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- EMDB-75580: Yeast co-transcriptional pre-60S assembly intermediate Nop12 RNP ... -

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Basic information

Entry
Database: EMDB / ID: EMD-75580
TitleYeast co-transcriptional pre-60S assembly intermediate Nop12 RNP - Composite map
Map data
Sample
  • Complex: Yeast co-transcriptional pre-60S assembly intermediate Nop12 RNP
    • RNA: x 3 types
    • Protein or peptide: x 14 types
  • Ligand: x 1 types
Keywordsribosome biogenesis / co-transcriptional assembly / RNA-binding protein / nucleolar RNP / RIBOSOME
Function / homology
Function and homology information


snoRNA release from pre-rRNA / nuclear division / exonucleolytic trimming to generate mature 5'-end of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage in ITS1 upstream of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / preribosome / PeBoW complex / ATP-dependent activity, acting on RNA / rRNA primary transcript binding / maturation of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / cytosolic large ribosomal subunit assembly ...snoRNA release from pre-rRNA / nuclear division / exonucleolytic trimming to generate mature 5'-end of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage in ITS1 upstream of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / preribosome / PeBoW complex / ATP-dependent activity, acting on RNA / rRNA primary transcript binding / maturation of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / cytosolic large ribosomal subunit assembly / Major pathway of rRNA processing in the nucleolus and cytosol / SRP-dependent cotranslational protein targeting to membrane / ribosomal large subunit binding / GTP hydrolysis and joining of the 60S ribosomal subunit / proteasome binding / preribosome, large subunit precursor / Formation of a pool of free 40S subunits / 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 / ribonucleoprotein complex binding / 90S preribosome / maturation of LSU-rRNA / nuclear periphery / proteasome complex / ribosomal large subunit biogenesis / protein catabolic process / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / small-subunit processome / rRNA processing / nuclear envelope / ribosomal small subunit biogenesis / 5S rRNA binding / ribosomal large subunit assembly / large ribosomal subunit rRNA binding / cytosolic large ribosomal subunit / protein-macromolecule adaptor activity / cytoplasmic translation / RNA helicase activity / rRNA binding / structural constituent of ribosome / RNA helicase / mRNA binding / nucleolus / ATP hydrolysis activity / DNA binding / RNA binding / nucleoplasm / ATP binding / identical protein binding / nucleus / cytoplasm / cytosol
Similarity search - Function
Nop12, RNA recognition motif 1 / : / Ribosome biogenesis protein 15, RNA recognition motif / Eukaryotic rRNA processing / Eukaryotic rRNA processing protein EBP2 / Ribosome biogenesis protein Nop16 / Ribosome biogenesis protein Nop16 / Ribosome biogenesis protein BRX1 / DDX18/Has1, DEAD-box helicase domain / Domain of unknown function DUF4217 ...Nop12, RNA recognition motif 1 / : / Ribosome biogenesis protein 15, RNA recognition motif / Eukaryotic rRNA processing / Eukaryotic rRNA processing protein EBP2 / Ribosome biogenesis protein Nop16 / Ribosome biogenesis protein Nop16 / Ribosome biogenesis protein BRX1 / DDX18/Has1, DEAD-box helicase domain / Domain of unknown function DUF4217 / ATP-dependent rRNA helicase SPB4-like, C-terminal extension / DUF4217 / BOP1, N-terminal domain / WD repeat BOP1/Erb1 / BOP1NT (NUC169) domain / BOP1NT (NUC169) domain / Pescadillo / Pescadillo N-terminus / BRCT domain / Brix domain / Brix domain / Brix domain profile. / Brix / DEAD-box subfamily ATP-dependent helicases signature. / ATP-dependent RNA helicase DEAD-box, conserved site / RNA helicase, DEAD-box type, Q motif / DEAD-box RNA helicase Q motif profile. / breast cancer carboxy-terminal domain / BRCT domain profile. / Ribosomal protein L13e, conserved site / Ribosomal protein L13e signature. / BRCT domain / Ribosomal protein L1-like / Ribosomal protein L1/ribosomal biogenesis protein / Ribosomal protein L1p/L10e family / BRCT domain superfamily / Ribosomal protein L13e / Ribosomal protein L13e / Ribosomal protein L36e signature. / Ribosomal protein L7A/L8 / Ribosomal protein L36e / Ribosomal protein L36e domain superfamily / Ribosomal protein L36e / Ribosomal protein L15e, conserved site / Ribosomal protein L15e signature. / DEAD/DEAH box helicase domain / DEAD/DEAH box helicase / Ribosomal protein L7Ae conserved site / Ribosomal protein L7Ae signature. / Ribosomal_L15e / Ribosomal protein L15e / Ribosomal protein L15e core domain superfamily / Ribosomal L15 / Ribosomal protein L7, eukaryotic/archaeal / Ribosomal protein L7/L30 / Ribosomal protein L7Ae/L8/Nhp2 family / RNA recognition motif / : / RNA recognition motif / Eukaryotic RNA Recognition Motif (RRM) profile. / RNA recognition motif domain / RNA-binding domain superfamily / Ribosomal protein L7Ae/L30e/S12e/Gadd45 / Ribosomal protein L7Ae/L30e/S12e/Gadd45 family / Helicase conserved C-terminal domain / 50S ribosomal protein L30e-like / Ribosomal protein L30, conserved site / Ribosomal protein L30 signature. / helicase superfamily c-terminal domain / Ribosomal protein L30, ferredoxin-like fold domain / Ribosomal protein L30p/L7e / Ribosomal protein L30, ferredoxin-like fold domain superfamily / WD domain, G-beta repeat / Superfamilies 1 and 2 helicase C-terminal domain profile. / Superfamilies 1 and 2 helicase ATP-binding type-1 domain profile. / DEAD-like helicases superfamily / Helicase, C-terminal / Helicase superfamily 1/2, ATP-binding domain / Ribosomal protein L23/L15e core domain superfamily / Nucleotide-binding alpha-beta plait domain superfamily / WD40 repeat, conserved site / Trp-Asp (WD) repeats signature. / Trp-Asp (WD) repeats profile. / Trp-Asp (WD) repeats circular profile. / WD40 repeats / WD40 repeat / WD40-repeat-containing domain superfamily / WD40/YVTN repeat-like-containing domain superfamily / P-loop containing nucleoside triphosphate hydrolase
Similarity search - Domain/homology
Large ribosomal subunit protein eL36A / Large ribosomal subunit protein eL15A / Large ribosomal subunit protein eL8A / rRNA-processing protein EBP2 / Proteasome-interacting protein CIC1 / Nucleolar protein 16 / Ribosome biogenesis protein RLP7 / Pescadillo homolog / Ribosome biogenesis protein 15 / ATP-dependent RNA helicase HAS1 ...Large ribosomal subunit protein eL36A / Large ribosomal subunit protein eL15A / Large ribosomal subunit protein eL8A / rRNA-processing protein EBP2 / Proteasome-interacting protein CIC1 / Nucleolar protein 16 / Ribosome biogenesis protein RLP7 / Pescadillo homolog / Ribosome biogenesis protein 15 / ATP-dependent RNA helicase HAS1 / Ribosome biogenesis protein ERB1 / Nucleolar protein 12 / Ribosome biogenesis protein BRX1 / Large ribosomal subunit protein eL13A
Similarity search - Component
Biological speciesSaccharomyces cerevisiae S288C (yeast)
Methodsingle particle reconstruction / cryo EM / Resolution: 2.78 Å
AuthorsPiwowarczyk R / Klinge S
Funding support United States, 2 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)5R01GM145950-03 United States
Chan Zuckerberg Initiative2023-332391 United States
CitationJournal: bioRxiv / Year: 2026
Title: Mechanism for the initiation of co-transcriptional pre-60 assembly.
Authors: Rafal Piwowarczyk / Sebastian Klinge
Abstract: Eukaryotic ribosomal large subunit (60 ) assembly requires an internal transcribed spacer 2 (ITS2) to license both nucleolar and nuclear pre-60 assembly intermediates. The underlying molecular ...Eukaryotic ribosomal large subunit (60 ) assembly requires an internal transcribed spacer 2 (ITS2) to license both nucleolar and nuclear pre-60 assembly intermediates. The underlying molecular mechanisms responsible for nucleation of pre-60 assembly, quality control, and installation of ITS2 during co-transcriptional stages remain unknown. Here we report the earliest co-transcriptional assembly intermediates of the eukaryotic 60 subunits. Together with biochemical assays, our data reveal the architecture of co-transcriptional pre-60 assembly initiation and progression, as well as the molecular logic of an assembly checkpoint. This study highlights an evolutionary solution by which complex RNA folding processes can be parallelized and integrated via biological AND-gating.
History
DepositionFeb 13, 2026-
Header (metadata) releaseJul 1, 2026-
Map releaseJul 1, 2026-
UpdateJul 1, 2026-
Current statusJul 1, 2026Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_75580.png

