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- EMDB-13029: The U1 part of Saccharomyces cerevisiae spliceosomal pre-A comple... -

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Basic information

Entry
Database: EMDB / ID: EMD-13029
TitleThe U1 part of Saccharomyces cerevisiae spliceosomal pre-A complex (delta BS-A ACT1)
Map dataU1 part of the pre-A complex
Sample
  • Complex: S. cerevisiae spliceosomal pre-A complex
    • Protein or peptide: x 16 types
    • RNA: x 2 types
Function / homology
Function and homology information


positive regulation of RNA binding / mRNA splice site recognition / splicing factor binding / U4/U6 snRNP / 7-methylguanosine cap hypermethylation / pICln-Sm protein complex / spliceosomal tri-snRNP complex / small nuclear ribonucleoprotein complex / SMN-Sm protein complex / positive regulation of mRNA splicing, via spliceosome ...positive regulation of RNA binding / mRNA splice site recognition / splicing factor binding / U4/U6 snRNP / 7-methylguanosine cap hypermethylation / pICln-Sm protein complex / spliceosomal tri-snRNP complex / small nuclear ribonucleoprotein complex / SMN-Sm protein complex / positive regulation of mRNA splicing, via spliceosome / U2-type prespliceosome assembly / commitment complex / U4 snRNP / U2 snRNP / poly(U) RNA binding / U1 snRNP / U2-type prespliceosome / pre-mRNA 5'-splice site binding / precatalytic spliceosome / spliceosomal complex assembly / mRNA 5'-splice site recognition / U5 snRNP / spliceosomal snRNP assembly / U1 snRNA binding / U4/U6 x U5 tri-snRNP complex / catalytic step 2 spliceosome / spliceosomal complex / mRNA splicing, via spliceosome / mRNA binding / RNA binding / zinc ion binding / nucleus / cytosol / cytoplasm
Similarity search - Function
Luc7-related / LUC7 N_terminus / Snu56-like U1 small nuclear ribonucleoprotein component / Snu56-like U1 small nuclear ribonucleoprotein component / U1 small nuclear ribonucleoprotein C / U1 small nuclear ribonucleoprotein of 70kDa N-terminal / U1 small nuclear ribonucleoprotein of 70kDa MW N terminal / U1-C, C2H2-type zinc finger / U1 zinc finger / PWI domain ...Luc7-related / LUC7 N_terminus / Snu56-like U1 small nuclear ribonucleoprotein component / Snu56-like U1 small nuclear ribonucleoprotein component / U1 small nuclear ribonucleoprotein C / U1 small nuclear ribonucleoprotein of 70kDa N-terminal / U1 small nuclear ribonucleoprotein of 70kDa MW N terminal / U1-C, C2H2-type zinc finger / U1 zinc finger / PWI domain / PWI, domain in splicing factors / Matrin/U1-C, C2H2-type zinc finger / Zinc finger matrin-type profile. / 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 / Matrin/U1-C-like, C2H2-type zinc finger / U1-like zinc finger / Sm-like protein Lsm6/SmF / LSM domain / LSM domain, eukaryotic/archaea-type / snRNP Sm proteins / HAT (Half-A-TPR) repeat / HAT (Half-A-TPR) repeats / : / Sm domain profile. / LSM domain superfamily / Zinc finger C2H2 superfamily / RNA recognition motif / RNA recognition motif / Eukaryotic RNA Recognition Motif (RRM) profile. / RNA recognition motif domain / RNA-binding domain superfamily / Tetratricopeptide-like helical domain superfamily / Nucleotide-binding alpha-beta plait domain superfamily
Similarity search - Domain/homology
