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- EMDB-16347: Enp1TAP-S21_A population of yeast small ribosomal subunit precurs... -

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Basic information

Entry
Database: EMDB / ID: EMD-16347
TitleEnp1TAP-S21_A population of yeast small ribosomal subunit precursors depleted of rpS21/eS21
Map dataEnp1TAP-S21_A, multibody refinement
Sample
  • Complex: Enp1-TAP associated immature ribosomal particles from S. cerevisiae depleted of rpS21/eS21
    • RNA: x 1 types
    • Protein or peptide: x 25 types
Function / homology
Function and homology information


positive regulation of RNA import into nucleus / endonucleolytic cleavage to generate mature 5'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / mTORC1-mediated signalling / Protein hydroxylation / : / U3 snoRNA binding / poly(A)+ mRNA export from nucleus / positive regulation of nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / Formation of the ternary complex, and subsequently, the 43S complex ...positive regulation of RNA import into nucleus / endonucleolytic cleavage to generate mature 5'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / mTORC1-mediated signalling / Protein hydroxylation / : / U3 snoRNA binding / poly(A)+ mRNA export from nucleus / positive regulation of nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / Formation of the ternary complex, and subsequently, the 43S complex / Translation initiation complex formation / Ribosomal scanning and start codon recognition / preribosome, small subunit precursor / snoRNA binding / Major pathway of rRNA processing in the nucleolus and cytosol / SRP-dependent cotranslational protein targeting to membrane / 90S preribosome / GTP hydrolysis and joining of the 60S ribosomal subunit / Formation of a pool of free 40S subunits / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / L13a-mediated translational silencing of Ceruloplasmin expression / proteasome assembly / endonucleolytic cleavage in ITS1 to separate SSU-rRNA from 5.8S rRNA and LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / regulation of translational fidelity / ribonucleoprotein complex binding / maturation of SSU-rRNA / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / small-subunit processome / maintenance of translational fidelity / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / ribosomal small subunit assembly / rRNA processing / cytoplasmic stress granule / cytosolic small ribosomal subunit / unfolded protein binding / ribosome biogenesis / cytoplasmic translation / small ribosomal subunit / rRNA binding / non-specific serine/threonine protein kinase / ribosome / protein kinase activity / structural constituent of ribosome / translation / phosphorylation / protein serine kinase activity / mRNA binding / GTPase activity / protein serine/threonine kinase activity / GTP binding / nucleolus / mitochondrion / RNA binding / nucleoplasm / ATP binding / metal ion binding / nucleus / cytosol / cytoplasm
Similarity search - Function
Serine/threonine-protein kinase Rio2 / RIO2 kinase winged helix domain, N-terminal / Rio2, N-terminal / RIO kinase / RIO-like kinase / RIO1 family / Krr1, KH1 domain / Krr1 KH1 domain / Bystin / Bystin ...Serine/threonine-protein kinase Rio2 / RIO2 kinase winged helix domain, N-terminal / Rio2, N-terminal / RIO kinase / RIO-like kinase / RIO1 family / Krr1, KH1 domain / Krr1 KH1 domain / Bystin / Bystin / Ribosome biogenesis protein BMS1/TSR1, C-terminal / AARP2CN / Bms1/Tsr1-type G domain / Ribosome biogenesis protein Bms1/Tsr1 / 40S ribosome biogenesis protein Tsr1 and BMS1 C-terminal / AARP2CN (NUC121) domain / Bms1-type guanine nucleotide-binding (G) domain profile. / AARP2CN (NUC121) domain / Protein of unknown function (DUF663) / K Homology domain, type 1 superfamily / Ribosomal protein S19e, conserved site / Ribosomal protein S25 / : / Ribosomal protein S30 / Ribosomal protein S8e subdomain, eukaryotes / S25 ribosomal protein / Ribosomal protein S19A/S15e / Ribosomal protein S30 / Ribosomal protein S3Ae, conserved site / Ribosomal protein S19e / Ribosomal_S19e / Ribosomal protein S8e, conserved site / 40S ribosomal protein S11, N-terminal / 40S ribosomal protein S1/3, eukaryotes / Ribosomal protein S6, eukaryotic / Ribosomal protein S7e / 40S ribosomal protein S4, C-terminal domain / Ribosomal protein S4e, N-terminal, conserved site / Ribosomal protein S19e signature. / Ribosomal protein S19e / Ribosomal protein S27, zinc-binding domain superfamily / Ribosomal protein S17, archaeal/eukaryotic / Ribosomal protein S27 / Ribosomal protein S28e conserved site / Ribosomal protein S6/S6e/A/B/2, conserved site / Ribosomal protein S28e / 40S ribosomal protein S4 C-terminus / Ribosomal protein S4e, N-terminal / Ribosomal protein S23, eukaryotic/archaeal / Ribosomal_S17 N-terminal / Ribosomal protein S3Ae / Ribosomal S3Ae family / Ribosomal protein S7e / Ribosomal protein S8e / Ribosomal protein S4, KOW domain / Ribosomal protein S5/S7, eukaryotic/archaeal / Ribosomal protein S4e / Ribosomal protein S4e, central region / Ribosomal protein S4e, central domain superfamily / Ribosomal protein S6e / Ribosomal protein S13/S15, N-terminal / Ribosomal protein S15P / Ribosomal S13/S15 N-terminal domain / Ribosomal protein S6e / Ribosomal protein S4/S9, eukaryotic/archaeal / RS4NT (NUC023) domain / Ribosomal protein S27 / Ribosomal S3Ae family / Ribosomal protein S28e / Ribosomal family S4e / Ribosomal S13/S15 N-terminal domain / Ribosomal protein S7e signature. / Ribosomal protein S6e / Ribosomal protein S3Ae signature. / Ribosomal protein S27e signature. / Ribosomal protein S4e signature. / Ribosomal protein S8e signature. / Ribosomal S24e conserved site / Ribosomal protein S24e signature. / Ribosomal protein S24e / Ribosomal protein S24e / Ribosomal protein S6e signature. / Ribosomal protein S28e signature. / Ribosomal protein S8e/ribosomal biogenesis NSA2 / Ribosomal protein S8e / : / Ribosomal protein S15/S19, conserved site / Ribosomal protein S19/S15 / Ribosomal protein S19/S15, superfamily / Ribosomal protein S7, conserved site / Ribosomal protein S17, conserved site / Ribosomal protein S19 / Ribosomal protein S13, conserved site / Ribosomal protein S13 / 30s ribosomal protein S13, C-terminal / Ribosomal protein S4/S9 N-terminal domain / Ribosomal protein S4/S9, N-terminal / Ribosomal protein S4, conserved site / Ribosomal protein S4/S9 N-terminal domain / Ribosomal protein S13/S18
Similarity search - Domain/homology
Small ribosomal subunit protein uS4A / Small ribosomal subunit protein uS15 / Small ribosomal subunit protein uS11A / Small ribosomal subunit protein eS19A / Small ribosomal subunit protein uS8A / Small ribosomal subunit protein uS12A / Small ribosomal subunit protein eS24A / Small ribosomal subunit protein eS30A / Small ribosomal subunit protein eS4A / Small ribosomal subunit protein eS6A ...Small ribosomal subunit protein uS4A / Small ribosomal subunit protein uS15 / Small ribosomal subunit protein uS11A / Small ribosomal subunit protein eS19A / Small ribosomal subunit protein uS8A / Small ribosomal subunit protein uS12A / Small ribosomal subunit protein eS24A / Small ribosomal subunit protein eS30A / Small ribosomal subunit protein eS4A / Small ribosomal subunit protein eS6A / Small ribosomal subunit protein eS8A / Small ribosomal subunit protein uS17A / Small ribosomal subunit protein uS9A / Small ribosomal subunit protein uS13A / Small ribosomal subunit protein uS7 / Small ribosomal subunit protein eS7A / Small ribosomal subunit protein eS1A / Small ribosomal subunit protein eS27A / Essential nuclear protein 1 / Serine/threonine-protein kinase RIO2 / Small ribosomal subunit protein uS19 / Ribosome biogenesis protein TSR1 / Small ribosomal subunit protein eS25A / Small ribosomal subunit protein eS28A / Pre-rRNA-processing protein PNO1
Similarity search - Component
Biological speciesSaccharomyces cerevisiae (brewer's yeast) / baker's yeast (brewer's yeast)
Methodsingle particle reconstruction / cryo EM / Resolution: 2.9 Å
AuthorsMilkereit P / Poell G
Funding support Germany, 1 items
OrganizationGrant numberCountry
German Research Foundation (DFG)SFB 960 Germany
CitationJournal: PLoS One / Year: 2023
Title: Impact of the yeast S0/uS2-cluster ribosomal protein rpS21/eS21 on rRNA folding and the architecture of small ribosomal subunit precursors.
Authors: Gisela Pöll / Joachim Griesenbeck / Herbert Tschochner / Philipp Milkereit /
Abstract: RpS0/uS2, rpS2/uS5, and rpS21/eS21 form a cluster of ribosomal proteins (S0-cluster) at the head-body junction near the central pseudoknot of eukaryotic small ribosomal subunits (SSU). Previous work ...RpS0/uS2, rpS2/uS5, and rpS21/eS21 form a cluster of ribosomal proteins (S0-cluster) at the head-body junction near the central pseudoknot of eukaryotic small ribosomal subunits (SSU). Previous work in yeast indicated that S0-cluster assembly is required for the stabilisation and maturation of SSU precursors at specific post-nucleolar stages. Here, we analysed the role of S0-cluster formation for rRNA folding. Structures of SSU precursors isolated from yeast S0-cluster expression mutants or control strains were analysed by cryogenic electron microscopy. The obtained resolution was sufficient to detect individual 2'-O-methyl RNA modifications using an unbiased scoring approach. The data show how S0-cluster formation enables the initial recruitment of the pre-rRNA processing factor Nob1 in yeast. Furthermore, they reveal hierarchical effects on the pre-rRNA folding pathway, including the final maturation of the central pseudoknot. Based on these structural insights we discuss how formation of the S0-cluster determines at this early cytoplasmic assembly checkpoint if SSU precursors further mature or are degraded.
History
DepositionDec 15, 2022-
Header (metadata) releaseDec 28, 2022-
Map releaseDec 28, 2022-
UpdateApr 12, 2023-
Current statusApr 12, 2023Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_16347.png