Downloads & links

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Map

FileDownload / File: emd_75580.map.gz / Format: CCP4 / Size: 103 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
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AxesX (Sec.)Y (Row.)Z (Col.)
1.06 Å/pix.
x 300 pix.
= 318.84 Å		1.06 Å/pix.
x 300 pix.
= 318.84 Å		1.06 Å/pix.
x 300 pix.
= 318.84 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.0628 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 6.0
Minimum - Maximum-28.453673999999999 - 51.204746
Average (Standard dev.)0.005792857 (±1.0791212)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderZYX
Origin000
Dimensions300300300
Spacing300300300
CellA=B=C: 318.84003 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : Yeast co-transcriptional pre-60S assembly intermediate Nop12 RNP

EntireName: Yeast co-transcriptional pre-60S assembly intermediate Nop12 RNP
Components
  • Complex: Yeast co-transcriptional pre-60S assembly intermediate Nop12 RNP
    • RNA: 25S ribosomal RNA
    • RNA: 5.8S ribosomal RNA
    • RNA: Internal transcribed spacer 2 ITS2
    • Protein or peptide: Nucleolar protein 16
    • Protein or peptide: 60S ribosomal protein L8-A
    • Protein or peptide: Nucleolar protein 12
    • Protein or peptide: Proteasome-interacting protein CIC1
    • Protein or peptide: 60S ribosomal protein L13-A
    • Protein or peptide: 60S ribosomal protein L15-A
    • Protein or peptide: Ribosome biogenesis protein BRX1
    • Protein or peptide: 60S ribosomal protein L36-A
    • Protein or peptide: rRNA-processing protein EBP2
    • Protein or peptide: Pescadillo homolog
    • Protein or peptide: Ribosome biogenesis protein 15
    • Protein or peptide: ATP-dependent RNA helicase HAS1
    • Protein or peptide: Ribosome biogenesis protein ERB1
    • Protein or peptide: Ribosome biogenesis protein RLP7
  • Ligand: MAGNESIUM ION

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Supramolecule #1: Yeast co-transcriptional pre-60S assembly intermediate Nop12 RNP

SupramoleculeName: Yeast co-transcriptional pre-60S assembly intermediate Nop12 RNP
type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#17
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)

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Macromolecule #1: 25S ribosomal RNA

