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- PDB-8q7n: cryo-EM structure of the human spliceosomal B complex protomer (t... -

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Entry
Database: PDB / ID: 8q7n
Titlecryo-EM structure of the human spliceosomal B complex protomer (tri-snRNP core region)
Components
  • (U4/U6 small nuclear ribonucleoprotein ...) x 3
  • 116 kDa U5 small nuclear ribonucleoprotein component
  • MINX pre-mRNA
  • Microfibrillar-associated protein 1
  • NHP2-like protein 1, N-terminally processed
  • Pre-mRNA-processing factor 6
  • Pre-mRNA-processing-splicing factor 8
  • Pre-mRNA-splicing factor 38A
  • Protein BUD31 homolog
  • Splicing factor 3A subunit 1
  • Thioredoxin-like protein 4A
  • Transcription elongation regulator 1
  • U4 snRNAU4 spliceosomal RNA
  • U4/U6.U5 tri-snRNP-associated protein 1
  • U5 snRNAU5 spliceosomal RNA
  • U6 snRNAU6 spliceosomal RNA
  • Ubiquitin-like protein 5
  • WW domain-binding protein 4
  • Zinc finger matrin-type protein 2
KeywordsSPLICING / spliceosome / pre-catalytic spliceosome / spliceosomal B complex
Function / homology
Function and homology information


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 ...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 / U6 snRNA binding / spliceosomal snRNP assembly / RNA processing / ribonucleoprotein complex binding / pre-mRNA intronic binding / maturation of SSU-rRNA / U1 snRNA binding / U4/U6 x U5 tri-snRNP complex / maturation of LSU-rRNA / catalytic step 2 spliceosome / small-subunit processome / mRNA Splicing - Major Pathway / RNA splicing / response to cocaine / nuclear receptor coactivator activity / 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 processing / mRNA splicing, via spliceosome / 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
Similarity search - Function
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 ...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 / PWI domain superfamily / PWI domain / PWI domain profile. / U1-C, C2H2-type zinc finger / U1 zinc finger / 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 / Small nuclear ribonucleoprotein Prp3, C-terminal domain / Small nuclear ribonucleoprotein Prp3, C-terminal domain / PWI domain / PWI, domain in splicing factors / 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
Similarity search - Domain/homology
: / : / RNA / RNA (> 10) / RNA (> 100) / 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 ...: / : / RNA / RNA (> 10) / RNA (> 100) / 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
Similarity search - Component
Biological speciesHomo sapiens (human)
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.1 Å
AuthorsZhang, Z. / Kumar, V. / Dybkov, O. / Will, C.L. / Urlaub, H. / Stark, H. / Luehrmann, R.
Funding support Germany, 1items
OrganizationGrant numberCountry
German Research Foundation (DFG) Germany
CitationJournal: 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, C.L. / Urlaub, H. / Stark, H. / Luehrmann, R.
History
DepositionAug 16, 2023Deposition site: PDBE / Processing site: PDBE
Revision 1.0Jan 24, 2024Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
5: U5 snRNA
6: U6 snRNA
7: Splicing factor 3A subunit 1
C: 116 kDa U5 small nuclear ribonucleoprotein component
D: Thioredoxin-like protein 4A
I: Pre-mRNA-splicing factor 38A
K: Microfibrillar-associated protein 1
M: NHP2-like protein 1, N-terminally processed
Q: Protein BUD31 homolog
X: WW domain-binding protein 4
Z: MINX pre-mRNA
r: Zinc finger matrin-type protein 2
s: Ubiquitin-like protein 5
L: U4/U6 small nuclear ribonucleoprotein Prp31
F: U4/U6 small nuclear ribonucleoprotein Prp4
N: Pre-mRNA-processing factor 6
A: Pre-mRNA-processing-splicing factor 8
S: U4/U6.U5 tri-snRNP-associated protein 1
T: Transcription elongation regulator 1
4: U4 snRNA
J: U4/U6 small nuclear ribonucleoprotein Prp3


Theoretical massNumber of molelcules
Total (without water)1,427,47921
Polymers1,427,47921
Non-polymers00
Water0
1


  • Idetical with deposited unit
  • defined by author&software
  • Evidence: electron microscopy, not applicable
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1

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Components

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RNA chain , 4 types, 4 molecules 56Z4

#1: RNA chain U5 snRNA / U5 spliceosomal RNA


Mass: 37254.855 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: GenBank: 20330981
#2: RNA chain U6 snRNA / U6 spliceosomal RNA


Mass: 34098.270 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: GenBank: NR_004394.1
#11: RNA chain MINX pre-mRNA


Mass: 111300.453 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human)
#20: RNA chain U4 snRNA / U4 spliceosomal RNA


Mass: 46528.465 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human)

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Protein , 14 types, 14 molecules 7CDIKMQXrsNAST

#3: Protein Splicing factor 3A subunit 1 / SF3a120 / Spliceosome-associated protein 114 / SAP 114


