EMDB-18225: cryo-EM structure of the human spliceosomal B complex protomer (t...

Basic information

Entry

Database: EMDB / ID: EMD-18225

• Title: cryo-EM structure of the human spliceosomal B complex protomer (tri-snRNP core region)

Sample

• Complex: human pre-catalytic spliceosome
  • RNA: x 4 types
  • Protein or peptide: x 17 types

• Keywords: spliceosome / pre-catalytic spliceosome / spliceosomal B complex / SPLICING

Function / homology

Function:

spliceosomal snRNP complex / microfibril / ribonucleoprotein complex localization / U4atac snRNP / positive regulation of cytotoxic T cell differentiation / maturation of 5S rRNA / RNA localization / U4atac snRNA binding / box C/D sno(s)RNA binding / snRNP binding / dense fibrillar component / box C/D methylation guide snoRNP complex / U4/U6 snRNP / transcription elongation factor activity / U2-type catalytic step 1 spliceosome / RNA splicing, via transesterification reactions / positive regulation of androgen receptor activity / spliceosomal tri-snRNP complex / U4 snRNA binding / mRNA cis splicing, via spliceosome / U2-type spliceosomal complex / U2-type precatalytic spliceosome / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / box C/D snoRNP assembly / U4 snRNP / proline-rich region binding / rRNA modification in the nucleus and cytosol / U2 snRNP / ubiquitin-like protein conjugating enzyme binding / RNA polymerase binding / U3 snoRNA binding / U2-type prespliceosome / K63-linked polyubiquitin modification-dependent protein binding / positive regulation of protein targeting to mitochondrion / precatalytic spliceosome / spliceosomal complex assembly / mRNA Splicing - Minor Pathway / negative regulation of transcription elongation by RNA polymerase II / single fertilization / MLL1 complex / mRNA 3'-splice site recognition / spliceosomal tri-snRNP complex assembly / Major pathway of rRNA processing in the nucleolus and cytosol / U5 snRNA binding / U5 snRNP / Cajal body / U2 snRNA binding / spliceosomal snRNP assembly / U6 snRNA binding / RNA processing / ribonucleoprotein complex binding / pre-mRNA intronic binding / maturation of SSU-rRNA / U1 snRNA binding / maturation of LSU-rRNA / U4/U6 x U5 tri-snRNP complex / catalytic step 2 spliceosome / mRNA Splicing - Major Pathway / small-subunit processome / RNA splicing / nuclear receptor coactivator activity / response to cocaine / nuclear receptor binding / positive regulation of transcription elongation by RNA polymerase II / transcription coregulator activity / spliceosomal complex / protein modification process / ribosomal small subunit biogenesis / protein tag activity / mRNA splicing, via spliceosome / mRNA processing / transcription corepressor activity / cellular response to xenobiotic stimulus / cellular response to tumor necrosis factor / cytosolic large ribosomal subunit / ATPase binding / cellular response to lipopolysaccharide / RNA polymerase II-specific DNA-binding transcription factor binding / transcription coactivator activity / nuclear speck / cell cycle / cell division / intracellular membrane-bounded organelle / GTPase activity / centrosome / chromatin / GTP binding / nucleolus / Golgi apparatus / negative regulation of transcription by RNA polymerase II / positive regulation of transcription by RNA polymerase II / protein-containing complex / DNA binding / RNA binding / zinc ion binding / nucleoplasm / membrane / identical protein binding / nucleus

Domain/homology:

