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- EMDB-43753: Yeast U1 snRNP with humanized U1C Zinc-Finger domain -

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Basic information

Entry
Database: EMDB / ID: EMD-43753
TitleYeast U1 snRNP with humanized U1C Zinc-Finger domain
Map data
Sample
  • Complex: U1snRNP
    • Protein or peptide: x 16 types
    • RNA: x 2 types
KeywordsU1snRNP / U1C humanization / SPLICING
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 / small nuclear ribonucleoprotein complex / SMN-Sm protein complex / spliceosomal tri-snRNP 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 / small nuclear ribonucleoprotein complex / SMN-Sm protein complex / spliceosomal tri-snRNP complex / positive regulation of mRNA splicing, via spliceosome / commitment complex / U2-type prespliceosome assembly / poly(U) RNA binding / U4 snRNP / U2 snRNP / U1 snRNP / pre-mRNA 5'-splice site binding / U2-type prespliceosome / 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 S288C (yeast)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.49 Å
AuthorsShi SS / Kuang ZL / Zhao R
Funding support United States, 2 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R35GM145289 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R01GM126157 (R.Z.) United States
CitationJournal: RNA / Year: 2024
Title: Selected humanization of yeast U1 snRNP leads to global suppression of pre-mRNA splicing and mitochondrial dysfunction in the budding yeast.
Authors: Subbaiah Chalivendra / Shasha Shi / Xueni Li / Zhiling Kuang / Joseph Giovinazzo / Lingdi Zhang / John Rossi / Jingxin Wang / Anthony Saviola / Robb Welty / Shiheng Liu / Katherine Vaeth / Z ...Authors: Subbaiah Chalivendra / Shasha Shi / Xueni Li / Zhiling Kuang / Joseph Giovinazzo / Lingdi Zhang / John Rossi / Jingxin Wang / Anthony Saviola / Robb Welty / Shiheng Liu / Katherine Vaeth / Z Hong Zhou / Kirk Hansen / J Matthew Taliaferro / Rui Zhao /
Abstract: The recognition of 5' splice site (5' ss) is one of the earliest steps of pre-mRNA splicing. To better understand the mechanism and regulation of 5' ss recognition, we selectively humanized ...The recognition of 5' splice site (5' ss) is one of the earliest steps of pre-mRNA splicing. To better understand the mechanism and regulation of 5' ss recognition, we selectively humanized components of the yeast U1 snRNP to reveal the function of these components in 5' ss recognition and splicing. We targeted U1C and Luc7, two proteins that interact with and stabilize the yeast U1 (yU1) snRNA and the 5' ss RNA duplex. We replaced the Zinc-Finger (ZnF) domain of yU1C with its human counterpart, which resulted in a cold-sensitive growth phenotype and moderate splicing defects. We next added an auxin-inducible degron to yLuc7 protein (to mimic the lack of Luc7Ls in human U1 snRNP) and found that Luc7-depleted yU1 snRNP resulted in the concomitant loss of PRP40 and Snu71 (two other essential yeast U1 snRNP proteins), and further biochemical analyses suggest a model of how these three proteins interact with each other in the U1 snRNP. The loss of these proteins resulted in a significant growth retardation accompanied by a global suppression of pre-mRNA splicing. The splicing suppression led to mitochondrial dysfunction as revealed by a release of Fe2+ into the growth medium and an induction of mitochondrial reactive oxygen species. Together, these observations indicate that the human U1C ZnF can substitute that of yeast, Luc7 is essential for the incorporation of the Luc7-Prp40-Snu71 trimer into yeast U1 snRNP, and splicing plays a major role in the regulation of mitochondrial function in yeast.
History
DepositionFeb 20, 2024-
Header (metadata) releaseMay 15, 2024-
Map releaseMay 15, 2024-
UpdateMay 15, 2024-
Current statusMay 15, 2024Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_43753.png

Downloads & links

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Map

FileDownload / File: emd_43753.map.gz / Format: CCP4 / Size: 1000 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Voxel sizeX=Y=Z: 0.83 Å
Density
Contour LevelBy AUTHOR: 0.08
Minimum - Maximum-1.2842387 - 2.1316335
Average (Standard dev.)0.00021066514 (±0.031006655)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderYXZ
Origin000
Dimensions640640640
Spacing640640640
CellA=B=C: 531.2 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #2

Fileemd_43753_half_map_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Half map: #1

Fileemd_43753_half_map_2.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
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Histogram

Histogram (log scale)

