EMDB-3539: Structure of a spliceosome remodeled for exon ligation

Basic information

• Entry: Database: EMDB / ID: EMD-3539
• Title: Structure of a spliceosome remodeled for exon ligation
• Map data: Saccharomyces cerevisiae C-star spliceosome, predisposed to carry out exon ligation, the second step of pre-mRNA splicing.

Sample

• Complex: Saccharomyces cerevisiae spliceosome. Complex C just after Prp16-mediated remodeling
  • RNA: x 5 types
  • Protein or peptide: x 25 types
• Ligand: x 5 types

Function / homology

Function:

U2-type post-spliceosomal complex / U2-type post-mRNA release spliceosomal complex / mRNA branch site recognition / cellular bud site selection / pre-mRNA 3'-splice site binding / post-mRNA release spliceosomal complex / cis assembly of pre-catalytic spliceosome / generation of catalytic spliceosome for first transesterification step / nuclear mRNA surveillance / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / splicing factor binding / U4/U6 snRNP / 7-methylguanosine cap hypermethylation / pre-mRNA binding / U2-type catalytic step 1 spliceosome / pICln-Sm protein complex / Prp19 complex / spliceosomal tri-snRNP complex / small nuclear ribonucleoprotein complex / SMN-Sm protein complex / mRNA cis splicing, via spliceosome / U2-type spliceosomal complex / U2-type prespliceosome assembly / commitment complex / U2-type catalytic step 2 spliceosome / U4 snRNP / U2 snRNP / poly(U) RNA binding / U1 snRNP / U2-type prespliceosome / precatalytic spliceosome / spliceosomal complex assembly / Dual incision in TC-NER / DNA replication origin binding / generation of catalytic spliceosome for second transesterification step / Gap-filling DNA repair synthesis and ligation in TC-NER / mRNA 3'-splice site recognition / mRNA 5'-splice site recognition / DNA replication initiation / spliceosomal tri-snRNP complex assembly / U5 snRNA binding / U5 snRNP / spliceosomal snRNP assembly / U2 snRNA binding / U6 snRNA binding / pre-mRNA intronic binding / positive regulation of cell cycle / U1 snRNA binding / U4/U6 x U5 tri-snRNP complex / catalytic step 2 spliceosome / nuclear periphery / positive regulation of RNA splicing / spliceosomal complex / mRNA splicing, via spliceosome / metallopeptidase activity / cell cycle / mRNA binding / GTPase activity / chromatin binding / chromatin / GTP binding / DNA binding / RNA binding / zinc ion binding / metal ion binding / nucleus / cytosol / cytoplasm

Domain/homology:

Prp18 / Pre-mRNA-splicing factor 18 / Prp18 domain / Pre-mRNA-splicing factor SLU7 domain / Pre-mRNA-splicing factor SLU7 / Pre-mRNA splicing Prp18-interacting factor / Slt11, RNA recognition motif / cwf21 / Torus domain / Pre-mRNA-splicing factor Cwc2, RNA recognition motif / Torus domain / mRNA splicing factor SYF2 / SYF2 splicing factor / mRNA splicing factor Cwf21 domain / cwf21 domain / Pre-mRNA-processing factor 17 / : / STL11, N-terminal / WD repeat Prp46/PLRG1-like / BUD31/G10-related, conserved site / : / : / G10 protein signature 1. / G10 protein signature 2. / SKI-interacting protein SKIP, SNW domain / SKI-interacting protein, SKIP / SKIP/SNW domain / Pre-mRNA-splicing factor Cwf15/Cwc15 / HAT (Half-A-TPR) repeat / Cwf15/Cwc15 cell cycle control protein / Pre-mRNA-splicing factor Cwc2/Slt11 / G10 protein / Pre-mRNA-splicing factor BUD31 / Pre-mRNA splicing factor component Cdc5p/Cef1, C-terminal / pre-mRNA splicing factor component / Initiation factor eIF-4 gamma, MA3 / MA3 domain / MI domain profile. / Domain in DAP-5, eIF4G, MA-3 and other proteins. / Pre-mRNA-splicing factor Syf1-like / Middle domain of eukaryotic initiation factor 4G (eIF4G) / MIF4G-like, type 3 / Snu114, GTP-binding domain / 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 / Small nuclear ribonucleoprotein Sm D3 / Small nuclear ribonucleoprotein Sm D2 / Small nuclear ribonucleoprotein E / Small nuclear ribonucleoprotein G / Small nuclear ribonucleoprotein F / Sm-like protein Lsm7/SmG / Like-Sm (LSM) domain containing protein, LSm4/SmD1/SmD3 / Sm-like protein Lsm6/SmF / Myb-type HTH DNA-binding domain profile. / Zinc finger, CCCH-type / Zinc finger C3H1-type profile. / LSM domain / LSM domain, eukaryotic/archaea-type / snRNP Sm proteins / HAT (Half-A-TPR) repeat / HAT (Half-A-TPR) repeats / Myb domain / : / Sm domain profile. / Myb-like DNA-binding domain / Translation elongation factor EFG/EF2, domain IV / Elongation factor G, domain IV / Elongation factor G, domain IV / LSM domain superfamily / Elongation factor G C-terminus / Elongation factor EFG, domain V-like / Elongation factor G C-terminus / EF-G domain III/V-like / SANT SWI3, ADA2, N-CoR and TFIIIB'' DNA-binding domains / SANT/Myb domain / 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 / JAB/MPN domain / JAB1/MPN/MOV34 metalloenzyme domain / MPN domain / MPN domain profile. / Translational (tr)-type GTP-binding domain / Elongation factor Tu GTP binding domain

Component:

Pre-mRNA-splicing factor BUD31 / Pre-mRNA-processing protein 45 / Pre-mRNA-splicing factor 8 / Pre-mRNA-splicing factor 18 / Pre-mRNA-splicing factor SNU114 / Pre-mRNA-splicing factor SLT11 / Small nuclear ribonucleoprotein-associated protein B / Small nuclear ribonucleoprotein G / Pre-mRNA-processing factor 17 / Small nuclear ribonucleoprotein Sm D3 / Pre-mRNA-splicing factor SYF2 / Pre-mRNA-splicing factor CWC22 / Small nuclear ribonucleoprotein F / Small nuclear ribonucleoprotein Sm D1 / Pre-mRNA-splicing factor SLU7 / Pre-mRNA-splicing factor CWC21 / Pre-mRNA-splicing factor CEF1 / Pre-mRNA-splicing factor CWC15 / Pre-mRNA-splicing factor SYF1 / Small nuclear ribonucleoprotein Sm D2 / Pre-mRNA-splicing factor CWC2 / Pre-mRNA-splicing factor CLF1 / Small nuclear ribonucleoprotein E / Pre-mRNA-splicing factor PRP46