WW domain-binding protein 4 / Pre-mRNA-splicing factor 38, C-terminal / Pre-mRNA-splicing factor 38-associated hydrophilic C-term / Micro-fibrillar-associated protein 1, C-terminal / Microfibrillar-associated protein 1 / Ubiquitin-like modifier Hub1/Ubl5 / Microfibril-associated/Pre-mRNA processing / PRP4-like superfamily / U4/U6.U5 small nuclear ribonucleoprotein component Snu23 / pre-mRNA processing factor 4 (PRP4) like / Pre-mRNA-splicing factor 38 / PRP38 family / Transcription elongation regulator 1-like / SNU66/SART1 family / HIND motif / SART-1 family / HIND motif / U1-C, C2H2-type zinc finger / U1 zinc finger / PWI domain superfamily / PWI domain / PWI domain profile. / Splicing factor 3A subunit 1, ubiquitin domain / Pre-mRNA processing factor 4 (PRP4)-like / Splicing Factor Motif, present in Prp18 and Pr04 / Prp31 C-terminal / U4/U6 small nuclear ribonucleoprotein Prp31 / Prp31 C terminal domain / Pre-mRNA-splicing factor 3 / U4/U6 small nuclear ribonucleoprotein Prp3 / pre-mRNA processing factor 3 domain / PWI domain / PWI, domain in splicing factors / Small nuclear ribonucleoprotein Prp3, C-terminal domain / Small nuclear ribonucleoprotein Prp3, C-terminal domain / FF domain / Dim1 family / Mitosis protein DIM1 / Mitosis protein DIM1 / PRP1 splicing factor, N-terminal / PRP1 splicing factor, N-terminal / Splicing factor 3A subunit 1 / FF domain / FF domain superfamily / FF domain profile. / Contains two conserved F residues / Splicing factor 3A subunit 1, conserved domain / Pre-mRNA splicing factor PRP21 like protein / SWAP/Surp / SWAP/Surp superfamily / Surp module / SURP motif repeat profile. / Suppressor-of-White-APricot splicing regulator / BUD31/G10-related, conserved site / G10 protein signature 1. / G10 protein signature 2. / G10 protein / Pre-mRNA-splicing factor BUD31 / Matrin/U1-C, C2H2-type zinc finger / Zinc finger matrin-type profile. / Pre-mRNA-splicing factor Syf1-like / NOSIC / Zinc-finger double-stranded RNA-binding / NOSIC (NUC001) domain / Nop domain / Zinc finger, double-stranded RNA binding / Nop domain superfamily / Nop, C-terminal domain / snoRNA binding domain, fibrillarin / Nop domain profile. / H/ACA ribonucleoprotein complex, subunit Nhp2-like / Snu114, GTP-binding domain / Tetratricopeptide repeat / 116kDa U5 small nuclear ribonucleoprotein component, N-terminal / 116kDa U5 small nuclear ribonucleoprotein component, C-terminal / 116 kDa U5 small nuclear ribonucleoprotein component N-terminus / Matrin/U1-C-like, C2H2-type zinc finger / U1-like zinc finger / Tetratricopeptide repeat / HAT (Half-A-TPR) repeat / HAT (Half-A-TPR) repeats / Elongation Factor G, domain II / Elongation Factor G, domain III / JAB1/Mov34/MPN/PAD-1 ubiquitin protease / Translation elongation factor EFG/EF2, domain IV / Elongation factor G, domain IV / Elongation factor G, domain IV / Elongation factor G C-terminus / Elongation factor EFG, domain V-like / Elongation factor G C-terminus / WW domain / WW/rsp5/WWP domain signature. / EF-G domain III/V-like / WW domain superfamily / WW/rsp5/WWP domain profile. / Domain with 2 conserved Trp (W) residues / WW domain / PROCT domain / Prp8 RNase domain IV, fingers region / PROCT (NUC072) domain

Component:

Transcription elongation regulator 1 / U4/U6 small nuclear ribonucleoprotein Prp4 / U4/U6.U5 tri-snRNP-associated protein 1 / U4/U6 small nuclear ribonucleoprotein Prp3 / WW domain-binding protein 4 / Pre-mRNA-processing factor 6 / Protein BUD31 homolog / Microfibrillar-associated protein 1 / NHP2-like protein 1 / Thioredoxin-like protein 4A / 116 kDa U5 small nuclear ribonucleoprotein component / Splicing factor 3A subunit 1 / Pre-mRNA-processing-splicing factor 8 / Pre-mRNA-splicing factor 38A / U4/U6 small nuclear ribonucleoprotein Prp31 / Zinc finger matrin-type protein 2 / Ubiquitin-like protein 5

