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Yorodumi- EMDB-9646: Cryo-EM structure of a human intron lariat spliceosome prior to P... -
+Open data
-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 information 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 / regulation of vitamin D receptor signaling pathway / histone pre-mRNA 3'end processing complex ...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 / regulation of vitamin D receptor signaling pathway / histone pre-mRNA 3'end processing complex / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / protein methylation / U12-type spliceosomal complex / embryonic brain development / methylosome / nuclear retinoic acid receptor binding / 7-methylguanosine cap hypermethylation / positive regulation of androgen receptor activity / Prp19 complex / poly(A) binding / U1 snRNP binding / pICln-Sm protein complex / mRNA 3'-end processing / sno(s)RNA-containing ribonucleoprotein complex / snRNP binding / U2-type catalytic step 1 spliceosome / RNA splicing, via transesterification reactions / small nuclear ribonucleoprotein complex / pre-mRNA binding / SMN-Sm protein complex / telomerase RNA binding / spliceosomal tri-snRNP complex / telomerase holoenzyme complex / U2-type spliceosomal complex / U2-type precatalytic spliceosome / positive regulation by host of viral transcription / mRNA cis splicing, via spliceosome / P granule / positive regulation of vitamin D receptor signaling pathway / commitment complex / U2-type prespliceosome assembly / nuclear vitamin D receptor binding / Transport of Mature mRNA derived from an Intron-Containing Transcript / U2-type catalytic step 2 spliceosome / Notch binding / Regulation of gene expression in late stage (branching morphogenesis) pancreatic bud precursor cells / U4 snRNP / positive regulation of mRNA splicing, via spliceosome / RUNX3 regulates NOTCH signaling / U2 snRNP / NOTCH4 Intracellular Domain Regulates Transcription / RNA Polymerase II Transcription Termination / U1 snRNP / NOTCH3 Intracellular Domain Regulates Transcription / ubiquitin-ubiquitin ligase activity / positive regulation of neurogenesis / U2-type prespliceosome / WD40-repeat domain binding / lipid biosynthetic process / nuclear androgen receptor binding / precatalytic spliceosome / K63-linked polyubiquitin modification-dependent protein binding / cyclosporin A binding / Notch-HLH transcription pathway / positive regulation of transforming growth factor beta receptor signaling pathway / Formation of paraxial mesoderm / mRNA Splicing - Minor Pathway / spliceosomal complex assembly / SMAD binding / mitotic G2 DNA damage checkpoint signaling / protein peptidyl-prolyl isomerization / blastocyst development / protein localization to nucleus / spliceosomal tri-snRNP complex assembly / protein K63-linked ubiquitination / intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / U5 snRNA binding / U5 snRNP / transcription-coupled nucleotide-excision repair / embryonic organ development / positive regulation of G1/S transition of mitotic cell cycle / proteasomal protein catabolic process / retinoic acid receptor signaling pathway / positive regulation of viral genome replication / U2 snRNA binding / U6 snRNA binding / spliceosomal snRNP assembly / Cajal body / RNA processing / pre-mRNA intronic binding / U1 snRNA binding / U4/U6 x U5 tri-snRNP complex / cellular response to retinoic acid / gastrulation / catalytic step 2 spliceosome / Gene and protein expression by JAK-STAT signaling after Interleukin-12 stimulation / mRNA Splicing - Major Pathway / RNA splicing / lipid droplet 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: 2019 Title: 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. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-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 (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-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 |
-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 |
-Sample preparation
Buffer | pH: 7.9 |
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Vitrification | Cryogen name: ETHANE |
-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 |
-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 |