Downloads & links

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Map

FileDownload / File: emd_16347.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationEnp1TAP-S21_A, multibody refinement
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.97 Å/pix.
x 400 pix.
= 387.2 Å		0.97 Å/pix.
x 400 pix.
= 387.2 Å		0.97 Å/pix.
x 400 pix.
= 387.2 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 0.968 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.007
Minimum - Maximum-0.009645895 - 0.047675118
Average (Standard dev.)0.0002905215 (±0.0016200148)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions400400400
Spacing400400400
CellA=B=C: 387.2 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : Enp1-TAP associated immature ribosomal particles from S. cerevisi...

EntireName: Enp1-TAP associated immature ribosomal particles from S. cerevisiae depleted of rpS21/eS21
Components
  • Complex: Enp1-TAP associated immature ribosomal particles from S. cerevisiae depleted of rpS21/eS21
    • RNA: 18S rRNA precursor
    • Protein or peptide: rpS5
    • Protein or peptide: 40S ribosomal protein S15
    • Protein or peptide: 40S ribosomal protein S16-ARibosome
    • Protein or peptide: 40S ribosomal protein S18-ARibosome
    • Protein or peptide: 40S ribosomal protein S19-ARibosome
    • Protein or peptide: 40S ribosomal protein S25-ARibosome
    • Protein or peptide: 40S ribosomal protein S28-ARibosome
    • Protein or peptide: 40S ribosomal protein S1-ARibosome
    • Protein or peptide: 40S ribosomal protein S4-ARibosome
    • Protein or peptide: 40S ribosomal protein S6-ARibosome
    • Protein or peptide: 40S ribosomal protein S7-ARibosome
    • Protein or peptide: 40S ribosomal protein S8-ARibosome
    • Protein or peptide: 40S ribosomal protein S9-ARibosome
    • Protein or peptide: 40S ribosomal protein S11-ARibosome
    • Protein or peptide: 40S ribosomal protein S13
    • Protein or peptide: 40S ribosomal protein S14-ARibosome
    • Protein or peptide: 40S ribosomal protein S22-ARibosome
    • Protein or peptide: 40S ribosomal protein S23-ARibosome
    • Protein or peptide: 40S ribosomal protein S24-ARibosome
    • Protein or peptide: Essential nuclear protein 1
    • Protein or peptide: 40S ribosomal protein S27-ARibosome
    • Protein or peptide: 40S ribosomal protein S30-ARibosome
    • Protein or peptide: Pre-rRNA-processing protein PNO1
    • Protein or peptide: Serine/threonine-protein kinase RIO2
    • Protein or peptide: Ribosome biogenesis protein TSR1

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Supramolecule #1: Enp1-TAP associated immature ribosomal particles from S. cerevisi...

SupramoleculeName: Enp1-TAP associated immature ribosomal particles from S. cerevisiae depleted of rpS21/eS21
type: complex / ID: 1 / Chimera: Yes / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast) / Strain: S288C-derivative laboratory strain

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Macromolecule #1: 18S rRNA precursor