• Biological species: Homo sapiens (human)
• Method: single particle reconstruction / cryo EM / Resolution: 3.1 Å
• Authors: Zhang Z / Kumar V / Dybkov O / Will CL / Urlaub H / Stark H / Luehrmann R

Funding support

Germany, 1 items

Organization	Grant number	Country
German Research Foundation (DFG)		Germany

• Citation: Journal: To Be Published; Title: New insights into the functions of B complex proteins revealed by cryo-EM of dimerized spliceosomes; Authors: Zhang Z / Kumar V / Dybkov O / Will CL / Urlaub H / Stark H / Luehrmann R

History

Deposition	Aug 16, 2023	-
Header (metadata) release	Jan 24, 2024	-
Map release	Jan 24, 2024	-
Update	Jan 24, 2024	-
Current status	Jan 24, 2024	Processing site: PDBe / Status: Released

Map

• File: Download / File: emd_18225.map.gz / Format: CCP4 / Size: 536.4 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
• Voxel size: X=Y=Z: 1.16 Å

Density

Contour Level	By AUTHOR: 0.03
Minimum - Maximum	-0.11173251 - 0.23956151
Average (Standard dev.)	0.00047533226 (±0.005526795)

• Symmetry: Space group: 1

Details

EMDB XML:

Map geometry	Axis order X Y Z Origin 0 0 0 Dimensions 520 520 520 Spacing 520 520 520
Cell	A=B=C: 603.2 Å α=β=γ: 90.0 °

Supplemental data

Half map: #2

• File: emd_18225_half_map_1.map

Half map: #1

• File: emd_18225_half_map_2.map

Sample components

Entire : human pre-catalytic spliceosome

• Entire: Name: human pre-catalytic spliceosome

Components

• Complex: human pre-catalytic spliceosome
  • RNA: U5 snRNAU5 spliceosomal RNA
  • RNA: U6 snRNAU6 spliceosomal RNA
  • Protein or peptide: Splicing factor 3A subunit 1
  • Protein or peptide: 116 kDa U5 small nuclear ribonucleoprotein component
  • Protein or peptide: Thioredoxin-like protein 4A
  • Protein or peptide: Pre-mRNA-splicing factor 38A
  • Protein or peptide: Microfibrillar-associated protein 1
  • Protein or peptide: NHP2-like protein 1, N-terminally processed
  • Protein or peptide: Protein BUD31 homolog
  • Protein or peptide: WW domain-binding protein 4
  • RNA: MINX pre-mRNA
  • Protein or peptide: Ubiquitin-like protein 5
  • Protein or peptide: U4/U6 small nuclear ribonucleoprotein Prp31
  • Protein or peptide: U4/U6 small nuclear ribonucleoprotein Prp4
  • Protein or peptide: Pre-mRNA-processing factor 6
  • Protein or peptide: Pre-mRNA-processing-splicing factor 8
  • Protein or peptide: U4/U6.U5 tri-snRNP-associated protein 1
  • Protein or peptide: Transcription elongation regulator 1
  • RNA: U4 snRNAU4 spliceosomal RNA
  • Protein or peptide: Zinc finger matrin-type protein 2
  • Protein or peptide: U4/U6 small nuclear ribonucleoprotein Prp3

Supramolecule #1: human pre-catalytic spliceosome

• Supramolecule: Name: human pre-catalytic spliceosome / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#11, #13-#20, #12, #21
• Source (natural): Organism: Homo sapiens (human)

Macromolecule #1: U5 snRNA

• Macromolecule: Name: U5 snRNA / type: rna / ID: 1 / Number of copies: 1
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 37.254855 KDa

Sequence

String:

AUACUCUGGU UUCUCUUCAG AUCGCAUAAA UCUUUCGCCU UUUACUAAAG AUUUCCGUGG AGAGGAACAA CUCUGAGUCU UAACCCAAU UUUUUGAGGC CUUGCUUUGG CAAGGCUA

GENBANK: GENBANK: AC105129.4

Macromolecule #2: U6 snRNA

• Macromolecule: Name: U6 snRNA / type: rna / ID: 2 / Number of copies: 1
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 34.09827 KDa

