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- EMDB-11696: Human pre-Bact-2 spliceosome (SF3b/U2 snRNP portion) -

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Basic information

Entry
Database: EMDB / ID: EMD-11696
TitleHuman pre-Bact-2 spliceosome (SF3b/U2 snRNP portion)
Map dataMasked/sharpened map of SF3b/U2 snRNP region of pre-Bact-2 spliceosome.
Sample
  • Complex: Human pre-Bact-2 spliceosome (SF3b/U2 snRNP portion)
    • Complex: Human pre-Bact-2 spliceosome (SF3b/U2 snRNP portion)
      • Protein or peptide: x 14 types
      • RNA: x 1 types
    • Complex: MINX M3 pre-mRNA
      • RNA: x 1 types
Function / homology
Function and homology information


U11/U12 snRNP / B-WICH complex / splicing factor binding / U12-type spliceosomal complex / miRNA processing / RNA splicing, via transesterification reactions / Prp19 complex / blastocyst formation / mRNA 3'-end processing / : ...U11/U12 snRNP / B-WICH complex / splicing factor binding / U12-type spliceosomal complex / miRNA processing / RNA splicing, via transesterification reactions / Prp19 complex / blastocyst formation / mRNA 3'-end processing / : / U2-type spliceosomal complex / positive regulation of mRNA splicing, via spliceosome / U2-type precatalytic spliceosome / regulation of mRNA splicing, via spliceosome / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / Transport of Mature mRNA derived from an Intron-Containing Transcript / U2 snRNP / SAGA complex / RNA Polymerase II Transcription Termination / positive regulation of transcription by RNA polymerase III / RHOBTB1 GTPase cycle / U2-type prespliceosome / WD40-repeat domain binding / 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 / localization / regulation of DNA repair / canonical NF-kappaB signal transduction / Protein methylation / U2 snRNA binding / RHOBTB2 GTPase cycle / catalytic step 2 spliceosome / mRNA Splicing - Major Pathway / RNA splicing / DNA damage checkpoint signaling / stem cell differentiation / RNA polymerase II transcription regulatory region sequence-specific DNA binding / spliceosomal complex / B-WICH complex positively regulates rRNA expression / negative regulation of protein catabolic process / positive regulation of neuron projection development / mRNA splicing, via spliceosome / mRNA processing / nuclear matrix / double-stranded DNA binding / DNA-binding transcription activator activity, RNA polymerase II-specific / nuclear membrane / DNA recombination / DNA replication / DNA-binding transcription factor activity, RNA polymerase II-specific / nuclear speck / chromatin remodeling / cell cycle / DNA repair / mRNA binding / intracellular membrane-bounded organelle / DNA damage response / protein-containing complex binding / nucleolus / regulation of transcription by RNA polymerase II / positive regulation of DNA-templated transcription / endoplasmic reticulum / positive regulation of transcription by RNA polymerase II / protein-containing complex / DNA binding / RNA binding / zinc ion binding / nucleoplasm / membrane / identical protein binding / metal ion binding / nucleus / cytosol / cytoplasm
Similarity search - Function
DNA/RNA-binding protein Kin17, WH-like domain / KIN17-like protein / DNA/RNA-binding protein KIN17, WH-like domain superfamily / KN17, SH3-like C-terminal domain / Kin17, KOW domain / Domain of Kin17 curved DNA-binding protein / KN17 SH3-like C-terminal domain / Domain of Kin17 curved DNA-binding protein / Ist3-like, RNA recognition motif / PWI domain superfamily ...DNA/RNA-binding protein Kin17, WH-like domain / KIN17-like protein / DNA/RNA-binding protein KIN17, WH-like domain superfamily / KN17, SH3-like C-terminal domain / Kin17, KOW domain / Domain of Kin17 curved DNA-binding protein / KN17 SH3-like C-terminal domain / Domain of Kin17 curved DNA-binding protein / Ist3-like, RNA recognition motif / PWI domain superfamily / PWI domain / PWI domain profile. / SF3B6, RNA recognition motif / SF3B4, RNA recognition motif 2 / Splicing factor SF3a60 binding domain / Splicing factor SF3a60 binding domain / PWI domain / : / Replication stress response SDE2 C-terminal / PWI, domain in splicing factors / Cactus-binding C-terminus of cactin 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 / 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 / Myb-like DNA-binding domain / proline-rich domain in spliceosome associated proteins / Splicing factor 3B subunit 5/RDS3 complex subunit 10 / Splicing factor 3B