U1 small nuclear ribonucleoprotein A / Pre-mRNA-processing factor 39 / Small nuclear ribonucleoprotein-associated protein B / Small nuclear ribonucleoprotein G / Small nuclear ribonucleoprotein Sm D3 / U1 small nuclear ribonucleoprotein component SNU71 / Small nuclear ribonucleoprotein F / Protein NAM8 / U1 small nuclear ribonucleoprotein 70 kDa homolog / Small nuclear ribonucleoprotein Sm D1 ...U1 small nuclear ribonucleoprotein A / Pre-mRNA-processing factor 39 / Small nuclear ribonucleoprotein-associated protein B / Small nuclear ribonucleoprotein G / Small nuclear ribonucleoprotein Sm D3 / U1 small nuclear ribonucleoprotein component SNU71 / Small nuclear ribonucleoprotein F / Protein NAM8 / U1 small nuclear ribonucleoprotein 70 kDa homolog / Small nuclear ribonucleoprotein Sm D1 / U1 small nuclear ribonucleoprotein component PRP42 / 56 kDa U1 small nuclear ribonucleoprotein component / U1 small nuclear ribonucleoprotein C / Small nuclear ribonucleoprotein Sm D2 / Protein LUC7 / Small nuclear ribonucleoprotein E
Similarity search - Component
Biological speciesSaccharomyces cerevisiae (brewer's yeast) / Baker's yeast (brewer's yeast)
Methodsingle particle reconstruction / cryo EM / Resolution: 4.1 Å
AuthorsZhang Z / Rigo N / Dybkov O / Fourmann J / Will CL / Kumar V / Urlaub H / Stark H / Luehrmann R
Funding support Germany, 1 items
OrganizationGrant numberCountry
German Research Foundation (DFG)SFB 860 Germany
CitationJournal: Nature / Year: 2021
Title: Structural insights into how Prp5 proofreads the pre-mRNA branch site.
Authors: Zhenwei Zhang / Norbert Rigo / Olexandr Dybkov / Jean-Baptiste Fourmann / Cindy L Will / Vinay Kumar / Henning Urlaub / Holger Stark / Reinhard Lührmann /
Abstract: During the splicing of introns from precursor messenger RNAs (pre-mRNAs), the U2 small nuclear ribonucleoprotein (snRNP) must undergo stable integration into the spliceosomal A complex-a poorly ...During the splicing of introns from precursor messenger RNAs (pre-mRNAs), the U2 small nuclear ribonucleoprotein (snRNP) must undergo stable integration into the spliceosomal A complex-a poorly understood, multistep process that is facilitated by the DEAD-box helicase Prp5 (refs. ). During this process, the U2 small nuclear RNA (snRNA) forms an RNA duplex with the pre-mRNA branch site (the U2-BS helix), which is proofread by Prp5 at this stage through an unclear mechanism. Here, by deleting the branch-site adenosine (BS-A) or mutating the branch-site sequence of an actin pre-mRNA, we stall the assembly of spliceosomes in extracts from the yeast Saccharomyces cerevisiae directly before the A complex is formed. We then determine the three-dimensional structure of this newly identified assembly intermediate by cryo-electron microscopy. Our structure indicates that the U2-BS helix has formed in this pre-A complex, but is not yet clamped by the HEAT domain of the Hsh155 protein (Hsh155), which exhibits an open conformation. The structure further reveals a large-scale remodelling/repositioning of the U1 and U2 snRNPs during the formation of the A complex that is required to allow subsequent binding of the U4/U6.U5 tri-snRNP, but that this repositioning is blocked in the pre-A complex by the presence of Prp5. Our data suggest that binding of Hsh155 to the bulged BS-A of the U2-BS helix triggers closure of Hsh155, which in turn destabilizes Prp5 binding. Thus, Prp5 proofreads the branch site indirectly, hindering spliceosome assembly if branch-site mutations prevent the remodelling of Hsh155. Our data provide structural insights into how a spliceosomal helicase enhances the fidelity of pre-mRNA splicing.
History
DepositionJun 3, 2021-
Header (metadata) releaseAug 11, 2021-
Map releaseAug 11, 2021-
UpdateSep 29, 2021-
Current statusSep 29, 2021Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.042
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by radius
  • Surface level: 0.042
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-7oqc
  • Surface level: 0.042
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_13029.png

