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Yorodumi- PDB-5z57: Cryo-EM structure of the human activated spliceosome (late Bact) ... -
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Basic information
| Entry | Database: PDB / ID: 5z57 | ||||||||||||
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| Title | Cryo-EM structure of the human activated spliceosome (late Bact) at 6.5 angstrom | ||||||||||||
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Keywords | SPLICING / spliceosome / cryo-EM structure / activated spliceosome / late Bact complex / pre-mRNA splicing | ||||||||||||
| Function / homology | Function and homology informationRES complex / U11/U12 snRNP / 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 ...RES complex / U11/U12 snRNP / 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 / cis assembly of pre-catalytic spliceosome / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / B-WICH complex / miRNA processing / embryonic brain development / 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 / C2H2 zinc finger domain binding / U2-type catalytic step 1 spliceosome / RNA splicing, via transesterification reactions / pre-mRNA binding / snRNP binding / positive regulation of mRNA splicing, via spliceosome / regulation of mRNA splicing, via spliceosome / blastocyst formation / small nuclear ribonucleoprotein complex / sno(s)RNA-containing ribonucleoprotein complex / splicing factor binding / SMN-Sm protein 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 prespliceosome assembly / U2-type spliceosomal complex / Transport of Mature mRNA derived from an Intron-Containing Transcript / U2-type catalytic step 2 spliceosome / U2 snRNP / RNA Polymerase II Transcription Termination / SAGA complex / U1 snRNP / U4 snRNP / U2-type prespliceosome / positive regulation of transcription by RNA polymerase III / inner cell mass cell proliferation / protein peptidyl-prolyl isomerization / K63-linked polyubiquitin modification-dependent protein binding / ubiquitin-ubiquitin ligase activity / cyclosporin A binding / WD40-repeat domain binding / precatalytic spliceosome / pattern recognition receptor activity / lipid biosynthetic process / regulation of RNA splicing / spliceosomal complex assembly / mRNA Splicing - Minor Pathway / mRNA 3'-splice site recognition / positive regulation of transcription by RNA polymerase I / spliceosomal tri-snRNP complex assembly / Prp19 complex / blastocyst development / U5 snRNA binding / protein K63-linked ubiquitination / U5 snRNP / protein localization to nucleus / 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 / regulation of DNA repair / transcription regulator inhibitor activity / ovarian follicle development / U4/U6 x U5 tri-snRNP complex / proteasomal protein catabolic process / spindle assembly / catalytic step 2 spliceosome / transcription-coupled nucleotide-excision repair / lipid droplet / negative regulation of canonical NF-kappaB signal transduction / antiviral innate immune response / mRNA Splicing - Major Pathway / Gene and protein expression by JAK-STAT signaling after Interleukin-12 stimulation Similarity search - Function | ||||||||||||
| Biological species | Homo sapiens (human) unidentified adenovirus | ||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 6.5 Å | ||||||||||||
Authors | Zhang, X. / Yan, C. / Zhan, X. / Li, L. / Lei, J. / Shi, Y. | ||||||||||||
| Funding support | China, 3items
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Citation | Journal: Cell Res / Year: 2018Title: Structure of the human activated spliceosome in three conformational states. Authors: Xiaofeng Zhang / Chuangye Yan / Xiechao Zhan / Lijia Li / Jianlin Lei / Yigong Shi / ![]() Abstract: During each cycle of pre-mRNA splicing, the pre-catalytic spliceosome (B complex) is converted into the activated spliceosome (B complex), which has a well-formed active site but cannot proceed to ...During each cycle of pre-mRNA splicing, the pre-catalytic spliceosome (B complex) is converted into the activated spliceosome (B complex), which has a well-formed active site but cannot proceed to the branching reaction. Here, we present the cryo-EM structure of the human B complex in three distinct conformational states. The EM map allows atomic modeling of nearly all protein components of the U2 small nuclear ribonucleoprotein (snRNP), including three of the SF3a complex and seven of the SF3b complex. The structure of the human B complex contains 52 proteins, U2, U5, and U6 small nuclear RNA (snRNA), and a pre-mRNA. Three distinct conformations have been captured, representing the early, mature, and late states of the human B complex. These complexes differ in the orientation of the Switch loop of Prp8, the splicing factors RNF113A and NY-CO-10, and most components of the NineTeen complex (NTC) and the NTC-related complex. Analysis of these three complexes and comparison with the B and C complexes reveal an ordered flux of components in the B-to-B and the B-to-B transitions, which ultimately prime the active site for the branching reaction. | ||||||||||||
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Structure visualization
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| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 5z57.cif.gz | 3.3 MB | Display | PDBx/mmCIF format |
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| PDB format | pdb5z57.ent.gz | Display | PDB format | |
| PDBx/mmJSON format | 5z57.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Summary document | 5z57_validation.pdf.gz | 2 MB | Display | wwPDB validaton report |
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| Full document | 5z57_full_validation.pdf.gz | 2.5 MB | Display | |
| Data in XML | 5z57_validation.xml.gz | 420.9 KB | Display | |
| Data in CIF | 5z57_validation.cif.gz | 677.7 KB | Display | |
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/z5/5z57 ftp://data.pdbj.org/pub/pdb/validation_reports/z5/5z57 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 6890MC ![]() 6889C ![]() 6891C ![]() 5z56C ![]() 5z58C C: citing same article ( M: map data used to model this data |
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| Similar structure data |
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Links
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Assembly
| Deposited unit | ![]()
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Components
+Protein , 15 types, 16 molecules ACbi6JLQNPRTUXYZ
+RNA chain , 4 types, 4 molecules BFGH
+U5 small nuclear ribonucleoprotein ... , 2 types, 2 molecules DE
+Small nuclear ribonucleoprotein ... , 6 types, 12 molecules ahcjdkfmelgn
+U2 small nuclear ribonucleoprotein ... , 2 types, 2 molecules op
+Splicing factor 3A subunit ... , 3 types, 3 molecules wuv
+Splicing factor 3B subunit ... , 6 types, 6 molecules 123457
+Pre-mRNA-processing factor ... , 2 types, 5 molecules qrstW
+Pre-mRNA-splicing factor ... , 5 types, 5 molecules KIOVx
+Peptidyl-prolyl cis-trans ... , 2 types, 2 molecules Sy
+Non-polymers , 5 types, 19 molecules 