• Biological species: Saccharomyces cerevisiae (brewer's yeast) / Baker's yeast (brewer's yeast)
• Method: single particle reconstruction / cryo EM / Resolution: 3.85 Å
• Authors: Fica SM / Oubridge C / Galej WP / Wilkinson ME / Newman AJ / Bai X-C / Nagai K

Funding support

United Kingdom, 3 items

Organization	Grant number	Country
Medical Research Council (United Kingdom)	MC-U105184330	United Kingdom
EMBO and Marie Sklodowska-Curie Fellowship	Ref. S.M.Fica
European Research Council	AdG-693087-SPLICE3D

• Citation: Journal: Nature / Year: 2016; Title: Cryo-EM structure of the spliceosome immediately after branching.; Authors: Wojciech P Galej / Max E Wilkinson / Sebastian M Fica / Chris Oubridge / Andrew J Newman / Kiyoshi Nagai / ; Abstract: Precursor mRNA (pre-mRNA) splicing proceeds by two consecutive transesterification reactions via a lariat-intron intermediate. Here we present the 3.8 Å cryo-electron microscopy structure of the spliceosome immediately after lariat formation. The 5'-splice site is cleaved but remains close to the catalytic Mg site in the U2/U6 small nuclear RNA (snRNA) triplex, and the 5'-phosphate of the intron nucleotide G(+1) is linked to the branch adenosine 2'OH. The 5'-exon is held between the Prp8 amino-terminal and linker domains, and base-pairs with U5 snRNA loop 1. Non-Watson-Crick interactions between the branch helix and 5'-splice site dock the branch adenosine into the active site, while intron nucleotides +3 to +6 base-pair with the U6 snRNA ACAGAGA sequence. Isy1 and the step-one factors Yju2 and Cwc25 stabilize docking of the branch helix. The intron downstream of the branch site emerges between the Prp8 reverse transcriptase and linker domains and extends towards the Prp16 helicase, suggesting a plausible mechanism of remodelling before exon ligation.

History

Deposition	Dec 18, 2016	-
Header (metadata) release	Jan 18, 2017	-
Map release	Jan 18, 2017	-
Update	Oct 7, 2020	-
Current status	Oct 7, 2020	Processing site: PDBe / Status: Released

Map

• File: Download / File: emd_3539.map.gz / Format: CCP4 / Size: 266.8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
• Annotation: Saccharomyces cerevisiae C-star spliceosome, predisposed to carry out exon ligation, the second step of pre-mRNA splicing.
• Voxel size: X=Y=Z: 1.43 Å

Density

Contour Level	By AUTHOR: 0.036 / Movie #1: 0.036
Minimum - Maximum	-0.07415286 - 0.17009385
Average (Standard dev.)	0.0000997267 (±0.0051415786)

• Symmetry: Space group: 1

Details

EMDB XML:

Map geometry	Axis order X Y Z Origin 0 0 0 Dimensions 412 412 412 Spacing 412 412 412
Cell	A=B=C: 589.16 Å α=β=γ: 90.0 °

CCP4 map header:

mode	Image stored as Reals
Å/pix. X/Y/Z	1.43	1.43	1.43
M x/y/z	412	412	412
origin x/y/z	0.000	0.000	0.000
length x/y/z	589.160	589.160	589.160
α/β/γ	90.000	90.000	90.000
MAP C/R/S	1	2	3
start NC/NR/NS	0	0	0
NC/NR/NS	412	412	412
D min/max/mean	-0.074	0.170	0.000

Supplemental data

Sample components

Entire : Saccharomyces cerevisiae spliceosome. Complex C just after Prp16-...

• Entire: Name: Saccharomyces cerevisiae spliceosome. Complex C just after Prp16-mediated remodeling

Components

• Complex: Saccharomyces cerevisiae spliceosome. Complex C just after Prp16-mediated remodeling
  • RNA: Yeast UBC4 gene for ubiquitin-conjugating enzyme
  • RNA: UBC4 gene exon
  • RNA: U2 snRNAU2 spliceosomal RNA
  • RNA: Saccharomyces cerevisiae strain T.52_2H chromosome XII sequence
  • RNA: U5 snRNAU5 spliceosomal RNA
  • Protein or peptide: Pre-mRNA-splicing factor 8
  • Protein or peptide: Pre-mRNA-splicing factor SNU114
  • Protein or peptide: Pre-mRNA-splicing factor CWC22
  • Protein or peptide: Pre-mRNA-splicing factor PRP46
  • Protein or peptide: Pre-mRNA-processing protein 45
  • Protein or peptide: Pre-mRNA-splicing factor BUD31
  • Protein or peptide: Pre-mRNA-splicing factor CWC2
  • Protein or peptide: Pre-mRNA-splicing factor SLT11
  • Protein or peptide: Pre-mRNA-splicing factor CEF1
  • Protein or peptide: Pre-mRNA-splicing factor CWC15
  • Protein or peptide: Pre-mRNA-splicing factor CWC21
  • Protein or peptide: Pre-mRNA-splicing factor CLF1
  • Protein or peptide: Pre-mRNA-splicing factor SYF1
  • Protein or peptide: Pre-mRNA-splicing factor 18
  • Protein or peptide: Pre-mRNA-splicing factor SLU7
  • Protein or peptide: Pre-mRNA-processing factor 17
  • Protein or peptide: Unknown
  • Protein or peptide: Pre-mRNA-splicing factor SYF2
  • Protein or peptide: Small nuclear ribonucleoprotein-associated protein B
  • 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
  • Protein or peptide: Small nuclear ribonucleoprotein Sm D1
  • Protein or peptide: Small nuclear ribonucleoprotein Sm D2
• Ligand: MAGNESIUM ION
• Ligand: POTASSIUM IONPotassium
• Ligand: INOSITOL HEXAKISPHOSPHATEPhytic acid
• Ligand: GUANOSINE-5'-TRIPHOSPHATEGuanosine triphosphate
• Ligand: ZINC ION

Supramolecule #1: Saccharomyces cerevisiae spliceosome. Complex C just after Prp16-...

• Supramolecule: Name: Saccharomyces cerevisiae spliceosome. Complex C just after Prp16-mediated remodeling; type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#30; Details: Splicing extract was prepared from Slu7-TAPS yeast strains. An in vitro transcribed yeast UBC4 pre-mRNA substrate (with 2 x MS2 bacteriophage coat protein-binding stem loops at the 5' end and with a 2'-deoxy substitution at the 3'-splice site sequence UAG sequence (UA-2'dG) was pre-bound to an MS2-maltose binding protein fusion protein. This substrate-protein complex was added to the splicing extract. The splicing reaction proceeded through the first step but the second step was blocked by the deoxy substitution. Substrate-bound spliceosomes from the splicing extract were purified on amylose resin and eluted with maltose. Subsequently the spliceosomes were captured on streptactin resin and eluted with desthiobiotin. Purified spliceosomes were concentrated in 20 mM HEPES KOH pH 7.9, 100 mM KCl, 0.25 mM EDTA.