Sequence

String:

GUGCUCGCUU CGGCAGCACA UAUACUAAAA UUGGAACGAU ACAGAGAAGA UUAGCAUGGC CCCUGCGCAA GGAUGACACG CAAAUUCGU GAAGCGUUCC AUAUUUU

GENBANK: GENBANK: NR_004394.1

Macromolecule #11: MINX pre-mRNA

• Macromolecule: Name: MINX pre-mRNA / type: rna / ID: 11 / Number of copies: 1
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 111.300453 KDa

Sequence

String:

GGUACCUAAU ACGACUCACU AUAGGGAGAC GGAAUUCGAG CUCGCCCACU CUUGGAUCGG AAACCCGUCG GCCUCCGAAC GGUAAGAGC CUAGCAUGUA GAACUGGUUA CCUGCAGCCC AAGCUUGCUG CACGUCUAGG GCGCAGUAGU CCAGGGUUUC C UUGAUGAU GUCAUACUUA UCCUGUCCCU UUUUUUUCCA CAGCUCGCGG UUGAGGACAA ACUCUUCGCG GUCUUUCCAG UG GGGAUCC AAUAUCCGUA CACCAUCAGG GUACGAGCUA GCCCAUGGCG UACACCAUCA GGGUACGACU AGUAGAUCUC GUA CACCAU CAGGGUACGG AAUUCUCUAG A

Macromolecule #20: U4 snRNA

• Macromolecule: Name: U4 snRNA / type: rna / ID: 20 / Number of copies: 1
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 46.528465 KDa

Sequence

String:

AGCUUUGCGC AGUGGCAGUA UCGUAGCCAA UGAGGUUUAU CCGAGGCGCG AUUAUUGCUA AUUGAAAACU UUUCCCAAUA CCCCGCCGU GACGACUUGC AAUAUAGUCG GCAUUGGCAA UUUUUGACAG UCUCUACGGA GACUGG

Macromolecule #3: Splicing factor 3A subunit 1

• Macromolecule: Name: Splicing factor 3A subunit 1 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 88.991094 KDa

Sequence

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

UniProtKB: Splicing factor 3A subunit 1

Macromolecule #4: 116 kDa U5 small nuclear ribonucleoprotein component

• Macromolecule: Name: 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 weight: Theoretical: 109.560625 KDa

Sequence

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

Macromolecule #5: Thioredoxin-like protein 4A

• Macromolecule: Name: Thioredoxin-like protein 4A / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 16.807346 KDa

Sequence

String:

MSYMLPHLHN GWQVDQAILS EEDRVVVIRF GHDWDPTCMK MDEVLYSIAE KVKNFAVIYL VDITEVPDFN KMYELYDPCT VMFFFRNKH IMIDLGTGNN NKINWAMEDK QEMVDIIETV YRGARKGRGL VVSPKDYSTK YRY

UniProtKB: Thioredoxin-like protein 4A

Macromolecule #6: Pre-mRNA-splicing factor 38A

• Macromolecule: Name: Pre-mRNA-splicing factor 38A / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 37.563863 KDa

Sequence

String:

MANRTVKDAH SIHGTNPQYL VEKIIRTRIY ESKYWKEECF GLTAELVVDK AMELRFVGGV YGGNIKPTPF LCLTLKMLQI QPEKDIIVE FIKNEDFKYV RMLGALYMRL TGTAIDCYKY LEPLYNDYRK IKSQNRNGEF ELMHVDEFID ELLHSERVCD I ILPRLQKR YVLEEAEQLE PRVSALEEDM DDVESSEEEE EEDEKLERVP SPDHRRRSYR DLDKPRRSPT LRYRRSRSRS PR RRSRSPK RRSPSPRRER HRSKSPRRHR SRSRDRRHRS RSKSPGHHRS HRHRSHSKSP ERSKKSHKKS RRGNE