subunit 10 (SF3b10) / Matrin/U1-C, C2H2-type zinc finger / Splicing factor 3B subunit 1-like / Zinc finger matrin-type profile. / Pre-mRNA splicing factor component Cdc5p/Cef1, C-terminal / pre-mRNA splicing factor component / SAP domain / SAP motif profile. / Putative DNA-binding (bihelical) motif predicted to be involved in chromosomal organisation / SAP domain / Matrin/U1-C-like, C2H2-type zinc finger / U1-like zinc finger / Myb-type HTH DNA-binding domain profile. / Forkhead associated domain / Forkhead-associated (FHA) domain profile. / FHA domain / Cleavage/polyadenylation specificity factor, A subunit, N-terminal / Mono-functional DNA-alkylating methyl methanesulfonate N-term / Myb domain / Cleavage/polyadenylation specificity factor, A subunit, C-terminal / CPSF A subunit region / Forkhead-associated (FHA) domain / SMAD/FHA domain superfamily / SANT SWI3, ADA2, N-CoR and TFIIIB'' DNA-binding domains / SANT/Myb domain / Zinc finger C2H2 superfamily / Zinc finger C2H2 type domain signature. / Zinc finger C2H2-type / RNA recognition motif / RNA recognition motif / Eukaryotic RNA Recognition Motif (RRM) profile. / RNA recognition motif domain / RNA-binding domain superfamily / Homeobox-like domain superfamily / Armadillo-like helical / Ribosomal protein L2, domain 2 / Armadillo-type fold / Nucleotide-binding alpha-beta plait domain superfamily / WD40-repeat-containing domain superfamily / WD40/YVTN repeat-like-containing domain superfamily
Similarity search - Domain/homology
DNA/RNA-binding protein KIN17 / Splicing factor 3B subunit 1 / 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 / PHD finger-like domain-containing protein 5A / Serine/arginine repetitive matrix protein 1 / Smad nuclear-interacting protein 1 ...DNA/RNA-binding protein KIN17 / Splicing factor 3B subunit 1 / 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 / PHD finger-like domain-containing protein 5A / Serine/arginine repetitive matrix protein 1 / Smad nuclear-interacting protein 1 / Cell division cycle 5-like protein / Splicing factor 3B subunit 5 / RNA-binding motif protein, X-linked 2 / Splicing factor 3B subunit 6
Similarity search - Component
Biological speciesHomo sapiens (human) / synthetic construct (others) / Human (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 4.5 Å
AuthorsTownsend C / Kastner B / Leelaram MN / Bertram K / Stark H / Luehrmann R
Funding support Germany, 1 items
OrganizationGrant numberCountry
German Research Foundation (DFG)860 Germany
CitationJournal: Science / Year: 2020
Title: Mechanism of protein-guided folding of the active site U2/U6 RNA during spliceosome activation.
Authors: Cole Townsend / Majety N Leelaram / Dmitry E Agafonov / Olexandr Dybkov / Cindy L Will / Karl Bertram / Henning Urlaub / Berthold Kastner / Holger Stark / Reinhard Lührmann /
Abstract: Spliceosome activation involves extensive protein and RNA rearrangements that lead to formation of a catalytically active U2/U6 RNA structure. At present, little is known about the assembly pathway ...Spliceosome activation involves extensive protein and RNA rearrangements that lead to formation of a catalytically active U2/U6 RNA structure. At present, little is known about the assembly pathway of the latter and the mechanism whereby proteins aid its proper folding. Here, we report the cryo-electron microscopy structures of two human, activated spliceosome precursors (that is, pre-B complexes) at core resolutions of 3.9 and 4.2 angstroms. These structures elucidate the order of the numerous protein exchanges that occur during activation, the mutually exclusive interactions that ensure the correct order of ribonucleoprotein rearrangements needed to form the U2/U6 catalytic RNA, and the stepwise folding pathway of the latter. Structural comparisons with mature B complexes reveal the molecular mechanism whereby a conformational change in the scaffold protein PRP8 facilitates final three-dimensional folding of the U2/U6 catalytic RNA.
History
DepositionSep 7, 2020-
Header (metadata) releaseDec 9, 2020-
Map releaseDec 9, 2020-
UpdateDec 30, 2020-
Current statusDec 30, 2020Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.035
  • Imaged by UCSF Chimera
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  • Surface view colored by radius
  • Surface level: 0.035
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-7abh
  • Surface level: 0.035
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_11696.png