MacromoleculeName: 18S rRNA precursor / type: rna / ID: 1 / Number of copies: 1
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 579.761938 KDa
SequenceString: UAUCUGGUUG AUCCUGCCAG UAGUCAUAUG CUUGUCUCAA AGAUUAAGCC AUGCAUGUCU AAGUAUAAGC AAUUUAUACA GUGAAACUG CGAAUGGCUC AUUAAAUCAG UUAUCGUUUA UUUGAUAGUU CCUUUACUAC AUGGUAUAAC UGUGGUAAUU C UAGAGCUA ...String:
UAUCUGGUUG AUCCUGCCAG UAGUCAUAUG CUUGUCUCAA AGAUUAAGCC AUGCAUGUCU AAGUAUAAGC AAUUUAUACA GUGAAACUG CGAAUGGCUC AUUAAAUCAG UUAUCGUUUA UUUGAUAGUU CCUUUACUAC AUGGUAUAAC UGUGGUAAUU C UAGAGCUA AUACAUGCUU AAAAUCUCGA CCCUUUGGAA GAGAUGUAUU UAUUAGAUAA AAAAUCAAUG UCUUCGGACU CU UUGAUGA UUCAUAAUAA CUUUUCGAAU CGCAUGGCCU UGUGCUGGCG AUGGUUCAUU CAAAUUUCUG CCCUAUCAAC UUU CGAUGG UAGGAUAGUG GCCUACCAUG GUUUCAACGG GUAACGGGGA AUAAGGGUUC GAUUCCGGAG AGGGAGCCUG AGAA ACGGC UACCACAUCC AAGGAAGGCA GCAGGCGCGC AAAUUACCCA AUCCUAAUUC AGGGAGGUAG UGACAAUAAA UAACG AUAC AGGGCCCAUU CGGGUCUUGU AAUUGGAAUG AGUACAAUGU AAAUACCUUA ACGAGGAACA AUUGGAGGGC AAGUCU GGU GCCAGCAGCC GCGGUAAUUC CAGCUCCAAU AGCGUAUAUU AAAGUUGUUG CAGUUAAAAA GCUCGUAGUU GAACUUU GG GCCCGGUUGG CCGGUCCGAU UUUUUCGUGU ACUGGAUUUC CAACGGGGCC UUUCCUUCUG GCUAACCUUG AGUCCUUG U GGCUCUUGGC GAACCAGGAC UUUUACUUUG AAAAAAUUAG AGUGUUCAAA GCAGGCGUAU UGCUCGAAUA UAUUAGCAU GGAAUAAUAG AAUAGGACGU UUGGUUCUAU UUUGUUGGUU UCUAGGACCA UCGUAAUGAU UAAUAGGGAC GGUCGGGGGC AUCAGUAUU CAAUUGUCAG AGGUGAAAUU CUUGGAUUUA UUGAAGACUA ACUACUGCGA AAGCAUUUGC CAAGGACGUU U UCAUUAAU CAAGAACGAA AGUUAGGGGA UCGAAGAUGA UCAGAUACCG UCGUAGUCUU AACCAUAAAC UAUGCCGACU AG GGAUCGG GUGGUGUUUU UUUAAUGACC CACUCGGCAC CUUACGAGAA AUCAAAGUCU UUGGGUUCUG GGGGGAGUAU GGU CGCAAG GCUGAAACUU AAAGGAAUUG ACGGAAGGGC ACCACCAGGA GUGGAGCCUG CGGCUUAAUU UGACUCAACA CGGG GAAAC UCACCAGGUC CAGACACAAU AAGGAUUGAC AGAUUGAGAG CUCUUUCUUG AUUUUGUGGG UGGUGGUGCA UGGCC GUUC UUAGUUGGUG GAGUGAUUUG UCUGCUUAAU UGCGAUAACG AACGAGACCU UAACCUACUA AAUAGUGGUG CUAGCA UUU GCUGGUUAUC CACUUCUUAG AGGGACUAUC GGUUUCAAGC CGAUGGAAGU UUGAGGCAAU AACAGGUCUG UGAUGCC CU UAGACGUUCU GGGCCGCACG CGCGCUACAC UGACGGAGCC AGCGAGUCUA ACCUUGGCCG AGAGGUCUUG GUAAUCUU G UGAAACUCCG UCGUGCUGGG GAUAGAGCAU UGUAAUUAUU GCUCUUCAAC GAGGAAUUCC UAGUAAGCGC AAGUCAUCA GCUUGCGUUG AUUACGUCCC UGCCCUUUGU ACACACCGCC CGUCGCUAGU ACCGAUUGAA UGGCUUAGUG AGGCCUCAGG AUCUGCUUA GAGAAGGGGG CAACUCCAUC UCAGAGCGGA GAAUUUGGAC AAACUUGGUC AUUUAGAGGA ACUAAAAGUC G UAACAAGG UUUCCGUAGG UGAACCUGCG GAAGGAUCAU UA

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Macromolecule #2: rpS5

MacromoleculeName: rpS5 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 25.0726 KDa
SequenceString: MSDTEAPVEV QEDFEVVEEF TPVVLATPIP EEVQQAQTEI KLFNKWSFEE VEVKDASLVD YVQVRQPIFV AHTAGRYANK RFRKAQCPI IERLTNSLMM NGRNNGKKLK AVRIIKHTLD IINVLTDQNP IQVVVDAITN TGPREDTTRV GGGGAARRQA V DVSPLRRV ...String:
MSDTEAPVEV QEDFEVVEEF TPVVLATPIP EEVQQAQTEI KLFNKWSFEE VEVKDASLVD YVQVRQPIFV AHTAGRYANK RFRKAQCPI IERLTNSLMM NGRNNGKKLK AVRIIKHTLD IINVLTDQNP IQVVVDAITN TGPREDTTRV GGGGAARRQA V DVSPLRRV NQAIALLTIG AREAAFRNIK TIAETLAEEL INAAKGSSTS YAIKKKDELE RVAKSNR

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Macromolecule #3: 40S ribosomal protein S15

MacromoleculeName: 40S ribosomal protein S15 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 16.031907 KDa
SequenceString:
MSQAVNAKKR VFKTHSYRGV DLEKLLEMST EDFVKLAPAR VRRRFARGMT SKPAGFMKKL RAAKLAAPEN EKPAPVRTHM RNMIIVPEM IGSVVGIYNG KAFNQVEIRP EMLGHYLGEF SITYTPVRHG RAGATTSRFI PLK

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Macromolecule #4: 40S ribosomal protein S16-A

MacromoleculeName: 40S ribosomal protein S16-A / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 15.87749 KDa
SequenceString:
MSAVPSVQTF GKKKSATAVA HVKAGKGLIK VNGSPITLVE PEILRFKVYE PLLLVGLDKF SNIDIRVRVT GGGHVSQVYA IRQAIAKGL VAYHQKYVDE QSKNELKKAF TSYDRTLLIA DSRRPEPKKF GGKGARSRFQ KSYR

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Macromolecule #5: 40S ribosomal protein S18-A

