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- EMDB-10689: human 17S U2 snRNP -

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Basic information

Entry
Database: EMDB / ID: EMD-10689
Titlehuman 17S U2 snRNP
Map data
Sample
  • Complex: human 17S U2 snRNP
    • Protein or peptide: x 18 types
    • RNA: x 1 types
Function / homology
Function and homology information


U11/U12 snRNP / snRNP binding / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / chromatin-protein adaptor activity / U7 snRNP / B-WICH complex / histone pre-mRNA 3'end processing complex / SLBP independent Processing of Histone Pre-mRNAs ...U11/U12 snRNP / snRNP binding / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / chromatin-protein adaptor activity / U7 snRNP / B-WICH complex / histone pre-mRNA 3'end processing complex / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / splicing factor binding / protein methylation / protein localization to site of double-strand break / U12-type spliceosomal complex / methylosome / 7-methylguanosine cap hypermethylation / pICln-Sm protein complex / U1 snRNP binding / RNA splicing, via transesterification reactions / blastocyst formation / poly-ADP-D-ribose modification-dependent protein binding / spliceosomal tri-snRNP complex / small nuclear ribonucleoprotein complex / P granule / SMN-Sm protein complex / : / mRNA cis splicing, via spliceosome / U2-type spliceosomal complex / telomerase RNA binding / telomerase holoenzyme complex / positive regulation of mRNA splicing, via spliceosome / U2-type precatalytic spliceosome / U2-type prespliceosome assembly / commitment complex / U2-type catalytic step 2 spliceosome / U4 snRNP / U2 snRNP / SAGA complex / RNA Polymerase II Transcription Termination / positive regulation of transcription by RNA polymerase III / U1 snRNP / U2-type prespliceosome / precatalytic spliceosome / spliceosomal complex assembly / positive regulation of transcription by RNA polymerase I / mRNA Splicing - Minor Pathway / regulation of RNA splicing / mRNA 3'-splice site recognition / U5 snRNP / regulation of DNA repair / Cajal body / U2 snRNA binding / spliceosomal snRNP assembly / U1 snRNA binding / U4/U6 x U5 tri-snRNP complex / catalytic step 2 spliceosome / Gene and protein expression by JAK-STAT signaling after Interleukin-12 stimulation / mRNA Splicing - Major Pathway / viral genome replication / RNA splicing / regulation of DNA-templated transcription elongation / stem cell differentiation / spliceosomal complex / double-strand break repair via homologous recombination / B-WICH complex positively regulates rRNA expression / negative regulation of protein catabolic process / positive regulation of neuron projection development / cytoplasmic ribonucleoprotein granule / mRNA splicing, via spliceosome / mRNA processing / fibrillar center / nuclear matrix / site of double-strand break / snRNP Assembly / SARS-CoV-2 modulates host translation machinery / spermatogenesis / RNA helicase activity / nuclear body / RNA helicase / nuclear speck / chromatin remodeling / mRNA binding / protein-containing complex binding / nucleolus / regulation of transcription by RNA polymerase II / positive regulation of DNA-templated transcription / enzyme binding / ATP hydrolysis activity / positive regulation of transcription by RNA polymerase II / DNA binding / RNA binding / extracellular exosome / zinc ion binding / nucleoplasm / ATP binding / nucleus / cytosol / cytoplasm
Similarity search - Function
TatSF1-like, RNA recognition motif 1 / TatSF1-like / SF3B6, RNA recognition motif / Splicing factor 3A subunit 1, ubiquitin domain / SF3B4, RNA recognition motif 2 / Splicing factor SF3a60 binding domain / Splicing factor SF3a60 binding domain / : / Replication stress response SDE2 C-terminal / Cactus-binding C-terminus of cactin protein ...TatSF1-like, RNA recognition motif 1 / TatSF1-like / SF3B6, RNA recognition motif / Splicing factor 3A subunit 1, ubiquitin domain / SF3B4, RNA recognition motif 2 / Splicing factor SF3a60 binding domain / Splicing factor SF3a60 binding domain / : / Replication stress response SDE2 C-terminal / Cactus-binding C-terminus of cactin protein / U2 small nuclear ribonucleoprotein B'', RNA recognition motif 2 / U2 small nuclear ribonucleoprotein B'', RNA recognition motif 1 / Splicing factor 3A subunit 1 / Splicing factor 3A subunit 1, conserved domain / Pre-mRNA splicing factor PRP21 like protein / SF3A2 domain / Pre-mRNA-splicing factor SF3a complex subunit 2 (Prp11) / Splicing factor SF3a60 /Prp9 subunit, C-terminal / SF3A3 domain / SF3B4, RNA recognition motif 1 / SF3a60/Prp9 C-terminal / Pre-mRNA-splicing factor SF3A3, of SF3a complex, Prp9 / Splicing factor 3B, subunit 5 / SWAP/Surp / SWAP/Surp superfamily / Surp module / SURP motif repeat profile. / Suppressor-of-White-APricot splicing regulator / Splicing factor 3B subunit 1 / Splicing factor 3B subunit 1 / Domain of unknown function DUF382 / Domain of unknown function (DUF382) / Zinc-finger of C2H2 type / PHF5-like / PHF5-like protein / PSP, proline-rich / PSP / proline-rich domain in spliceosome associated proteins / Splicing factor 3B subunit 5/RDS3 complex subunit 10 / Splicing factor 3B subunit 10 (SF3b10) / Small ribonucleoprotein associated, SmB/SmN / Matrin/U1-C, C2H2-type zinc finger / Splicing factor 3B subunit 1-like / Zinc finger matrin-type profile. / Small nuclear ribonucleoprotein D1 / U2A'/phosphoprotein 32 family A, C-terminal / occurring C-terminal to leucine-rich repeats / Leucine-rich repeat / SAP domain / SAP motif profile. / Putative DNA-binding (bihelical) motif predicted to be involved in chromosomal organisation / SAP domain / Small nuclear ribonucleoprotein Sm D3 / Small nuclear ribonucleoprotein Sm D2 / Small nuclear ribonucleoprotein E / Small nuclear ribonucleoprotein G / Small nuclear ribonucleoprotein F / Sm-like protein Lsm7/SmG / Like-Sm (LSM) domain containing protein, LSm4/SmD1/SmD3 / Matrin/U1-C-like, C2H2-type zinc finger / U1-like zinc finger / Sm-like protein Lsm6/SmF / LSM domain / LSM domain, eukaryotic/archaea-type / snRNP Sm proteins / Cleavage/polyadenylation specificity factor, A subunit, N-terminal / Mono-functional DNA-alkylating methyl methanesulfonate N-term / Cleavage/polyadenylation specificity factor, A subunit, C-terminal / CPSF A subunit region / : / DEAD-box subfamily ATP-dependent helicases signature. / Sm domain profile. / ATP-dependent RNA helicase DEAD-box, conserved site / RNA helicase, DEAD-box type, Q motif / DEAD-box RNA helicase Q motif profile. / LSM domain superfamily / Zinc finger C2H2 superfamily / Leucine-rich repeat profile. / DEAD/DEAH box helicase / DEAD/DEAH box helicase domain / Zinc finger C2H2-type / Leucine-rich repeat / RNA recognition motif / RNA recognition motif / Eukaryotic RNA Recognition Motif (RRM) profile. / RNA recognition motif domain / Helicase conserved C-terminal domain / Leucine-rich repeat domain superfamily / RNA-binding domain superfamily / Armadillo-like helical / Ubiquitin family / Ubiquitin homologues / Ubiquitin-like domain / helicase superfamily c-terminal domain / Ubiquitin domain profile. / Superfamilies 1 and 2 helicase C-terminal domain profile. / Superfamilies 1 and 2 helicase ATP-binding type-1 domain profile. / DEAD-like helicases superfamily / Helicase, C-terminal / Helicase superfamily 1/2, ATP-binding domain
Similarity search - Domain/homology
17S U2 SnRNP complex component HTATSF1 / Splicing factor 3B subunit 1 / U2 small nuclear ribonucleoprotein B'' / U2 small nuclear ribonucleoprotein A' / Small nuclear ribonucleoprotein-associated proteins B and B' / Small nuclear ribonucleoprotein E / Small nuclear ribonucleoprotein F / Small nuclear ribonucleoprotein G / Small nuclear ribonucleoprotein Sm D1 / Small nuclear ribonucleoprotein Sm D2 ...17S U2 SnRNP complex component HTATSF1 / Splicing factor 3B subunit 1 / U2 small nuclear ribonucleoprotein B'' / U2 small nuclear ribonucleoprotein A' / Small nuclear ribonucleoprotein-associated proteins B and B' / Small nuclear ribonucleoprotein E / Small nuclear ribonucleoprotein F / Small nuclear ribonucleoprotein G / Small nuclear ribonucleoprotein Sm D1 / Small nuclear ribonucleoprotein Sm D2 / Small nuclear ribonucleoprotein Sm D3 / Splicing factor 3A subunit 3 / Splicing factor 3B subunit 2 / Splicing factor 3B subunit 3 / Splicing factor 3B subunit 4 / Splicing factor 3A subunit 2 / Splicing factor 3A subunit 1 / Probable ATP-dependent RNA helicase DDX46 / PHD finger-like domain-containing protein 5A / Splicing factor 3B subunit 5 / Splicing factor 3B subunit 6
Similarity search - Component
Biological speciesHomo sapiens (human) / Human (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 7.1 Å
AuthorsZhang Z / Will CL / Bertram K / Luehrmann R / Stark H
Funding support Germany, 1 items
OrganizationGrant numberCountry
German Research Foundation (DFG) Germany
CitationJournal: Nature / Year: 2020
Title: Molecular architecture of the human 17S U2 snRNP.
Authors: Zhenwei Zhang / Cindy L Will / Karl Bertram / Olexandr Dybkov / Klaus Hartmuth / Dmitry E Agafonov / Romina Hofele / Henning Urlaub / Berthold Kastner / Reinhard Lührmann / Holger Stark /
Abstract: The U2 small nuclear ribonucleoprotein (snRNP) has an essential role in the selection of the precursor mRNA branch-site adenosine, the nucleophile for the first step of splicing. Stable addition of ...The U2 small nuclear ribonucleoprotein (snRNP) has an essential role in the selection of the precursor mRNA branch-site adenosine, the nucleophile for the first step of splicing. Stable addition of U2 during early spliceosome formation requires the DEAD-box ATPase PRP5. Yeast U2 small nuclear RNA (snRNA) nucleotides that form base pairs with the branch site are initially sequestered in a branchpoint-interacting stem-loop (BSL), but whether the human U2 snRNA folds in a similar manner is unknown. The U2 SF3B1 protein, a common mutational target in haematopoietic cancers, contains a HEAT domain (SF3B1) with an open conformation in isolated SF3b, but a closed conformation in spliceosomes, which is required for stable interaction between U2 and the branch site. Here we report a 3D cryo-electron microscopy structure of the human 17S U2 snRNP at a core resolution of 4.1 Å and combine it with protein crosslinking data to determine the molecular architecture of this snRNP. Our structure reveals that SF3B1 interacts with PRP5 and TAT-SF1, and maintains its open conformation in U2 snRNP, and that U2 snRNA forms a BSL that is sandwiched between PRP5, TAT-SF1 and SF3B1. Thus, substantial remodelling of the BSL and displacement of BSL-interacting proteins must occur to allow formation of the U2-branch-site helix. Our studies provide a structural explanation of why TAT-SF1 must be displaced before the stable addition of U2 to the spliceosome, and identify RNP rearrangements facilitated by PRP5 that are required for stable interaction between U2 and the branch site.
History
DepositionFeb 24, 2020-
Header (metadata) releaseJun 17, 2020-
Map releaseJun 17, 2020-
UpdateJul 22, 2020-
Current statusJul 22, 2020Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.013
  • Imaged by UCSF Chimera
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  • Surface view colored by radius
  • Surface level: 0.013
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-6y53
  • Surface level: 0.013
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-6y5q
  • Surface level: 0.013
  • Imaged by UCSF Chimera
  • Download
  • Simplified surface model + fitted atomic model
  • Atomic modelsPDB-6y53
  • Imaged by Jmol
  • Download
  • Simplified surface model + fitted atomic model
  • Atomic modelsPDB-6y5q
  • Imaged by Jmol
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_10689.png