Downloads & links

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Map

FileDownload / File: emd_13029.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationU1 part of the pre-A complex
Voxel sizeX=Y=Z: 1.16 Å
Density
Contour LevelBy AUTHOR: 0.042 / Movie #1: 0.042
Minimum - Maximum-0.11696728 - 0.18834156
Average (Standard dev.)0.0007718664 (±0.0057305587)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions400400400
Spacing400400400
CellA=B=C: 464.0 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.161.161.16
M x/y/z400400400
origin x/y/z0.0000.0000.000
length x/y/z464.000464.000464.000
α/β/γ90.00090.00090.000
start NX/NY/NZ000
NX/NY/NZ400400400
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS400400400
D min/max/mean-0.1170.1880.001

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Supplemental data

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Half map: half map 2

Fileemd_13029_half_map_1.map
Annotationhalf map 2
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: half map1

Fileemd_13029_half_map_2.map
Annotationhalf map1
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : S. cerevisiae spliceosomal pre-A complex

EntireName: S. cerevisiae spliceosomal pre-A complex
Components
  • Complex: S. cerevisiae spliceosomal pre-A complex
    • Protein or peptide: Protein NAM8
    • RNA: ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A)
    • Protein or peptide: U1 small nuclear ribonucleoprotein component PRP42
    • Protein or peptide: U1 small nuclear ribonucleoprotein component SNU71
    • RNA: U1 snRNAU1 spliceosomal RNA
    • Protein or peptide: 56 kDa U1 small nuclear ribonucleoprotein component
    • Protein or peptide: U1 small nuclear ribonucleoprotein A
    • Protein or peptide: U1 small nuclear ribonucleoprotein C
    • 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
    • Protein or peptide: Protein LUC7
    • Protein or peptide: Pre-mRNA-processing factor 39
    • Protein or peptide: U1 small nuclear ribonucleoprotein 70 kDa homolog

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Supramolecule #1: S. cerevisiae spliceosomal pre-A complex

SupramoleculeName: S. cerevisiae spliceosomal pre-A complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#18
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)

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Macromolecule #1: Protein NAM8

MacromoleculeName: Protein NAM8 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 57.01084 KDa
SequenceString: MSYKQTTYYP SRGNLVRNDS SPYTNTISSE TNNSSTSVLS LQGASNVSLG TTGNQLYMGD LDPTWDKNTV RQIWASLGEA NINVRMMWN NTLNNGSRSS MGPKNNQGYC FVDFPSSTHA ANALLKNGML IPNFPNKKLK LNWATSSYSN SNNSLNNVKS G NNCSIFVG ...String:
MSYKQTTYYP SRGNLVRNDS SPYTNTISSE TNNSSTSVLS LQGASNVSLG TTGNQLYMGD LDPTWDKNTV RQIWASLGEA NINVRMMWN NTLNNGSRSS MGPKNNQGYC FVDFPSSTHA ANALLKNGML IPNFPNKKLK LNWATSSYSN SNNSLNNVKS G NNCSIFVG DLAPNVTESQ LFELFINRYA STSHAKIVHD QVTGMSKGYG FVKFTNSDEQ QLALSEMQGV FLNGRAIKVG PT SGQQQHV SGNNDYNRSS SSLNNENVDS RFLSKGQSFL SNGNNNMGFK RNHMSQFIYP VQQQPSLNHF TDPNNTTVFI GGL SSLVTE DELRAYFQPF GTIVYVKIPV GKCCGFVQYV DRLSAEAAIA GMQGFPIANS RVRLSWGRSA KQTALLQQAM LSNS LQVQQ QQPGLQQPNY GYIPSSTCEA PVLPDNNVSS TMLPGCQILN YSNPYANANG LGSNNFSFYS NNNATNTQAT SLLAD TSSM DLSGTGGQQV IMQGSEAVVN STNAMLNRLE QGSNGFMFA

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Macromolecule #3: U1 small nuclear ribonucleoprotein component PRP42