UniProtKB: Pre-mRNA-splicing factor 38A

Macromolecule #7: Microfibrillar-associated protein 1

• Macromolecule: Name: Microfibrillar-associated protein 1 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 52.050527 KDa

Sequence

String:

MSVPSALMKQ PPIQSTAGAV PVRNEKGEIS MEKVKVKRYV SGKRPDYAPM ESSDEEDEEF QFIKKAKEQE AEPEEQEEDS SSDPRLRRL QNRISEDVEE RLARHRKIVE PEVVGESDSE VEGDAWRMER EDSSEEEEEE IDDEEIERRR GMMRQRAQER K NEEMEVME VEDEGRSGEE SESESEYEEY TDSEDEMEPR LKPVFIRKKD RVTVQEREAE ALKQKELEQE AKRMAEERRK YT LKIVEEE TKKELEENKR SLAALDALNT DDENDEEEYE AWKVRELKRI KRDREDREAL EKEKAEIERM RNLTEEERRA ELR ANGKVI TNKAVKGKYK FLQKYYHRGA FFMDEDEEVY KRDFSAPTLE DHFNKTILPK VMQVKNFGRS GRTKYTHLVD QDTT SFDSA WGQESAQNTK FFKQKAAGVR DVFERPSAKK RKTT

UniProtKB: Microfibrillar-associated protein 1

Macromolecule #8: NHP2-like protein 1, N-terminally processed

• Macromolecule: Name: NHP2-like protein 1, N-terminally processed / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 14.191524 KDa

Sequence

String:

MTEADVNPKA YPLADAHLTK KLLDLVQQSC NYKQLRKGAN EATKTLNRGI SEFIVMAADA EPLEIILHLP LLCEDKNVPY VFVRSKQAL GRACGVSRPV IACSVTIKEG SQLKQQIQSI QQSIERLLV

UniProtKB: NHP2-like protein 1

Macromolecule #9: Protein BUD31 homolog

• Macromolecule: Name: Protein BUD31 homolog / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 17.03285 KDa

Sequence

String:

MPKVKRSRKA PPDGWELIEP TLDELDQKMR EAETEPHEGK RKVESLWPIF RIHHQKTRYI FDLFYKRKAI SRELYEYCIK EGYADKNLI AKWKKQGYEN LCCLRCIQTR DTNFGTNCIC RVPKSKLEVG RIIECTHCGC RGCSG

UniProtKB: Protein BUD31 homolog

Macromolecule #10: WW domain-binding protein 4

• Macromolecule: Name: WW domain-binding protein 4 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 42.575801 KDa

Sequence

String:

MADYWKSQPK KFCDYCKCWI ADNRPSVEFH ERGKNHKENV AKRISEIKQK SLDKAKEEEK ASKEFAAMEA AALKAYQEDL KRLGLESEI LEPSITPVTS TIPPTSTSNQ QKEKKEKKKR KKDPSKGRWV EGITSEGYHY YYDLISGASQ WEKPEGFQGD L KKTAVKTV WVEGLSEDGF TYYYNTETGE SRWEKPDDFI PHTSDLPSSK VNENSLGTLD ESKSSDSHSD SDGEQEAEEG GV STETEKP KIKFKEKNKN SDGGSDPETQ KEKSIQKQNS LGSNEEKSKT LKKSNPYGEW QEIKQEVESH EEVDLELPST ENE YVSTSE ADGGGEPKVV FKEKTVTSLG VMADGVAPVF KKRRTENGKS RNLRQRGDDQ

UniProtKB: WW domain-binding protein 4

Macromolecule #12: Zinc finger matrin-type protein 2

• Macromolecule: Name: Zinc finger matrin-type protein 2 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 23.664047 KDa

Sequence

String:

MASGSGTKNL DFRRKWDKDE YEKLAEKRLT EEREKKDGKP VQPVKRELLR HRDYKVDLES KLGKTIVITK TTPQSEMGGY YCNVCDCVV KDSINFLDHI NGKKHQRNLG MSMRVERSTL DQVKKRFEVN KKKMEEKQKD YDFEERMKEL REEEEKAKAY K KEKQKEKK RRAEEDLTFE EDDEMAAVMG FSGFGSTKKS Y

