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Yorodumi- EMDB-9646: Cryo-EM structure of a human intron lariat spliceosome prior to P... -
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Basic information
| Entry | Database: EMDB / ID: EMD-9646 | |||||||||
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| Title | Cryo-EM structure of a human intron lariat spliceosome prior to Prp43 loaded (ILS1 complex) at 2.9 angstrom resolution | |||||||||
Map data | The 2.9 angstrom map of the human ILS1 complex | |||||||||
Sample |
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Keywords | Human Intron Lariat Spliceosome / SPLICING | |||||||||
| Function / homology | Function and homology informationregulation of retinoic acid receptor signaling pathway / post-mRNA release spliceosomal complex / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / 3'-5' RNA helicase activity / U7 snRNP / generation of catalytic spliceosome for first transesterification step / histone pre-mRNA 3'end processing complex / regulation of vitamin D receptor signaling pathway ...regulation of retinoic acid receptor signaling pathway / post-mRNA release spliceosomal complex / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / 3'-5' RNA helicase activity / U7 snRNP / generation of catalytic spliceosome for first transesterification step / histone pre-mRNA 3'end processing complex / regulation of vitamin D receptor signaling pathway / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / embryonic brain development / nuclear retinoic acid receptor binding / protein methylation / oocyte development / U12-type spliceosomal complex / alternative mRNA splicing, via spliceosome / 7-methylguanosine cap hypermethylation / poly(A) binding / U1 snRNP binding / methylosome / mRNA 3'-end processing / pICln-Sm protein complex / U2-type catalytic step 1 spliceosome / RNA splicing, via transesterification reactions / pre-mRNA binding / snRNP binding / positive regulation of mRNA splicing, via spliceosome / small nuclear ribonucleoprotein complex / sno(s)RNA-containing ribonucleoprotein complex / SMN-Sm protein complex / spliceosomal tri-snRNP complex / host-mediated activation of viral transcription / P granule / telomerase holoenzyme complex / U2-type precatalytic spliceosome / commitment complex / mRNA cis splicing, via spliceosome / telomerase RNA binding / positive regulation of vitamin D receptor signaling pathway / U2-type prespliceosome assembly / U2-type spliceosomal complex / nuclear vitamin D receptor binding / Notch binding / Transport of Mature mRNA derived from an Intron-Containing Transcript / U2-type catalytic step 2 spliceosome / Regulation of gene expression in late stage (branching morphogenesis) pancreatic bud precursor cells / RUNX3 regulates NOTCH signaling / NOTCH4 Intracellular Domain Regulates Transcription / U2 snRNP / RNA Polymerase II Transcription Termination / positive regulation of neurogenesis / U1 snRNP / U4 snRNP / NOTCH3 Intracellular Domain Regulates Transcription / U2-type prespliceosome / inner cell mass cell proliferation / protein peptidyl-prolyl isomerization / K63-linked polyubiquitin modification-dependent protein binding / ubiquitin-ubiquitin ligase activity / nuclear androgen receptor binding / cyclosporin A binding / WD40-repeat domain binding / precatalytic spliceosome / lipid biosynthetic process / Notch-HLH transcription pathway / Formation of paraxial mesoderm / SMAD binding / positive regulation of transforming growth factor beta receptor signaling pathway / mitotic G2 DNA damage checkpoint signaling / spliceosomal complex assembly / mRNA Splicing - Minor Pathway / spliceosomal tri-snRNP complex assembly / Prp19 complex / blastocyst development / U5 snRNA binding / protein K63-linked ubiquitination / intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / U5 snRNP / protein localization to nucleus / embryonic organ development / U2 snRNA binding / positive regulation of G1/S transition of mitotic cell cycle / U6 snRNA binding / pre-mRNA intronic binding / spliceosomal snRNP assembly / RNA processing / U1 snRNA binding / positive regulation of viral genome replication / Cajal body / ovarian follicle development / retinoic acid receptor signaling pathway / U4/U6 x U5 tri-snRNP complex / proteasomal protein catabolic process / spindle assembly / cellular response to retinoic acid / gastrulation / catalytic step 2 spliceosome / transcription-coupled nucleotide-excision repair Similarity search - Function | |||||||||
| Biological species | Homo sapiens (human) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 2.9 Å | |||||||||
Authors | Zhang X / Zhan X | |||||||||
| Funding support | China, 2 items
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Citation | Journal: Cell Res / Year: 2019Title: Structures of the human spliceosomes before and after release of the ligated exon. Authors: Xiaofeng Zhang / Xiechao Zhan / Chuangye Yan / Wenyu Zhang / Dongliang Liu / Jianlin Lei / Yigong Shi / ![]() Abstract: Pre-mRNA splicing is executed by the spliceosome, which has eight major functional states each with distinct composition. Five of these eight human spliceosomal complexes, all preceding exon ...Pre-mRNA splicing is executed by the spliceosome, which has eight major functional states each with distinct composition. Five of these eight human spliceosomal complexes, all preceding exon ligation, have been structurally characterized. In this study, we report the cryo-electron microscopy structures of the human post-catalytic spliceosome (P complex) and intron lariat spliceosome (ILS) at average resolutions of 3.0 and 2.9 Å, respectively. In the P complex, the ligated exon remains anchored to loop I of U5 small nuclear RNA, and the 3'-splice site is recognized by the junction between the 5'-splice site and the branch point sequence. The ATPase/helicase Prp22, along with the ligated exon and eight other proteins, are dissociated in the P-to-ILS transition. Intriguingly, the ILS complex exists in two distinct conformations, one with the ATPase/helicase Prp43 and one without. Comparison of these three late-stage human spliceosomes reveals mechanistic insights into exon release and spliceosome disassembly. | |||||||||