Downloads & links

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Map

FileDownload / File: emd_10689.map.gz / Format: CCP4 / Size: 103 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Voxel sizeX=Y=Z: 1.16 Å
Density
Contour LevelBy AUTHOR: 0.013 / Movie #1: 0.013
Minimum - Maximum-0.012199608 - 0.05390597
Average (Standard dev.)0.00074574474 (±0.0040977914)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions300300300
Spacing300300300
CellA=B=C: 348.0 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.161.161.16
M x/y/z300300300
origin x/y/z0.0000.0000.000
length x/y/z348.000348.000348.000
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS300300300
D min/max/mean-0.0120.0540.001

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Supplemental data

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Sample components

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Entire : human 17S U2 snRNP

EntireName: human 17S U2 snRNP
Components
  • Complex: human 17S U2 snRNP
    • Protein or peptide: U2 small nuclear ribonucleoprotein A'
    • Protein or peptide: U2 small nuclear ribonucleoprotein B''
    • Protein or peptide: Splicing factor 3B subunit 6
    • Protein or peptide: Probable ATP-dependent RNA helicase DDX46
    • Protein or peptide: Splicing factor 3A subunit 1
    • Protein or peptide: Splicing factor 3A subunit 2
    • Protein or peptide: Splicing factor 3A subunit 3
    • Protein or peptide: Splicing factor 3B subunit 4
    • Protein or peptide: Small nuclear ribonucleoprotein-associated proteins B and B'
    • Protein or peptide: Small nuclear ribonucleoprotein G
    • Protein or peptide: Small nuclear ribonucleoprotein Sm D1
    • Protein or peptide: Small nuclear ribonucleoprotein Sm D2
    • Protein or peptide: Small nuclear ribonucleoprotein Sm D3
    • Protein or peptide: Small nuclear ribonucleoprotein E
    • Protein or peptide: Small nuclear ribonucleoprotein F
    • Protein or peptide: HIV Tat-specific factor 1
    • Protein or peptide: Splicing factor 3B subunit 2
    • Protein or peptide: Splicing factor 3B subunit 1
    • RNA: U2 snRNAU2 spliceosomal RNA

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Supramolecule #1: human 17S U2 snRNP

SupramoleculeName: human 17S U2 snRNP / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Homo sapiens (human)

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Macromolecule #1: U2 small nuclear ribonucleoprotein A'

MacromoleculeName: U2 small nuclear ribonucleoprotein A' / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 28.456584 KDa
SequenceString: MVKLTAELIE QAAQYTNAVR DRELDLRGYK IPVIENLGAT LDQFDAIDFS DNEIRKLDGF PLLRRLKTLL VNNNRICRIG EGLDQALPC LTELILTNNS LVELGDLDPL ASLKSLTYLS ILRNPVTNKK HYRLYVIYKV PQVRVLDFQK VKLKERQEAE K MFKGKRGA ...String:
MVKLTAELIE QAAQYTNAVR DRELDLRGYK IPVIENLGAT LDQFDAIDFS DNEIRKLDGF PLLRRLKTLL VNNNRICRIG EGLDQALPC LTELILTNNS LVELGDLDPL ASLKSLTYLS ILRNPVTNKK HYRLYVIYKV PQVRVLDFQK VKLKERQEAE K MFKGKRGA QLAKDIARRS KTFNPGAGLP TDKKKGGPSP GDVEAIKNAI ANASTLAEVE RLKGLLQSGQ IPGRERRSGP TD DGEEEME EDTVTNGS