UniProtKB: Zinc finger matrin-type protein 2

Macromolecule #13: Ubiquitin-like protein 5

• Macromolecule: Name: Ubiquitin-like protein 5 / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 8.560945 KDa

Sequence

String:

MIEVVCNDRL GKKVRVKCNT DDTIGDLKKL IAAQTGTRWN KIVLKKWYTI FKDHVSLGDY EIHDGMNLEL YYQ

UniProtKB: Ubiquitin-like protein 5

Macromolecule #14: U4/U6 small nuclear ribonucleoprotein Prp31

• Macromolecule: Name: U4/U6 small nuclear ribonucleoprotein Prp31 / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 55.528969 KDa

Sequence

String:

MSLADELLAD LEEAAEEEEG GSYGEEEEEP AIEDVQEETQ LDLSGDSVKT IAKLWDSKMF AEIMMKIEEY ISKQAKASEV MGPVEAAPE YRVIVDANNL TVEIENELNI IHKFIRDKYS KRFPELESLV PNALDYIRTV KELGNSLDKC KNNENLQQIL T NATIMVVS VTASTTQGQQ LSEEELERLE EACDMALELN ASKHRIYEYV ESRMSFIAPN LSIIIGASTA AKIMGVAGGL TN LSKMPAC NIMLLGAQRK TLSGFSSTSV LPHTGYIYHS DIVQSLPPDL RRKAARLVAA KCTLAARVDS FHESTEGKVG YEL KDEIER KFDKWQEPPP VKQVKPLPAP LDGQRKKRGG RRYRKMKERL GLTEIRKQAN RMSFGEIEED AYQEDLGFSL GHLG KSGSG RVRQTQVNEA TKARISKTLQ RTLQKQSVVY GGKSTIRDRS SGTASSVAFT PLQGLEIVNP QAAEKKVAEA NQKYF SSMA EFLKVKGEKS GLMST

UniProtKB: U4/U6 small nuclear ribonucleoprotein Prp31

Macromolecule #15: U4/U6 small nuclear ribonucleoprotein Prp4

• Macromolecule: Name: U4/U6 small nuclear ribonucleoprotein Prp4 / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 58.536105 KDa

Sequence

String:

MASSRASSTQ ATKTKAPDDL VAPVVKKPHI YYGSLEEKER ERLAKGESGI LGKDGLKAGI EAGNINITSG EVFEIEEHIS ERQAEVLAE FERRKRARQI NVSTDDSEVK ACLRALGEPI TLFGEGPAER RERLRNILSV VGTDALKKTK KDDEKSKKSK E EYQQTWYH EGPNSLKVAR LWIANYSLPR AMKRLEEARL HKEIPETTRT SQMQELHKSL RSLNNFCSQI GDDRPISYCH FS PNSKMLA TACWSGLCKL WSVPDCNLLH TLRGHNTNVG AIVFHPKSTV SLDPKDVNLA SCAADGSVKL WSLDSDEPVA DIE GHTVRV ARVMWHPSGR FLGTTCYDRS WRLWDLEAQE EILHQEGHSM GVYDIAFHQD GSLAGTGGLD AFGRVWDLRT GRCI MFLEG HLKEIYGINF SPNGYHIATG SGDNTCKVWD LRQRRCVYTI PAHQNLVTGV KFEPIHGNFL LTGAYDNTAK IWTHP GWSP LKTLAGHEGK VMGLDISSDG QLIATCSYDR TFKLWMAE

UniProtKB: U4/U6 small nuclear ribonucleoprotein Prp4

Macromolecule #16: Pre-mRNA-processing factor 6

• Macromolecule: Name: Pre-mRNA-processing factor 6 / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 107.092242 KDa

Sequence

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

Macromolecule #17: Pre-mRNA-processing-splicing factor 8

• Macromolecule: Name: Pre-mRNA-processing-splicing factor 8 / type: protein_or_peptide / ID: 17 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 273.97425 KDa

