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- EMDB-18229: Structure of the recycling U5 snRNP bound to chaperones CD2BP2 an... -

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Basic information

Entry
Database: EMDB / ID: EMD-18229
TitleStructure of the recycling U5 snRNP bound to chaperones CD2BP2 and TSSC4 (State 2, Map 2)
Map dataSharpened map
Sample
  • Complex: Structure of the recycling U5 snRNP bound to chaperones CD2BP2 and TSSC4 (State , Map 2)
    • RNA: x 1 types
    • Protein or peptide: x 14 types
  • Ligand: x 2 types
KeywordsU5 snRNP / CD2BP2 / TSSC4 / spliceosome / splicing
Function / homology
Function and homology information


RNA localization / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / U7 snRNP / 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 / protein methylation ...RNA localization / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / U7 snRNP / 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 / protein methylation / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / molecular sequestering activity / U12-type spliceosomal complex / methylosome / 7-methylguanosine cap hypermethylation / U1 snRNP binding / pICln-Sm protein complex / sno(s)RNA-containing ribonucleoprotein complex / snRNP binding / U2-type catalytic step 1 spliceosome / RNA splicing, via transesterification reactions / small nuclear ribonucleoprotein complex / SMN-Sm protein complex / telomerase RNA binding / spliceosomal tri-snRNP complex / telomerase holoenzyme complex / U2-type spliceosomal complex / U2-type precatalytic spliceosome / P granule / commitment complex / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / U4 snRNP / U2 snRNP / RNA Polymerase II Transcription Termination / U1 snRNP / U2-type prespliceosome / precatalytic spliceosome / K63-linked polyubiquitin modification-dependent protein binding / mRNA Splicing - Minor Pathway / spliceosomal complex assembly / spliceosomal tri-snRNP complex assembly / U5 snRNA binding / U5 snRNP / U2 snRNA binding / U6 snRNA binding / ribonucleoprotein complex binding / spliceosomal snRNP assembly / Cajal body / RNA processing / pre-mRNA intronic binding / U1 snRNA binding / U4/U6 x U5 tri-snRNP complex / catalytic step 2 spliceosome / mRNA Splicing - Major Pathway / RNA splicing / response to cocaine / helicase activity / spliceosomal complex / mRNA splicing, via spliceosome / fibrillar center / mRNA processing / osteoblast differentiation / cellular response to xenobiotic stimulus / cellular response to tumor necrosis factor / snRNP Assembly / protein-macromolecule adaptor activity / SARS-CoV-2 modulates host translation machinery / cellular response to lipopolysaccharide / RNA helicase activity / nuclear body / RNA helicase / nuclear speck / intracellular membrane-bounded organelle / GTPase activity / protein-containing complex binding / GTP binding / enzyme binding / positive regulation of transcription by RNA polymerase II / ATP hydrolysis activity / RNA binding / extracellular exosome / nucleoplasm / ATP binding / identical protein binding / membrane / nucleus / cytosol / cytoplasm
Similarity search - Function
Tumour suppressing sub-chromosomal transferable candidate 4 / Tumour suppressing sub-chromosomal transferable candidate 4 / CD2 antigen cytoplasmic tail-binding protein 2/Lin1 / GYF domain / GYF-like domain superfamily / GYF domain / GYF domain profile. / Contains conserved Gly-Tyr-Phe residues / PRP1 splicing factor, N-terminal / PRP1 splicing factor, N-terminal ...Tumour suppressing sub-chromosomal transferable candidate 4 / Tumour suppressing sub-chromosomal transferable candidate 4 / CD2 antigen cytoplasmic tail-binding protein 2/Lin1 / GYF domain / GYF-like domain superfamily / GYF domain / GYF domain profile. / Contains conserved Gly-Tyr-Phe residues / PRP1 splicing factor, N-terminal / PRP1 splicing factor, N-terminal / : / Small ribonucleoprotein associated, SmB/SmN / Small nuclear ribonucleoprotein D1 / Brr2, N-terminal helicase PWI domain / : / N-terminal helicase PWI domain / Pre-mRNA-splicing helicase BRR2 plug domain / Tetratricopeptide repeat / Sec63 Brl domain / : / Pre-mRNA-splicing factor Syf1-like / 116kDa U5 small nuclear ribonucleoprotein component, N-terminal / 116kDa U5 small nuclear ribonucleoprotein component, C-terminal / Snu114, GTP-binding domain / 116 kDa U5 small nuclear ribonucleoprotein component N-terminus / Sec63 domain / Sec63 Brl domain / Small nuclear ribonucleoprotein Sm D3 / : / Small nuclear ribonucleoprotein Sm D2 / Small nuclear ribonucleoprotein E / Small nuclear ribonucleoprotein G / Small nuclear ribonucleoprotein F / Like-Sm (LSM) domain containing protein, LSm4/SmD1/SmD3 / Sm-like