MacromoleculeName: 40S ribosomal protein S18-A / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 17.071641 KDa
SequenceString:
MSLVVQEQGS FQHILRLLNT NVDGNIKIVY ALTTIKGVGR RYSNLVCKKA DVDLHKRAGE LTQEELERIV QIMQNPTHYK IPAWFLNRQ NDITDGKDYH TLANNVESKL RDDLERLKKI RAHRGIRHFW GLRVRGQHTK TTGRRRA

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Macromolecule #6: 40S ribosomal protein S19-A

MacromoleculeName: 40S ribosomal protein S19-A / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 15.942125 KDa
SequenceString:
MPGVSVRDVA AQDFINAYAS FLQRQGKLEV PGYVDIVKTS SGNEMPPQDA EGWFYKRAAS VARHIYMRKQ VGVGKLNKLY GGAKSRGVR PYKHIDASGS INRKVLQALE KIGIVEISPK GGRRISENGQ RDLDRIAAQT LEEDE

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Macromolecule #7: 40S ribosomal protein S25-A

MacromoleculeName: 40S ribosomal protein S25-A / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 12.067272 KDa
SequenceString:
MPPKQQLSKA AKAAAALAGG KKSKKKWSKK SMKDRAQHAV ILDQEKYDRI LKEVPTYRYV SVSVLVDRLK IGGSLARIAL RHLEKEGII KPISKHSKQA IYTRATASE

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Macromolecule #8: 40S ribosomal protein S28-A

MacromoleculeName: 40S ribosomal protein S28-A / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 7.605847 KDa
SequenceString:
MDNKTPVTLA KVIKVLGRTG SRGGVTQVRV EFLEDTSRTI VRNVKGPVRE NDILVLMESE REARRLR

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Macromolecule #9: 40S ribosomal protein S1-A

MacromoleculeName: 40S ribosomal protein S1-A / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 28.798467 KDa
SequenceString: MAVGKNKRLS KGKKGQKKRV VDPFTRKEWF DIKAPSTFEN RNVGKTLVNK STGLKSASDA LKGRVVEVCL ADLQGSEDHS FRKIKLRVD EVQGKNLLTN FHGMDFTTDK LRSMVRKWQT LIEANVTVKT SDDYVLRIFA IAFTRKQANQ VKRHSYAQSS H IRAIRKVI ...String:
MAVGKNKRLS KGKKGQKKRV VDPFTRKEWF DIKAPSTFEN RNVGKTLVNK STGLKSASDA LKGRVVEVCL ADLQGSEDHS FRKIKLRVD EVQGKNLLTN FHGMDFTTDK LRSMVRKWQT LIEANVTVKT SDDYVLRIFA IAFTRKQANQ VKRHSYAQSS H IRAIRKVI SEILTKEVQG STLAQLTSKL IPEVINKEIE NATKDIFPLQ NIHVRKVKLL KQPKFDVGAL MALHGEGSGE EK GKKVTGF KDEVLETV

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Macromolecule #10: 40S ribosomal protein S4-A

MacromoleculeName: 40S ribosomal protein S4-A / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 29.46933 KDa
SequenceString: MARGPKKHLK RLAAPHHWLL DKLSGCYAPR PSAGPHKLRE SLPLIVFLRN RLKYALNGRE VKAILMQRHV KVDGKVRTDT TYPAGFMDV ITLDATNENF RLVYDVKGRF AVHRITDEEA SYKLGKVKKV QLGKKGVPYV VTHDGRTIRY PDPNIKVNDT V KIDLASGK ...String:
MARGPKKHLK RLAAPHHWLL DKLSGCYAPR PSAGPHKLRE SLPLIVFLRN RLKYALNGRE VKAILMQRHV KVDGKVRTDT TYPAGFMDV ITLDATNENF RLVYDVKGRF AVHRITDEEA SYKLGKVKKV QLGKKGVPYV VTHDGRTIRY PDPNIKVNDT V KIDLASGK ITDFIKFDAG KLVYVTGGRN LGRIGTIVHK ERHDGGFDLV HIKDSLDNTF VTRLNNVFVI GEQGKPYISL PK GKGIKLS IAEERDRRRA QQGL

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Macromolecule #11: 40S ribosomal protein S6-A

MacromoleculeName: 40S ribosomal protein S6-A / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 27.054486 KDa
SequenceString: MKLNISYPVN GSQKTFEIDD EHRIRVFFDK RIGQEVDGEA VGDEFKGYVF KISGGNDKQG FPMKQGVLLP TRIKLLLTKN VSCYRPRRD GERKRKSVRG AIVGPDLAVL ALVIVKKGEQ ELEGLTDTTV PKRLGPKRAN NIRKFFGLSK EDDVRDFVIR R EVTKGEKT ...String:
MKLNISYPVN GSQKTFEIDD EHRIRVFFDK RIGQEVDGEA VGDEFKGYVF KISGGNDKQG FPMKQGVLLP TRIKLLLTKN VSCYRPRRD GERKRKSVRG AIVGPDLAVL ALVIVKKGEQ ELEGLTDTTV PKRLGPKRAN NIRKFFGLSK EDDVRDFVIR R EVTKGEKT YTKAPKIQRL VTPQRLQRKR HQRALKVRNA QAQREAAAEY AQLLAKRLSE RKAEKAEIRK RRASSLKA