Macromolecule #1: Yeast UBC4 gene for ubiquitin-conjugating enzyme

• Macromolecule: Name: Yeast UBC4 gene for ubiquitin-conjugating enzyme / type: rna / ID: 1 ; Details: Saccharomyces cerevisiae UBC4 pre-mRNA intron, in vitro transcribed.; Number of copies: 1
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 30.20073 KDa

Sequence

String:

GUAUGUCUAA AGUUAUGGCC ACGUUUCAAA UGCGUGCUUU UUUUUUAAAA CUUAUGCUCU UAUUUACUAA CAAAAUCAAC AUGCUAUUG AACUAG

Macromolecule #2: UBC4 gene exon

• Macromolecule: Name: UBC4 gene exon / type: rna / ID: 2 ; Details: Saccharomyces cerevisiae UBC4 pre-mRNA exon, in vitro transcribed.; Number of copies: 1
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 6.518976 KDa

Sequence

String:

GAAGUAAGUG AUCUAGAAAG

Macromolecule #3: U2 snRNA

• Macromolecule: Name: U2 snRNA / type: rna / ID: 3 / Details: Saccharomyces cerevisiae U2 snRNA / Number of copies: 1
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 376.267406 KDa

Sequence

String:

ACGAAUCUCU UUGCCUUUUG GCUUAGAUCA AGUGUAGUAU CUGUUCUUUU CAGUGUAACA ACUGAAAUGA CCUCAAUGAG GCUCAUUAC CUUUUAAUUU GUUACAAUAC ACAUUUUUUG GCACCCAAAA UAAUAAAAUG GACGGGAAGA GACUUUUUAA G CAAGUUGU UUUCCGCUAA UGUCAGGUCU CACUACUUUU UGCUGCUAUU UUUCUUCGCU CAUGGUUUCU UCAUAAGGCG UU UUUAUGA UGGUUUUUCG AAAUUGGUUU UUGAGACGAC GGUUGCUCAA GGUUAUUGUU UUUGUUUUCU UCUGGUUGUU UUC UAUUUU CUUUUUUUUA GCUUUCUGUU UCUCCCUUAG UUUGGCUUUU UGCUUCAUAC UCUUCCCUGU CUUUCCGAGC CGUU UAUGU CCAACGCGGG AUUUGGUUUU UCUUUAUCGA UGGGAAGAAA UGGUGCUAUA GUAGGUUGGG AGAUAAUAUU UAUGG UAUG GGGUGCUAGU GCGGAUGGGG CGCUCUUAUU GUUGAUUUCU UCGCUCGUCU UCUUUUUCUG GUGGCGCUGC AAGAGG AAG UUUUUCGACU UUGUUAUGAU UUUUGGUUUG CAAGGAAAGG UGUCUUACGA UUCUUUUUUU GAUGUAAUAG GAUAAGC UU GCUUAUCCCC CAAGUAUCGG CCAAAGUUGU UGAUUUUCCU UUUGAAGUGU CCUCGGUUUG AGGGGGUGUA GGGUGGGG U UGGUCUACAA UAAGAGUGUU CCAUUGUUAA CGUGCUGGCG UCUUUUACUA UAUUUUUUUU CCCAGUUUAU UUUGUGCUU AUUUUCUCAU UGAGGAGAAG GAGCUCUUCU CGCAGGAUAU AAAUGGAGGU UUGCUAAAGG GGAGGAGAUG UGUUUGUGAG AAUACUGCU GAGAGAGUUC UGGAAGAGAA AAAAAGGAGG CAAUGGAAGG CGUUUGCUGG GAAAAGAGAA GAGCCAUGAC U GCAUCUGU UGUUUCAAGG CCAGUUUUAU UAACCGCCUA UGUCAUAGAG GCGUUUUUUU UGGAGGGAUU UGAAGAAUGC CG GCGGCAU CAAGAAACGG ACUUGAUGGU UGACGCCUGU UUUUAAAGUU AGAGACGUCG CGACCCUCGC ACUUGUGGAG UCG UUCUUG ACUUUUACUU UGGUCGCUUG AUGUUUCUCU CGUCUUCCCG UUCGCUCUU

Macromolecule #4: Saccharomyces cerevisiae strain T.52_2H chromosome XII sequence

• Macromolecule: Name: Saccharomyces cerevisiae strain T.52_2H chromosome XII sequence; type: rna / ID: 4 / Details: Saccharomyces cerevisiae U6 snRNA / Number of copies: 1
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 35.883176 KDa

Sequence

String:

GUUCGCGAAG UAACCCUUCG UGGACAUUUG GUCAAUUUGA AACAAUACAG AGAUGAUCAG CAGUUCCCCU GCAUAAGGAU GAACCGUUU UACAAAGAGA UUUAUUUCGU UUU

Macromolecule #5: U5 snRNA

• Macromolecule: Name: U5 snRNA / type: rna / ID: 5 / Details: Saccharomyces cerevisiae U5 snRNA / Number of copies: 1
• Source (natural): Organism: Saccharomyces cerevisiae (brewer's yeast)
• Molecular weight: Theoretical: 57.444875 KDa

Sequence

String:

AAGCAGCUUU ACAGAUCAAU GGCGGAGGGA GGUCAACAUC AAGAACUGUG GGCCUUUUAU UGCCUAUAGA ACUUAUAACG AACAUGGUU CUUGCCUUUU ACCAGAACCA UCCGGGUGUU GUCUCCAUAG AAACAGGUAA AGCUGUCCGU UACUGUGGGC U UGCCAUAU UUUUUGGAAC U

Macromolecule #6: Pre-mRNA-splicing factor 8

• Macromolecule: Name: Pre-mRNA-splicing factor 8 / type: protein_or_peptide / ID: 6 / Details: Saccharomyces cerevisiae Prp8 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 279.867469 KDa

Sequence

String:

MSGLPPPPPG FEEDSDLALP PPPPPPPGYE IEELDNPMVP SSVNEDTFLP PPPPPPSNFE INAEEIVDFT LPPPPPPPGL DELETKAEK KVELHGKRKL DIGKDTFVTR KSRKRAKKMT KKAKRSNLYT PKAEMPPEHL RKIINTHSDM ASKMYNTDKK A FLGALKYL PHAILKLLEN MPHPWEQAKE VKVLYHTSGA ITFVNETPRV IEPVYTAQWS ATWIAMRREK RDRTHFKRMR FP PFDDDEP PLSYEQHIEN IEPLDPINLP LDSQDDEYVK DWLYDSRPLE EDSKKVNGTS YKKWSFDLPE MSNLYRLSTP LRD EVTDKN YYYLFDKKSF FNGKALNNAI PGGPKFEPLY PREEEEDYNE FNSIDRVIFR VPIRSEYKVA FPHLYNSRPR SVRI PWYNN PVSCIIQNDE EYDTPALFFD PSLNPIPHFI DNNSSLNVSN TKENGDFTLP EDFAPLLAEE EELILPNTKD AMSLY HSPF PFNRTKGKMV RAQDVALAKK WFLQHPDEEY PVKVKVSYQK LLKNYVLNEL HPTLPTNHNK TKLLKSLKNT KYFQQT TID WVEAGLQLCR QGHNMLNLLI HRKGLTYLHL DYNFNLKPTK TLTTKERKKS RLGNSFHLMR ELLKMMKLIV DTHVQFR LG NVDAFQLADG IHYILNHIGQ LTGIYRYKYK VMHQIRACKD LKHIIYYKFN KNLGKGPGCG FWQPAWRVWL NFLRGTIP L LERYIGNLIT RQFEGRSNEI VKTTTKQRLD AYYDLELRNS VMDDILEMMP ESIRQKKART ILQHLSEAWR CWKANIPWD VPGMPAPIKK IIERYIKSKA DAWVSAAHYN RERIKRGAHV EKTMVKKNLG RLTRLWIKNE QERQRQIQKN GPEITPEEAT TIFSVMVEW LESRSFSPIP FPPLTYKNDT KILVLALEDL KDVYASKVRL NASEREELAL IEEAYDNPHD TLNRIKKYLL T QRVFKPVD ITMMENYQNI SPVYSVDPLE KITDAYLDQY LWYEADQRKL FPNWIKPSDS EIPPLLVYKW TQGINNLSEI WD VSRGQSA VLLETTLGEM AEKIDFTLLN RLLRLIVDPN IADYITAKNN VVINFKDMSH VNKYGLIRGL KFASFIFQYY GLV IDLLLL GQERATDLAG PANNPNEFMQ FKSKEVEKAH PIRLYTRYLD RIYMLFHFEE DEGEELTDEY LAENPDPNFE NSIG YNNRK CWPKDSRMRL IRQDVNLGRA VFWEIQSRVP TSLTSIKWEN AFVSVYSKNN PNLLFSMCGF EVRILPRQRM EEVVS NDEG VWDLVDERTK QRTAKAYLKV SEEEIKKFDS RIRGILMASG STTFTKVAAK WNTSLISLFT YFREAIVATE PLLDIL VKG ETRIQNRVKL GLNSKMPTRF PPAVFYTPKE LGGLGMISAS HILIPASDLS WSKQTDTGIT HFRAGMTHED EKLIPTI FR YITTWENEFL DSQRVWAEYA TKRQEAIQQN RRLAFEELEG SWDRGIPRIS TLFQRDRHTL AYDRGHRIRR EFKQYSLE R NSPFWWTNSH HDGKLWNLNA YRTDVIQALG GIETILEHTL FKGTGFNSWE GLFWEKASGF EDSMQFKKLT HAQRTGLSQ IPNRRFTLWW SPTINRANVY VGFLVQLDLT GIFLHGKIPT LKISLIQIFR AHLWQKIHES IVFDICQILD GELDVLQIES VTKETVHPR KSYKMNSSAA DITMESVHEW EVSKPSLLHE TNDSFKGLIT NKMWFDVQLR YGDYDSHDIS RYVRAKFLDY T TDNVSMYP SPTGVMIGID LAYNMYDAYG NWFNGLKPLI QNSMRTIMKA NPALYVLRER IRKGLQIYQS SVQEPFLNSS NY AELFNND IKLFVDDTNV YRVTVHKTFE GNVATKAING CIFTLNPKTG HLFLKIIHTS VWAGQKRLSQ LAKWKTAEEV SAL VRSLPK EEQPKQIIVT RKAMLDPLEV HMLDFPNIAI RPTELRLPFS AAMSIDKLSD VVMKATEPQM VLFNIYDDWL DRIS SYTAF SRLTLLLRAL KTNEESAKMI LLSDPTITIK SYHLWPSFTD EQWITIESQM RDLILTEYGR KYNVNISALT QTEIK DIIL GQNIKAPSVK RQKMAELEAA RSEKQNDEEA AGASTVMKTK TINAQGEEIV VVASADYESQ TFSSKNEWRK SAIANT LLY LRLKNIYVSA DDFVEEQNVY VLPKNLLKKF IEISDVKIQV AAFIYGMSAK DHPKVKEIKT VVLVPQLGHV GSVQISN IP DIGDLPDTEG LELLGWIHTQ TEELKFMAAS EVATHSKLFA DKKRDCIDIS IFSTPGSVSL SAYNLTDEGY QWGEENKD I MNVLSEGFEP TFSTHAQLLL SDRITGNFII PSGNVWNYTF MGTAFNQEGD YNFKYGIPLE FYNEMHRPVH FLQFSELAG DEELEAEQID VFS

Macromolecule #7: Pre-mRNA-splicing factor SNU114

• Macromolecule: Name: Pre-mRNA-splicing factor SNU114 / type: protein_or_peptide / ID: 7 / Details: c Snu114 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 114.174008 KDa

Sequence

String:

MEGDDLFDEF GNLIGVDPFD SDEEESVLDE QEQYQTNTFE GSGNNNEIES RQLTSLGSKK ELGISLEHPY GKEVEVLMET KNTQSPQTP LVEPVTERTK LQEHTIFTQL KKNIPKTRYN RDYMLSMANI PERIINVGVI GPLHSGKTSL MDLLVIDSHK R IPDMSKNV ELGWKPLRYL DNLKQEIDRG LSIKLNGSTL LCTDLESKSR MINFLDAPGH VNFMDETAVA LAASDLVLIV ID VVEGVTF VVEQLIKQSI KNNVAMCFVI NKLDRLILDL KLPPMDAYLK LNHIIANINS FTKGNVFSPI DNNIIFASTK LGF TFTIKE FVSYYYAHSI PSSKIDDFTT RLWGSVYYHK GNFRTKPFEN VEKYPTFVEF ILIPLYKIFS YALSMEKDKL KNLL RSNFR VNLSQEALQY DPQPFLKHVL QLIFRQQTGL VDAITRCYQP FELFDNKTAH LSIPGKSTPE GTLWAHVLKT VDYGG AEWS LVRIYSGLLK RGDTVRILDT SQSESRQKRQ LHDISKTETS NEDEDEDDET PSCEVEEIGL LGGRYVYPVH EAHKGQ IVL IKGISSAYIK SATLYSVKSK EDMKQLKFFK PLDYITEAVF KIVLQPLLPR ELPKLLDALN KISKYYPGVI IKVEESG EH VILGNGELYM DCLLYDLRAS YAKIEIKISD PLTVFSESCS NESFASIPVS NSISRLGEEN LPGLSISVAA EPMDSKMI Q DLSRNTLGKG QNCLDIDGIM DNPRKLSKIL RTEYGWDSLA SRNVWSFYNG NVLINDTLPD EISPELLSKY KEQIIQGFY WAVKEGPLAE EPIYGVQYKL LSISVPSDVN IDVMKSQIIP LMKKACYVGL LTAIPILLEP IYEVDITVHA PLLPIVEELM KKRRGSRIY KTIKVAGTPL LEVRGQVPVI ESAGFETDLR LSTNGLGMCQ LYFWHKIWRK VPGDVLDKDA FIPKLKPAPI N SLSRDFVM KTRRRKGIST GGFMSNDGPT LEKYISAELY AQLRENGLVP