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Sample components

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Entire : U1snRNP

EntireName: U1snRNP
Components
  • Complex: U1snRNP
    • Protein or peptide: U1 small nuclear ribonucleoprotein 70 kDa homolog
    • Protein or peptide: U1 small nuclear ribonucleoprotein C
    • Protein or peptide: U1 small nuclear ribonucleoprotein A
    • Protein or peptide: U1 small nuclear ribonucleoprotein component PRP42
    • Protein or peptide: Pre-mRNA-processing factor 39
    • Protein or peptide: Protein NAM8
    • Protein or peptide: 56 kDa U1 small nuclear ribonucleoprotein component
    • Protein or peptide: U1 small nuclear ribonucleoprotein component SNU71
    • Protein or peptide: Protein LUC7
    • Protein or peptide: Small nuclear ribonucleoprotein-associated protein B
    • Protein or peptide: Small nuclear ribonucleoprotein Sm D1
    • Protein or peptide: Small nuclear ribonucleoprotein Sm D2
    • 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
    • RNA: ACT1 pre-mRNA
    • RNA: U1 snRNAU1 spliceosomal RNA

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Supramolecule #1: U1snRNP

SupramoleculeName: U1snRNP / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)

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Macromolecule #1: U1 small nuclear ribonucleoprotein 70 kDa homolog

MacromoleculeName: U1 small nuclear ribonucleoprotein 70 kDa homolog / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 34.506148 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
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

UniProtKB: U1 small nuclear ribonucleoprotein 70 kDa homolog

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Macromolecule #2: U1 small nuclear ribonucleoprotein C

MacromoleculeName: U1 small nuclear ribonucleoprotein C / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 27.084949 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString: MPKFYCDYCD TYLTHDSPSV RKTHCSGRKH KENVKDYYRN KARDIINKHN HKRRHIGKRG RKERENSSQN ETLKVTCLSN KEKRHIMHV KKMNQKELAQ TSIDTLKLLY DGSPGYSKVF VDANRFDIGD LVKASKLPQR ANEKSAHHSF KQTSRSRDET C ESNPFPRL ...String:
MPKFYCDYCD TYLTHDSPSV RKTHCSGRKH KENVKDYYRN KARDIINKHN HKRRHIGKRG RKERENSSQN ETLKVTCLSN KEKRHIMHV KKMNQKELAQ TSIDTLKLLY DGSPGYSKVF VDANRFDIGD LVKASKLPQR ANEKSAHHSF KQTSRSRDET C ESNPFPRL NNPKKLEPPK ILSQWSNTIP KTSIFYSVDI LQTTIKESKK RMHSDGIRKP SSANGYKRRR YGN

UniProtKB: U1 small nuclear ribonucleoprotein C

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Macromolecule #3: U1 small nuclear ribonucleoprotein A

MacromoleculeName: U1 small nuclear ribonucleoprotein A / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 40.413961 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
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 KGRRIPGLIN PWKRRWKKNF IAV SAANRF KKISSSGALD YDIPTTASEN LYFQ

UniProtKB: U1 small nuclear ribonucleoprotein A

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Macromolecule #4: U1 small nuclear ribonucleoprotein component PRP42

MacromoleculeName: U1 small nuclear ribonucleoprotein component PRP42 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 65.22202 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
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

UniProtKB: U1 small nuclear ribonucleoprotein component PRP42

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Macromolecule #5: Pre-mRNA-processing factor 39

MacromoleculeName: Pre-mRNA-processing factor 39 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 74.834742 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
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

UniProtKB: Pre-mRNA-processing factor 39

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Macromolecule #6: Protein NAM8

MacromoleculeName: Protein NAM8 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 57.01084 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
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

UniProtKB: Protein NAM8

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Macromolecule #7: 56 kDa U1 small nuclear ribonucleoprotein component

MacromoleculeName: 56 kDa U1 small nuclear ribonucleoprotein component / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 56.575277 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
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

UniProtKB: 56 kDa U1 small nuclear ribonucleoprotein component

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Macromolecule #8: U1 small nuclear ribonucleoprotein component SNU71

MacromoleculeName: U1 small nuclear ribonucleoprotein component SNU71 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 5.97476 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString:
MRDIVFVSPQ LYLSSQEGWK SDSAKSGFIP ILKNDLQRFQ DSLKHIVDAR NS

UniProtKB: U1 small nuclear ribonucleoprotein component SNU71

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Macromolecule #9: Protein LUC7

MacromoleculeName: Protein LUC7 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 30.245885 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
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

UniProtKB: Protein LUC7

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Macromolecule #10: Small nuclear ribonucleoprotein-associated protein B

MacromoleculeName: Small nuclear ribonucleoprotein-associated protein B / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 22.42699 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
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

UniProtKB: Small nuclear ribonucleoprotein-associated protein B

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Macromolecule #11: Small nuclear ribonucleoprotein Sm D1