Downloads & links

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Map

FileDownload / File: emd_11696.map.gz / Format: CCP4 / Size: 216 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationMasked/sharpened map of SF3b/U2 snRNP region of pre-Bact-2 spliceosome.
Voxel sizeX=Y=Z: 1.16 Å
Density
Contour LevelBy AUTHOR: 0.047 / Movie #1: 0.035
Minimum - Maximum-0.052662313 - 0.10835708
Average (Standard dev.)0.00020037475 (±0.0027742009)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions384384384
Spacing384384384
CellA=B=C: 445.44 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.161.161.16
M x/y/z384384384
origin x/y/z0.0000.0000.000
length x/y/z445.440445.440445.440
α/β/γ90.00090.00090.000
start NX/NY/NZ000
NX/NY/NZ240240240
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS384384384
D min/max/mean-0.0530.1080.000

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Supplemental data

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Sample components

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Entire : Human pre-Bact-2 spliceosome (SF3b/U2 snRNP portion)

EntireName: Human pre-Bact-2 spliceosome (SF3b/U2 snRNP portion)
Components
  • Complex: Human pre-Bact-2 spliceosome (SF3b/U2 snRNP portion)
    • Complex: Human pre-Bact-2 spliceosome (SF3b/U2 snRNP portion)
      • Protein or peptide: Cell division cycle 5-like protein
      • Protein or peptide: PHD finger-like domain-containing protein 5A
      • Protein or peptide: Splicing factor 3A subunit 2
      • Protein or peptide: Splicing factor 3A subunit 3
      • Protein or peptide: Splicing factor 3B subunit 1
      • Protein or peptide: Splicing factor 3B subunit 2
      • Protein or peptide: Splicing factor 3B subunit 3
      • Protein or peptide: Splicing factor 3B subunit 4
      • Protein or peptide: Splicing factor 3B subunit 5
      • Protein or peptide: Splicing factor 3B subunit 6
      • Protein or peptide: Smad nuclear-interacting protein 1
      • Protein or peptide: RNA-binding motif protein, X-linked 2
      • Protein or peptide: Serine/arginine repetitive matrix protein 1
      • RNA: U2 snRNAU2 spliceosomal RNA
      • Protein or peptide: DNA/RNA-binding protein KIN17
    • Complex: MINX M3 pre-mRNA
      • RNA: MINX M3 pre-mRNA

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Supramolecule #1: Human pre-Bact-2 spliceosome (SF3b/U2 snRNP portion)

SupramoleculeName: Human pre-Bact-2 spliceosome (SF3b/U2 snRNP portion) / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#16

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Supramolecule #2: Human pre-Bact-2 spliceosome (SF3b/U2 snRNP portion)

SupramoleculeName: Human pre-Bact-2 spliceosome (SF3b/U2 snRNP portion) / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1-#2, #4-#16
Source (natural)Organism: Homo sapiens (human)

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Supramolecule #3: MINX M3 pre-mRNA

SupramoleculeName: MINX M3 pre-mRNA / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #3
Source (natural)Organism: synthetic construct (others)
Recombinant expressionOrganism: synthetic construct (others)

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Macromolecule #1: Cell division cycle 5-like protein