Macromolecule #8: Pre-mRNA-splicing factor CWC22

• Macromolecule: Name: Pre-mRNA-splicing factor CWC22 / type: protein_or_peptide / ID: 8 / Details: Saccharomyces cerevisiae Cwc22 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 67.386062 KDa

Sequence

String:

MSTATIQDED IKFQRENWEM IRSHVSPIIS NLTMDNLQES HRDLFQVNIL IGRNIICKNV VDFTLNKQNG RLIPALSALI ALLNSDIPD IGETLAKELM LMFVQQFNRK DYVSCGNILQ CLSILFLYDV IHEIVILQIL LLLLEKNSLR LVIAVMKICG W KLALVSKK THDMIWEKLR YILQTQELSS TLRESLETLF EIRQKDYKSG SQGLFILDPT SYTVHTHSYI VSDEDEANKE LG NFEKCEN FNELTMAFDT LRQKLLINNT SDTNEGSNSQ LQIYDMTSTN DVEFKKKIYL VLKSSLSGDE AAHKLLKLKI ANN LKKSVV DIIIKSSLQE STFSKFYSIL SERMITFHRS WQTAYNETFE QNYTQDIEDY ETDQLRILGK FWGHLISYEF LPMD CLKII KLTEEESCPQ GRIFIKFLFQ ELVNELGLDE LQLRLNSSKL DGMFPLEGDA EHIRYSINFF TAIGLGLLTE DMRSR LTII QEVEDAEEEE KKLREEEELE KLRKKARESQ PTQGPKIHES RLFLQKDTRE NSRSRSPFTV ETRKRARSRT PPRGSR NHR NRSRTPPARR QRHR

Macromolecule #9: Pre-mRNA-splicing factor PRP46

• Macromolecule: Name: Pre-mRNA-splicing factor PRP46 / type: protein_or_peptide / ID: 9 / Details: Saccharomyces cerevisiae Prp46 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 50.771289 KDa

Sequence

String:

MDGNDHKVEN LGDVDKFYSR IRWNNQFSYM ATLPPHLQSE MEGQKSLLMR YDTYRKESSS FSGEGKKVTL QHVPTDFSEA SQAVISKKD HDTHASAFVN KIFQPEVAEE LIVNRYEKLL SQRPEWHAPW KLSRVINGHL GWVRCVAIDP VDNEWFITGS N DTTMKVWD LATGKLKTTL AGHVMTVRDV AVSDRHPYLF SVSEDKTVKC WDLEKNQIIR DYYGHLSGVR TVSIHPTLDL IA TAGRDSV IKLWDMRTRI PVITLVGHKG PINQVQCTPV DPQVVSSSTD ATVRLWDVVA GKTMKVLTHH KRSVRATALH PKE FSVASA CTDDIRSWGL AEGSLLTNFE SEKTGIINTL SINQDDVLFA GGDNGVLSFY DYKSGHKYQS LATREMVGSL EGER SVLCS TFDKTGLRLI TGEADKSIKI WKQDETATKE SEPGLAWNPN LSAKRF

Macromolecule #10: Pre-mRNA-processing protein 45

• Macromolecule: Name: Pre-mRNA-processing protein 45 / type: protein_or_peptide / ID: 10 / Details: Saccharomyces cerevisiae Prp45 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 42.548727 KDa

Sequence

String:

MFSNRLPPPK HSQGRVSTAL SSDRVEPAIL TDQIAKNVKL DDFIPKRQSN FELSVPLPTK AEIQECTART KSYIQRLVNA KLANSNNRA SSRYVTETHQ APANLLLNNS HHIEVVSKQM DPLLPRFVGK KARKVVAPTE NDEVVPVLHM DGSNDRGEAD P NEWKIPAA VSNWKNPNGY TVALERRVGK ALDNENNTIN DGFMKLSEAL ENADKKARQE IRSKMELKRL AMEQEMLAKE SK LKELSQR ARYHNGTPQT GAIVKPKKQT STVARLKELA YSQGRDVSEK IILGAAKRSE QPDLQYDSRF FTRGANASAK RHE DQVYDN PLFVQQDIES IYKTNYEKLD EAVNVKSEGA SGSHGPIQFT KAESDDKSDN YGA

Macromolecule #11: Pre-mRNA-splicing factor BUD31

• Macromolecule: Name: Pre-mRNA-splicing factor BUD31 / type: protein_or_peptide / ID: 11 / Details: Saccharomyces cerevisiae Bud31 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 18.484502 KDa

Sequence

String:

MPRIKTRRSK PAPDGFEKIK PTLTDFEIQL RDAQKDKSSK LAAKSNEQLW EIMQLHHQRS RYIYTLYYKR KAISKDLYDW LIKEKYADK LLIAKWRKTG YEKLCCLRCI QKNETNNGST CICRVPRAQL EEEARKKGTQ VSFHQCVHCG CRGCASTD

Macromolecule #12: Pre-mRNA-splicing factor CWC2

• Macromolecule: Name: Pre-mRNA-splicing factor CWC2 / type: protein_or_peptide / ID: 12 / Details: Saccharomyces cerevisiae Cwc2 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 38.486562 KDa

Sequence

String:

MTSWRDKSAK VQVKESELPS SIPAQTGLTF NIWYNKWSQG FAGNTRFVSP FALQPQLHSG KTRGDNDGQL FFCLFFAKGM CCLGPKCEY LHHIPDEEDI GKLALRTEVL DCFGREKFAD YREDMGGIGS FRKKNKTLYV GGIDGALNSK HLKPAQIESR I RFVFSRLG DIDRIRYVES KNCGFVKFKY QANAEFAKEA MSNQTLLLPS DKEWDDRREG TGLLVKWANE DPDPAAQKRL QE ELKLESL NMMVHLINNN TNSAGTEVNN KNNERLDRTF PEASVDNVKK RLLPLDNGME SDDFIEKLKK VKKNISRENI SSK PSVGKL GGPLLDYLSS DED

Macromolecule #13: Pre-mRNA-splicing factor SLT11

• Macromolecule: Name: Pre-mRNA-splicing factor SLT11 / type: protein_or_peptide / ID: 13 / Details: Saccharomyces cerevisiae Ecm2 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 40.98859 KDa

Sequence

String:

MNDEINEPPP NICEQCLGDE ANIRMTKIPQ GSECKICTLP FTLYHFKTSK RSNNIIKTLI CVRCATQRNI CQCCMLDSRW HIPIQLRDH LISLVNEENV MTEEAKNDMM KRFLSLKNVK LGGAQITSDP SEADNIVDKL KNILLRATSD GPSTPLIKNT T ALYKNEKG ANEVKNLEKY ASVDISHILK KLPLNESFLK NPSTKSFFLY NIDASIPEWK ITDTVSQLLG IKKWKDGNSL SL IVNHKAK CGGLRFQSSE LGERFVSKIS ETLVTPKGLK RGVLLIDRFR IFIIPWSSGF SAASFGTNTA ENIKLSLSLN KLI QLELGL SFPTKSTDNA KNDKKKTSKK VHKDRSKKSK PRANKLTI

Macromolecule #14: Pre-mRNA-splicing factor CEF1

• Macromolecule: Name: Pre-mRNA-splicing factor CEF1 / type: protein_or_peptide / ID: 14 / Details: Saccharomyces cerevisiae Cef1 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 67.837773 KDa

Sequence

String:

MPPVPIYVKG GVWTNVEDQI LKAAVQKYGT HQWSKVASLL QKKTARQSEL RWNEYLNPKL NFTEFSKEED AQLLDLAREL PNQWRTIAD MMARPAQVCV ERYNRLLESE DSGGAALSTG VTDLKAGDIN PNAETQMARP DNGDLEDEEK EMLAEARARL L NTQGKKAT RKIRERMLEE SKRIAELQKR RELKQAGINV AIKKPKKKYG TDIDYNEDIV YEQAPMPGIY DTSTEDRQIK KK FEQFERK VNRKGLDGNK DKPSKKNKDK KRKHDENEHV EKAALGESTT LTDEYKKPKL ILSAPGTKQG KVTYKKKLES KRQ KLIEAQ ATGTVLTPKE LLPHDSGQED NERSNIKSGK QLKSRIRKFL VQMFASLPSP KNDFEIVLSE DEKEEDAEIA EYEK EFENE RAMNEEDNFI EPPSQNDAPR VSLVAVPLAY STLPIPEFKN NPQSAIDNKY NLLVANAINK EPHMVPEDTV DFLKE VESR MQHITQGRTS MKIQFKTAMP PTEVLLESIQ SKVESIEQLQ RKLQHVQPLE QQNNEMCSTL CHHSLPALIE GQRKYY ADY YAYRQEIRSL EGRRKRLQAM LNSSSSI

Macromolecule #15: Pre-mRNA-splicing factor CWC15

• Macromolecule: Name: Pre-mRNA-splicing factor CWC15 / type: protein_or_peptide / ID: 15 / Details: Saccharomyces cerevisiae Cwc15 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 19.975195 KDa

Sequence

String:

MTTSHRPQLE ARSGAKAAAY TPTGIEHARL LPGHTTLKYR KFKEEENLRA NCAQEDRSND KSLEEAVMNE EKQDVVGSGN LQETRSEKD QKDSLQELLV TQKNKVEDKA ELEGNEQLKG GNSSRRSWRK GTAFGRHKVT KETNIKEHAT KKSASGYIND M TKSEYHQE FLHKHVR

Macromolecule #16: Pre-mRNA-splicing factor CWC21

• Macromolecule: Name: Pre-mRNA-splicing factor CWC21 / type: protein_or_peptide / ID: 16 / Details: Saccharomyces cerevisiae Cwc21 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 15.793596 KDa

Sequence

String:

MSYNGIGLKS AKGSSTSGHV QRSLASNNRR RPQGSQQQRQ QRQNAIKKAS HDKASRPLAV QKQIETHMEK REIEVQVSEL RDRLEEEET LSEEQIDKKC EALRAKLTNE WQEQQRMSSL YTPRKARLTE EQHRHE

Macromolecule #17: Pre-mRNA-splicing factor CLF1

• Macromolecule: Name: Pre-mRNA-splicing factor CLF1 / type: protein_or_peptide / ID: 17 / Details: Saccharomyces cerevisiae Clf1 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 82.555859 KDa

Sequence

String:

MDTLEPTAVD THVSAEQILR DVYKKGQKAR GSTNIDILDL EELREYQRRK RTEYEGYLKR NRLDMGQWIR YAQFEIEQHD MRRARSIFE RALLVDSSFI PLWIRYIDAE LKVKCINHAR NLMNRAISTL PRVDKLWYKY LIVEESLNNV EIVRSLYTKW C SLEPGVNA WNSFVDFEIR QKNWNGVREI YSKYVMAHPQ MQTWLKWVRF ENRHGNTEFT RSVYSLAIDT VANLQNLQIW SD MEVAKLV NSFAHWEAAQ QEYERSSALY QIAIEKWPSN QLLKAGLLDF EKQFGDINSI EETISYKRKM EYETILSNNA YDY DTWWLY LDLISESFPK QIMQTFEKAI VDSRPKELSK NVQWKRYIYL WMRYICYVEL ELENSLLEEE LFQRLIDDII PHKH FTFSK IWLMYAKFLI RHDDVPKARK ILGKAIGLCP KAKTFKGYIE LEVKLKEFDR VRKIYEKFIE FQPSDLQIWS QYGEL EENL GDWDRVRGIY TIALDENSDF LTKEAKIVLL QKYITFETES QEFEKARKLY RRYLELNQYS PQSWIEFAMY QTSTPT EQQ LLDLAKLQSE NVDEDIEFEI TDENKLEARK VFEEAIVFFK EKDDKQGRLS ILEALKDYEE TYGTELDQET VKKRFPK VI KKVRLQNGVE EEFVDYIFPD DIDDDKPKPS KFLELAKKWK QEQAL

Macromolecule #18: Pre-mRNA-splicing factor SYF1

• Macromolecule: Name: Pre-mRNA-splicing factor SYF1 / type: protein_or_peptide / ID: 18 / Details: Saccharomyces cerevisiae Syf1 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 101.875852 KDa

Sequence

String:

MSAYIAMKGV ITNVDENIRN DEDVAFEYEI QKTPQNILTW KRYIEYWKEE GRTDKQIRWL YERFCSQFVT DTSIWEDYIR WESTKEVVE TSRIFWLFQR CLKSCVRDCD RICLSYLELA IEQYDLAMIR HALASSLMKM EREMHRKVWD PVIKFVEEKV L PLTQLDST QEDEEESTDE AELINVLLVK GFTKGGFISE EISENGSRGD IWSSHILERY LKVAPQQKRN ESLATLALTR DN ITIKSVY EKYLPQDENS GKYLPSSELP FELNFNYLAS LEKLGLDNQY EEFMRQMNGI YPDKWLFLIL SLAKYYISRG RLD SCGDLL KKSLQQTLRY SDFDRIYNFY LLFEQECSQF ILGKLKENDS KFFNQKDWTE KLQAHMATFE SLINLYDIYL NDVA LRQDS NLVETWMKRV SLQKSAAEKC NVYSEAILKI DPRKVGTPGS FGRLWCSYGD LYWRSNAIST ARELWTQSLK VPYPY IEDL EEIYLNWADR ELDKEGVERA FSILEDALHV PTNPEILLEK YKNGHRKIPA QTVLFNSLRI WSKYIDYLEA YCPKDA NSS DKIFNKTKMA YNTVIDLRLI TPAMAENFAL FLQNHYEVME SFQVYEKTIP LFPPEIQYEL WIEYLEVATS HQLSSLS PE HIRFLFEKAL KNLCSNGIDC KTIFIAYSVF EERISGLISK SIEILRRGAV IGTVSVSTHL ESRLQLWRMC ISKAESTL G PSVTRELYQE CIQILPNSKA VEFVIKFSDF ESSIGETIRA REILAYGAKL LPPSRNTELW DSFEIFELKH GDKETYKDM LKMKKVLESN MLIDSASVSH EEGNINFVAA ATSHAPNSHT LTQSTSSYSI NPDEIELDI(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)

Macromolecule #19: Pre-mRNA-splicing factor 18

• Macromolecule: Name: Pre-mRNA-splicing factor 18 / type: protein_or_peptide / ID: 19 / Details: Saccharomyces cerevisiae Prp18 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 28.414391 KDa

Sequence

String:

MDLDLASILK GEISKKKKEL ANSKGVQPPC TEKFQPHESA NIDETPRQVE QESTDEENLS DNQSDDIRTT ISKLENRPER IQEAIAQDK TISVIIDPSQ IGSTEGKPLL SMKCNLYIHE ILSRWKASLE AYHPELFLDT KKALFPLLLQ LRRNQLAPDL L ISLATVLY HLQQPKEINL AVQSYMKLSI GNVAWPIGVT SVGIHARSAH SKIQGGRNAA NIMIDERTRL WITSIKRLIT FE EWYTSNH DSLA

Macromolecule #20: Pre-mRNA-splicing factor SLU7

• Macromolecule: Name: Pre-mRNA-splicing factor SLU7 / type: protein_or_peptide / ID: 20 / Details: Saccharomyces cerevisiae Slu7 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 44.722875 KDa

Sequence

String:

MNNNSRNNEN RSTINRNKRQ LQQAKEKNEN IHIPRYIRNQ PWYYKDTPKE QEGKKPGNDD TSTAEGGEKS DYLVHHRQKA KGGALDIDN NSEPKIGMGI KDEFKLIRPQ KMSVRDSHSL SFCRNCGEAG HKEKDCMEKP RKMQKLVPDL NSQKNNGTVL V RATDDDWD SRKDRWYGYS GKEYNELISK WERDKRNKIK GKDKSQTDET LWDTDEEIEL MKLELYKDSV GSLKKDDADN SQ LYRTSTR LREDKAAYLN DINSTESNYD PKSRLYKTET LGAVDEKSKM FRRHLTGEGL KLNELNQFAR SHAKEMGIRD EIE DKEKVQ HVLVANPTKY EYLKKKREQE ETKQPKIVSI GDLEARKVDG TKQSEEQRNH LKDLYG

Macromolecule #21: Pre-mRNA-processing factor 17

• Macromolecule: Name: Pre-mRNA-processing factor 17 / type: protein_or_peptide / ID: 21 / Details: Saccharomyces cerevisiae Prp17 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 52.128762 KDa

Sequence

String:

MGLVDGYDTS SDSDLNFDEG KSVHEKKNGN LHEDTSYEPS SNNIHKRKSH FTKSELKRRR KTRKGDGPWG SWSSSDDETS QASETQKED QDIFVHALAE DNLDSEQIEV EEVSHFYGKS EKDYQGRGYL YPPNDVDVDL REERISFRCY LPKKVIRNYP G HPEGTTAL KFLPKTGHLI LSGGNDHTIK IWDFYHDYEC LRDFQGHNKP IKALRFTEDC QSFLSSSFDR SVKIWDTETG KV KTRLHLN STPADVESRP TNPHEFIVGL SNSKILHYDD RVSENQGLVQ TYDHHLSSIL ALKYFPDGSK FISSSEDKTV RIW ENQINV PIKQISDTAQ HSMPFLNVHP SQNYFCAQSM DNRIYSFSLK PKYKRHPKKI FKGHSSAGYG ISLAFSGDGR YICS GDSKS RLFTWDWNTS RLLNNIKIPG NKPITQVDWH PQETSKVICS GAAGKIYVCD

Macromolecule #22: Unknown

• Macromolecule: Name: Unknown / type: protein_or_peptide / ID: 22 / Details: Unknown protein helices within C-star complex / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 5.805147 KDa

Sequence

String:

(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK) (UNK)(UNK)

Macromolecule #23: Pre-mRNA-splicing factor SYF2

• Macromolecule: Name: Pre-mRNA-splicing factor SYF2 / type: protein_or_peptide / ID: 23 / Details: Saccharomyces cerevisiae Syf2 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 24.850719 KDa

Sequence

String:

MDFYKLDEKL KELKRKRVDV SIKSRKLADR EIQEVSANRK PRVYSMEDVN DADESVGDTE SPEKEKAFHY TVQEYDAWER RHPQGKTGQ SQRGGISYDQ LAKLSYEKTL RNLATQTQNS SKQDSSADEE DNKNVPKKGR IGKVQKDTKT GKITIADDDK L VNKLAVSL QSESKKRYEA RKRQMQNAKT LYGVESFIND KNKQFNEKLS RESKGSE

Macromolecule #24: Small nuclear ribonucleoprotein-associated protein B

• Macromolecule: Name: Small nuclear ribonucleoprotein-associated protein B / type: protein_or_peptide / ID: 24 / Details: Saccharomyces cerevisiae SmB protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 22.42699 KDa

Sequence

String:

MSKIQVAHSS RLANLIDYKL RVLTQDGRVY IGQLMAFDKH MNLVLNECIE ERVPKTQLDK LRPRKDSKDG TTLNIKVEKR VLGLTILRG EQILSTVVED KPLLSKKERL VRDKKEKKQA QKQTKLRKEK EKKPGKIAKP NTANAKHTSS NSREIAQPSS S RYNGGNDN IGANRSRFNN EAPPQTRKFQ PPPGFKRK

Macromolecule #25: Small nuclear ribonucleoprotein Sm D3

• Macromolecule: Name: Small nuclear ribonucleoprotein Sm D3 / type: protein_or_peptide / ID: 25 / Details: Saccharomyces cerevisiae SmD3 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 11.240139 KDa

Sequence

String:

MTMNGIPVKL LNEAQGHIVS LELTTGATYR GKLVESEDSM NVQLRDVIAT EPQGAVTHMD QIFVRGSQIK FIVVPDLLKN APLFKKNSS RPMPPIRGPK RR

Macromolecule #26: Small nuclear ribonucleoprotein E

• Macromolecule: Name: Small nuclear ribonucleoprotein E / type: protein_or_peptide / ID: 26 / Details: Saccharomyces cerevisiae SmE protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 10.385098 KDa