protein Lsm7/SmG / Sm-like protein Lsm6/SmF / LSM domain / LSM domain, eukaryotic/archaea-type / snRNP Sm proteins / Tetratricopeptide repeat / HAT (Half-A-TPR) repeat / HAT (Half-A-TPR) repeats / : / Sm domain profile. / Elongation Factor G, domain II / Elongation Factor G, domain III / LSM domain superfamily / Translation elongation factor EFG/EF2, domain IV / Elongation factor G, domain IV / Elongation factor G, domain IV / Elongation factor G C-terminus / JAB1/Mov34/MPN/PAD-1 ubiquitin protease / Elongation factor EFG, domain V-like / Elongation factor G C-terminus / EF-G domain III/V-like / PROCT domain / Prp8 RNase domain IV, fingers region / PROCT (NUC072) domain / PRO8NT domain / PROCN domain / Pre-mRNA-processing-splicing factor 8, U6-snRNA-binding / Pre-mRNA-processing-splicing factor 8, U5-snRNA-binding / RNA recognition motif, spliceosomal PrP8 / PRP8 domain IV core / Pre-mRNA-processing-splicing factor 8, U5-snRNA-binding domain superfamily / Prp8 RNase domain IV, palm region / PRO8NT (NUC069), PrP8 N-terminal domain / PROCN (NUC071) domain / U6-snRNA interacting domain of PrP8 / U5-snRNA binding site 2 of PrP8 / RNA recognition motif of the spliceosomal PrP8 / PRP8 domain IV core / Pre-mRNA-processing-splicing factor 8 / Translation elongation factor EFTu-like, domain 2 / Elongation factor Tu domain 2 / JAB/MPN domain / JAB1/MPN/MOV34 metalloenzyme domain / MPN domain / MPN domain profile. / Tetratricopeptide repeats / Translational (tr)-type GTP-binding domain / Elongation factor Tu GTP binding domain / Translational (tr)-type guanine nucleotide-binding (G) domain profile. / DEAD/DEAH box helicase domain / DEAD/DEAH box helicase / C2 domain superfamily / Tetratricopeptide repeat / Helicase conserved C-terminal domain / Small GTP-binding protein domain / Tetratricopeptide-like helical domain superfamily / helicase superfamily c-terminal domain / Immunoglobulin E-set / 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 / Translation protein, beta-barrel domain superfamily / Ribosomal protein S5 domain 2-type fold, subgroup / G-protein beta WD-40 repeat
Similarity search - Domain/homology
U5 small nuclear ribonucleoprotein 200 kDa helicase / Pre-mRNA-processing factor 6 / CD2 antigen cytoplasmic tail-binding protein 2 / 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 ...U5 small nuclear ribonucleoprotein 200 kDa helicase / Pre-mRNA-processing factor 6 / CD2 antigen cytoplasmic tail-binding protein 2 / 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 / 116 kDa U5 small nuclear ribonucleoprotein component / Pre-mRNA-processing-splicing factor 8 / U5 small nuclear ribonucleoprotein 40 kDa protein / U5 small nuclear ribonucleoprotein TSSC4
Similarity search - Component
Biological speciesHomo sapiens (human)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.2 Å
AuthorsRiabov Bassat D / Plaschka C / Vorlaender MK
Funding supportEuropean Union, 2 items
OrganizationGrant numberCountry
European Research Council (ERC)ERC- 2020-STG 949081European Union
H2020 Marie Curie Actions of the European Commission101028744European Union
CitationJournal: Nat Struct Mol Biol / Year: 2024
Title: Structural basis of human U5 snRNP late biogenesis and recycling.
Authors: Daria Riabov Bassat / Supapat Visanpattanasin / Matthias K Vorländer / Laura Fin / Alexander W Phillips / Clemens Plaschka /
Abstract: Pre-mRNA splicing by the spliceosome requires the biogenesis and recycling of its small nuclear ribonucleoprotein (snRNP) complexes, which are consumed in each round of splicing. The human U5 snRNP ...Pre-mRNA splicing by the spliceosome requires the biogenesis and recycling of its small nuclear ribonucleoprotein (snRNP) complexes, which are consumed in each round of splicing. The human U5 snRNP is the ~1 MDa 'heart' of the spliceosome and is recycled through an unknown mechanism involving major architectural rearrangements and the dedicated chaperones CD2BP2 and TSSC4. Late steps in U5 snRNP biogenesis similarly involve these chaperones. Here we report cryo-electron microscopy structures of four human U5 snRNP-CD2BP2-TSSC4 complexes, revealing how a series of molecular events primes the U5 snRNP to generate the ~2 MDa U4/U6.U5 tri-snRNP, the largest building block of the spliceosome.
History
DepositionAug 16, 2023-
Header (metadata) releaseApr 17, 2024-
Map releaseApr 17, 2024-
UpdateMay 29, 2024-
Current statusMay 29, 2024Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_18229.png

Downloads & links

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Map

FileDownload / File: emd_18229.map.gz / Format: CCP4 / Size: 178 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationSharpened map
Projections & slices