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Macromolecule #2: U2 small nuclear ribonucleoprotein B''

MacromoleculeName: U2 small nuclear ribonucleoprotein B'' / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 25.524367 KDa
SequenceString: MDIRPNHTIY INNMNDKIKK EELKRSLYAL FSQFGHVVDI VALKTMKMRG QAFVIFKELG SSTNALRQLQ GFPFYGKPMR IQYAKTDSD IISKMRGTFA DKEKKKEKKK AKTVEQTATT TNKKPGQGTP NSANTQGNST PNPQVPDYPP NYILFLNNLP E ETNEMMLS ...String:
MDIRPNHTIY INNMNDKIKK EELKRSLYAL FSQFGHVVDI VALKTMKMRG QAFVIFKELG SSTNALRQLQ GFPFYGKPMR IQYAKTDSD IISKMRGTFA DKEKKKEKKK AKTVEQTATT TNKKPGQGTP NSANTQGNST PNPQVPDYPP NYILFLNNLP E ETNEMMLS MLFNQFPGFK EVRLVPGRHD IAFVEFENDG QAGAARDALQ GFKITPSHAM KITYAKK

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Macromolecule #3: Splicing factor 3B subunit 6

MacromoleculeName: Splicing factor 3B subunit 6 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 14.6069 KDa
SequenceString:
MAMQAAKRAN IRLPPEVNRI LYIRNLPYKI TAEEMYDIFG KYGPIRQIRV GNTPETRGTA YVVYEDIFDA KNACDHLSGF NVCNRYLVV LYYNANRAFQ KMDTKKKEEQ LKLLKEKYGI NTDPPK

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Macromolecule #4: Probable ATP-dependent RNA helicase DDX46

MacromoleculeName: Probable ATP-dependent RNA helicase DDX46 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO / EC number: RNA helicase
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 117.576484 KDa
SequenceString: MGRESRHYRK RSASRGRSGS RSRSRSPSDK RSKRGDDRRS RSRDRDRRRE RSRSRDKRRS RSRDRKRLRR SRSRERDRSR ERRRSRSRD RRRSRSRSRG RRSRSSSPGN KSKKTENRSR SKEKTDGGES SKEKKKDKDD KEDEKEKDAG NFDQNKLEEE M RKRKERVE ...String:
MGRESRHYRK RSASRGRSGS RSRSRSPSDK RSKRGDDRRS RSRDRDRRRE RSRSRDKRRS RSRDRKRLRR SRSRERDRSR ERRRSRSRD RRRSRSRSRG RRSRSSSPGN KSKKTENRSR SKEKTDGGES SKEKKKDKDD KEDEKEKDAG NFDQNKLEEE M RKRKERVE KWREEQRKKA MENIGELKKE IEEMKQGKKW SLEDDDDDED DPAEAEKEGN EMEGEELDPL DAYMEEVKEE VK KFNMRSV KGGGGNEKKS GPTVTKVVTV VTTKKAVVDS DKKKGELMEN DQDAMEYSSE EEEVDLQTAL TGYQTKQRKL LEP VDHGKI EYEPFRKNFY VEVPELAKMS QEEVNVFRLE MEGITVKGKG CPKPIKSWVQ CGISMKILNS LKKHGYEKPT PIQT QAIPA IMSGRDLIGI AKTGSGKTIA FLLPMFRHIM DQRSLEEGEG PIAVIMTPTR ELALQITKEC KKFSKTLGLR VVCVY GGTG ISEQIAELKR GAEIIVCTPG RMIDMLAANS GRVTNLRRVT YVVLDEADRM FDMGFEPQVM RIVDNVRPDR QTVMFS ATF PRAMEALARR ILSKPIEVQV GGRSVVCSDV EQQVIVIEEE KKFLKLLELL GHYQESGSVI IFVDKQEHAD GLLKDLM RA SYPCMSLHGG IDQYDRDSII NDFKNGTCKL LVATSVAARG LDVKHLILVV NYSCPNHYED YVHRAGRTGR AGNKGYAY T FITEDQARYA GDIIKALELS GTAVPPDLEK LWSDFKDQQK AEGKIIKKSS GFSGKGFKFD ETEQALANER KKLQKAALG LQDSDDEDAA VDIDEQIESM FNSKKRVKDM AAPGTSSVPA PTAGNAEKLE IAKRLALRIN AQKNLGIESQ DVMQQATNAI LRGGTILAP TVSAKTIAEQ LAEKINAKLN YVPLEKQEEE RQDGGQNESF KRYEEELEIN DFPQTARWKV TSKEALQRIS E YSEAAITI RGTYFPPGKE PKEGERKIYL AIESANELAV QKAKAEITRL IKEELIRLQN SYQPTNKGRY KVL

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Macromolecule #5: Splicing factor 3A subunit 1