Sequence

String:

MSNKVKTKAM VPPINCIFNF LQQQTPVTIW LFEQIGIRIK GKIVGFDEFM NVVIDEAVEI PVNSADGKED VEKGTPLGKI LLKGDNITL ITSAD

Macromolecule #27: Small nuclear ribonucleoprotein F

• Macromolecule: Name: Small nuclear ribonucleoprotein F / type: protein_or_peptide / ID: 27 / Details: Saccharomyces cerevisiae SmF protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 9.669945 KDa

Sequence

String:

MSESSDISAM QPVNPKPFLK GLVNHRVGVK LKFNSTEYRG TLVSTDNYFN LQLNEAEEFV AGVSHGTLGE IFIRCNNVLY IRELPN

Macromolecule #28: Small nuclear ribonucleoprotein G

• Macromolecule: Name: Small nuclear ribonucleoprotein G / type: protein_or_peptide / ID: 28 / Details: Saccharomyces cerevisiae SmG protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 8.490809 KDa

Sequence

String:

MVSTPELKKY MDKKILLNIN GSRKVAGILR GYDIFLNVVL DDAMEINGED PANNHQLGLQ TVIRGNSIIS LEALDAI

Macromolecule #29: Small nuclear ribonucleoprotein Sm D1

• Macromolecule: Name: Small nuclear ribonucleoprotein Sm D1 / type: protein_or_peptide / ID: 29 / Details: Saccharomyces cerevisiae SmD1 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 16.296798 KDa

Sequence

String:

MKLVNFLKKL RNEQVTIELK NGTTVWGTLQ SVSPQMNAIL TDVKLTLPQP RLNKLNSNGI AMASLYLTGG QQPTASDNIA SLQYINIRG NTIRQIILPD SLNLDSLLVD QKQLNSLRRS GQIANDPSKK RRRDFGAPAN KRPRRGL

Macromolecule #30: Small nuclear ribonucleoprotein Sm D2

• Macromolecule: Name: Small nuclear ribonucleoprotein Sm D2 / type: protein_or_peptide / ID: 30 / Details: Saccharomyces cerevisiae SmD2 protein / Number of copies: 1 / Enantiomer: LEVO
• Source (natural): Organism: Baker's yeast (brewer's yeast)
• Molecular weight: Theoretical: 12.876066 KDa

Sequence

String:

MSSQIIDRPK HELSRAELEE LEEFEFKHGP MSLINDAMVT RTPVIISLRN NHKIIARVKA FDRHCNMVLE NVKELWTEKK GKNVINRER FISKLFLRGD SVIVVLKTPV E

Macromolecule #31: MAGNESIUM ION

• Macromolecule: Name: MAGNESIUM ION / type: ligand / ID: 31 / Number of copies: 3 / Formula: MG
• Molecular weight: Theoretical: 24.305 Da

Macromolecule #32: POTASSIUM ION

• Macromolecule: Name: POTASSIUM ION / type: ligand / ID: 32 / Number of copies: 2 / Formula: K
• Molecular weight: Theoretical: 39.098 Da

Macromolecule #33: INOSITOL HEXAKISPHOSPHATE

• Macromolecule: Name: INOSITOL HEXAKISPHOSPHATE / type: ligand / ID: 33 / Number of copies: 1 / Formula: IHP
• Molecular weight: Theoretical: 660.035 Da

Chemical component information

ChemComp-IHP:
INOSITOL HEXAKISPHOSPHATE / Phytic acid

Macromolecule #34: GUANOSINE-5'-TRIPHOSPHATE

• Macromolecule: Name: GUANOSINE-5'-TRIPHOSPHATE / type: ligand / ID: 34 / Number of copies: 1 / Formula: GTP
• Molecular weight: Theoretical: 523.18 Da

Chemical component information

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

Macromolecule #35: ZINC ION

• Macromolecule: Name: ZINC ION / type: ligand / ID: 35 / Number of copies: 6 / Formula: ZN
• Molecular weight: Theoretical: 65.409 Da

Experimental details

Structure determination

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

Sample preparation

• Concentration: 0.3 mg/mL

Buffer

pH: 7.9
Component:

Concentration	Name	Formula
20.0 millimolar	Hepes.KOH pH 7.9
100.0 millimolar	potassium chloride	KCl
250.0 micromolar	EDTAEthylenediaminetetraacetic acid
1.0 % v/v	glycerol
0.0025 % v/v	Nonidet P-40 (NP-40)

Details: NP-40 is also called IGEPAL CA-630

• Grid: Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: HOLEY ARRAY / Support film - Film thickness: 6.0 nm / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Atmosphere: AIR / Pretreatment - Pressure: 20.0 kPa
• Vitrification: Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK III; Details: 3.5 microlitres sample were applied to the grid, left for 25 seconds and then blotted for 3.0-3.5 seconds before plunging..

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: 4.5 µm / Nominal defocus min: 0.5 µm / Nominal magnification: 81000
• Specialist optics: Energy filter - Name: GIF Quantum
• Sample stage: Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
• Details: GIF Quantum energy filter, 20 eV slit width
• Image recording: Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Number real images: 3596 / Average exposure time: 0.8 sec. / Average electron dose: 2.0 e/Ų; Details: Total dose: 40 electrons/Angstrom^2 over 16 seconds. 20 movie frames collected at 1.25 frames per second.
• Experimental equipment: Model: Titan Krios / Image courtesy: FEI Company

Image processing

• Particle selection: Number selected: 350000 ; Details: Selected initial particles automatically using C complex 2D class averages, low-pass filtered to 20 Angstrom for automatic particle picking.
• CTF correction: Software - Name: CTFFIND (ver. 4)
• Startup model: Type of model: INSILICO MODEL; In silico model: An initial set of 30000 particles selected using 2D class averages from the C complex (EMDB-4055) and used for 3D classification with the C complex map (EMDB-4055) as a model. A class significantly different from C complex was obtained following this procedure and used as an initial C-star model
• Initial angle assignment: Type: PROJECTION MATCHING / Software - Name: RELION (ver. 1.4)
• Final 3D classification: Software - Name: RELION (ver. 1.4)
• Final angle assignment: Type: PROJECTION MATCHING / Software - Name: RELION (ver. 1.4)
• Final reconstruction: Number classes used: 4 / Resolution.type: BY AUTHOR / Resolution: 3.85 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 1.4) / Number images used: 65824

Atomic model buiding 1

• Details: Used secondary structure restraints generated in ProSMART and LibG.
• Refinement: Space: RECIPROCAL / Protocol: FLEXIBLE FIT / Overall B value: 330 / Target criteria: Fourier Shell Correlation

Output model

PDB-5mps:
Structure of a spliceosome remodeled for exon ligation