Image control

Size
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AxesZ (Sec.)Y (Row.)X (Col.)
1.24 Å/pix.
x 360 pix.
= 446.688 Å		1.24 Å/pix.
x 360 pix.
= 446.688 Å		1.24 Å/pix.
x 360 pix.
= 446.688 Å

Surface

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Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.2408 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.2
Minimum - Maximum-0.8164386 - 1.9644549
Average (Standard dev.)0.002519824 (±0.04367764)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions360360360
Spacing360360360
CellA=B=C: 446.68802 Å
α=β=γ: 90.0 °

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Supplemental data

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Additional map: Unsharpened map

Fileemd_18229_additional_1.map
AnnotationUnsharpened map
Projections & Slices
AxesZYX

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Slices (1/2)
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Half map: Half map 1

Fileemd_18229_half_map_1.map
AnnotationHalf map 1
Projections & Slices
AxesZYX

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Histogram

Histogram (log scale)

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Half map: Half map 1

Fileemd_18229_half_map_2.map
AnnotationHalf map 1
Projections & Slices
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Sample components

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Entire : Structure of the recycling U5 snRNP bound to chaperones CD2BP2 an...

EntireName: Structure of the recycling U5 snRNP bound to chaperones CD2BP2 and TSSC4 (State , Map 2)
Components
  • Complex: Structure of the recycling U5 snRNP bound to chaperones CD2BP2 and TSSC4 (State , Map 2)
    • RNA: U5 snRNA
    • Protein or peptide: Pre-mRNA-processing-splicing factor 8
    • Protein or peptide: U5 small nuclear ribonucleoprotein 200 kDa helicase
    • Protein or peptide: 116 kDa U5 small nuclear ribonucleoprotein component
    • Protein or peptide: U5 small nuclear ribonucleoprotein 40 kDa protein
    • Protein or peptide: Pre-mRNA-processing factor 6
    • Protein or peptide: CD2 antigen cytoplasmic tail-binding protein 2
    • Protein or peptide: Protein TSSC4
    • Protein or peptide: Small nuclear ribonucleoprotein Sm D1
    • Protein or peptide: Small nuclear ribonucleoprotein-associated proteins B and B'
    • 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: Small nuclear ribonucleoprotein G
  • Ligand: MAGNESIUM ION
  • Ligand: GUANOSINE-5'-TRIPHOSPHATE

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Supramolecule #1: Structure of the recycling U5 snRNP bound to chaperones CD2BP2 an...

SupramoleculeName: Structure of the recycling U5 snRNP bound to chaperones CD2BP2 and TSSC4 (State , Map 2)
type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#15
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 1 MDa

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Macromolecule #1: U5 snRNA

MacromoleculeName: U5 snRNA / type: rna / ID: 1 / Number of copies: 1
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 37.254855 KDa
SequenceString:
AUACUCUGGU UUCUCUUCAG AUCGCAUAAA UCUUUCGCCU UUUACUAAAG AUUUCCGUGG AGAGGAACAA CUCUGAGUCU UAACCCAAU UUUUUGAGGC CUUGCUUUGG CAAGGCUA

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Macromolecule #2: Pre-mRNA-processing-splicing factor 8