MacromoleculeName: Splicing factor 3A subunit 1 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 88.991094 KDa
SequenceString: MPAGPVQAVP PPPPVPTEPK QPTEEEASSK EDSAPSKPVV GIIYPPPEVR NIVDKTASFV ARNGPEFEAR IRQNEINNPK FNFLNPNDP YHAYYRHKVS EFKEGKAQEP SAAIPKVMQQ QQQTTQQQLP QKVQAQVIQE TIVPKEPPPE FEFIADPPSI S AFDLDVVK ...String:
MPAGPVQAVP PPPPVPTEPK QPTEEEASSK EDSAPSKPVV GIIYPPPEVR NIVDKTASFV ARNGPEFEAR IRQNEINNPK FNFLNPNDP YHAYYRHKVS EFKEGKAQEP SAAIPKVMQQ QQQTTQQQLP QKVQAQVIQE TIVPKEPPPE FEFIADPPSI S AFDLDVVK LTAQFVARNG RQFLTQLMQK EQRNYQFDFL RPQHSLFNYF TKLVEQYTKI LIPPKGLFSK LKKEAENPRE VL DQVCYRV EWAKFQERER KKEEEEKEKE RVAYAQIDWH DFVVVETVDF QPNEQGNFPP PTTPEELGAR ILIQERYEKF GES EEVEME VESDEEDDKQ EKAEEPPSQL DQDTQVQDMD EGSDDEEEGQ KVPPPPETPM PPPLPPTPDQ VIVRKDYDPK ASKP LPPAP APDEYLVSPI TGEKIPASKM QEHMRIGLLD PRWLEQRDRS IREKQSDDEV YAPGLDIESS LKQLAERRTD IFGVE ETAI GKKIGEEEIQ KPEEKVTWDG HSGSMARTQQ AAQANITLQE QIEAIHKAKG LVPEDDTKEK IGPSKPNEIP QQPPPP SSA TNIPSSAPPI TSVPRPPTMP PPVRTTVVSA VPVMPRPPMA SVVRLPPGSV IAPMPPIIHA PRINVVPMPP SAPPIMA PR PPPMIVPTAF VPAPPVAPVP APAPMPPVHP PPPMEDEPTS KKLKTEDSLM PEEEFLRRNK GPVSIKVQVP NMQDKTEW K LNGQVLVFTL PLTDQVSVIK VKIHEATGMP AGKQKLQYEG IFIKDSNSLA YYNMANGAVI HLALKERGGR KK

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Macromolecule #6: Splicing factor 3A subunit 2

MacromoleculeName: Splicing factor 3A subunit 2 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 49.327355 KDa
SequenceString: MDFQHRPGGK TGSGGVASSS ESNRDRRERL RQLALETIDI NKDPYFMKNH LGSYECKLCL TLHNNEGSYL AHTQGKKHQT NLARRAAKE AKEAPAQPAP EKVKVEVKKF VKIGRPGYKV TKQRDSEMGQ QSLLFQIDYP EIAEGIMPRH RFMSAYEQRI E PPDRRWQY ...String:
MDFQHRPGGK TGSGGVASSS ESNRDRRERL RQLALETIDI NKDPYFMKNH LGSYECKLCL TLHNNEGSYL AHTQGKKHQT NLARRAAKE AKEAPAQPAP EKVKVEVKKF VKIGRPGYKV TKQRDSEMGQ QSLLFQIDYP EIAEGIMPRH RFMSAYEQRI E PPDRRWQY LLMAAEPYET IAFKVPSREI DKAEGKFWTH WNRETKQFFL QFHFKMEKPP APPSLPAGPP GVKRPPPPLM NG LPPRPPL PESLPPPPPG GLPLPPMPPT GPAPSGPPGP PQLPPPAPGV HPPAPVVHPP ASGVHPPAPG VHPPAPGVHP PAP GVHPPT SGVHPPAPGV HPPAPGVHPP APGVHPPAPG VHPPAPGVHP PPSAGVHPQA PGVHPAAPAV HPQAPGVHPP APGM HPQAP GVHPQPPGVH PSAPGVHPQP PGVHPSNPGV HPPTPMPPML RPPLPSEGPG NIPPPPPTN

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Macromolecule #7: Splicing factor 3A subunit 3

MacromoleculeName: Splicing factor 3A subunit 3 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 58.934844 KDa
SequenceString: METILEQQRR YHEEKERLMD VMAKEMLTKK STLRDQINSD HRTRAMQDRY MEVSGNLRDL YDDKDGLRKE ELNAISGPNE FAEFYNRLK QIKEFHRKHP NEICVPMSVE FEELLKAREN PSEEAQNLVE FTDEEGYGRY LDLHDCYLKY INLKASEKLD Y ITYLSIFD ...String:
METILEQQRR YHEEKERLMD VMAKEMLTKK STLRDQINSD HRTRAMQDRY MEVSGNLRDL YDDKDGLRKE ELNAISGPNE FAEFYNRLK QIKEFHRKHP NEICVPMSVE FEELLKAREN PSEEAQNLVE FTDEEGYGRY LDLHDCYLKY INLKASEKLD Y ITYLSIFD QLFDIPKERK NAEYKRYLEM LLEYLQDYTD RVKPLQDQNE LFGKIQAEFE KKWENGTFPG WPKETSSALT HA GAHLDLS AFSSWEELAS LGLDRLKSAL LALGLKCGGT LEERAQRLFS TKGKSLESLD TSLFAKNPKS KGTKRDTERN KDI AFLEAQ IYEYVEILGE QRHLTHENVQ RKQARTGEER EEEEEEQISE SESEDEENEI IYNPKNLPLG WDGKPIPYWL YKLH GLNIN YNCEICGNYT YRGPKAFQRH FAEWRHAHGM RCLGIPNTAH FANVTQIEDA VSLWAKLKLQ KASERWQPDT EEEYE DSSG NVVNKKTYED LKRQGLL