Mass: 88991.094 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q15459
#4: Protein 116 kDa U5 small nuclear ribonucleoprotein component / Elongation factor Tu GTP-binding domain-containing protein 2 / SNU114 homolog / hSNU114 / U5 snRNP- ...Elongation factor Tu GTP-binding domain-containing protein 2 / SNU114 homolog / hSNU114 / U5 snRNP-specific protein / 116 kDa / U5-116 kDa


Mass: 109560.625 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q15029
#5: Protein Thioredoxin-like protein 4A / DIM1 protein homolog / Spliceosomal U5 snRNP-specific 15 kDa protein / Thioredoxin-like U5 snRNP ...DIM1 protein homolog / Spliceosomal U5 snRNP-specific 15 kDa protein / Thioredoxin-like U5 snRNP protein U5-15kD


Mass: 16807.346 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P83876
#6: Protein Pre-mRNA-splicing factor 38A


Mass: 37563.863 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q8NAV1
#7: Protein Microfibrillar-associated protein 1 / Spliceosome B complex protein MFAP1


Mass: 52050.527 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P55081
#8: Protein NHP2-like protein 1, N-terminally processed


Mass: 14191.524 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P55769
#9: Protein Protein BUD31 homolog / Protein EDG-2 / Protein G10 homolog


Mass: 17032.850 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P41223
#10: Protein WW domain-binding protein 4 / WBP-4 / Formin-binding protein 21 / WW domain-containing-binding protein 4


Mass: 42575.801 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O75554
#12: Protein Zinc finger matrin-type protein 2


Mass: 23664.047 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q96NC0
#13: Protein Ubiquitin-like protein 5


Mass: 8560.945 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9BZL1
#16: Protein Pre-mRNA-processing factor 6 / Androgen receptor N-terminal domain-transactivating protein 1 / ANT-1 / PRP6 homolog / U5 snRNP- ...Androgen receptor N-terminal domain-transactivating protein 1 / ANT-1 / PRP6 homolog / U5 snRNP-associated 102 kDa protein / U5-102 kDa protein


Mass: 107092.242 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O94906
#17: Protein Pre-mRNA-processing-splicing factor 8 / 220 kDa U5 snRNP-specific protein / PRP8 homolog / Splicing factor Prp8 / p220


Mass: 273974.250 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q6P2Q9
#18: Protein U4/U6.U5 tri-snRNP-associated protein 1 / SNU66 homolog / hSnu66 / Squamous cell carcinoma antigen recognized by T-cells 1 / SART-1 / hSART-1 ...SNU66 homolog / hSnu66 / Squamous cell carcinoma antigen recognized by T-cells 1 / SART-1 / hSART-1 / U4/U6.U5 tri-snRNP-associated 110 kDa protein


Mass: 90414.117 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O43290
#19: Protein Transcription elongation regulator 1 / / TATA box-binding protein-associated factor 2S / Transcription factor CA150


Mass: 124083.000 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O14776

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U4/U6 small nuclear ribonucleoprotein ... , 3 types, 3 molecules LFJ

#14: Protein U4/U6 small nuclear ribonucleoprotein Prp31 / Pre-mRNA-processing factor 31 / Serologically defined breast cancer antigen NY-BR-99 / U4/U6 snRNP ...Pre-mRNA-processing factor 31 / Serologically defined breast cancer antigen NY-BR-99 / U4/U6 snRNP 61 kDa protein / Protein 61K / hPrp31


Mass: 55528.969 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q8WWY3
#15: Protein U4/U6 small nuclear ribonucleoprotein Prp4 / PRP4 homolog / hPrp4 / U4/U6 snRNP 60 kDa protein / WD splicing factor Prp4


Mass: 58536.105 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O43172
#21: Protein U4/U6 small nuclear ribonucleoprotein Prp3 / Pre-mRNA-splicing factor 3 / hPrp3 / U4/U6 snRNP 90 kDa protein


Mass: 77669.188 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O43395

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Experimental details

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Experiment

ExperimentMethod: ELECTRON MICROSCOPY
EM experimentAggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction

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Sample preparation

ComponentName: human pre-catalytic spliceosome / Type: COMPLEX / Entity ID: #1-#11, #13-#20, #12, #21 / Source: NATURAL
Source (natural)Organism: Homo sapiens (human)
Buffer solutionpH: 7.9
SpecimenEmbedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
VitrificationCryogen name: ETHANE

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
MicroscopyModel: FEI TITAN KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM
Electron lensMode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 3000 nm / Nominal defocus min: 1500 nm
Image recordingElectron dose: 48 e/Å2 / Detector mode: INTEGRATING / Film or detector model: FEI FALCON III (4k x 4k)

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Processing

CTF correctionType: PHASE FLIPPING AND AMPLITUDE CORRECTION
3D reconstructionResolution: 3.1 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 251564 / Symmetry type: POINT

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