MacromoleculeName: U1 small nuclear ribonucleoprotein component PRP42 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 65.22202 KDa
SequenceString: MDKYTALIHD ENFSTLTLNV SRYPKSLAYW EKLLNYIVKA SAPICKSTEP QLLKLIRCTY SSMLNEFPYL ENYYIDFALL EYKLGNVSM SHKIFQRGLQ AFNQRSLLLW TSYLKFCNNV ISHQKQLFKK YETAEEYVGL HFFSGEFWDL YLEQISSRCT S SKKYWNVL ...String:
MDKYTALIHD ENFSTLTLNV SRYPKSLAYW EKLLNYIVKA SAPICKSTEP QLLKLIRCTY SSMLNEFPYL ENYYIDFALL EYKLGNVSM SHKIFQRGLQ AFNQRSLLLW TSYLKFCNNV ISHQKQLFKK YETAEEYVGL HFFSGEFWDL YLEQISSRCT S SKKYWNVL RKILEIPLHS FSKFYALWLQ RIDDIMDLKQ LSQLTSKDEL LKKLKIDINY SGRKGPYLQD AKKKLKKITK EM YMVVQYQ VLEIYSIFES KIYINYYTSP ETLVSSDEIE TWIKYLDYTI TLQTDSLTHL NFQRALLPLA HYDLVWIKYS KWL INSKND LLGAKNVLLM GLKFSLKKTE IIKLLYSVIC KLNEYVLLRN LLEKIESSYS DNVENVDDFE IFWDYLQFKT FCQN SLYSS RYSDSQSNGL LNKELFDKVW KRLSCKEKKS GQEILLNNLV QFYSKDTVEF VEKNIFQKII EFGWEYYLQN GMFWN CYCR LIYFDTSRSY LDKRQYIVRK IWPQIDKKFA QSVLPSLTEF CESYFPEEMD TLEEMFTEEP

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Macromolecule #4: U1 small nuclear ribonucleoprotein component SNU71

MacromoleculeName: U1 small nuclear ribonucleoprotein component SNU71 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 71.484555 KDa
SequenceString: MRDIVFVSPQ LYLSSQEGWK SDSAKSGFIP ILKNDLQRFQ DSLKHIVDAR NSLSETLLNS NDDGSIHNSD QNTGLNKDKE ASIADNNSA NKCATSSSRY QELKQFLPIS LDQQIHTVSL QGVSSSFSRG QIESLLDHCL NLALTETQSN SALKVEAWSS F SSFLDTQD ...String:
MRDIVFVSPQ LYLSSQEGWK SDSAKSGFIP ILKNDLQRFQ DSLKHIVDAR NSLSETLLNS NDDGSIHNSD QNTGLNKDKE ASIADNNSA NKCATSSSRY QELKQFLPIS LDQQIHTVSL QGVSSSFSRG QIESLLDHCL NLALTETQSN SALKVEAWSS F SSFLDTQD IFIRFSKVDE DEAFVNTLNY CKALFAFIRK LHEDFKIELH LDLNTKEYVE DRTGTIPSVK PEKASEFYSV FK NIEDQTD ERNSKKEQLD DSSTQYKVDT NTLSDLPSDA LDQLCKDIIE FRTKVVSIEK EKKMKSTYEE SRRQRHQMQK VFD QIRKNH SGAKGSANTE EEDTNMEDED EEDDTEDDLA LEKRKEERDL EESNRRYEDM LHQLHSNTEP KIKSIRADIM SAEN YEEHL EKNRSLYLKE LLHLANDVHY DHHRSFKEQE ERRDEEDRAK NGNAKELAPI QLSDGKAISA GKAAAITLPE GTVKS ENYN ADKNVSESSE HVKIKFDFKK AIDHSVESSS EDEGYRESEL PPTKPSERSA AEDRLPFTAD ELNIRLTNLK ESRYVD ELV REFLGVYEDE LVEYILENIR VNQSKQALLN ELRETFDEDG ETIADRLWSR KEFRLGT

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Macromolecule #6: 56 kDa U1 small nuclear ribonucleoprotein component