MacromoleculeName: Cell division cycle 5-like protein / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 92.406883 KDa
SequenceString: MPRIMIKGGV WRNTEDEILK AAVMKYGKNQ WSRIASLLHR KSAKQCKARW YEWLDPSIKK TEWSREEEEK LLHLAKLMPT QWRTIAPII GRTAAQCLEH YEFLLDKAAQ RDNEEETTDD PRKLKPGEID PNPETKPARP DPIDMDEDEL EMLSEARARL A NTQGKKAK ...String:
MPRIMIKGGV WRNTEDEILK AAVMKYGKNQ WSRIASLLHR KSAKQCKARW YEWLDPSIKK TEWSREEEEK LLHLAKLMPT QWRTIAPII GRTAAQCLEH YEFLLDKAAQ RDNEEETTDD PRKLKPGEID PNPETKPARP DPIDMDEDEL EMLSEARARL A NTQGKKAK RKAREKQLEE ARRLAALQKR RELRAAGIEI QKKRKRKRGV DYNAEIPFEK KPALGFYDTS EENYQALDAD FR KLRQQDL DGELRSEKEG RDRKKDKQHL KRKKESDLPS AILQTSGVSE FTKKRSKLVL PAPQISDAEL QEVVKVGQAS EIA RQTAEE SGITNSASST LLSEYNVTNN SVALRTPRTP ASQDRILQEA QNLMALTNVD TPLKGGLNTP LHESDFSGVT PQRQ VVQTP NTVLSTPFRT PSNGAEGLTP RSGTTPKPVI NSTPGRTPLR DKLNINPEDG MADYSDPSYV KQMERESREH LRLGL LGLP APKNDFEIVL PENAEKELEE REIDDTYIED AADVDARKQA IRDAERVKEM KRMHKAVQKD LPRPSEVNET ILRPLN VEP PLTDLQKSEE LIKKEMITML HYDLLHHPYE PSGNKKGKTV GFGTNNSEHI TYLEHNPYEK FSKEELKKAQ DVLVQEM EV VKQGMSHGEL SSEAYNQVWE ECYSQVLYLP GQSRYTRANL ASKKDRIESL EKRLEINRGH MTTEAKRAAK MEKKMKIL L GGYQSRAMGL MKQLNDLWDQ IEQAHLELRT FEELKKHEDS AIPRRLECLK EDVQRQQERE KELQHRYADL LLEKETLKS KF

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Macromolecule #2: PHD finger-like domain-containing protein 5A

MacromoleculeName: PHD finger-like domain-containing protein 5A / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 12.427524 KDa
SequenceString:
MAKHHPDLIF CRKQAGVAIG RLCEKCDGKC VICDSYVRPC TLVRICDECN YGSYQGRCVI CGGPGVSDAY YCKECTIQEK DRDGCPKIV NLGSSKTDLF YERKKYGFKK R

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Macromolecule #4: Splicing factor 3A subunit 2

MacromoleculeName: Splicing factor 3A subunit 2 / type: protein_or_peptide / ID: 4 / 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 #5: Splicing factor 3A subunit 3

MacromoleculeName: Splicing factor 3A subunit 3 / type: protein_or_peptide / ID: 5 / 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 #6: Splicing factor 3B subunit 1

MacromoleculeName: Splicing factor 3B subunit 1 / type: protein_or_peptide / ID: 6 / 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

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Macromolecule #7: Splicing factor 3B subunit 2

MacromoleculeName: Splicing factor 3B subunit 2 / type: protein_or_peptide / ID: 7 / 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

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Macromolecule #8: Splicing factor 3B subunit 3