MacromoleculeName: 25S ribosomal RNA / type: rna / ID: 1 / Number of copies: 1
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 1.0974938749999998 MDa
SequenceString: GUUUGACCUC AAAUCAGGUA GGAGUACCCG CUGAACUUAA GCAUAUCAAU AAGCGGAGGA AAAGAAACCA ACCGGGAUUG CCUUAGUAA CGGCGAGUGA AGCGGCAAAA GCUCAAAUUU GAAAUCUGGU ACCUUCGGUG CCCGAGUUGU AAUUUGGAGA G GGCAACUU ...String:
GUUUGACCUC AAAUCAGGUA GGAGUACCCG CUGAACUUAA GCAUAUCAAU AAGCGGAGGA AAAGAAACCA ACCGGGAUUG CCUUAGUAA CGGCGAGUGA AGCGGCAAAA GCUCAAAUUU GAAAUCUGGU ACCUUCGGUG CCCGAGUUGU AAUUUGGAGA G GGCAACUU UGGGGCCGUU CCUUGUCUAU GUUCCUUGGA ACAGGACGUC AUAGAGGGUG AGAAUCCCGU GUGGCGAGGA GU GCGGUUC UUUGUAAAGU GCCUUCGAAG AGUCGAGUUG UUUGGGAAUG CAGCUCUAAG UGGGUGGUAA AUUCCAUCUA AAG CUAAAU AUUGGCGAGA GACCGAUAGC GAACAAGUAC AGUGAUGGAA AGAUGAAAAG AACUUUGAAA AGAGAGUGAA AAAG UACGU GAAAUUGUUG AAAGGGAAGG GCAUUUGAUC AGACAUGGUG UUUUGUGCCC UCUGCUCCUU GUGGGUAGGG GAAUC UCGC AUUUCACUGG GCCAGCAUCA GUUUUGGUGG CAGGAUAAAU CCAUAGGAAU GUAGCUUGCC UCGGUAAGUA UUAUAG CCU GUGGGAAUAC UGCCAGCUGG GACUGAGGAC UGCGACGUAA GUCAAGGAUG CUGGCAUAAU GGUUAUAUGC CGCCCGU CU UGAAACACGG ACCAAGGAGU CUAACGUCUA UGCGAGUGUU UGGGUGUAAA ACCCAUACGC GUAAUGAAAG UGAACGUA G GUUGGGGCCU CGCAAGAGGU GCACAAUCGA CCGAUCCUGA UGUCUUCGGA UGGAUUUGAG UAAGAGCAUA GCUGUUGGG ACCCGAAAGA UGGUGAACUA UGCCUGAAUA GGGUGAAGCC AGAGGAAACU CUGGUGGAGG CUCGUAGCGG UUCUGACGUG CAAAUCGAU CGUCGAAUUU GGGUAUAGGG GCGAAAGACU AAUCGAACCA UCUAGUAGCU GGUUCCUGCC GAAGUUUCCC U CAGGAUAG CAGAAGCUCG UAUCAGUUUU AUGAGGUAAA GCGAAUGAUU AGAGGUUCCG GGGUCGAAAU GACCUUGACC UA UUCUCAA ACUUUAAAUA UGUAAGAAGU CCUUGUUACU UAAUUGAACG UGGACAUUUG AAUGAAGAGC UUUUAGUGGG CCA UUUUUG GUAAGCAGAA CUGGCGAUGC GGGAUGAACC GAACGUAGAG UUAAGGUGCC GGAAUACACG CUCAUCAGAC ACCA CAAAA GGUGUUAGUU CAUCUAGACA GCCGGACGGU GGCCAUGGAA GUCGGAAUCC GCUAAGGAGU GUGUAACAAC UCACC GGCC GAAUGAACUA GCCCUGAAAA UGGAUGGCGC UCAAGCGUGU UACCUAUACU CUACCGUCAG GGUUGAUAUG AUGCCC UGA CGAGUAGGCA GGCGUGGAGG UCAGUGACGA AGCCUAGACC GUAAGGUCGG GUCGAACGGC CUCUAGUGCA GAUCUUG GU GGUAGUAGCA AAUAUUCAAA UGAGAACUUU GAAGACUGAA GUGGGGAAAG GUUCCACGUC AACAGCAGUU GGACGUGG G UUAGUCGAUC CUAAGAGAUG GGGAAGCUCC GUUUCAAAGG CCUGAUUUUA UGCAGGCCAC CAUCGAAAGG GAAUCCGGU UAAGAUUCCG GAACCUGGAU AUGGAUUCUU CACGGUAACG UAACUGAAUG UGGAGACGUC GGCGCGAGCC CUGGGAGGAG UUAUCUUUU CUUCUUAACA GCUUAUCACC CCGGAAUUGG UUUAUCCGGA GAUGGGGUCU UAUGGCUGGA AGAGGCCAGC A CCUUUGCU GGCUCCGGUG CGCUUGUGAC GGCCCGUGAA AAUCCACAGG AAGGAAUAGU UUUCAUGCCA GGUCGUACUG AU AACCGCA GCAGGUCUCC AAGGUGAACA GCCUCUAGUU GAUAGAAUAA UGUAGAUAAG GGAAGUCGGC AAAAUAGAUC CGU AACUUC GGGAUAAGGA UUGGCUCUAA GGGUCGGGUA GUGAGGGCCU UGGUCAGACG CAGCGGGCGU GCUUGUGGAC UGCU UGGUG GGGCUUGCUC UGCUAGGCGG ACUACUUGCG UGCCUUGUUG UAGACGGCCU UGGUAGGUCU CUUGUAGACC GUCGC UUGC UACAAUUAAC GAUCAACUUA GAACUGGUAC GGACAAGGGG AAUCUGACUG UCUAAUUAAA ACAUAGCAUU GCGAUG GUC AGAAAGUGAU GUUGACGCAA UGUGAUUUCU GCCCAGUGCU CUGAAUGUCA AAGUGAAGAA AUUCAACCAA GCGCGGG UA AACGGCGGGA GUAACUAUGA CUCUCUUAAG GUAGCCAAAU GCCUCGUCAU CUAAUUAGUG ACGCGCAUGA AUGGAUUA A CGAGAUUCCC ACUGUCCCUA UCUACUAUCU AGCGAAACCA CAGCCAAGGG AACGGGCUUG GCAGAAUCAG CGGGGAAAG AAGACCCUGU UGAGCUUGAC UCUAGUUUGA CAUUGUGAAG AGACAUAGAG GGUGUAGAAU AAGUGGGAGC UUCGGCGCCA GUGAAAUAC CACUACCUUU AUAGUUUCUU UACUUAUUCA AUGAAGCGGA GCUGGAAUUC AUUUUCCACG UUCUAGCAUU C AAGGUCCC AUUCGGGGCU GAUCCGGGUU GAAGACAUUG UCAGGUGGGG AGUUUGGCUG GGGCGGCACA UCUGUUAAAC GA UAACGCA GAUGUCCUAA GGGGGGCUCA UGGAGAACAG AAAUCUCCAG UAGAACAAAA GGGUAAAAGC CCCCUUGAUU UUG AUUUUC AGUGUGAAUA CAAACCAUGA AAGUGUGGCC UAUCGAUCCU UUAGUCCCUC GGAAUUUGAG GCUAGAGGUG CCAG AAAAG UUACCACAGG GAUAACUGGC UUGUGGCAGU CAAGCGUUCA UAGCGACAUU GCUUUUUGAU UCUUCGAUGU CGGCU CUUC CUAUCAUACC GAAGCAGAAU UCGGUAAGCG UUGGAUUGUU CACCCACUAA UAGGGAACGU GAGCUGGGUU UAGACC GUC GUGAGACAGG UUAGUUUUAC CCUACUGAUG AAUGUUACCG CAAUAGUAAU UGAACUUAGU ACGAGAGGAA CAGUUCA UU CGGAUAAUUG GUUUUUGCGG CUGUCUGAUC AGGCAUUGCC GCGAAGCUAC CAUCCGCUGG AUUAUGGCUG AACGCCUC U AAGUCAGAAU CCAUGCUAGA ACGCGGUGAU UUCUUUGCUC CACACAAUAU AGAUGGAUAC GAAUAAGGCG UCCUUGUGG CGUCGCUGAA CCAUAGCAGG CUAGCAACGG UGCACUUGGC GGAAAGGCCU UGGGUGCUUG CUGGCGAAUU GCAAUGUCAU UUUGCGUGG GGAUAAAUCA UUUGUAUACG ACUUAGAUGU ACAACGGGGU AUUGUAAGCA GUAGAGUAGC CUUGUUGUUA C GAUCUGCU GAGAUUAAGC CUUUGUUGUC UGAUUUGU