MacromoleculeName: Small nuclear ribonucleoprotein Sm D1 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 16.296798 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString:
MKLVNFLKKL RNEQVTIELK NGTTVWGTLQ SVSPQMNAIL TDVKLTLPQP RLNKLNSNGI AMASLYLTGG QQPTASDNIA SLQYINIRG NTIRQIILPD SLNLDSLLVD QKQLNSLRRS GQIANDPSKK RRRDFGAPAN KRPRRGL

UniProtKB: Small nuclear ribonucleoprotein Sm D1

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Macromolecule #12: Small nuclear ribonucleoprotein Sm D2

MacromoleculeName: Small nuclear ribonucleoprotein Sm D2 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 12.876066 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString:
MSSQIIDRPK HELSRAELEE LEEFEFKHGP MSLINDAMVT RTPVIISLRN NHKIIARVKA FDRHCNMVLE NVKELWTEKK GKNVINRER FISKLFLRGD SVIVVLKTPV E

UniProtKB: Small nuclear ribonucleoprotein Sm D2

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Macromolecule #13: Small nuclear ribonucleoprotein Sm D3

MacromoleculeName: Small nuclear ribonucleoprotein Sm D3 / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 11.240139 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString:
MTMNGIPVKL LNEAQGHIVS LELTTGATYR GKLVESEDSM NVQLRDVIAT EPQGAVTHMD QIFVRGSQIK FIVVPDLLKN APLFKKNSS RPMPPIRGPK RR

UniProtKB: Small nuclear ribonucleoprotein Sm D3

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Macromolecule #14: Small nuclear ribonucleoprotein E

MacromoleculeName: Small nuclear ribonucleoprotein E / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 10.385098 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString:
MSNKVKTKAM VPPINCIFNF LQQQTPVTIW LFEQIGIRIK GKIVGFDEFM NVVIDEAVEI PVNSADGKED VEKGTPLGKI LLKGDNITL ITSAD

UniProtKB: Small nuclear ribonucleoprotein E

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Macromolecule #15: Small nuclear ribonucleoprotein F

MacromoleculeName: Small nuclear ribonucleoprotein F / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 9.669945 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString:
MSESSDISAM QPVNPKPFLK GLVNHRVGVK LKFNSTEYRG TLVSTDNYFN LQLNEAEEFV AGVSHGTLGE IFIRCNNVLY IRELPN

UniProtKB: Small nuclear ribonucleoprotein F

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Macromolecule #16: Small nuclear ribonucleoprotein G

MacromoleculeName: Small nuclear ribonucleoprotein G / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 8.490809 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae S288C (yeast)
SequenceString:
MVSTPELKKY MDKKILLNIN GSRKVAGILR GYDIFLNVVL DDAMEINGED PANNHQLGLQ TVIRGNSIIS LEALDAI

UniProtKB: Small nuclear ribonucleoprotein G

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Macromolecule #17: ACT1 pre-mRNA

MacromoleculeName: ACT1 pre-mRNA / type: rna / ID: 17 / Number of copies: 1
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 59.778191 KDa
SequenceString:
GGAUCCGAUA UCCGUACACC AUCAGGGUAC GAGCUAGCCC AUGGCGUACA CCAUCAGGGU ACGACUAGUA GAUCUCGUAC ACCAUCAGG GUACGGAAUU GUCUAGACUU UUAGAUUUUU CACGCUUACU GCUUUUUUCU UCCCAAGAUC GAAAAUUUAC U GAAUUAAC AAUGGAUUCU GGUAUGUUC

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Macromolecule #18: U1 snRNA

MacromoleculeName: U1 snRNA / type: rna / ID: 18 / Number of copies: 1
Source (natural)Organism: Saccharomyces cerevisiae S288C (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

Concentration1.5 mg/mL
BufferpH: 7.9
Component:
ConcentrationFormulaName
120.0 mMNaClSodium chloridesodium chloride
20.0 mMC8H18N2O4SHepes
2.0 mMC14H24N2O10EGTA

Details: 20 mM Hepes, pH7.9, 120 mM KCl, 2 mM EGTA
GridModel: EMS Lacey Carbon / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: HOLEY / Support film - Film thickness: 20 / Pretreatment - Type: PLASMA CLEANING / Pretreatment - Time: 20 sec. / Pretreatment - Atmosphere: OTHER
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV
Details: The grids were obtained with the chamber at 100% humidity, 2.5 s blotting time, -6 blotting force and 15 s wait time and flash-frozen into liquid ethane with a Vitrobot Mark IV (Thermo Fisher Scientific).
DetailsThis sample is monodisperse

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal defocus max: 2.2 µm / Nominal defocus min: 0.8 µm
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 1.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: NONE
Initial angle assignmentType: OTHER
Final angle assignmentType: OTHER
Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.49 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 138997
FSC plot (resolution estimation)

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