MacromoleculeName: Splicing factor 3B subunit 3 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 135.718844 KDa
SequenceString: MFLYNLTLQR ATGISFAIHG NFSGTKQQEI VVSRGKILEL LRPDPNTGKV HTLLTVEVFG VIRSLMAFRL TGGTKDYIVV GSDSGRIVI LEYQPSKNMF EKIHQETFGK SGCRRIVPGQ FLAVDPKGRA VMISAIEKQK LVYILNRDAA ARLTISSPLE A HKANTLVY ...String:
MFLYNLTLQR ATGISFAIHG NFSGTKQQEI VVSRGKILEL LRPDPNTGKV HTLLTVEVFG VIRSLMAFRL TGGTKDYIVV GSDSGRIVI LEYQPSKNMF EKIHQETFGK SGCRRIVPGQ FLAVDPKGRA VMISAIEKQK LVYILNRDAA ARLTISSPLE A HKANTLVY HVVGVDVGFE NPMFACLEMD YEEADNDPTG EAAANTQQTL TFYELDLGLN HVVRKYSEPL EEHGNFLITV PG GSDGPSG VLICSENYIT YKNFGDQPDI RCPIPRRRND LDDPERGMIF VCSATHKTKS MFFFLAQTEQ GDIFKITLET DED MVTEIR LKYFDTVPVA AAMCVLKTGF LFVASEFGNH YLYQIAHLGD DDEEPEFSSA MPLEEGDTFF FQPRPLKNLV LVDE LDSLS PILFCQIADL ANEDTPQLYV ACGRGPRSSL RVLRHGLEVS EMAVSELPGN PNAVWTVRRH IEDEFDAYII VSFVN ATLV LSIGETVEEV TDSGFLGTTP TLSCSLLGDD ALVQVYPDGI RHIRADKRVN EWKTPGKKTI VKCAVNQRQV VIALTG GEL VYFEMDPSGQ LNEYTERKEM SADVVCMSLA NVPPGEQRSR FLAVGLVDNT VRIISLDPSD CLQPLSMQAL PAQPESL CI VEMGGTEKQD ELGERGSIGF LYLNIGLQNG VLLRTVLDPV TGDLSDTRTR YLGSRPVKLF RVRMQGQEAV LAMSSRSW L SYSYQSRFHL TPLSYETLEF ASGFASEQCP EGIVAISTNT LRILALEKLG AVFNQVAFPL QYTPRKFVIH PESNNLIII ETDHNAYTEA TKAQRKQQMA EEMVEAAGED ERELAAEMAA AFLNENLPES IFGAPKAGNG QWASVIRVMN PIQGNTLDLV QLEQNEAAF SVAVCRFSNT GEDWYVLVGV AKDLILNPRS VAGGFVYTYK LVNNGEKLEF LHKTPVEEVP AAIAPFQGRV L IGVGKLLR VYDLGKKKLL RKCENKHIAN YISGIQTIGH RVIVSDVQES FIWVRYKRNE NQLIIFADDT YPRWVTTASL LD YDTVAGA DKFGNICVVR LPPNTNDEVD EDPTGNKALW DRGLLNGASQ KAEVIMNYHV GETVLSLQKT TLIPGGSESL VYT TLSGGI GILVPFTSHE DHDFFQHVEM HLRSEHPPLC GRDHLSFRSY YFPVKNVIDG DLCEQFNSME PNKQKNVSEE LDRT PPEVS KKLEDIRTRY AF

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Macromolecule #9: Splicing factor 3B subunit 4

MacromoleculeName: Splicing factor 3B subunit 4 / type: protein_or_peptide / ID: 9 / 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 #10: Splicing factor 3B subunit 5

MacromoleculeName: Splicing factor 3B subunit 5 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 10.149369 KDa
SequenceString:
MTDRYTIHSQ LEHLQSKYIG TGHADTTKWE WLVNQHRDSY CSYMGHFDLL NYFAIAENES KARVRFNLME KMLQPCGPPA DKPEEN

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Macromolecule #11: Splicing factor 3B subunit 6

MacromoleculeName: Splicing factor 3B subunit 6 / type: protein_or_peptide / ID: 11 / 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 #12: Smad nuclear-interacting protein 1

MacromoleculeName: Smad nuclear-interacting protein 1 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 45.880738 KDa
SequenceString: MKAVKSERER GSRRRHRDGD VVLPAGVVVK QERLSPEVAP PAHRRPDHSG GSPSPPTSEP ARSGHRGNRA RGVSRSPPKK KNKASGRRS KSPRSKRNRS PHHSTVKVKQ EREDHPRRGR EDRQHREPSE QEHRRARNSD RDRHRGHSHQ RRTSNERPGS G QGQGRDRD ...String:
MKAVKSERER GSRRRHRDGD VVLPAGVVVK QERLSPEVAP PAHRRPDHSG GSPSPPTSEP ARSGHRGNRA RGVSRSPPKK KNKASGRRS KSPRSKRNRS PHHSTVKVKQ EREDHPRRGR EDRQHREPSE QEHRRARNSD RDRHRGHSHQ RRTSNERPGS G QGQGRDRD TQNLQAQEEE REFYNARRRE HRQRNDVGGG GSESQELVPR PGGNNKEKEV PAKEKPSFEL SGALLEDTNT FR GVVIKYS EPPEARIPKK RWRLYPFKND EVLPVMYIHR QSAYLLGRHR RIADIPIDHP SCSKQHAVFQ YRLVEYTRAD GTV GRRVKP YIIDLGSGNG TFLNNKRIEP QRYYELKEKD VLKFGFSSRE YVLLHESSDT SEIDRKDDED EEEEEEVSDS