GENBANK: GENBANK: NR_132207.1

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Macromolecule #2: 5.8S ribosomal RNA

MacromoleculeName: 5.8S ribosomal RNA / type: rna / ID: 2 / Number of copies: 1
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 51.012129 KDa
SequenceString:
AAAACUUUCA ACAACGGAUC UCUUGGUUCU CGCAUCGAUG AAGAACGCAG CGAAAUGCGA UACGUAAUGU GAAUUGCAGA AUUCCGUGA AUCAUCGAAU CUUUGAACGC ACAUUGCGCC CCUUGGUAUU CCAGGGGGCA UGCCUGUUUG AGCGUCAUUU

GENBANK: GENBANK: MK973014.1

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Macromolecule #3: Internal transcribed spacer 2 ITS2

MacromoleculeName: Internal transcribed spacer 2 ITS2 / type: rna / ID: 3 / Number of copies: 1
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 74.308391 KDa
SequenceString: CCUUCUCAAA CAUUCUGUUU GGUAGUGAGU GAUACUCUUU GGAGUUAACU UGAAAUUGCU GGCCUUUUCA UUGGAUGUUU UUUUUCCAA AGAGAGGUUU CUCUGCGUGC UUGAGGUAUA AUGCAAGUAC GGUCGUUUUA GGUUUUACCA ACUGCGGCUA A UCUUUUUU ...String:
CCUUCUCAAA CAUUCUGUUU GGUAGUGAGU GAUACUCUUU GGAGUUAACU UGAAAUUGCU GGCCUUUUCA UUGGAUGUUU UUUUUCCAA AGAGAGGUUU CUCUGCGUGC UUGAGGUAUA AUGCAAGUAC GGUCGUUUUA GGUUUUACCA ACUGCGGCUA A UCUUUUUU UAUACUGAGC GUAUUGGAAC GUUAUCGAUA AGAAGAGAGC GUCUAGGCGA ACAAUGUUCU UAAA

GENBANK: GENBANK: CP159571.1

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Macromolecule #4: Nucleolar protein 16

