
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Keywords | U2 snRNP / PRP5 / SF3B1 / Splicing | |||||||||
| Function / homology | Function and homology informationU11/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 / poly-ADP-D-ribose modification-dependent protein binding ...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 / poly-ADP-D-ribose modification-dependent protein binding / 7-methylguanosine cap hypermethylation / U12-type spliceosomal complex / U1 snRNP binding / pICln-Sm protein complex / methylosome / RNA splicing, via transesterification reactions / small nuclear ribonucleoprotein complex / splicing factor binding / SMN-Sm protein complex / protein localization to site of double-strand break / 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 catalytic step 2 spliceosome / U2-type spliceosomal complex / telomerase holoenzyme complex / telomerase RNA binding / pre-mRNA binding / U1 snRNP / U2 snRNP / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / chromatin-protein adaptor activity / RNA Polymerase II Transcription Termination / protein methylation / U2-type prespliceosome / precatalytic spliceosome / 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 / U2 snRNA binding / U4/U6 x U5 tri-snRNP complex / Cajal body / U1 snRNA binding / spliceosomal snRNP assembly / stem cell differentiation / catalytic step 2 spliceosome / spliceosomal complex / mRNA Splicing - Major Pathway / Gene and protein expression by JAK-STAT signaling after Interleukin-12 stimulation / RNA splicing / regulation of DNA repair / positive regulation of neuron projection development / negative regulation of protein catabolic process / mRNA splicing, via spliceosome / double-strand break repair via homologous recombination / B-WICH complex positively regulates rRNA expression / mRNA processing / mRNA Polyadenylation / nuclear matrix / site of double-strand break / Dengue Virus-Host Interactions / snRNP Assembly / spermatogenesis / SARS-CoV-2 modulates host translation machinery / nuclear body / RNA helicase activity / nuclear speck / RNA helicase / chromatin remodeling / hydrolase activity / mRNA binding / nucleolus / positive regulation of DNA-templated transcription / protein-containing complex binding / chromatin / enzyme binding / positive regulation of transcription by RNA polymerase II / DNA binding / DNA-templated transcription / RNA binding / extracellular exosome / nucleoplasm / zinc ion binding / ATP binding / nucleus / cytosol / cytoplasm Similarity search - Function | |||||||||
| Biological species | Homo sapiens (human) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 2.5 Å | |||||||||
Authors | Zhang X / Zhan X | |||||||||
| 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. | |||||||||
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_31334.map.gz | 116.7 MB | EMDB map data format | |
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| Header (meta data) | emd-31334-v30.xml emd-31334.xml | 40.3 KB 40.3 KB | Display Display | EMDB header |
| Images | emd_31334.png | 35.5 KB | ||
| Filedesc metadata | emd-31334.cif.gz | 12.9 KB | ||
| Archive directory | https://data.pdbj.org/pub/emdb/structures/EMD-31334 ftp://data.pdbj.org/pub/emdb/structures/EMD-31334 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 7evoMC ![]() 7vpxC 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_31334.map.gz / Format: CCP4 / Size: 125 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | The EM map of U2 snRNP | ||||||||||||||||||||||||||||||||||||
| 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 : The human U2 snRNP
+Supramolecule #1: The human U2 snRNP
+Macromolecule #1: U2 snRNA
+Macromolecule #2: Splicing factor 3B subunit 1
+Macromolecule #3: Splicing factor 3B subunit 2
+Macromolecule #4: Splicing factor 3B subunit 3
+Macromolecule #5: Splicing factor 3B subunit 4
+Macromolecule #6: Splicing factor 3B subunit 5
+Macromolecule #7: PHD finger-like domain-containing protein 5A
+Macromolecule #8: Splicing factor 3A subunit 1
+Macromolecule #9: Splicing factor 3A subunit 2
+Macromolecule #10: Splicing factor 3A subunit 3
+Macromolecule #11: HIV Tat-specific factor 1
+Macromolecule #12: RNA helicase
+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 #22: [(2~{S},3~{S},4~{E},6~{S},7~{R},10~{R})-3,7-dimethyl-2-[(2~{E},4~...
+Macromolecule #23: ZINC ION
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Concentration | 0.5 mg/mL |
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| Buffer | pH: 7.9 |
| Grid | Model: Quantifoil R1.2/1.3 / Material: GOLD / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE |
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Instrument: FEI VITROBOT MARK IV |
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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 / Cs: 2.7 mm |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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About Yorodumi




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



















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Y (Row.)
X (Col.)





















Processing
FIELD EMISSION GUN