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Macromolecule #12: 40S ribosomal protein S7-A

MacromoleculeName: 40S ribosomal protein S7-A / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 21.658209 KDa
SequenceString:
MSAPQAKILS QAPTELELQV AQAFVELENS SPELKAELRP LQFKSIREID VAGGKKALAI FVPVPSLAGF HKVQTKLTRE LEKKFQDRH VIFLAERRIL PKPSRTSRQV QKRPRSRTLT AVHDKILEDL VFPTEIVGKR VRYLVGGNKI QKVLLDSKDV Q QIDYKLES FQAVYNKLTG KQIVFEIPSE TH

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Macromolecule #13: 40S ribosomal protein S8-A

MacromoleculeName: 40S ribosomal protein S8-A / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 22.537803 KDa
SequenceString: MGISRDSRHK RSATGAKRAQ FRKKRKFELG RQPANTKIGA KRIHSVRTRG GNKKYRALRI ETGNFSWASE GISKKTRIAG VVYHPSNNE LVRTNTLTKA AIVQIDATPF RQWFEAHYGQ TLGKKKNVKE EETVAKSKNA ERKWAARAAS AKIESSVESQ F SAGRLYAC ...String:
MGISRDSRHK RSATGAKRAQ FRKKRKFELG RQPANTKIGA KRIHSVRTRG GNKKYRALRI ETGNFSWASE GISKKTRIAG VVYHPSNNE LVRTNTLTKA AIVQIDATPF RQWFEAHYGQ TLGKKKNVKE EETVAKSKNA ERKWAARAAS AKIESSVESQ F SAGRLYAC ISSRPGQSGR CDGYILEGEE LAFYLRRLTA KK

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Macromolecule #14: 40S ribosomal protein S9-A

MacromoleculeName: 40S ribosomal protein S9-A / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 22.487893 KDa
SequenceString: MPRAPRTYSK TYSTPKRPYE SSRLDAELKL AGEFGLKNKK EIYRISFQLS KIRRAARDLL TRDEKDPKRL FEGNALIRRL VRVGVLSED KKKLDYVLAL KVEDFLERRL QTQVYKLGLA KSVHHARVLI TQRHIAVGKQ IVNIPSFMVR LDSEKHIDFA P TSPFGGAR ...String:
MPRAPRTYSK TYSTPKRPYE SSRLDAELKL AGEFGLKNKK EIYRISFQLS KIRRAARDLL TRDEKDPKRL FEGNALIRRL VRVGVLSED KKKLDYVLAL KVEDFLERRL QTQVYKLGLA KSVHHARVLI TQRHIAVGKQ IVNIPSFMVR LDSEKHIDFA P TSPFGGAR PGRVARRNAA RKAEASGEAA DEADEADEE

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Macromolecule #15: 40S ribosomal protein S11-A

MacromoleculeName: 40S ribosomal protein S11-A / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 17.785934 KDa
SequenceString:
MSTELTVQSE RAFQKQPHIF NNPKVKTSKR TKRWYKNAGL GFKTPKTAIE GSYIDKKCPF TGLVSIRGKI LTGTVVSTKM HRTIVIRRA YLHYIPKYNR YEKRHKNVPV HVSPAFRVQV GDIVTVGQCR PISKTVRFNV VKVSAAAGKA NKQFAKF

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Macromolecule #16: 40S ribosomal protein S13

MacromoleculeName: 40S ribosomal protein S13 / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 17.059945 KDa
SequenceString:
MGRMHSAGKG ISSSAIPYSR NAPAWFKLSS ESVIEQIVKY ARKGLTPSQI GVLLRDAHGV TQARVITGNK IMRILKSNGL APEIPEDLY YLIKKAVSVR KHLERNRKDK DAKFRLILIE SRIHRLARYY RTVAVLPPNW KYESATASAL VN

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Macromolecule #17: 40S ribosomal protein S14-A

MacromoleculeName: 40S ribosomal protein S14-A / type: protein_or_peptide / ID: 17 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 14.562655 KDa
SequenceString:
MSNVVQARDN SQVFGVARIY ASFNDTFVHV TDLSGKETIA RVTGGMKVKA DRDESSPYAA MLAAQDVAAK CKEVGITAVH VKIRATGGT RTKTPGPGGQ AALRALARSG LRIGRIEDVT PVPSDSTRKK GGRRGRRL

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Macromolecule #18: 40S ribosomal protein S22-A

MacromoleculeName: 40S ribosomal protein S22-A / type: protein_or_peptide / ID: 18 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 14.650062 KDa
SequenceString:
MTRSSVLADA LNAINNAEKT GKRQVLIRPS SKVIIKFLQV MQKHGYIGEF EYIDDHRSGK IVVQLNGRLN KCGVISPRFN VKIGDIEKW TANLLPARQF GYVILTTSAG IMDHEEARRK HVSGKILGFV Y

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Macromolecule #19: 40S ribosomal protein S23-A