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Macromolecule #13: RNA-binding motif protein, X-linked 2

MacromoleculeName: RNA-binding motif protein, X-linked 2 / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 37.425984 KDa
SequenceString: MNPLTKVKLI NELNEREVQL GVADKVSWHS EYKDSAWIFL GGLPYELTEG DIICVFSQYG EIVNINLVRD KKTGKSKGFC FLCYEDQRS TILAVDNFNG IKIKGRTIRV DHVSNYRAPK DSEEIDDVTR QLQEKGCGAR TPSPSLSESS EDEKPTKKHK K DKKEKKKK ...String:
MNPLTKVKLI NELNEREVQL GVADKVSWHS EYKDSAWIFL GGLPYELTEG DIICVFSQYG EIVNINLVRD KKTGKSKGFC FLCYEDQRS TILAVDNFNG IKIKGRTIRV DHVSNYRAPK DSEEIDDVTR QLQEKGCGAR TPSPSLSESS EDEKPTKKHK K DKKEKKKK KKEKEKADRE VQAEQPSSSS PRRKTVKEKD DTGPKKHSSK NSERAQKSEP REGQKLPKSR TAYSGGAEDL ER ELKKEKP KHEHKSSSRR EAREEKTRIR DRGRSSDAHS SWYNGRSEGR SYRSRSRSRD KSHRHKRARR SRERESSNPS DRW RH

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Macromolecule #14: Serine/arginine repetitive matrix protein 1

MacromoleculeName: Serine/arginine repetitive matrix protein 1 / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 102.600539 KDa
SequenceString: MDAGFFRGTS AEQDNRFSNK QKKLLKQLKF AECLEKKVDM SKVNLEVIKP WITKRVTEIL GFEDDVVIEF IFNQLEVKNP DSKMMQINL TGFLNGKNAR EFMGELWPLL LSAQENIAGI PSAFLELKKE EIKQRQIEQE KLASMKKQDE DKDKRDKEEK E SSREKRER ...String:
MDAGFFRGTS AEQDNRFSNK QKKLLKQLKF AECLEKKVDM SKVNLEVIKP WITKRVTEIL GFEDDVVIEF IFNQLEVKNP DSKMMQINL TGFLNGKNAR EFMGELWPLL LSAQENIAGI PSAFLELKKE EIKQRQIEQE KLASMKKQDE DKDKRDKEEK E SSREKRER SRSPRRRKSR SPSPRRRSSP VRRERKRSHS RSPRHRTKSR SPSPAPEKKE KTPELPEPSV KVKEPSVQEA TS TSDILKV PKPEPIPEPK EPSPEKNSKK EKEKEKTRPR SRSRSKSRSR TRSRSPSHTR PRRRHRSRSR SYSPRRRPSP RRR PSPRRR TPPRRMPPPP RHRRSRSPVR RRRRSSASLS GSSSSSSSSR SRSPPKKPPK RTSSPPRKTR RLSPSASPPR RRHR PSPPA TPPPKTRHSP TPQQSNRTRK SRVSVSPGRT SGKVTKHKGT EKRESPSPAP KPRKVELSES EEDKGGKMAA ADSVQ QRRQ YRRQNQQSSS DSGSSSSSED ERPKRSHVKN GEVGRRRRHS PSRSASPSPR KRQKETSPRG RRRRSPSPPP TRRRRS PSP APPPRRRRTP TPPPRRRTPS PPPRRRSPSP RRYSPPIQRR YSPSPPPKRR TASPPPPPKR RASPSPPPKR RVSHSPP PK QRSSPVTKRR SPSLSSKHRK GSSPSRSTRE ARSPQPNKRH SPSPRPRAPQ TSSSPPPVRR GASSSPQRRQ SPSPSTRP I RRVSRTPEPK KIKKAASPSP QSVRRVSSSR SVSGSPEPAA KKPPAPPSPV QSQSPSTNWS PAVPVKKAKS PTPSPSPPR NSDQEGGGKK KKKKKDKKHK KDKKHKKHKK HKKEKAVAAA AAAAVTPAAI AAATTTLAQE EPVAAPEPKK ETESEAEDNL DDLEKHLRE KALRSMRKAQ VSPQS