MacromoleculeName: 56 kDa U1 small nuclear ribonucleoprotein component / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 56.575277 KDa
SequenceString: MRPRRRGLAY HHTKPKGQLS QGHYPTTSND GQRRKVGNSE AFQSFDIWKN LDRIRSTKKN AGQFIKGSLL ILPMRTEDKQ QFDECMDEL HKYISKDILR CYPQKEQKDE GMLFYIVLKD FNILDSCFVL SVLLAFQKRL WMAPSEKSYF RVPKNINLTG S FYLPKNIE ...String:
MRPRRRGLAY HHTKPKGQLS QGHYPTTSND GQRRKVGNSE AFQSFDIWKN LDRIRSTKKN AGQFIKGSLL ILPMRTEDKQ QFDECMDEL HKYISKDILR CYPQKEQKDE GMLFYIVLKD FNILDSCFVL SVLLAFQKRL WMAPSEKSYF RVPKNINLTG S FYLPKNIE TGRGHIITSY RREQPSSSIV EVGFNVVPDF QQFQVKACHV SKFMNELSNF FSQVEFGKCE ANVINYFKRE YN RTYSQIS LALYELPLIG DGLFDIKSYI SKTRPIIETS KAQMIKHISE MKAYNEISGL QGDQFPRQQR PLSNSPSSNS ISS SQTIEA GATSYQTQPQ RHAVNKPSNV LNSSNRHSGP KTFEDGRYSE GNKPGFMTQD EIKQHCIGTI KASMDAVKKK SSYQ ILKTY VRCPRQNYID IVYQNLNDLR SKTNCNIVVL NLNNLHESQM WLESLNTTNY TIFAQAPHPS TIRVISIGGV GEYIV KALE LILNILEH

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Macromolecule #7: U1 small nuclear ribonucleoprotein A

MacromoleculeName: U1 small nuclear ribonucleoprotein A / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 34.438105 KDa
SequenceString: MSALYFQNLP SRPANKENYT RLLLKHINPN NKYAINPSLP LPHNKLQISS QPLMLLDDQM GLLEVSISRS SKMTNQAFLT FVTQEEADR FLEKYTTTAL KVQGRKVRMG KARTNSLLGL SIEMQKKKGN DETYNLDIKK VLKARKLKRK LRSDDICAKK F RLKRQIRR ...String:
MSALYFQNLP SRPANKENYT RLLLKHINPN NKYAINPSLP LPHNKLQISS QPLMLLDDQM GLLEVSISRS SKMTNQAFLT FVTQEEADR FLEKYTTTAL KVQGRKVRMG KARTNSLLGL SIEMQKKKGN DETYNLDIKK VLKARKLKRK LRSDDICAKK F RLKRQIRR LKHKLRSRKV EEAEIDRIVK EFETRRLENM KSQQENLKQS QKPLKRAKVS NTMENPPNKV LLIQNLPSGT TE QLLSQIL GNEALVEIRL VSVRNLAFVE YETVADATKI KNQLGSTYKL QNNDVTIGFA K

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Macromolecule #8: U1 small nuclear ribonucleoprotein C

MacromoleculeName: U1 small nuclear ribonucleoprotein C / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 27.106016 KDa
SequenceString: MTRYYCEYCH SYLTHDTLSV RKSHLVGKNH LRITADYYRN KARDIINKHN HKRRHIGKRG RKERENSSQN ETLKVTCLSN KEKRHIMHV KKMNQKELAQ TSIDTLKLLY DGSPGYSKVF VDANRFDIGD LVKASKLPQR ANEKSAHHSF KQTSRSRDET C ESNPFPRL ...String:
MTRYYCEYCH SYLTHDTLSV RKSHLVGKNH LRITADYYRN KARDIINKHN HKRRHIGKRG RKERENSSQN ETLKVTCLSN KEKRHIMHV KKMNQKELAQ TSIDTLKLLY DGSPGYSKVF VDANRFDIGD LVKASKLPQR ANEKSAHHSF KQTSRSRDET C ESNPFPRL NNPKKLEPPK ILSQWSNTIP KTSIFYSVDI LQTTIKESKK RMHSDGIRKP SSANGYKRRR YGN

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Macromolecule #9: Small nuclear ribonucleoprotein-associated protein B