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Structure visualization
| Movie |
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| Structure viewer | EM map: SurfView Molmil Jmol/JSmol |
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_9646.map.gz | 226.7 MB | EMDB map data format | |
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| Header (meta data) | emd-9646-v30.xml emd-9646.xml | 52.2 KB 52.2 KB | Display Display | EMDB header |
| Images | emd_9646.png | 91.3 KB | ||
| Filedesc metadata | emd-9646.cif.gz | 16.2 KB | ||
| Others | emd_9646_additional.map.gz | 193.4 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-9646 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-9646 | HTTPS FTP |
-Validation report
| Summary document | emd_9646_validation.pdf.gz | 537.4 KB | Display | EMDB validaton report |
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| Full document | emd_9646_full_validation.pdf.gz | 536.9 KB | Display | |
| Data in XML | emd_9646_validation.xml.gz | 7 KB | Display | |
| Data in CIF | emd_9646_validation.cif.gz | 8.1 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9646 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9646 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 6id0MC ![]() 9645C ![]() 9647C ![]() 6iczC ![]() 6id1C M: atomic model generated by this map C: citing same article ( |
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| Similar structure data |
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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_9646.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Annotation | The 2.9 angstrom map of the human ILS1 complex | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.338 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Additional map: The 4.2 angstrom map of the human ILS1 complex
| File | emd_9646_additional.map | ||||||||||||
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| Annotation | The 4.2 angstrom map of the human ILS1 complex | ||||||||||||
| Projections & Slices |
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| Density Histograms |
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Sample components
+Entire : Human Intron Lariat Spliceosome
+Supramolecule #1: Human Intron Lariat Spliceosome
+Macromolecule #1: Pre-mRNA-processing-splicing factor 8
+Macromolecule #3: 116 kDa U5 small nuclear ribonucleoprotein component
+Macromolecule #4: U5 small nuclear ribonucleoprotein 40 kDa protein
+Macromolecule #6: Crooked neck-like protein 1
+Macromolecule #7: Cell division cycle 5-like protein
+Macromolecule #8: Pre-mRNA-splicing factor SYF2
+Macromolecule #9: Protein BUD31 homolog
+Macromolecule #10: Pre-mRNA-splicing factor RBM22
+Macromolecule #11: Spliceosome-associated protein CWC15 homolog
+Macromolecule #12: SNW domain-containing protein 1
+Macromolecule #13: Peptidyl-prolyl cis-trans isomerase-like 1
+Macromolecule #14: Pleiotropic regulator 1
+Macromolecule #15: Pre-mRNA-processing factor 17
+Macromolecule #18: CWF19-like protein 2
+Macromolecule #19: Pre-mRNA-splicing factor SYF1
+Macromolecule #20: Small nuclear ribonucleoprotein Sm D3
+Macromolecule #21: Small nuclear ribonucleoprotein-associated protein
+Macromolecule #22: Small nuclear ribonucleoprotein Sm D1
+Macromolecule #23: Small nuclear ribonucleoprotein Sm D2
+Macromolecule #24: Small nuclear ribonucleoprotein F
+Macromolecule #25: Small nuclear ribonucleoprotein E
+Macromolecule #26: Small nuclear ribonucleoprotein G
+Macromolecule #27: Pre-mRNA-processing factor 19
+Macromolecule #28: Pre-mRNA-splicing factor SPF27
+Macromolecule #29: U2 small nuclear ribonucleoprotein A'
+Macromolecule #30: U2 small nuclear ribonucleoprotein B''
+Macromolecule #31: RNA helicase aquarius
+Macromolecule #32: Peptidyl-prolyl cis-trans isomerase E
+Macromolecule #2: U5snRNA
+Macromolecule #5: U6snRNA
+Macromolecule #16: pre-mRNA
+Macromolecule #17: U2snRNA
+Macromolecule #33: INOSITOL HEXAKISPHOSPHATE
+Macromolecule #34: GUANOSINE-5'-TRIPHOSPHATE
+Macromolecule #35: MAGNESIUM ION
+Macromolecule #36: 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
| 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 K2 SUMMIT (4k x 4k) / Average electron dose: 45.0 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
| Startup model | Type of model: PDB ENTRY |
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| Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 2.9 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 390072 |
| Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
| Final angle assignment | Type: MAXIMUM LIKELIHOOD |
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About Yorodumi


Keywords
Homo sapiens (human)
Authors
China, 2 items
Citation
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