+Details
-Experimental details
-Experiment
| Experiment | Method: ELECTRON MICROSCOPY |
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| EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
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Sample preparation
| Component | Name: late Bact splieosome / Type: COMPLEX / Entity ID: #1-#47 / Source: NATURAL |
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| Source (natural) | Organism: Homo sapiens (human) |
| Buffer solution | pH: 7.9 / Details: 20 mM HEPES-KOH, pH 7.9, 150 mM NaCl, 1.5 mM MgCl2 |
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
| Vitrification | Cryogen name: ETHANE |
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Electron microscopy imaging
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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| Microscopy | Model: FEI TITAN KRIOS |
| Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
| Electron lens | Mode: BRIGHT FIELD / Cs: 2.7 mm / Alignment procedure: BASIC |
| Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
| Image recording | Electron dose: 48 e/Å2 / Detector mode: SUPER-RESOLUTION / Film or detector model: GATAN K2 SUMMIT (4k x 4k) |
| EM imaging optics | Energyfilter name: GIF Quantum LS |
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Processing
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| CTF correction | Type: NONE | ||||||||||||||||||||||||||||
| Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||||||
| 3D reconstruction | Resolution: 6.5 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 14316 / Algorithm: FOURIER SPACE / Symmetry type: POINT | ||||||||||||||||||||||||||||
| Refinement | Highest resolution: 6.5 Å |
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About Yorodumi



Homo sapiens (human)
unidentified adenovirus
China, 3items
Citation
UCSF Chimera













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