MacromoleculeName: Pre-mRNA-processing-splicing factor 8 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 273.97425 KDa
SequenceString: MAGVFPYRGP GNPVPGPLAP LPDYMSEEKL QEKARKWQQL QAKRYAEKRK FGFVDAQKED MPPEHVRKII RDHGDMTNRK FRHDKRVYL GALKYMPHAV LKLLENMPMP WEQIRDVPVL YHITGAISFV NEIPWVIEPV YISQWGSMWI MMRREKRDRR H FKRMRFPP ...String:
MAGVFPYRGP GNPVPGPLAP LPDYMSEEKL QEKARKWQQL QAKRYAEKRK FGFVDAQKED MPPEHVRKII RDHGDMTNRK FRHDKRVYL GALKYMPHAV LKLLENMPMP WEQIRDVPVL YHITGAISFV NEIPWVIEPV YISQWGSMWI MMRREKRDRR H FKRMRFPP FDDEEPPLDY ADNILDVEPL EAIQLELDPE EDAPVLDWFY DHQPLRDSRK YVNGSTYQRW QFTLPMMSTL YR LANQLLT DLVDDNYFYL FDLKAFFTSK ALNMAIPGGP KFEPLVRDIN LQDEDWNEFN DINKIIIRQP IRTEYKIAFP YLY NNLPHH VHLTWYHTPN VVFIKTEDPD LPAFYFDPLI NPISHRHSVK SQEPLPDDDE EFELPEFVEP FLKDTPLYTD NTAN GIALL WAPRPFNLRS GRTRRALDIP LVKNWYREHC PAGQPVKVRV SYQKLLKYYV LNALKHRPPK AQKKRYLFRS FKATK FFQS TKLDWVEVGL QVCRQGYNML NLLIHRKNLN YLHLDYNFNL KPVKTLTTKE RKKSRFGNAF HLCREVLRLT KLVVDS HVQ YRLGNVDAFQ LADGLQYIFA HVGQLTGMYR YKYKLMRQIR MCKDLKHLIY YRFNTGPVGK GPGCGFWAAG WRVWLFF MR GITPLLERWL GNLLARQFEG RHSKGVAKTV TKQRVESHFD LELRAAVMHD ILDMMPEGIK QNKARTILQH LSEAWRCW K ANIPWKVPGL PTPIENMILR YVKAKADWWT NTAHYNRERI RRGATVDKTV CKKNLGRLTR LYLKAEQERQ HNYLKDGPY ITAEEAVAVY TTTVHWLESR RFSPIPFPPL SYKHDTKLLI LALERLKEAY SVKSRLNQSQ REELGLIEQA YDNPHEALSR IKRHLLTQR AFKEVGIEFM DLYSHLVPVY DVEPLEKITD AYLDQYLWYE ADKRRLFPPW IKPADTEPPP LLVYKWCQGI N NLQDVWET SEGECNVMLE SRFEKMYEKI DLTLLNRLLR LIVDHNIADY MTAKNNVVIN YKDMNHTNSY GIIRGLQFAS FI VQYYGLV MDLLVLGLHR ASEMAGPPQM PNDFLSFQDI ATEAAHPIRL FCRYIDRIHI FFRFTADEAR DLIQRYLTEH PDP NNENIV GYNNKKCWPR DARMRLMKHD VNLGRAVFWD IKNRLPRSVT TVQWENSFVS VYSKDNPNLL FNMCGFECRI LPKC RTSYE EFTHKDGVWN LQNEVTKERT AQCFLRVDDE SMQRFHNRVR QILMASGSTT FTKIVNKWNT ALIGLMTYFR EAVVN TQEL LDLLVKCENK IQTRIKIGLN SKMPSRFPPV VFYTPKELGG LGMLSMGHVL IPQSDLRWSK QTDVGITHFR SGMSHE EDQ LIPNLYRYIQ PWESEFIDSQ RVWAEYALKR QEAIAQNRRL TLEDLEDSWD RGIPRINTLF QKDRHTLAYD KGWRVRT DF KQYQVLKQNP FWWTHQRHDG KLWNLNNYRT DMIQALGGVE GILEHTLFKG TYFPTWEGLF WEKASGFEES MKWKKLTN A QRSGLNQIPN RRFTLWWSPT INRANVYVGF QVQLDLTGIF MHGKIPTLKI SLIQIFRAHL WQKIHESIVM DLCQVFDQE LDALEIETVQ KETIHPRKSY KMNSSCADIL LFASYKWNVS RPSLLADSKD VMDSTTTQKY WIDIQLRWGD YDSHDIERYA RAKFLDYTT DNMSIYPSPT GVLIAIDLAY NLHSAYGNWF PGSKPLIQQA MAKIMKANPA LYVLRERIRK GLQLYSSEPT E PYLSSQNY GELFSNQIIW FVDDTNVYRV TIHKTFEGNL TTKPINGAIF IFNPRTGQLF LKIIHTSVWA GQKRLGQLAK WK TAEEVAA LIRSLPVEEQ PKQIIVTRKG MLDPLEVHLL DFPNIVIKGS ELQLPFQACL KVEKFGDLIL KATEPQMVLF NLY DDWLKT ISSYTAFSRL ILILRALHVN NDRAKVILKP DKTTITEPHH IWPTLTDEEW IKVEVQLKDL ILADYGKKNN VNVA SLTQS EIRDIILGME ISAPSQQRQQ IAEIEKQTKE QSQLTATQTR TVNKHGDEII TSTTSNYETQ TFSSKTEWRV RAISA ANLH LRTNHIYVSS DDIKETGYTY ILPKNVLKKF ICISDLRAQI AGYLYGVSPP DNPQVKEIRC IVMVPQWGTH QTVHLP GQL PQHEYLKEME PLGWIHTQPN ESPQLSPQDV TTHAKIMADN PSWDGEKTII ITCSFTPGSC TLTAYKLTPS GYEWGRQ NT DKGNNPKGYL PSHYERVQML LSDRFLGFFM VPAQSSWNYN FMGVRHDPNM KYELQLANPK EFYHEVHRPS HFLNFALL Q EGEVYSADRE DLYA

UniProtKB: Pre-mRNA-processing-splicing factor 8

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Macromolecule #3: U5 small nuclear ribonucleoprotein 200 kDa helicase

