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- 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 | |||||||||
![]() | The 2.9 angstrom map of the human ILS1 complex | |||||||||
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![]() | Human Intron Lariat Spliceosome / SPLICING | |||||||||
Function / homology | ![]() 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 / regulation of vitamin D receptor signaling pathway / generation of catalytic spliceosome for first transesterification step / alternative mRNA splicing, via spliceosome ...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 / regulation of vitamin D receptor signaling pathway / generation of catalytic spliceosome for first transesterification step / alternative mRNA splicing, via spliceosome / histone pre-mRNA 3'end processing complex / positive regulation of androgen receptor activity / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / nuclear retinoic acid receptor binding / protein methylation / embryonic brain development / U12-type spliceosomal complex / poly(A) binding / 7-methylguanosine cap hypermethylation / methylosome / mRNA 3'-end processing / sno(s)RNA-containing ribonucleoprotein complex / U1 snRNP binding / RNA splicing, via transesterification reactions / oocyte development / pICln-Sm protein complex / positive regulation of mRNA splicing, via spliceosome / U2-type catalytic step 1 spliceosome / pre-mRNA binding / snRNP binding / small nuclear ribonucleoprotein complex / telomerase RNA binding / telomerase holoenzyme complex / SMN-Sm protein complex / P granule / positive regulation by host of viral transcription / spliceosomal tri-snRNP complex / positive regulation of vitamin D receptor signaling pathway / U2-type precatalytic spliceosome / nuclear vitamin D receptor binding / U2-type spliceosomal complex / mRNA cis splicing, via spliceosome / commitment complex / U2-type prespliceosome assembly / Transport of Mature mRNA derived from an Intron-Containing Transcript / Notch binding / Regulation of gene expression in late stage (branching morphogenesis) pancreatic bud precursor cells / RUNX3 regulates NOTCH signaling / U2-type catalytic step 2 spliceosome / NOTCH4 Intracellular Domain Regulates Transcription / U4 snRNP / RNA Polymerase II Transcription Termination / U2 snRNP / U1 snRNP / NOTCH3 Intracellular Domain Regulates Transcription / WD40-repeat domain binding / positive regulation of neurogenesis / U2-type prespliceosome / inner cell mass cell proliferation / ubiquitin-ubiquitin ligase activity / K63-linked polyubiquitin modification-dependent protein binding / nuclear androgen receptor binding / precatalytic spliceosome / lipid biosynthetic process / cyclosporin A binding / Notch-HLH transcription pathway / protein peptidyl-prolyl isomerization / positive regulation of transforming growth factor beta receptor signaling pathway / Formation of paraxial mesoderm / SMAD binding / spliceosomal complex assembly / mitotic G2 DNA damage checkpoint signaling / mRNA Splicing - Minor Pathway / blastocyst development / spliceosomal tri-snRNP complex assembly / protein K63-linked ubiquitination / Prp19 complex / protein localization to nucleus / intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / positive regulation of G1/S transition of mitotic cell cycle / U5 snRNA binding / U5 snRNP / embryonic organ development / U2 snRNA binding / U6 snRNA binding / RNA processing / positive regulation of viral genome replication / pre-mRNA intronic binding / spindle assembly / spliceosomal snRNP assembly / proteasomal protein catabolic process / transcription-coupled nucleotide-excision repair / Cajal body / retinoic acid receptor signaling pathway / U1 snRNA binding / ovarian follicle development / U4/U6 x U5 tri-snRNP complex / cellular response to retinoic acid / gastrulation Similarity search - Function | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.9 Å | |||||||||
![]() | Zhang X / Zhan X | |||||||||
Funding support | ![]()
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![]() | ![]() 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 |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 226.7 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 52.2 KB 52.2 KB | Display Display | ![]() |
Images | ![]() | 91.3 KB | ||
Filedesc metadata | ![]() | 16.2 KB | ||
Others | ![]() | 193.4 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 537.4 KB | Display | ![]() |
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Full document | ![]() | 536.9 KB | Display | |
Data in XML | ![]() | 7 KB | Display | |
Data in CIF | ![]() | 8.1 KB | Display | |
Arichive directory | ![]() ![]() | 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 | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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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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![]() | 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: ![]() |
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 |