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- PDB-7a5p: Human C Complex Spliceosome - Medium-resolution PERIPHERY -

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Basic information

Entry
Database: PDB / ID: 7a5p
TitleHuman C Complex Spliceosome - Medium-resolution PERIPHERY
Components
  • (Pre-mRNA-processing factor ...) x 2
  • (Pre-mRNA-splicing factor ...) x 7
  • (Small nuclear ribonucleoprotein ...SnRNP) x 6
  • (U2 small nuclear ribonucleoprotein ...) x 2
  • Cell division cycle 5-like protein
  • Crooked neck-like protein 1
  • Eukaryotic initiation factor 4A-III
  • Intron-binding protein aquarius
  • Peptidyl-prolyl cis-trans isomerase E
  • Peptidylprolyl isomerase domain and WD repeat-containing protein 1
  • Pre-mRNA-processing-splicing factor 8
  • Protein FRG1
  • Protein mago nashi homolog
  • RNA-binding protein 8A
  • SNW domain-containing protein 1
  • Serine/arginine repetitive matrix protein 2
  • Small nuclear ribonucleoprotein-associated proteins B and B'
  • Splicing factor YJU2
  • U2 snRNAU2 spliceosomal RNA
  • U5 small nuclear ribonucleoprotein 200 kDa helicase
  • U5 snRNAU5 spliceosomal RNA
  • U6 snRNAU6 spliceosomal RNA
  • UNKNOWN
  • WD repeat-containing protein 70
  • pre-mRNAPrimary transcript
KeywordsSPLICING / human C complex / spliceosome / hC / human step 1 spliceosome
Function / homology
Function and homology information