MacromoleculeName: U5 small nuclear ribonucleoprotein 200 kDa helicase / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO / EC number: RNA helicase
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 244.823422 KDa
SequenceString: MADVTARSLQ YEYKANSNLV LQADRSLIDR TRRDEPTGEV LSLVGKLEGT RMGDKAQRTK PQMQEERRAK RRKRDEDRHD INKMKGYTL LSEGIDEMVG IIYKPKTKET RETYEVLLSF IQAALGDQPR DILCGAADEV LAVLKNEKLR DKERRKEIDL L LGQTDDTR ...String:
MADVTARSLQ YEYKANSNLV LQADRSLIDR TRRDEPTGEV LSLVGKLEGT RMGDKAQRTK PQMQEERRAK RRKRDEDRHD INKMKGYTL LSEGIDEMVG IIYKPKTKET RETYEVLLSF IQAALGDQPR DILCGAADEV LAVLKNEKLR DKERRKEIDL L LGQTDDTR YHVLVNLGKK ITDYGGDKEI QNMDDNIDET YGVNVQFESD EEEGDEDVYG EVREEASDDD MEGDEAVVRC TL SANLVAS GELMSSKKKD LHPRDIDAFW LQRQLSRFYD DAIVSQKKAD EVLEILKTAS DDRECENQLV LLLGFNTFDF IKV LRQHRM MILYCTLLAS AQSEAEKERI MGKMEADPEL SKFLYQLHET EKEDLIREER SRRERVRQSR MDTDLETMDL DQGG EALAP RQVLDLEDLV FTQGSHFMAN KRCQLPDGSF RRQRKGYEEV HVPALKPKPF GSEEQLLPVE KLPKYAQAGF EGFKT LNRI QSKLYRAALE TDENLLLCAP TGAGKTNVAL MCMLREIGKH INMDGTINVD DFKIIYIAPM RSLVQEMVGS FGKRLA TYG ITVAELTGDH QLCKEEISAT QIIVCTPEKW DIITRKGGER TYTQLVRLII LDEIHLLHDD RGPVLEALVA RAIRNIE MT QEDVRLIGLS ATLPNYEDVA TFLRVDPAKG LFYFDNSFRP VPLEQTYVGI TEKKAIKRFQ IMNEIVYEKI MEHAGKNQ V LVFVHSRKET GKTARAIRDM CLEKDTLGLF LREGSASTEV LRTEAEQCKN LELKDLLPYG FAIHHAGMTR VDRTLVEDL FADKHIQVLV STATLAWGVN LPAHTVIIKG TQVYSPEKGR WTELGALDIL QMLGRAGRPQ YDTKGEGILI TSHGELQYYL SLLNQQLPI ESQMVSKLPD MLNAEIVLGN VQNAKDAVNW LGYAYLYIRM LRSPTLYGIS HDDLKGDPLL DQRRLDLVHT A ALMLDKNN LVKYDKKTGN FQVTELGRIA SHYYITNDTV QTYNQLLKPT LSEIELFRVF SLSSEFKNIT VREEEKLELQ KL LERVPIP VKESIEEPSA KINVLLQAFI SQLKLEGFAL MADMVYVTQS AGRLMRAIFE IVLNRGWAQL TDKTLNLCKM IDK RMWQSM CPLRQFRKLP EEVVKKIEKK NFPFERLYDL NHNEIGELIR MPKMGKTIHK YVHLFPKLEL SVHLQPITRS TLKV ELTIT PDFQWDEKVH GSSEAFWILV EDVDSEVILH HEYFLLKAKY AQDEHLITFF VPVFEPLPPQ YFIRVVSDRW LSCET QLPV SFRHLILPEK YPPPTELLDL QPLPVSALRN SAFESLYQDK FPFFNPIQTQ VFNTVYNSDD NVFVGAPTGS GKTICA EFA ILRMLLQSSE GRCVYITPME ALAEQVYMDW YEKFQDRLNK KVVLLTGETS TDLKLLGKGN IIISTPEKWD ILSRRWK QR KNVQNINLFV VDEVHLIGGE NGPVLEVICS RMRYISSQIE RPIRIVALSS SLSNAKDVAH WLGCSATSTF NFHPNVRP V PLELHIQGFN ISHTQTRLLS MAKPVYHAIT KHSPKKPVIV FVPSRKQTRL TAIDILTTCA ADIQRQRFLH CTEKDLIPY LEKLSDSTLK ETLLNGVGYL HEGLSPMERR LVEQLFSSGA IQVVVASRSL CWGMNVAAHL VIIMDTQYYN GKIHAYVDYP IYDVLQMVG HANRPLQDDE GRCVIMCQGS KKDFFKKFLY EPLPVESHLD HCMHDHFNAE IVTKTIENKQ DAVDYLTWTF L YRRMTQNP NYYNLQGISH RHLSDHLSEL VEQTLSDLEQ SKCISIEDEM DVAPLNLGMI AAYYYINYTT IELFSMSLNA KT KVRGLIE IISNAAEYEN IPIRHHEDNL LRQLAQKVPH KLNNPKFNDP HVKTNLLLQA HLSRMQLSAE LQSDTEEILS KAI RLIQAC VDVLSSNGWL SPALAAMELA QMVTQAMWSK DSYLKQLPHF TSEHIKRCTD KGVESVFDIM EMEDEERNAL LQLT DSQIA DVARFCNRYP NIELSYEVVD KDSIRSGGPV VVLVQLEREE EVTGPVIAPL FPQKREEGWW VVIGDAKSNS LISIK RLTL QQKAKVKLDF VAPATGAHNY TLYFMSDAYM GCDQEYKFSV DVKEAETDSD SD