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Macromolecule #8: Splicing factor 3B subunit 4

MacromoleculeName: Splicing factor 3B subunit 4 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 44.43657 KDa
SequenceString: MAAGPISERN QDATVYVGGL DEKVSEPLLW ELFLQAGPVV NTHMPKDRVT GQHQGYGFVE FLSEEDADYA IKIMNMIKLY GKPIRVNKA SAHNKNLDVG ANIFIGNLDP EIDEKLLYDT FSAFGVILQT PKIMRDPDTG NSKGYAFINF ASFDASDAAI E AMNGQYLC ...String:
MAAGPISERN QDATVYVGGL DEKVSEPLLW ELFLQAGPVV NTHMPKDRVT GQHQGYGFVE FLSEEDADYA IKIMNMIKLY GKPIRVNKA SAHNKNLDVG ANIFIGNLDP EIDEKLLYDT FSAFGVILQT PKIMRDPDTG NSKGYAFINF ASFDASDAAI E AMNGQYLC NRPITVSYAF KKDSKGERHG SAAERLLAAQ NPLSQADRPH QLFADAPPPP SAPNPVVSSL GSGLPPPGMP PP GSFPPPV PPPGALPPGI PPAMPPPPMP PGAAGHGPPS AGTPGAGHPG HGHSHPHPFP PGGMPHPGMS QMQLAHHGPH GLG HPHAGP PGSGGQPPPR PPPGMPHPGP PPMGMPPRGP PFGSPMGHPG PMPPHGMRGP PPLMPPHGYT GPPRPPPYGY QRGP LPPPR PTPRPPVPPR GPLRGPLPQ

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Macromolecule #9: Small nuclear ribonucleoprotein-associated proteins B and B'

MacromoleculeName: Small nuclear ribonucleoprotein-associated proteins B and B'
type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 24.642131 KDa
SequenceString: MTVGKSSKML QHIDYRMRCI LQDGRIFIGT FKAFDKHMNL ILCDCDEFRK IKPKNSKQAE REEKRVLGLV LLRGENLVSM TVEGPPPKD TGIARVPLAG AAGGPGIGRA AGRGIPAGVP MPQAPAGLAG PVRGVGGPSQ QVMTPQGRGT VAAAAAAATA S IAGAPTQY ...String:
MTVGKSSKML QHIDYRMRCI LQDGRIFIGT FKAFDKHMNL ILCDCDEFRK IKPKNSKQAE REEKRVLGLV LLRGENLVSM TVEGPPPKD TGIARVPLAG AAGGPGIGRA AGRGIPAGVP MPQAPAGLAG PVRGVGGPSQ QVMTPQGRGT VAAAAAAATA S IAGAPTQY PPGRGGPPPP MGRGAPPPGM MGPPPGMRPP MGPPMGIPPG RGTPMGMPPP GMRPPPPGMR GPPPPGMRPP RP

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Macromolecule #10: Small nuclear ribonucleoprotein G

MacromoleculeName: Small nuclear ribonucleoprotein G / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 8.508084 KDa
SequenceString:
MSKAHPPELK KFMDKKLSLK LNGGRHVQGI LRGFDPFMNL VIDECVEMAT SGQQNNIGMV VIRGNSIIML EALERV

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Macromolecule #11: Small nuclear ribonucleoprotein Sm D1

MacromoleculeName: Small nuclear ribonucleoprotein Sm D1 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 13.310653 KDa
SequenceString:
MKLVRFLMKL SHETVTIELK NGTQVHGTIT GVDVSMNTHL KAVKMTLKNR EPVQLETLSI RGNNIRYFIL PDSLPLDTLL VDVEPKVKS KKREAVAGRG RGRGRGRGRG RGRGRGGPRR

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Macromolecule #12: Small nuclear ribonucleoprotein Sm D2

MacromoleculeName: Small nuclear ribonucleoprotein Sm D2 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 13.551928 KDa
SequenceString:
MSLLNKPKSE MTPEELQKRE EEEFNTGPLS VLTQSVKNNT QVLINCRNNK KLLGRVKAFD RHCNMVLENV KEMWTEVPKS GKGKKKSKP VNKDRYISKM FLRGDSVIVV LRNPLIAGK

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Macromolecule #13: Small nuclear ribonucleoprotein Sm D3

MacromoleculeName: Small nuclear ribonucleoprotein Sm D3 / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 13.940308 KDa
SequenceString:
MSIGVPIKVL HEAEGHIVTC ETNTGEVYRG KLIEAEDNMN CQMSNITVTY RDGRVAQLEQ VYIRGSKIRF LILPDMLKNA PMLKSMKNK NQGSGAGRGK AAILKAQVAA RGRGRGMGRG NIFQKRR

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Macromolecule #14: Small nuclear ribonucleoprotein E

MacromoleculeName: Small nuclear ribonucleoprotein E / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 10.817601 KDa
SequenceString:
MAYRGQGQKV QKVMVQPINL IFRYLQNRSR IQVWLYEQVN MRIEGCIIGF DEYMNLVLDD AEEIHSKTKS RKQLGRIMLK GDNITLLQS VSN