striated muscle dense body / regulation of histone H2B conserved C-terminal lysine ubiquitination / regulation of DNA double-strand break processing / negative regulation of selenocysteine incorporation / negative regulation of selenocysteine insertion sequence binding / cellular response to selenite ion / positive regulation of vitamin D receptor signaling pathway / post-spliceosomal complex / cellular response to interleukin-2 / cellular response to wortmannin ...striated muscle dense body / regulation of histone H2B conserved C-terminal lysine ubiquitination / regulation of DNA double-strand break processing / negative regulation of selenocysteine incorporation / negative regulation of selenocysteine insertion sequence binding / cellular response to selenite ion / positive regulation of vitamin D receptor signaling pathway / post-spliceosomal complex / cellular response to interleukin-2 / cellular response to wortmannin / cellular response to prolactin / exon-exon junction complex / snRNP binding / U2 snRNP binding / U7 snRNA binding / leucine zipper domain binding / signal transduction involved in DNA damage checkpoint / histone pre-mRNA DCP binding / regulation of translation at postsynapse, modulating synaptic transmission / transcription corepressor binding / U7 snRNP / histone pre-mRNA 3'end processing complex / negative regulation of excitatory postsynaptic potential / cis assembly of pre-catalytic spliceosome / methylosome / regulation of vitamin D receptor signaling pathway / histone mRNA metabolic process / exploration behavior / mitotic G2 DNA damage checkpoint / post-mRNA release spliceosomal complex / RNA splicing, via transesterification reactions / inner cell mass cell proliferation / generation of catalytic spliceosome for first transesterification step / RNA stem-loop binding / hair cycle / selenocysteine insertion sequence binding / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / positive regulation of neurogenesis / positive regulation of astrocyte differentiation / telomerase holoenzyme complex / U1 snRNP binding / protein phosphatase 1 binding / U2-type catalytic step 1 spliceosome / WD40-repeat domain binding / U2-type precatalytic spliceosome / positive regulation of mRNA splicing, via spliceosome / regulation of retinoic acid receptor signaling pathway / ubiquitin-ubiquitin ligase activity / P granule / C2H2 zinc finger domain binding / pICln-Sm protein complex / retinoic acid receptor binding / U4 snRNP / poly(A) binding / telomerase RNA binding / import into nucleus / small nuclear ribonucleoprotein complex / pre-mRNA binding / spliceosomal tri-snRNP complex / protein methylation / SMN-Sm protein complex / positive regulation by host of viral transcription / embryonic cranial skeleton morphogenesis / regulation of mRNA splicing, via spliceosome / Prp19 complex / mRNA cis splicing, via spliceosome / vitamin D receptor binding / U2-type catalytic step 2 spliceosome / Notch binding / K63-linked polyubiquitin modification-dependent protein binding / U2 snRNP / commitment complex / U12-type spliceosomal complex / mRNA 3'-end processing / generation of catalytic spliceosome for second transesterification step / spliceosomal complex assembly / blastocyst development / cellular response to fibroblast growth factor stimulus / cellular response to brain-derived neurotrophic factor stimulus / muscle organ development / spliceosomal snRNP assembly / U2-type prespliceosome / small nucleolar ribonucleoprotein complex / positive regulation of transcription of Notch receptor target / U1 snRNP / mRNA 3'-splice site recognition / precatalytic spliceosome / translation regulator activity / lipid biosynthetic process / protein K63-linked ubiquitination / RNA export from nucleus / cyclosporin A binding / spliceosomal tri-snRNP complex assembly / retinoic acid receptor signaling pathway / positive regulation of transforming growth factor beta receptor signaling pathway / U5 snRNP / U5 snRNA binding / U2 snRNA binding / positive regulation of histone H3-K4 methylation / regulation of alternative mRNA splicing, via spliceosome
RNA-binding motif protein 8 / Pre-mRNA splicing factor component Cdc5p/Cef1 / RNA recognition motif domain / Pre-mRNA-processing-splicing factor 8, U6-snRNA-binding / Pre-mRNA-processing-splicing factor 8, U5-snRNA-binding / Tetratricopeptide repeat / WD40 repeat, conserved site / JAB1/MPN/MOV34 metalloenzyme domain / Zinc finger, CCCH-type / G-protein beta WD-40 repeat ...RNA-binding motif protein 8 / Pre-mRNA splicing factor component Cdc5p/Cef1 / RNA recognition motif domain / Pre-mRNA-processing-splicing factor 8, U6-snRNA-binding / Pre-mRNA-processing-splicing factor 8, U5-snRNA-binding / Tetratricopeptide repeat / WD40 repeat, conserved site / JAB1/MPN/MOV34 metalloenzyme domain / Zinc finger, CCCH-type / G-protein beta WD-40 repeat / ATP-dependent RNA helicase DEAD-box, conserved site / Ricin B, lectin domain / SANT/Myb domain / Cyclophilin-type peptidyl-prolyl cis-trans isomerase, conserved site / LSM domain, eukaryotic/archaea-type / Myb domain / PRP8 domain IV core / Spt5 C-terminal domain / CWF11 family / Small nuclear ribonucleoprotein E / Leucine-rich repeat / Helicase, C-terminal / Like-Sm (LSM) domain containing protein, LSm4/SmD1/SmD3 / WD40 repeat / Small nuclear ribonucleoprotein Sm D2 / P-loop containing nucleoside triphosphate hydrolase / Pre-mRNA-processing-splicing factor 8 / Cyclophilin-like domain superfamily / WD40-repeat-containing domain / SKI-interacting protein, SKIP / Intron-binding protein aquarius, N-terminal / mRNA splicing factor Cwf21 domain / DEAD/DEAH box helicase domain / Domain of unknown function DUF1605 / Tetratricopeptide-like helical domain superfamily / Ribonuclease H-like superfamily / LSM domain superfamily / PRO8NT domain / Protein FRG1 / PROCN domain / Nucleotide-binding alpha-beta plait domain superfamily / PROCT domain / Tetratricopeptide repeat-containing domain / Zinc finger, RING/FYVE/PHD-type / Pre-mRNA-splicing factor Isy1 / Small ribonucleoprotein associated, SmB/SmN / mRNA splicing factor SYF2 / Pre-mRNA-splicing factor 19 / Helicase superfamily 1/2, ATP-binding domain / RNA helicase, DEAD-box type, Q motif / Immunoglobulin E-set / WD40/YVTN repeat-like-containing domain superfamily / MIF4G-like domain superfamily / Pre-mRNA-splicing factor SPF27 / Armadillo-type fold / Homeobox-like domain superfamily / Peptidyl-prolyl cis-trans isomerase E / Actin-crosslinking / Sm-like protein Lsm6/SmF / Helix hairpin bin domain superfamily / RNA recognition motif, spliceosomal PrP8 / Leucine-rich repeat domain superfamily / RNA-binding domain superfamily / Peptidyl-prolyl cis-trans isomerase E, RNA recognition motif / Pre-mRNA-processing factor 17 / U2 small nuclear ribonucleoprotein B'', RNA recognition motif 2 / Brr2, N-terminal helicase PWI domain / U2 small nuclear ribonucleoprotein B'', RNA recognition motif 1 / C2 domain superfamily / Pre-mRNA-splicing factor Cwc2/Slt11 / U2A'/phosphoprotein 32 family A, C-terminal / Pre-mRNA-processing factor 19 / U box domain / Initiation factor eIF-4 gamma, MA3 / WD40-repeat-containing domain superfamily / Winged helix DNA-binding domain superfamily / Mago nashi superfamily / Zinc finger, CCCH-type superfamily / Pre-mRNA-splicing factor Isy1 superfamily / MPN domain / HAT (Half-A-TPR) repeat / DNA2/NAM7 helicase, helicase domain / MIF4G-like, type 3 / Prp8 RNase domain IV, fingers region / RBM8, RNA recognition motif / Small nuclear ribonucleoprotein G / Small nuclear ribonucleoprotein Sm D3 / Cyclophilin-type peptidyl-prolyl cis-trans isomerase domain / Small nuclear ribonucleoprotein F / Saf4/Yju2 protein / Small nuclear ribonucleoprotein D1 / Helicase-associated domain / SKI-interacting protein SKIP, SNW domain / Prp8 RNase domain IV, palm region / Sec63 domain / Pre-mRNA-processing-splicing factor 8, U5-snRNA-binding domain superfamily / Mago nashi protein / DNA2/NAM7 helicase-like, C-terminal / DNA/RNA helicase, ATP-dependent, DEAH-box type, conserved site
gb:36516: / RNA-binding protein 8A / Serine/arginine repetitive matrix protein 2 / Peptidyl-prolyl cis-trans isomerase E / Pre-mRNA-processing factor 19 / Pre-mRNA-splicing factor ISY1 homolog / WD repeat-containing protein 70 / Pre-mRNA-processing-splicing factor 8 / Pre-mRNA-splicing factor RBM22 / Pre-mRNA-splicing factor SYF1 ...gb:36516: / RNA-binding protein 8A / Serine/arginine repetitive matrix protein 2 / Peptidyl-prolyl cis-trans isomerase E / Pre-mRNA-processing factor 19 / Pre-mRNA-splicing factor ISY1 homolog / WD repeat-containing protein 70 / Pre-mRNA-processing-splicing factor 8 / Pre-mRNA-splicing factor RBM22 / Pre-mRNA-splicing factor SYF1 / Pre-mRNA-splicing factor CWC22 homolog / Crooked neck-like protein 1 / Splicing factor YJU2 / Cell division cycle 5-like protein / Peptidylprolyl isomerase domain and WD repeat-containing protein 1 / Pre-mRNA-splicing factor ATP-dependent RNA helicase PRP16 / Small nuclear ribonucleoprotein-associated proteins B and B' / Protein FRG1 / U2 small nuclear ribonucleoprotein A' / RNA helicase aquarius / Pre-mRNA-processing factor 17 / U5 small nuclear ribonucleoprotein 200 kDa helicase / Pre-mRNA-splicing factor SPF27 / Pre-mRNA-splicing factor SYF2 / U2 small nuclear ribonucleoprotein B'' / Eukaryotic initiation factor 4A-III / SNW domain-containing protein 1 / Protein mago nashi homolog / 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 / gb:36515:
Biological speciesHomo sapiens (human)
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 5 Å
AuthorsBertram, K. / Kastner, B.
Funding support Germany, 2items
OrganizationGrant numberCountry
Max Planck Society Germany
German Research Foundation (DFG)SFB 860 Germany
CitationJournal: Mol Cell / Year: 2020
Title: Structural Insights into the Roles of Metazoan-Specific Splicing Factors in the Human Step 1 Spliceosome.
Authors: Karl Bertram / Leyla El Ayoubi / Olexandr Dybkov / Dmitry E Agafonov / Cindy L Will / Klaus Hartmuth / Henning Urlaub / Berthold Kastner / Holger Stark / Reinhard Lührmann /
Abstract: Human spliceosomes contain numerous proteins absent in yeast, whose functions remain largely unknown. Here we report a 3D cryo-EM structure of the human spliceosomal C complex at 3.4 Å core ...Human spliceosomes contain numerous proteins absent in yeast, whose functions remain largely unknown. Here we report a 3D cryo-EM structure of the human spliceosomal C complex at 3.4 Å core resolution and 4.5-5.7 Å at its periphery, and aided by protein crosslinking we determine its molecular architecture. Our structure provides additional insights into the spliceosome's architecture between the catalytic steps of splicing, and how proteins aid formation of the spliceosome's catalytically active RNP (ribonucleoprotein) conformation. It reveals the spatial organization of the metazoan-specific proteins PPWD1, WDR70, FRG1, and CIR1 in human C complexes, indicating they stabilize functionally important protein domains and RNA structures rearranged/repositioned during the B to C transition. Structural comparisons with human B, C, and P complexes reveal an intricate cascade of RNP rearrangements during splicing catalysis, with intermediate RNP conformations not found in yeast, and additionally elucidate the structural basis for the sequential recruitment of metazoan-specific spliceosomal proteins.
Validation Report
SummaryFull reportAbout validation report
History
DepositionAug 21, 2020Deposition site: PDBE / Processing site: PDBE
Revision 1.0Oct 14, 2020Provider: repository / Type: Initial release