UniProtKB: U5 small nuclear ribonucleoprotein 200 kDa helicase

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Macromolecule #4: 116 kDa U5 small nuclear ribonucleoprotein component

MacromoleculeName: 116 kDa U5 small nuclear ribonucleoprotein component / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 109.560625 KDa
SequenceString: MDTDLYDEFG NYIGPELDSD EDDDELGRET KDLDEMDDDD DDDDVGDHDD DHPGMEVVLH EDKKYYPTAE EVYGPEVETI VQEEDTQPL TEPIIKPVKT KKFTLMEQTL PVTVYEMDFL ADLMDNSELI RNVTLCGHLH HGKTCFVDCL IEQTHPEIRK R YDQDLCYT ...String:
MDTDLYDEFG NYIGPELDSD EDDDELGRET KDLDEMDDDD DDDDVGDHDD DHPGMEVVLH EDKKYYPTAE EVYGPEVETI VQEEDTQPL TEPIIKPVKT KKFTLMEQTL PVTVYEMDFL ADLMDNSELI RNVTLCGHLH HGKTCFVDCL IEQTHPEIRK R YDQDLCYT DILFTEQERG VGIKSTPVTV VLPDTKGKSY LFNIMDTPGH VNFSDEVTAG LRISDGVVLF IDAAEGVMLN TE RLIKHAV QERLAVTVCI NKIDRLILEL KLPPTDAYYK LRHIVDEVNG LISMYSTDEN LILSPLLGNV CFSSSQYSIC FTL GSFAKI YADTFGDINY QEFAKRLWGD IYFNPKTRKF TKKAPTSSSQ RSFVEFILEP LYKILAQVVG DVDTSLPRTL DELG IHLTK EELKLNIRPL LRLVCKKFFG EFTGFVDMCV QHIPSPKVGA KPKIEHTYTG GVDSDLGEAM SDCDPDGPLM CHTTK MYST DDGVQFHAFG RVLSGTIHAG QPVKVLGENY TLEDEEDSQI CTVGRLWISV ARYHIEVNRV PAGNWVLIEG VDQPIV KTA TITEPRGNEE AQIFRPLKFN TTSVIKIAVE PVNPSELPKM LDGLRKVNKS YPSLTTKVEE SGEHVILGTG ELYLDCV MH DLRKMYSEID IKVADPVVTF CETVVETSSL KCFAETPNKK NKITMIAEPL EKGLAEDIEN EVVQITWNRK KLGEFFQT K YDWDLLAARS IWAFGPDATG PNILVDDTLP SEVDKALLGS VKDSIVQGFQ WGTREGPLCD ELIRNVKFKI LDAVVAQEP LHRGGGQIIP TARRVVYSAF LMATPRLMEP YYFVEVQAPA DCVSAVYTVL ARRRGHVTQD APIPGSPLYT IKAFIPAIDS FGFETDLRT HTQGQAFSLS VFHHWQIVPG DPLDKSIVIR PLEPQPAPHL AREFMIKTRR RKGLSEDVSI SKFFDDPMLL E LAKQDVVL NYPM

UniProtKB: 116 kDa U5 small nuclear ribonucleoprotein component

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Macromolecule #5: U5 small nuclear ribonucleoprotein 40 kDa protein

MacromoleculeName: U5 small nuclear ribonucleoprotein 40 kDa protein / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 39.359492 KDa
SequenceString: MIEQQKRKGP ELPLVPVKRQ RHELLLGAGS GPGAGQQQAT PGALLQAGPP RCSSLQAPIM LLSGHEGEVY CCKFHPNGST LASAGFDRL ILLWNVYGDC DNYATLKGHS GAVMELHYNT DGSMLFSAST DKTVAVWDSE TGERVKRLKG HTSFVNSCYP A RRGPQLVC ...String:
MIEQQKRKGP ELPLVPVKRQ RHELLLGAGS GPGAGQQQAT PGALLQAGPP RCSSLQAPIM LLSGHEGEVY CCKFHPNGST LASAGFDRL ILLWNVYGDC DNYATLKGHS GAVMELHYNT DGSMLFSAST DKTVAVWDSE TGERVKRLKG HTSFVNSCYP A RRGPQLVC TGSDDGTVKL WDIRKKAAIQ TFQNTYQVLA VTFNDTSDQI ISGGIDNDIK VWDLRQNKLT YTMRGHADSV TG LSLSSEG SYLLSNAMDN TVRVWDVRPF APKERCVKIF QGNVHNFEKN LLRCSWSPDG SKIAAGSADR FVYVWDTTSR RIL YKLPGH AGSINEVAFH PDEPIIISAS SDKRLYMGEI Q

UniProtKB: U5 small nuclear ribonucleoprotein 40 kDa protein

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Macromolecule #6: Pre-mRNA-processing factor 6