+
Macromolecule #15: Small nuclear ribonucleoprotein F

MacromoleculeName: Small nuclear ribonucleoprotein F / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 9.734171 KDa
SequenceString:
MSLPLNPKPF LNGLTGKPVM VKLKWGMEYK GYLVSVDGYM NMQLANTEEY IDGALSGHLG EVLIRCNNVL YIRGVEEEEE DGEMRE

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Macromolecule #16: HIV Tat-specific factor 1

MacromoleculeName: HIV Tat-specific factor 1 / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 85.965961 KDa
SequenceString: MSGTNLDGND EFDEQLRMQE LYGDGKDGDT QTDAGGEPDS LGQQPTDTPY EWDLDKKAWF PKITEDFIAT YQANYGFSND GASSSTANV EDVHARTAEE PPQEKAPEPT DARKKGEKRK AESGWFHVEE DRNTNVYVSG LPPDITVDEF IQLMSKFGII M RDPQTEEF ...String:
MSGTNLDGND EFDEQLRMQE LYGDGKDGDT QTDAGGEPDS LGQQPTDTPY EWDLDKKAWF PKITEDFIAT YQANYGFSND GASSSTANV EDVHARTAEE PPQEKAPEPT DARKKGEKRK AESGWFHVEE DRNTNVYVSG LPPDITVDEF IQLMSKFGII M RDPQTEEF KVKLYKDNQG NLKGDGLCCY LKRESVELAL KLLDEDEIRG YKLHVEVAKF QLKGEYDASK KKKKCKDYKK KL SMQQKQL DWRPERRAGP SRMRHERVVI IKNMFHPMDF EDDPLVLNEI REDLRVECSK FGQIRKLLLF DRHPDGVASV SFR DPEEAD YCIQTLDGRW FGGRQITAQA WDGTTDYQVE ETSREREERL RGWEAFLNAP EANRGLRRSD SVSASERAGP SRAR HFSEH PSTSKMNAQE TATGMAFEEP IDEKKFEKTE DGGEFEEGAS ENNAKESSPE KEAEEGCPEK ESEEGCPKRG FEGSC SQKE SEEGNPVRGS EEDSPKKESK KKTLKNDCEE NGLAKESEDD LNKESEEEVG PTKESEEDDS EKESDEDCSE KQSEDG SER EFEENGLEKD LDEEGSEKEL HENVLDKELE ENDSENSEFE DDGSEKVLDE EGSEREFDED SDEKEEEEDT YEKVFDD ES DEKEDEEYAD EKGLEAADKK AEEGDADEKL FEESDDKEDE DADGKEVEDA DEKLFEDDDS NEKLFDEEED SSEKLFDD S DERGTLGGFG SVEEGPLSTG SSFILSSDDD DDDI

+
Macromolecule #17: Splicing factor 3B subunit 2

MacromoleculeName: Splicing factor 3B subunit 2 / type: protein_or_peptide / ID: 17 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 100.377812 KDa
SequenceString: MATEHPEPPK AELQLPPPPP PGHYGAWAAQ ELQAKLAEIG APIQGNREEL VERLQSYTRQ TGIVLNRPVL RGEDGDKAAP PPMSAQLPG IPMPPPPLGL PPLQPPPPPP PPPPGLGLGF PMAHPPNLGP PPPLRVGEPV ALSEEERLKL AQQQAALLMQ Q EERAKQQG ...String:
MATEHPEPPK AELQLPPPPP PGHYGAWAAQ ELQAKLAEIG APIQGNREEL VERLQSYTRQ TGIVLNRPVL RGEDGDKAAP PPMSAQLPG IPMPPPPLGL PPLQPPPPPP PPPPGLGLGF PMAHPPNLGP PPPLRVGEPV ALSEEERLKL AQQQAALLMQ Q EERAKQQG DHSLKEHELL EQQKRAAVLL EQERQQEIAK MGTPVPRPPQ DMGQIGVRTP LGPRVAAPVG PVGPTPTVLP MG APVPRPR GPPPPPGDEN REMDDPSVGP KIPQALEKIL QLKESRQEEM NSQQEEEEME TDARSSLGQS ASETEEDTVS VSK KEKNRK RRNRKKKKKP QRVRGVSSES SGDREKDSTR SRGSDSPAAD VEIEYVTEEP EIYEPNFIFF KRIFEAFKLT DDVK KEKEK EPEKLDKLEN SAAPKKKGFE EEHKDSDDDS SDDEQEKKPE APKLSKKKLR RMNRFTVAEL KQLVARPDVV EMHDV TAQD PKLLVHLKAT RNSVPVPRHW CFKRKYLQGK RGIEKPPFEL PDFIKRTGIQ EMREALQEKE EQKTMKSKMR EKVRPK MGK IDIDYQKLHD AFFKWQTKPK LTIHGDLYYE GKEFETRLKE KKPGDLSDEL RISLGMPVGP NAHKVPPPWL IAMQRYG PP PSYPNLKIPG LNSPIPESCS FGYHAGGWGK PPVDETGKPL YGDVFGTNAA EFQTKTEEEE IDRTPWGELE PSDEESSE E EEEEESDEDK PDETGFITPA DSGLITPGGF SSVPAGMETP ELIELRKKKI EEAMDGSETP QLFTVLPEKR TATVGGAMM GSTHIYDMST VMSRKGPAPE LQGVEVALAP EELELDPMAM TQKYEEHVRE QQAQVEKEDF SDMVAEHAAK QKQKKRKAQP QDSRGGSKK YKEFKF