Sequence

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

Macromolecule #18: U4/U6.U5 tri-snRNP-associated protein 1

• Macromolecule: Name: U4/U6.U5 tri-snRNP-associated protein 1 / type: protein_or_peptide / ID: 18 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 90.414117 KDa

Sequence

String:

MGSSKKHRGE KEAAGTTAAA GTGGATEQPP RHREHKKHKH RSGGSGGSGG ERRKRSRERG GERGSGRRGA EAEARSSTHG RERSQAEPS ERRVKREKRD DGYEAAASSK TSSGDASSLS IEETNKLRAK LGLKPLEVNA IKKEAGTKEE PVTADVINPM A LRQREELR EKLAAAKEKR LLNQKLGKIK TLGEDDPWLD DTAAWIERSR QLQKEKDLAE KRAKLLEEMD QEFGVSTLVE EE FGQRRQD LYSARDLQGL TVEHAIDSFR EGETMILTLK DKGVLQEEED VLVNVNLVDK ERAEKNVELR KKKPDYLPYA EDE SVDDLA QQKPRSILSK YDEELEGERP HSFRLEQGGT ADGLRERELE EIRAKLRLQA QSLSTVGPRL ASEYLTPEEM VTFK KTKRR VKKIRKKEKE VVVRADDLLP LGDQTQDGDF GSRLRGRGRR RVSEVEEEKE PVPQPLPSDD TRVENMDISD EEEGG APPP GSPQVLEEDE AELELQKQLE KGRRLRQLQQ LQQLRDSGEK VVEIVKKLES RQRGWEEDED PERKGAIVFN ATSEFC RTL GEIPTYGLAG NREEQEELMD FERDEERSAN GGSESDGEEN IGWSTVNLDE EKQQQDFSAS STTILDEEPI VNRGLAA AL LLCQNKGLLE TTVQKVARVK APNKSLPSAV YCIEDKMAID DKYSRREEYR GFTQDFKEKD GYKPDVKIEY VDETGRKL T PKEAFRQLSH RFHGKGSGKM KTERRMKKLD EEALLKKMSS SDTPLGTVAL LQEKQKAQKT PYIVLSGSGK SMNANTITK

UniProtKB: U4/U6.U5 tri-snRNP-associated protein 1

Macromolecule #19: Transcription elongation regulator 1

• Macromolecule: Name: Transcription elongation regulator 1 / type: protein_or_peptide / ID: 19 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 124.083 KDa

Sequence

String:

MAERGGDGGE SERFNPGELR MAQQQALRFR GPAPPPNAVM RGPPPLMRPP PPFGMMRGPP PPPRPPFGRP PFDPNMPPMP PPGGIPPPM GPPHLQRPPF MPPPMSSMPP PPGMMFPPGM PPVTAPGTPA LPPTEEIWVE NKTPDGKVYY YNARTRESAW T KPDGVKVI QQSELTPMLA AQAQVQAQAQ AQAQAQAQAQ AQAQAQAQAQ AQAQAQAQAQ AQAQAQAQAQ AQAQAQAQAQ AQ AQVQAQV QAQVQAQAVG ASTPTTSSPA PAVSTSTSSS TPSSTTSTTT TATSVAQTVS TPTTQDQTPS SAVSVATPTV SVS TPAPTA TPVQTVPQPH PQTLPPAVPH SVPQPTTAIP AFPPVMVPPF RVPLPGMPIP LPGVAMMQIV SCPYVKTVAT TKTG VLPGM APPIVPMIHP QVAIAASPAT LAGATAVSEW TEYKTADGKT YYYNNRTLES TWEKPQELKE KEKLEEKIKE PIKEP SEEP LPMETEEEDP KEEPIKEIKE EPKEEEMTEE EKAAQKAKPV ATAPIPGTPW CVVWTGDERV FFYNPTTRLS MWDRPD DLI GRADVDKIIQ EPPHKKGMEE LKKLRHPTPT MLSIQKWQFS MSAIKEEQEL MEEINEDEPV KAKKRKRDDN KDIDSEK EA AMEAEIKAAR ERAIVPLEAR MKQFKDMLLE RGVSAFSTWE KELHKIVFDP RYLLLNPKER KQVFDQYVKT RAEEERRE K KNKIMQAKED FKKMMEEAKF NPRATFSEFA AKHAKDSRFK AIEKMKDREA LFNEFVAAAR KKEKEDSKTR GEKIKSDFF ELLSNHHLDS QSRWSKVKDK VESDPRYKAV DSSSMREDLF KQYIEKIAKN LDSEKEKELE RQARIEASLR EREREVQKAR SEQTKEIDR EREQHKREEA IQNFKALLSD MVRSSDVSWS DTRRTLRKDH RWESGSLLER EEKEKLFNEH IEALTKKKRE H FRQLLDET SAITLTSTWK EVKKIIKEDP RCIKFSSSDR KKQREFEEYI RDKYITAKAD FRTLLKETKF ITYRSKKLIQ ES DQHLKDV EKILQNDKRY LVLDCVPEER RKLIVAYVDD LDRRGPPPPP TASEPTRRST K