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Assembly

Deposited unit
2: U2 snRNA
5: U5 snRNA
6: U6 snRNA
8: UNKNOWN
A: Pre-mRNA-processing-splicing factor 8
C: SNW domain-containing protein 1
E: Pre-mRNA-processing factor 17
G: Pre-mRNA-processing factor 19
H: Pre-mRNA-processing factor 19
I: Pre-mRNA-processing factor 19
J: Pre-mRNA-processing factor 19
K: Pre-mRNA-splicing factor SPF27
L: Cell division cycle 5-like protein
M: Pre-mRNA-splicing factor SYF1
N: Pre-mRNA-splicing factor SYF2
O: Crooked neck-like protein 1
P: Pre-mRNA-splicing factor RBM22
S: Serine/arginine repetitive matrix protein 2
T: Pre-mRNA-splicing factor CWC22 homolog
U: Intron-binding protein aquarius
W: U2 small nuclear ribonucleoprotein A'
X: U2 small nuclear ribonucleoprotein B''
Y: pre-mRNA
a: Small nuclear ribonucleoprotein Sm D3
b: Small nuclear ribonucleoprotein-associated proteins B and B'
c: Small nuclear ribonucleoprotein Sm D1
d: Small nuclear ribonucleoprotein Sm D2
e: Small nuclear ribonucleoprotein E
f: Small nuclear ribonucleoprotein F
g: Small nuclear ribonucleoprotein G
h: Small nuclear ribonucleoprotein Sm D2
i: Small nuclear ribonucleoprotein F
j: Small nuclear ribonucleoprotein E
k: Small nuclear ribonucleoprotein G
l: Small nuclear ribonucleoprotein Sm D3
m: Small nuclear ribonucleoprotein-associated proteins B and B'
n: Small nuclear ribonucleoprotein Sm D1
o: Peptidyl-prolyl cis-trans isomerase E
p: WD repeat-containing protein 70
q: U5 small nuclear ribonucleoprotein 200 kDa helicase
r: Pre-mRNA-splicing factor ATP-dependent RNA helicase PRP16
s: Pre-mRNA-splicing factor ISY1 homolog
u: Splicing factor YJU2
v: Protein mago nashi homolog
w: RNA-binding protein 8A
x: Protein FRG1
y: Eukaryotic initiation factor 4A-III
z: Peptidylprolyl isomerase domain and WD repeat-containing protein 1


Theoretical massNumber of molelcules
Total (without water)3,093,74048
Polymers3,093,74048
Non-polymers00
Water0
1


TypeNameSymmetry operationNumber
identity operation1_5551

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Components

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RNA chain , 4 types, 4 molecules 256Y

#1: RNA chain U2 snRNA / U2 spliceosomal RNA


Mass: 312349.656 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: GenBank: 36516
#2: RNA chain U5 snRNA / U5 spliceosomal RNA