MacromoleculeName: 40S ribosomal protein S23-A / type: protein_or_peptide / ID: 19 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 16.073896 KDa
SequenceString:
MGKGKPRGLN SARKLRVHRR NNRWAENNYK KRLLGTAFKS SPFGGSSHAK GIVLEKLGIE SKQPNSAIRK CVRVQLIKNG KKVTAFVPN DGCLNFVDEN DEVLLAGFGR KGKAKGDIPG VRFKVVKVSG VSLLALWKEK KEKPRS

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Macromolecule #20: 40S ribosomal protein S24-A

MacromoleculeName: 40S ribosomal protein S24-A / type: protein_or_peptide / ID: 20 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 15.362848 KDa
SequenceString:
MSDAVTIRTR KVISNPLLAR KQFVVDVLHP NRANVSKDEL REKLAEVYKA EKDAVSVFGF RTQFGGGKSV GFGLVYNSVA EAKKFEPTY RLVRYGLAEK VEKASRQQRK QKKNRDKKIF GTGKRLAKKV ARRNAD

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Macromolecule #21: Essential nuclear protein 1

MacromoleculeName: Essential nuclear protein 1 / type: protein_or_peptide / ID: 21 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 55.207422 KDa
SequenceString: MARASSTKAR KQRHDPLLKD LDAAQGTLKK INKKKLAQND AANHDAANEE DGYIDSKASR KILQLAKEQQ DEIEGEELAE SERNKQFEA RFTTMSYDDE DEDEDEDEEA FGEDISDFEP EGDYKEEEEI VEIDEEDAAM FEQYFKKSDD FNSLSGSYNL A DKIMASIR ...String:
MARASSTKAR KQRHDPLLKD LDAAQGTLKK INKKKLAQND AANHDAANEE DGYIDSKASR KILQLAKEQQ DEIEGEELAE SERNKQFEA RFTTMSYDDE DEDEDEDEEA FGEDISDFEP EGDYKEEEEI VEIDEEDAAM FEQYFKKSDD FNSLSGSYNL A DKIMASIR EKESQVEDMQ DDEPLANEQN TSRGNISSGL KSGEGVALPE KVIKAYTTVG SILKTWTHGK LPKLFKVIPS LR NWQDVIY VTNPEEWSPH VVYEATKLFV SNLTAKESQK FINLILLERF RDNIETSEDH SLNYHIYRAV KKSLYKPSAF FKG FLFPLV ETGCNVREAT IAGSVLAKVS VPALHSSAAL SYLLRLPFSP PTTVFIKILL DKKYALPYQT VDDCVYYFMR FRIL DDGSN GEDATRVLPV IWHKAFLTFA QRYKNDITQD QRDFLLETVR QRGHKDIGPE IRRELLAGAS REFVDPQEAN DDLMI DVN

+
Macromolecule #22: 40S ribosomal protein S27-A

MacromoleculeName: 40S ribosomal protein S27-A / type: protein_or_peptide / ID: 22 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 8.893391 KDa
SequenceString:
MVLVQDLLHP TAASEARKHK LKTLVQGPRS YFLDVKCPGC LNITTVFSHA QTAVTCESCS TILCTPTGGK AKLSEGTSFR RK

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Macromolecule #23: 40S ribosomal protein S30-A

MacromoleculeName: 40S ribosomal protein S30-A / type: protein_or_peptide / ID: 23 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 7.137541 KDa
SequenceString:
MAKVHGSLAR AGKVKSQTPK VEKTEKPKKP KGRAYKRLLY TRRFVNVTLV NGKRRMNPGP SVQ

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Macromolecule #24: Pre-rRNA-processing protein PNO1

MacromoleculeName: Pre-rRNA-processing protein PNO1 / type: protein_or_peptide / ID: 24 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 30.380623 KDa
SequenceString: MVAPTALKKA TVTPVSGQDG GSSRIIGINN TESIDEDDDD DVLLDDSDNN TAKEEVEGEE GSRKTHESKT VVVDDQGKPR FTSASKTQG NKIKFESRKI MVPPHRMTPL RNSWTKIYPP LVEHLKLQVR MNLKTKSVEL RTNPKFTTDP GALQKGADFI K AFTLGFDL ...String:
MVAPTALKKA TVTPVSGQDG GSSRIIGINN TESIDEDDDD DVLLDDSDNN TAKEEVEGEE GSRKTHESKT VVVDDQGKPR FTSASKTQG NKIKFESRKI MVPPHRMTPL RNSWTKIYPP LVEHLKLQVR MNLKTKSVEL RTNPKFTTDP GALQKGADFI K AFTLGFDL DDSIALLRLD DLYIETFEVK DVKTLTGDHL SRAIGRIAGK DGKTKFAIEN ATRTRIVLAD SKIHILGGFT HI RMARESV VSLILGSPPG KVYGNLRTVA SRLKERY

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Macromolecule #25: Serine/threonine-protein kinase RIO2