+
Macromolecule #18: Splicing factor 3B subunit 1

MacromoleculeName: Splicing factor 3B subunit 1 / type: protein_or_peptide / ID: 18 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 146.024938 KDa
SequenceString: MAKIAKTHED IEAQIREIQG KKAALDEAQG VGLDSTGYYD QEIYGGSDSR FAGYVTSIAA TELEDDDDDY SSSTSLLGQK KPGYHAPVA LLNDIPQSTE QYDPFAEHRP PKIADREDEY KKHRRTMIIS PERLDPFADG GKTPDPKMNA RTYMDVMREQ H LTKEEREI ...String:
MAKIAKTHED IEAQIREIQG KKAALDEAQG VGLDSTGYYD QEIYGGSDSR FAGYVTSIAA TELEDDDDDY SSSTSLLGQK KPGYHAPVA LLNDIPQSTE QYDPFAEHRP PKIADREDEY KKHRRTMIIS PERLDPFADG GKTPDPKMNA RTYMDVMREQ H LTKEEREI RQQLAEKAKA GELKVVNGAA ASQPPSKRKR RWDQTADQTP GATPKKLSSW DQAETPGHTP SLRWDETPGR AK GSETPGA TPGSKIWDPT PSHTPAGAAT PGRGDTPGHA TPGHGGATSS ARKNRWDETP KTERDTPGHG SGWAETPRTD RGG DSIGET PTPGASKRKS RWDETPASQM GGSTPVLTPG KTPIGTPAMN MATPTPGHIM SMTPEQLQAW RWEREIDERN RPLS DEELD AMFPEGYKVL PPPAGYVPIR TPARKLTATP TPLGGMTGFH MQTEDRTMKS VNDQPSGNLP FLKPDDIQYF DKLLV DVDE STLSPEEQKE RKIMKLLLKI KNGTPPMRKA ALRQITDKAR EFGAGPLFNQ ILPLLMSPTL EDQERHLLVK VIDRIL YKL DDLVRPYVHK ILVVIEPLLI DEDYYARVEG REIISNLAKA AGLATMISTM RPDIDNMDEY VRNTTARAFA VVASALG IP SLLPFLKAVC KSKKSWQARH TGIKIVQQIA ILMGCAILPH LRSLVEIIEH GLVDEQQKVR TISALAIAAL AEAATPYG I ESFDSVLKPL WKGIRQHRGK GLAAFLKAIG YLIPLMDAEY ANYYTREVML ILIREFQSPD EEMKKIVLKV VKQCCGTDG VEANYIKTEI LPPFFKHFWQ HRMALDRRNY RQLVDTTVEL ANKVGAAEII SRIVDDLKDE AEQYRKMVME TIEKIMGNLG AADIDHKLE EQLIDGILYA FQEQTTEDSV MLNGFGTVVN ALGKRVKPYL PQICGTVLWR LNNKSAKVRQ QAADLISRTA V VMKTCQEE KLMGHLGVVL YEYLGEEYPE VLGSILGALK AIVNVIGMHK MTPPIKDLLP RLTPILKNRH EKVQENCIDL VG RIADRGA EYVSAREWMR ICFELLELLK AHKKAIRRAT VNTFGYIAKA IGPHDVLATL LNNLKVQERQ NRVCTTVAIA IVA ETCSPF TVLPALMNEY RVPELNVQNG VLKSLSFLFE YIGEMGKDYI YAVTPLLEDA LMDRDLVHRQ TASAVVQHMS LGVY GFGCE DSLNHLLNYV WPNVFETSPH VIQAVMGALE GLRVAIGPCR MLQYCLQGLF HPARKVRDVY WKIYNSIYIG SQDAL IAHY PRIYNDDKNT YIRYELDYIL

+
Macromolecule #19: U2 snRNA

MacromoleculeName: U2 snRNA / type: rna / ID: 19 / Number of copies: 1
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 60.186445 KDa
SequenceString:
AUCGCUUCUC GGCCUUUUGG CUAAGAUCAA GUGUAGUAUC UGUUCUUAUC AGUUUAAUAU CUGAUACGUC CUCUAUCCGA GGACAAUAU AUUAAAUGGA UUUUUGGAGC AGGGAGAUGG AAUAGGAGCU UGCUCCGUCC ACUCCACGCA UCGACCUGGU A UUGCAGUA CCUCCAGGAA CGGUGCACCC

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

-
Sample preparation

BufferpH: 7.9
GridModel: Quantifoil R3.5/1 / Material: COPPER / Support film - Material: CARBON / Support film - topology: CONTINUOUS
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK I

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 0.01 mm
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Image recordingFilm or detector model: FEI FALCON III (4k x 4k) / Detector mode: INTEGRATING / Average exposure time: 1.0 sec. / Average electron dose: 72.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.0)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.0)
Final reconstructionResolution.type: BY AUTHOR / Resolution: 7.1 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.0) / Number images used: 120070

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Atomic model buiding 1

RefinementSpace: REAL
Output model

PDB-6y53:
human 17S U2 snRNP low resolution part

PDB-6y5q:
human 17S U2 snRNP

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