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Macromolecule #16: DNA/RNA-binding protein KIN17

MacromoleculeName: DNA/RNA-binding protein KIN17 / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Human (human)
Molecular weightTheoretical: 45.453801 KDa
SequenceString: MGKSDFLTPK AIANRIKSKG LQKLRWYCQM CQKQCRDENG FKCHCMSESH QRQLLLASEN PQQFMDYFSE EFRNDFLELL RRRFGTKRV HNNIVYNEYI SHREHIHMNA TQWETLTDFT KWLGREGLCK VDETPKGWYI QYIDRDPETI RRQLELEKKK K QDLDDEEK ...String:
MGKSDFLTPK AIANRIKSKG LQKLRWYCQM CQKQCRDENG FKCHCMSESH QRQLLLASEN PQQFMDYFSE EFRNDFLELL RRRFGTKRV HNNIVYNEYI SHREHIHMNA TQWETLTDFT KWLGREGLCK VDETPKGWYI QYIDRDPETI RRQLELEKKK K QDLDDEEK TAKFIEEQVR RGLEGKEQEV PTFTELSREN DEEKVTFNLS KGACSSSGAT SSKSSTLGPS ALKTIGSSAS VK RKESSQS STQSKEKKKK KSALDEIMEI EEEKKRTART DYWLQPEIIV KIITKKLGEK YHKKKAIVKE VIDKYTAVVK MID SGDKLK LDQTHLETVI PAPGKRILVL NGGYRGNEGT LESINEKTFS ATIVIETGPL KGRRVEGIQY EDISKLA

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Macromolecule #3: MINX M3 pre-mRNA

MacromoleculeName: MINX M3 pre-mRNA / type: rna / ID: 3 / Number of copies: 1
Source (natural)Organism: synthetic construct (others)
Molecular weightTheoretical: 73.712359 KDa
SequenceString: GGGAGACGGA AUUCGAGCUC GCCCACUCUU GGAUCGGAAA CCCGUCGGCC UCCGAACGGU AAGAGCCUAG CAUGUAGAAC UGGUUACCU GCAGCCCAAG CUUGCUGCAC GUCUAGGGCG CAGUAGUCCA GGGUUUCCUU GAUGAUGUCA UACUUAUCCU G UCCCUUUU ...String:
GGGAGACGGA AUUCGAGCUC GCCCACUCUU GGAUCGGAAA CCCGUCGGCC UCCGAACGGU AAGAGCCUAG CAUGUAGAAC UGGUUACCU GCAGCCCAAG CUUGCUGCAC GUCUAGGGCG CAGUAGUCCA GGGUUUCCUU GAUGAUGUCA UACUUAUCCU G UCCCUUUU UUUUCCACAG CUCGCGGUUG AGGACAAACU CUUCGCGGUC UUUCCAGUGG GGAUCCAAUA UC

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Macromolecule #15: U2 snRNA

MacromoleculeName: U2 snRNA / type: rna / ID: 15 / 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

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Sample preparation

BufferpH: 7.9
GridModel: Quantifoil R3.5/1 / Material: COPPER / Support film - Material: CARBON / Support film - topology: CONTINUOUS
VitrificationCryogen name: ETHANE / Instrument: FEI VITROBOT MARK IV

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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
Image recordingFilm or detector model: FEI FALCON III (4k x 4k) / Detector mode: INTEGRATING / Digitization - Dimensions - Width: 4096 pixel / Digitization - Dimensions - Height: 4096 pixel / Average exposure time: 1.0 sec. / Average electron dose: 2.25 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: OTHER / Details: cryoSPARC ab initio reconstruction
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
Final reconstructionResolution.type: BY AUTHOR / Resolution: 4.5 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.0) / Number images used: 39336

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Atomic model buiding 1

RefinementSpace: REAL / Protocol: RIGID BODY FIT
Output model

PDB-7abh:
Human pre-Bact-2 spliceosome (SF3b/U2 snRNP portion)

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