Mass: 36908.668 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: GenBank: 36515
#3: 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) / Strain: HeLa S3
#20: RNA chain pre-mRNA / Primary transcript


Mass: 103979.148 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / Strain: HeLa S3

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Protein , 17 types, 18 molecules 8ACLOSUbmopquvwxyz

#4: Protein UNKNOWN


Mass: 17379.371 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / Strain: HeLa S3
#5: 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
#6: Protein SNW domain-containing protein 1 / Nuclear protein SkiP / Nuclear receptor coactivator NCoA-62 / Ski-interacting protein


Mass: 61610.703 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q13573
#10: Protein Cell division cycle 5-like protein / Cdc5-like protein / Pombe cdc5-related protein


Mass: 92406.883 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q99459
#13: Protein Crooked neck-like protein 1 / Crooked neck homolog / hCrn


Mass: 100610.008 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9BZJ0
#15: Protein Serine/arginine repetitive matrix protein 2 / 300 kDa nuclear matrix antigen / Serine/arginine-rich splicing factor-related nuclear matrix ...300 kDa nuclear matrix antigen / Serine/arginine-rich splicing factor-related nuclear matrix protein of 300 kDa / Ser/Arg-related nuclear matrix protein of 300 kDa / Splicing coactivator subunit SRm300 / Tax-responsive enhancer element-binding protein 803 / TaxREB803


Mass: 300255.312 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9UQ35
#17: Protein Intron-binding protein aquarius / Intron-binding protein of 160 kDa / IBP160


Mass: 173331.328 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O60306, RNA helicase
#22: Protein Small nuclear ribonucleoprotein-associated proteins B and B' / snRNP-B / Sm protein B/B' / SmB/B'


Mass: 24642.131 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P14678
#28: Protein Peptidyl-prolyl cis-trans isomerase E / PPIase E / Cyclophilin E / Cyclophilin-33 / Rotamase E


Mass: 33475.773 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9UNP9, peptidylprolyl isomerase
#29: Protein WD repeat-containing protein 70


Mass: 73306.172 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9NW82
#30: Protein U5 small nuclear ribonucleoprotein 200 kDa helicase / Activating signal cointegrator 1 complex subunit 3-like 1 / BRR2 homolog / U5 snRNP-specific 200 ...Activating signal cointegrator 1 complex subunit 3-like 1 / BRR2 homolog / U5 snRNP-specific 200 kDa protein / U5-200KD


Mass: 244823.422 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O75643, RNA helicase
#33: Protein Splicing factor YJU2 / Coiled-coil domain-containing protein 94


Mass: 37141.816 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9BW85
#34: Protein Protein mago nashi homolog


Mass: 17189.625 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P61326
#35: Protein RNA-binding protein 8A / Binder of OVCA1-1 / BOV-1 / RNA-binding motif protein 8A / RNA-binding protein Y14 / Ribonucleoprotein RBM8A


Mass: 19925.070 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9Y5S9
#36: Protein Protein FRG1 / FSHD region gene 1 protein


Mass: 29227.492 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q14331
#37: Protein Eukaryotic initiation factor 4A-III / eIF4A-III / ATP-dependent RNA helicase DDX48 / ATP-dependent RNA helicase eIF4A-3 / DEAD box ...eIF4A-III / ATP-dependent RNA helicase DDX48 / ATP-dependent RNA helicase eIF4A-3 / DEAD box protein 48 / Eukaryotic initiation factor 4A-like NUK-34 / Eukaryotic translation initiation factor 4A isoform 3 / Nuclear matrix protein 265 / hNMP 265


Mass: 46930.961 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P38919, RNA helicase
#38: Protein Peptidylprolyl isomerase domain and WD repeat-containing protein 1 / Spliceosome-associated cyclophilin


Mass: 73679.891 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q96BP3, peptidylprolyl isomerase

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Pre-mRNA-processing factor ... , 2 types, 5 molecules EGHIJ

#7: Protein Pre-mRNA-processing factor 17 / Cell division cycle 40 homolog / EH-binding protein 3 / Ehb3 / PRP17 homolog / hPRP17