MacromoleculeName: Small nuclear ribonucleoprotein-associated protein B / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 22.42699 KDa
SequenceString: MSKIQVAHSS RLANLIDYKL RVLTQDGRVY IGQLMAFDKH MNLVLNECIE ERVPKTQLDK LRPRKDSKDG TTLNIKVEKR VLGLTILRG EQILSTVVED KPLLSKKERL VRDKKEKKQA QKQTKLRKEK EKKPGKIAKP NTANAKHTSS NSREIAQPSS S RYNGGNDN ...String:
MSKIQVAHSS RLANLIDYKL RVLTQDGRVY IGQLMAFDKH MNLVLNECIE ERVPKTQLDK LRPRKDSKDG TTLNIKVEKR VLGLTILRG EQILSTVVED KPLLSKKERL VRDKKEKKQA QKQTKLRKEK EKKPGKIAKP NTANAKHTSS NSREIAQPSS S RYNGGNDN IGANRSRFNN EAPPQTRKFQ PPPGFKRK

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Macromolecule #10: Small nuclear ribonucleoprotein Sm D3

MacromoleculeName: Small nuclear ribonucleoprotein Sm D3 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 11.240139 KDa
SequenceString:
MTMNGIPVKL LNEAQGHIVS LELTTGATYR GKLVESEDSM NVQLRDVIAT EPQGAVTHMD QIFVRGSQIK FIVVPDLLKN APLFKKNSS RPMPPIRGPK RR

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Macromolecule #11: Small nuclear ribonucleoprotein E

MacromoleculeName: Small nuclear ribonucleoprotein E / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 10.385098 KDa
SequenceString:
MSNKVKTKAM VPPINCIFNF LQQQTPVTIW LFEQIGIRIK GKIVGFDEFM NVVIDEAVEI PVNSADGKED VEKGTPLGKI LLKGDNITL ITSAD

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Macromolecule #12: Small nuclear ribonucleoprotein F

MacromoleculeName: Small nuclear ribonucleoprotein F / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 9.669945 KDa
SequenceString:
MSESSDISAM QPVNPKPFLK GLVNHRVGVK LKFNSTEYRG TLVSTDNYFN LQLNEAEEFV AGVSHGTLGE IFIRCNNVLY IRELPN

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Macromolecule #13: Small nuclear ribonucleoprotein G

MacromoleculeName: Small nuclear ribonucleoprotein G / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 8.490809 KDa
SequenceString:
MVSTPELKKY MDKKILLNIN GSRKVAGILR GYDIFLNVVL DDAMEINGED PANNHQLGLQ TVIRGNSIIS LEALDAI

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Macromolecule #14: Small nuclear ribonucleoprotein Sm D1

MacromoleculeName: Small nuclear ribonucleoprotein Sm D1 / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 16.296798 KDa
SequenceString:
MKLVNFLKKL RNEQVTIELK NGTTVWGTLQ SVSPQMNAIL TDVKLTLPQP RLNKLNSNGI AMASLYLTGG QQPTASDNIA SLQYINIRG NTIRQIILPD SLNLDSLLVD QKQLNSLRRS GQIANDPSKK RRRDFGAPAN KRPRRGL

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Macromolecule #15: Small nuclear ribonucleoprotein Sm D2

MacromoleculeName: Small nuclear ribonucleoprotein Sm D2 / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 12.876066 KDa
SequenceString:
MSSQIIDRPK HELSRAELEE LEEFEFKHGP MSLINDAMVT RTPVIISLRN NHKIIARVKA FDRHCNMVLE NVKELWTEKK GKNVINRER FISKLFLRGD SVIVVLKTPV E

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Macromolecule #16: Protein LUC7

MacromoleculeName: Protein LUC7 / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 30.245885 KDa
SequenceString: MSTMSTPAAE QRKLVEQLMG RDFSFRHNRY SHQKRDLGLH DPKICKSYLV GECPYDLFQG TKQSLGKCPQ MHLTKHKIQY EREVKQGKT FPEFEREYLA ILSRFVNECN GQISVALQNL KHTAEERMKI QQVTEELDVL DVRIGLMGQE IDSLIRADEV S MGMLQSVK ...String:
MSTMSTPAAE QRKLVEQLMG RDFSFRHNRY SHQKRDLGLH DPKICKSYLV GECPYDLFQG TKQSLGKCPQ MHLTKHKIQY EREVKQGKT FPEFEREYLA ILSRFVNECN GQISVALQNL KHTAEERMKI QQVTEELDVL DVRIGLMGQE IDSLIRADEV S MGMLQSVK LQELISKRKE VAKRVRNITE NVGQSAQQKL QVCEVCGAYL SRLDTDRRLA DHFLGKIHLG YVKMREDYDR LM KNNRTTN ASKTATTLPG RRFV