MacromoleculeName: Pre-mRNA-processing factor 6 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 107.092242 KDa
SequenceString: MNKKKKPFLG MPAPLGYVPG LGRGATGFTT RSDIGPARDA NDPVDDRHAP PGKRTVGDQM KKNQAADDDD EDLNDTNYDE FNGYAGSLF SSGPYEKDDE EADAIYAALD KRMDERRKER REQREKEEIE KYRMERPKIQ QQFSDLKRKL AEVTEEEWLS I PEVGDARN ...String:
MNKKKKPFLG MPAPLGYVPG LGRGATGFTT RSDIGPARDA NDPVDDRHAP PGKRTVGDQM KKNQAADDDD EDLNDTNYDE FNGYAGSLF SSGPYEKDDE EADAIYAALD KRMDERRKER REQREKEEIE KYRMERPKIQ QQFSDLKRKL AEVTEEEWLS I PEVGDARN KRQRNPRYEK LTPVPDSFFA KHLQTGENHT SVDPRQTQFG GLNTPYPGGL NTPYPGGMTP GLMTPGTGEL DM RKIGQAR NTLMDMRLSQ VSDSVSGQTV VDPKGYLTDL NSMIPTHGGD INDIKKARLL LKSVRETNPH HPPAWIASAR LEE VTGKLQ VARNLIMKGT EMCPKSEDVW LEAARLQPGD TAKAVVAQAV RHLPQSVRIY IRAAELETDI RAKKRVLRKA LEHV PNSVR LWKAAVELEE PEDARIMLSR AVECCPTSVE LWLALARLET YENARKVLNK ARENIPTDRH IWITAAKLEE ANGNT QMVE KIIDRAITSL RANGVEINRE QWIQDAEECD RAGSVATCQA VMRAVIGIGI EEEDRKHTWM EDADSCVAHN ALECAR AIY AYALQVFPSK KSVWLRAAYF EKNHGTRESL EALLQRAVAH CPKAEVLWLM GAKSKWLAGD VPAARSILAL AFQANPN SE EIWLAAVKLE SENDEYERAR RLLAKARSSA PTARVFMKSV KLEWVQDNIR AAQDLCEEAL RHYEDFPKLW MMKGQIEE Q KEMMEKAREA YNQGLKKCPH STPLWLLLSR LEEKIGQLTR ARAILEKSRL KNPKNPGLWL ESVRLEYRAG LKNIANTLM AKALQECPNS GILWSEAIFL EARPQRRTKS VDALKKCEHD PHVLLAVAKL FWSQRKITKA REWFHRTVKI DSDLGDAWAF FYKFELQHG TEEQQEEVRK RCESAEPRHG ELWCAVSKDI ANWQKKIGDI LRLVAGRIKN TF

UniProtKB: Pre-mRNA-processing factor 6

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Macromolecule #7: CD2 antigen cytoplasmic tail-binding protein 2

MacromoleculeName: CD2 antigen cytoplasmic tail-binding protein 2 / type: protein_or_peptide / ID: 7 / Details: the N-terminal GP comes from a cleaved 3C tag / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 37.841523 KDa
Recombinant expressionOrganism: Homo sapiens (human)
SequenceString: GPMPKRKVTF QGVGDEEDED EIIVPKKKLV DPVAGSGGPG SRFKGKHSLD SDEEEDDDDG GSSKYDILAS EDVEGQEAAT LPSEGGVRI TPFNLQEEME EGHFDADGNY FLNRDAQIRD SWLDNIDWVK IRERPPGQRQ ASDSEEEDSL GQTSMSAQAL L EGLLELLL ...String:
GPMPKRKVTF QGVGDEEDED EIIVPKKKLV DPVAGSGGPG SRFKGKHSLD SDEEEDDDDG GSSKYDILAS EDVEGQEAAT LPSEGGVRI TPFNLQEEME EGHFDADGNY FLNRDAQIRD SWLDNIDWVK IRERPPGQRQ ASDSEEEDSL GQTSMSAQAL L EGLLELLL PRETVAGALR RLGARGGGKG RKGPGQPSSP QRLDRLSGLA DQMVARGNLG VYQETRERLA MRLKGLGCQT LG PHNPTPP PSLDMFAEEL AEEELETPTP TQRGEAESRG DGLVDVMWEY KWENTGDAEL YGPFTSAQMQ TWVSEGYFPD GVY CRKLDP PGGQFYNSKR IDFDLYT

UniProtKB: CD2 antigen cytoplasmic tail-binding protein 2

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Macromolecule #8: Protein TSSC4