MacromoleculeName: Serine/threonine-protein kinase RIO2 / type: protein_or_peptide / ID: 25 / Number of copies: 1 / Enantiomer: LEVO / EC number: non-specific serine/threonine protein kinase
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 49.192852 KDa
SequenceString: MKLDTSHMRY LTTDDFRVLQ AVEQGSRSHE VVPTPLIHQI SGMRSQSGTN RAISDLAKLS LISKMRNVKY DGYRLTYNGI DYLALKTML NRDTVYSVGN TIGVGKESDI YKVSDKNGNP RVMKIHRLGR TSFHSVRNNR DYLKKSNQGA NWMHLSRLAA N KEYQFMSM ...String:
MKLDTSHMRY LTTDDFRVLQ AVEQGSRSHE VVPTPLIHQI SGMRSQSGTN RAISDLAKLS LISKMRNVKY DGYRLTYNGI DYLALKTML NRDTVYSVGN TIGVGKESDI YKVSDKNGNP RVMKIHRLGR TSFHSVRNNR DYLKKSNQGA NWMHLSRLAA N KEYQFMSM LYSKGFKVPE PFDNSRHIVV MELIEGYPMR RLRKHKNIPK LYSDLMCFIV DLANSGLIHC DFNEFNIMIK DK LEDENDC GFVVIDFPQC ISIQHQDADY YFQRDVDCIR RFFKKKLKYE PKPDSSMLDT EGFGDGYKYA YPDFKRDVKR TDN LDELVQ ASGFSKKHPG DRGLETAVES MRNAVYNSDD DMSNDEAEEE NGEGDYSEED EYYDSELDNE SSEDDSEDAQ EEEN ERIIE ALSSGVENLK MDKLGNYILE

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Macromolecule #26: Ribosome biogenesis protein TSR1

MacromoleculeName: Ribosome biogenesis protein TSR1 / type: protein_or_peptide / ID: 26 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: baker's yeast (brewer's yeast)
Molecular weightTheoretical: 90.876539 KDa
SequenceString: MAGHSHRSSL KNGHKSYKSK HASKGALKRL YKGKVEKEPV GTGKPDKQVS KLQRKNKAKQ LRAQRILDSI ENRKLFEGKN GAAKIITIV PLVNDLDPLD ILYKLLKCAD DEGIMVQEVD SKRIFNVHIK KFKSNLKIII PDMTNFLNIL DCAKVADFVV F GLSGVQEV ...String:
MAGHSHRSSL KNGHKSYKSK HASKGALKRL YKGKVEKEPV GTGKPDKQVS KLQRKNKAKQ LRAQRILDSI ENRKLFEGKN GAAKIITIV PLVNDLDPLD ILYKLLKCAD DEGIMVQEVD SKRIFNVHIK KFKSNLKIII PDMTNFLNIL DCAKVADFVV F GLSGVQEV DEEFGEQIIR ALELQGIASY IGVISNLSAV HEKEKFQLDV KQSLESYFKH FFPSEERVYN LEKNSDALNV LR TLCQRLP RSINWRDNRG YVVADFVDFV ETSPDSGDLV IEGTVRGIGF NANRLVHIPD FGDFQLNKIE KISESSQKRK IIK EKATDS LSLELDLQTV FESNMNRDTL DEYAPEGTED WSDYDEDFEY DGLTTARYDD HGFLPGREQT SKKAAVPKGT SDYQ AKWYL DDVIDANEEE EAEQTNGKDE TMMEIDDEMM VEQDNEEVAG DEEYDIEDNE GFEELSPEEE ERQLREFRDM EKEDR EFPD EIELEPSESA IERLKRYRGL KNLYNCDWQV DEKDPSSPAE WKRLLRIGNY KNTKNRIIKE TKNEAQAIAG DRIRMF IRF PKFLLEKIQD PKQLLFAVYG LLLHEHKNAV VNFSLQRWEQ YDKPVPSQEP IVVQYGVRRY TIQPLFSQGS NSPNNVH KY ERFLHPDTVS VATCIAPVDF TQSPAIFFKP SPTDAKNIEL IGHGTFLNAD HSRILAKRAI LTGHPFRFHK TVVTVRYM F FRPEDVEWFK SIPLFTKSGR SGFIKESLGT HGYFKATFDG KLSAQDVVAM SLYKRMWPMP SLPWNGM

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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: 8
Component:
ConcentrationName
200.0 mMpotassium chloride
5.0 mMmagnesium acetate
20.0 mMTris pH8
GridModel: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 200 sec. / Pretreatment - Atmosphere: AIR / Pretreatment - Pressure: 0.4 kPa
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeJEOL CRYO ARM 200
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 70.0 µm / Illumination 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 / Nominal magnification: 50000
Sample stageSpecimen holder model: JEOL CRYOSPECPORTER / Cooling holder cryogen: NITROGEN
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Number grids imaged: 1 / Number real images: 8575 / Average exposure time: 4.8 sec. / Average electron dose: 40.0 e/Å2

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Image processing

Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 4.0)
Final 3D classificationSoftware - Name: RELION (ver. 4.0)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 4.0)
Final reconstructionNumber classes used: 1 / Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 2.9 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 4.0) / Number images used: 132563

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Atomic model buiding 1

RefinementProtocol: FLEXIBLE FIT
Output model

PDB-8c00:
Enp1TAP-S21_A population of yeast small ribosomal subunit precursors depleted of rpS21/eS21

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