UniProtKB: Transcription elongation regulator 1

Macromolecule #21: U4/U6 small nuclear ribonucleoprotein Prp3

• Macromolecule: Name: U4/U6 small nuclear ribonucleoprotein Prp3 / type: protein_or_peptide / ID: 21 / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Homo sapiens (human)
• Molecular weight: Theoretical: 77.669188 KDa

Sequence

String:

MALSKRELDE LKPWIEKTVK RVLGFSEPTV VTAALNCVGK GMDKKKAADH LKPFLDDSTL RFVDKLFEAV EEGRSSRHSK SSSDRSRKR ELKEVFGDDS EISKESSGVK KRRIPRFEEV EEEPEVIPGP PSESPGMLTK LQIKQMMEAA TRQIEERKKQ L SFISPPTP QPKTPSSSQP ERLPIGNTIQ PSQAATFMND AIEKARKAAE LQARIQAQLA LKPGLIGNAN MVGLANLHAM GI APPKVEL KDQTKPTPLI LDEQGRTVDA TGKEIELTHR MPTLKANIRA VKREQFKQQL KEKPSEDMES NTFFDPRVSI APS QRQRRT FKFHDKGKFE KIAQRLRTKA QLEKLQAEIS QAARKTGIHT STRLALIAPK KELKEGDIPE IEWWDSYIIP NGFD LTEEN PKREDYFGIT NLVEHPAQLN PPVDNDTPVT LGVYLTKKEQ KKLRRQTRRE AQKELQEKVR LGLMPPPEPK VRISN LMRV LGTEAVQDPT KVEAHVRAQM AKRQKAHEEA NAARKLTAEQ RKVKKIKKLK EDISQGVHIS VYRVRNLSNP AKKFKI EAN AGQLYLTGVV VLHKDVNVVV VEGGPKAQKK FKRLMLHRIK WDEQTSNTKG DDDEESDEEA VKKTNKCVLV WEGTAKD RS FGEMKFKQCP TENMAREHFK KHGAEHYWDL ALSESVLEST D

UniProtKB: U4/U6 small nuclear ribonucleoprotein Prp3

Experimental details

Structure determination

• Method: cryo EM
• Processing: single particle reconstruction
• Aggregation state: particle

Sample preparation

• Buffer: pH: 7.9
• Vitrification: Cryogen name: ETHANE

Electron microscopy

• Microscope: FEI TITAN KRIOS
• Electron beam: Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
• Electron optics: Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.5 µm
• Image recording: Film or detector model: FEI FALCON III (4k x 4k) / Detector mode: INTEGRATING / Average electron dose: 48.0 e/Ų
• Experimental equipment: Model: Titan Krios / Image courtesy: FEI Company

Image processing

• Startup model: Type of model: OTHER / Details: ab initio 3D reconstruction
• Initial angle assignment: Type: MAXIMUM LIKELIHOOD
• Final angle assignment: Type: MAXIMUM LIKELIHOOD
• Final reconstruction: Resolution.type: BY AUTHOR / Resolution: 3.1 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 251564