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Macromolecule #17: Pre-mRNA-processing factor 39

MacromoleculeName: Pre-mRNA-processing factor 39 / type: protein_or_peptide / ID: 17 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 74.834742 KDa
SequenceString: MPDETNFTIE DIEPRPDALR GLDTQFLQDN TALVQAYRGL DWSDISSLTQ MVDVIEQTVV KYGNPNDSIK LALETILWQI LRKYPLLFG FWKRFATIEY QLFGLKKSIA VLATSVKWFP TSLELWCDYL NVLCVNNPNE TDFIRNNFEI AKDLIGKQFL S HPFWDKFI ...String:
MPDETNFTIE DIEPRPDALR GLDTQFLQDN TALVQAYRGL DWSDISSLTQ MVDVIEQTVV KYGNPNDSIK LALETILWQI LRKYPLLFG FWKRFATIEY QLFGLKKSIA VLATSVKWFP TSLELWCDYL NVLCVNNPNE TDFIRNNFEI AKDLIGKQFL S HPFWDKFI EFEVGQKNWH NVQRIYEYII EVPLHQYARF FTSYKKFLNE KNLKTTRNID IVLRKTQTTV NEIWQFESKI KQ PFFNLGQ VLNDDLENWS RYLKFVTDPS KSLDKEFVMS VFDRCLIPCL YHENTWMMYI KWLTKKNISD EVVVDIYQKA NTF LPLDFK TLRYDFLRFL KRKYRSNNTL FNNIFNETVS RYLKIWPNDI LLMTEYLCML KRHSFKNSLD QSPKEILEKQ TSFT KILET SITNYINNQI DAKVHLQTLI NDKNLSIVVV ELIKTTWLVL KNNMQTRKYF NLYQKNILIK NSVPFWLTYY KFEKS NVNF TKLNKFIREL GVEIYLPTTV MNDILTDYKT FYLTHSNIVT YESSIIDSNT FDPILYPELK MSNPKYDPVL NTTANV DWH KKTEWKEAGH IGITTERPQI SNSIIECNSG TLIQKPISLP NFRNLEKINQ VKINDLYTEE FLKEGK

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Macromolecule #18: U1 small nuclear ribonucleoprotein 70 kDa homolog

MacromoleculeName: U1 small nuclear ribonucleoprotein 70 kDa homolog / type: protein_or_peptide / ID: 18 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 34.506148 KDa
SequenceString: MNYNLSKYPD DVSRLFKPRP PLSYKRPTDY PYAKRQTNPN ITGVANLLST SLKHYMEEFP EGSPNNHLQR YEDIKLSKIK NAQLLDRRL QNWNPNVDPH IKDTDPYRTI FIGRLPYDLD EIELQKYFVK FGEIEKIRIV KDKITQKSKG YAFIVFKDPI S SKMAFKEI ...String:
MNYNLSKYPD DVSRLFKPRP PLSYKRPTDY PYAKRQTNPN ITGVANLLST SLKHYMEEFP EGSPNNHLQR YEDIKLSKIK NAQLLDRRL QNWNPNVDPH IKDTDPYRTI FIGRLPYDLD EIELQKYFVK FGEIEKIRIV KDKITQKSKG YAFIVFKDPI S SKMAFKEI GVHRGIQIKD RICIVDIERG RTVKYFKPRR LGGGLGGRGY SNRDSRLPGR FASASTSNPA ERNYAPRLPR RE TSSSAYS ADRYGSSTLD ARYRGNRPLL SAATPTAAVT SVYKSRNSRT RESQPAPKEA PDY

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Macromolecule #2: ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mR...