MacromoleculeName: Protein TSSC4 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 34.366465 KDa
SequenceString: MAEAGTGEPS PSVEGEHGTE YDTLPSDTVS LSDSDSDLSL PGGAEVEALS PMGLPGEEDS GPDEPPSPPS GLLPATVQPF HLRGMSSTF SQRSRDIFDC LEGAARRAPS SVAHTSMSDN GGFKRPLAPS GRSPVEGLGR AHRSPASPRV PPVPDYVAHP E RWTKYSLE ...String:
MAEAGTGEPS PSVEGEHGTE YDTLPSDTVS LSDSDSDLSL PGGAEVEALS PMGLPGEEDS GPDEPPSPPS GLLPATVQPF HLRGMSSTF SQRSRDIFDC LEGAARRAPS SVAHTSMSDN GGFKRPLAPS GRSPVEGLGR AHRSPASPRV PPVPDYVAHP E RWTKYSLE DVTEVSEQSN QATALAFLGS QSLAAPTDCV SSFNQDPSSC GEGRVIFTKP VRGVEARHER KRVLGKVGEP GR GGLGNPA TDRGEGPVEL AHLAGPGSPE AEEWGSHHGG LQEVEALSGS VHSGSVPGLP PVETVGFHGS RKRSRDHFRN KSS SPEDPG AEV

UniProtKB: U5 small nuclear ribonucleoprotein TSSC4

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Macromolecule #9: Small nuclear ribonucleoprotein Sm D1

MacromoleculeName: Small nuclear ribonucleoprotein Sm D1 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 13.310653 KDa
SequenceString:
MKLVRFLMKL SHETVTIELK NGTQVHGTIT GVDVSMNTHL KAVKMTLKNR EPVQLETLSI RGNNIRYFIL PDSLPLDTLL VDVEPKVKS KKREAVAGRG RGRGRGRGRG RGRGRGGPRR

UniProtKB: Small nuclear ribonucleoprotein Sm D1

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Macromolecule #10: Small nuclear ribonucleoprotein-associated proteins B and B'

MacromoleculeName: Small nuclear ribonucleoprotein-associated proteins B and B'
type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (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

UniProtKB: Small nuclear ribonucleoprotein-associated proteins B and B'

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Macromolecule #11: Small nuclear ribonucleoprotein Sm D2

MacromoleculeName: Small nuclear ribonucleoprotein Sm D2 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 13.551928 KDa
SequenceString:
MSLLNKPKSE MTPEELQKRE EEEFNTGPLS VLTQSVKNNT QVLINCRNNK KLLGRVKAFD RHCNMVLENV KEMWTEVPKS GKGKKKSKP VNKDRYISKM FLRGDSVIVV LRNPLIAGK

UniProtKB: Small nuclear ribonucleoprotein Sm D2

+
Macromolecule #12: Small nuclear ribonucleoprotein Sm D3

MacromoleculeName: Small nuclear ribonucleoprotein Sm D3 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 13.940308 KDa
SequenceString:
MSIGVPIKVL HEAEGHIVTC ETNTGEVYRG KLIEAEDNMN CQMSNITVTY RDGRVAQLEQ VYIRGSKIRF LILPDMLKNA PMLKSMKNK NQGSGAGRGK AAILKAQVAA RGRGRGMGRG NIFQKRR

UniProtKB: Small nuclear ribonucleoprotein Sm D3

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Macromolecule #13: Small nuclear ribonucleoprotein E

MacromoleculeName: Small nuclear ribonucleoprotein E / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 10.817601 KDa
SequenceString:
MAYRGQGQKV QKVMVQPINL IFRYLQNRSR IQVWLYEQVN MRIEGCIIGF DEYMNLVLDD AEEIHSKTKS RKQLGRIMLK GDNITLLQS VSN

UniProtKB: Small nuclear ribonucleoprotein E

+
Macromolecule #14: Small nuclear ribonucleoprotein F

MacromoleculeName: Small nuclear ribonucleoprotein F / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 9.734171 KDa
SequenceString:
MSLPLNPKPF LNGLTGKPVM VKLKWGMEYK GYLVSVDGYM NMQLANTEEY IDGALSGHLG EVLIRCNNVL YIRGVEEEEE DGEMRE

UniProtKB: Small nuclear ribonucleoprotein F

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Macromolecule #15: Small nuclear ribonucleoprotein G

MacromoleculeName: Small nuclear ribonucleoprotein G / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Homo sapiens (human)
Molecular weightTheoretical: 8.508084 KDa
SequenceString:
MSKAHPPELK KFMDKKLSLK LNGGRHVQGI LRGFDPFMNL VIDECVEMAT SGQQNNIGMV VIRGNSIIML EALERV

UniProtKB: Small nuclear ribonucleoprotein G

+
Macromolecule #16: MAGNESIUM ION

MacromoleculeName: MAGNESIUM ION / type: ligand / ID: 16 / Number of copies: 1 / Formula: MG
Molecular weightTheoretical: 24.305 Da

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Macromolecule #17: GUANOSINE-5'-TRIPHOSPHATE

MacromoleculeName: GUANOSINE-5'-TRIPHOSPHATE / type: ligand / ID: 17 / Number of copies: 1 / Formula: GTP
Molecular weightTheoretical: 523.18 Da
Chemical component information

ChemComp-GTP:
GUANOSINE-5'-TRIPHOSPHATE / GTP, energy-carrying molecule*YM

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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
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: FEI FALCON IV (4k x 4k) / Average electron dose: 40.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.5 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: PDB ENTRY
PDB model - PDB ID:

6qw6

Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.2 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 19393
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
FSC plot (resolution estimation)

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