Mass: 65612.180 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O60508
#8: Protein
Pre-mRNA-processing factor 19 / Nuclear matrix protein 200 / PRP19/PSO4 homolog / hPso4 / RING-type E3 ubiquitin transferase PRP19 ...Nuclear matrix protein 200 / PRP19/PSO4 homolog / hPso4 / RING-type E3 ubiquitin transferase PRP19 / Senescence evasion factor


Mass: 55245.547 Da / Num. of mol.: 4 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human)
References: UniProt: Q9UMS4, RING-type E3 ubiquitin transferase

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Pre-mRNA-splicing factor ... , 7 types, 7 molecules KMNPTrs

#9: Protein Pre-mRNA-splicing factor SPF27 / Breast carcinoma-amplified sequence 2 / DNA amplified in mammary carcinoma 1 protein / Spliceosome- ...Breast carcinoma-amplified sequence 2 / DNA amplified in mammary carcinoma 1 protein / Spliceosome-associated protein SPF 27


Mass: 26163.420 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O75934
#11: Protein Pre-mRNA-splicing factor SYF1 / Protein HCNP / XPA-binding protein 2


Mass: 100148.711 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9HCS7
#12: Protein Pre-mRNA-splicing factor SYF2 / CCNDBP1-interactor / p29


Mass: 28780.518 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O95926
#14: Protein Pre-mRNA-splicing factor RBM22 / RNA-binding motif protein 22 / Zinc finger CCCH domain-containing protein 16


Mass: 46959.555 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9NW64
#16: Protein Pre-mRNA-splicing factor CWC22 homolog / Nucampholin homolog / fSAPb


Mass: 105646.578 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9HCG8
#31: Protein Pre-mRNA-splicing factor ATP-dependent RNA helicase PRP16 / ATP-dependent RNA helicase DHX38 / DEAH box protein 38


Mass: 140777.688 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q92620, RNA helicase
#32: Protein Pre-mRNA-splicing factor ISY1 homolog


Mass: 33046.254 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9ULR0

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U2 small nuclear ribonucleoprotein ... , 2 types, 2 molecules WX

#18: Protein U2 small nuclear ribonucleoprotein A' / U2 snRNP A'


Mass: 28484.592 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P09661
#19: Protein U2 small nuclear ribonucleoprotein B'' / U2 snRNP B''


Mass: 25524.367 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P08579

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Small nuclear ribonucleoprotein ... , 6 types, 12 molecules alcndhejfigk

#21: Protein Small nuclear ribonucleoprotein Sm D3 / Sm-D3 / snRNP core protein D3


Mass: 13940.308 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62318
#23: Protein Small nuclear ribonucleoprotein Sm D1 / Sm-D1 / Sm-D autoantigen / snRNP core protein D1


Mass: 13310.653 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62314
#24: Protein Small nuclear ribonucleoprotein Sm D2 / Sm-D2 / snRNP core protein D2


Mass: 13551.928 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62316
#25: Protein Small nuclear ribonucleoprotein E / snRNP-E / Sm protein E / SmE


Mass: 10817.601 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62304
#26: Protein Small nuclear ribonucleoprotein F / snRNP-F / Sm protein F / SmF


Mass: 9734.171 Da / Num. of mol.: 2 / Source method: isolated from a natural source
Details: MSLPLNPKPFLNGLTGKPVMVKLKWGMEYKGYLVSVDGYMNMQLANTEEYIDGALSGHLGEVLIRCNNVLYIRGVEEEEEDGEMRE
Source: (natural) Homo sapiens (human) / References: UniProt: P62306
#27: Protein Small nuclear ribonucleoprotein G / snRNP-G / Sm protein G / SmG


Mass: 8508.084 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62308

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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 C Complex Spliceosome / Type: RIBOSOME / Entity ID: #1-#38 / Source: NATURAL
Molecular weightExperimental value: NO
Source (natural)Organism: Homo sapiens (human) / Strain: HeLa S3
Buffer solutionpH: 7.9
SpecimenEmbedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
VitrificationCryogen name: ETHANE / Humidity: 100 %

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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: SPOT SCAN
Electron lensMode: BRIGHT FIELDBright-field microscopy
Image recordingElectron dose: 6 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: 5 Å / Resolution method: OTHER / Num. of particles: 69000 / Symmetry type: POINT

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