MacromoleculeName: ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 ...Name: ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A),ACT1 pre-mRNA (delta BS-A)
type: rna / ID: 2 / Number of copies: 1
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 111.601883 KDa
SequenceString: AUGGAUUCUG GUAUGUUC(N)(N) (N)(N)(N)(N)(N)(N)(N)(N)(N)(N) (N)(N)(N)(N)(N)(N)(N)(N) (N) (N)(N)(N)(N)(N)(N)(N)(N)(N)(N) (N)(N)(N)(N)(N)(N)(N)(N)(N)(N) (N)(N)(N)(N)(N) (N)(N)(N)(N) (N)(N)(N) ...String:
AUGGAUUCUG GUAUGUUC(N)(N) (N)(N)(N)(N)(N)(N)(N)(N)(N)(N) (N)(N)(N)(N)(N)(N)(N)(N) (N) (N)(N)(N)(N)(N)(N)(N)(N)(N)(N) (N)(N)(N)(N)(N)(N)(N)(N)(N)(N) (N)(N)(N)(N)(N) (N)(N)(N)(N) (N)(N)(N)UAGCGCU UGCACCAUCC CAUUUAACUG UAAGAAGAAU UGCACGGUCC CAAUUGCUCG AG AGAUUUC UCUUUUACCU UUUUUUACUA UUUUUCACUC UCCCAUAACC UCCUAUAUUG ACUGAUCUGU AAUAACCACG AUA UUAUUG GAAUAAAUAG GGGCUUGAAA UUUGGAAAAA AAAAAAAAAC UGAAAUAUUU UCGUGAUAAG UGAUAGUGAU AUUC UUCUU UUAUUUGCUA CUGUUACUAA GUCUCAUGUA CUACAUCGAU UGCUUCAUUC UUUUUGUUGC UAUAUUAUAU GUUUA G

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Macromolecule #5: U1 snRNA

MacromoleculeName: U1 snRNA / type: rna / ID: 5 / Number of copies: 1
Source (natural)Organism: Baker's yeast (brewer's yeast)
Molecular weightTheoretical: 182.114516 KDa
SequenceString: AUACUUACCU UAAGAUAUCA GAGGAGAUCA AGAAGUCCUA CUGAUCAAAC AUGCGCUUCC AAUAGUAGAA GGACGUUAAG CAUUUAUCA UUGAACUAUA AUUGUUCAUU GAAGUCAUUG AUGCAAACUC CUUGGUCACA CACACAUACG GCGCGGAAGG C GUGUUUGC ...String:
AUACUUACCU UAAGAUAUCA GAGGAGAUCA AGAAGUCCUA CUGAUCAAAC AUGCGCUUCC AAUAGUAGAA GGACGUUAAG CAUUUAUCA UUGAACUAUA AUUGUUCAUU GAAGUCAUUG AUGCAAACUC CUUGGUCACA CACACAUACG GCGCGGAAGG C GUGUUUGC UGACGUUUCC AUUCCCUUGU UUCAAUCAUU GGUUAAUCCC UUGAUUCCUU UGGGGAUUUU UGGGUUAAAC UG AUUUUUG GGGCCCUUUG UUUCUUCUGC CUGGAGAAGU UUGACACCAA AUUCAAAUUG GUGUUAGGGG AGCUGGGGCC UUU CAAAAG AGAGCUUUGU AGAGGCAUUC UUUUUGACUA CUUUUCUCUA GCGUGCCAUU UUAGUUUUUG ACGGCAGAUU CGAA UGAAC UUAAGUUUAU GAUGAAGGUA UGGCUGUUGA GAUUAUUUGG UCGGGAUUGU AGUUUGAAGA UGUGCUCUUU UGAGC AGUC UCAACUUUGC UCGUUCCCGU UAUGGGAAAA AUUUUGGAAG GUCUUGGUAG GAACGGGUGG AUCUUAUAAU UUUUGA UUU AUUUU

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

BufferpH: 7.9
GridModel: Quantifoil R3.5/1 / Material: COPPER / Support film - Material: CARBON / Support film - topology: CONTINUOUS
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK I

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 100.0 µm / Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 0.01 mm
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Image recordingFilm or detector model: FEI FALCON III (4k x 4k) / Detector mode: INTEGRATING / Average exposure time: 1.02 sec. / Average electron dose: 44.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: OTHER / Details: Ab initio model
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.0)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.0)
Final reconstructionAlgorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 4.1 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.0) / Number images used: 217460
FSC plot (resolution estimation)

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