MacromoleculeName: Nucleolar protein 16 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 26.954447 KDa
SequenceString: MTSVRKRKMN RSSVGKATRR NKDKQRKINI QSNPIIAANW DYSLTMAQNY KKLGLRAKLQ TPAGGKEADL SKVVKRIPLT KPVLDEDED EDEGEDEQND YNAATVELDE NEIPEGGARI QRDKNGDVVR VVYGKKKNFD ADEDVNEIKA RDTTEETEVV K KLEELASR ...String:
MTSVRKRKMN RSSVGKATRR NKDKQRKINI QSNPIIAANW DYSLTMAQNY KKLGLRAKLQ TPAGGKEADL SKVVKRIPLT KPVLDEDED EDEGEDEQND YNAATVELDE NEIPEGGARI QRDKNGDVVR VVYGKKKNFD ADEDVNEIKA RDTTEETEVV K KLEELASR PVIRKERSQS EREEEWLEKL YKKHGDDYKK MFFDKKLNIY QQSEGDLKRR LLRWKKRNGI ASK

UniProtKB: Nucleolar protein 16

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Macromolecule #5: 60S ribosomal protein L8-A

MacromoleculeName: 60S ribosomal protein L8-A / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 28.17582 KDa
SequenceString: MAPGKKVAPA PFGAKSTKSN KTRNPLTHST PKNFGIGQAV QPKRNLSRYV KWPEYVRVQR QKKILSIRLK VPPTIAQFQY TLDRNTAAE TFKLFNKYRP ETAAEKKERL TKEAAAVAEG KSKQDASPKP YAVKYGLNHV VALIENKKAK LVLIANDVDP I ELVVFLPA ...String:
MAPGKKVAPA PFGAKSTKSN KTRNPLTHST PKNFGIGQAV QPKRNLSRYV KWPEYVRVQR QKKILSIRLK VPPTIAQFQY TLDRNTAAE TFKLFNKYRP ETAAEKKERL TKEAAAVAEG KSKQDASPKP YAVKYGLNHV VALIENKKAK LVLIANDVDP I ELVVFLPA LCKKMGVPYA IVKGKARLGT LVNQKTSAVA ALTEVRAEDE AALAKLVSTI DANFADKYDE VKKHWGGGIL GN KAQAKMD KRAKNSDSA

UniProtKB: Large ribosomal subunit protein eL8A

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Macromolecule #6: Nucleolar protein 12

MacromoleculeName: Nucleolar protein 12 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 52.046086 KDa
SequenceString: MSSAIDNLFG NIDEKKIESS VDKLFSSSCG PINKLEVKSK TRTVLPDSKK RERAAEADQE EKEASKPDVS DEQTEEVALP KVKKAKKSK RNDEDEDLEA RYYAKLLNEE AEAEDDKPTV TKTDETSVPL TSAAKKVDFK EDELEKAERT VFIGNILSTV I TSKKVYKE ...String:
MSSAIDNLFG NIDEKKIESS VDKLFSSSCG PINKLEVKSK TRTVLPDSKK RERAAEADQE EKEASKPDVS DEQTEEVALP KVKKAKKSK RNDEDEDLEA RYYAKLLNEE AEAEDDKPTV TKTDETSVPL TSAAKKVDFK EDELEKAERT VFIGNILSTV I TSKKVYKE FKKLFGTNPI AETEESGNEK EEESSKKSDN NEFAIESIRF RSISFDEALP RKVAFVQQKF HKSRDTINAY IV YKNKSAV RKICSNLNAV VFQDHHLRVD SVAHPAPHDK KRSIFVGNLD FEEIEESLWK HFEPCGDIEY VRIIRDSKTN MGK GFAYVQ FKDLQSVNKA LLLNEKPMKS QKQEDENTKK PTKKARKLRV SRCKNMKKGT TIGTGLDRNG LTDSQRTRAG RAKK ILGKA DRATLGQEIT IEGLRAKKGE GSTHLKKKKQ RSATGRVTKR SIAFKKAQAE KSKK

UniProtKB: Nucleolar protein 12

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Macromolecule #7: Proteasome-interacting protein CIC1

MacromoleculeName: Proteasome-interacting protein CIC1 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 42.596691 KDa
SequenceString: MAKKSNSKKS TPVSTPSKEK KKVIEKKSST AIPRERVIKA VNELIKFTSK PQDENNEEGN NGKKNLLEDD EEELKKDLQL IVVNNKSFT GTSKSFKLKL LNVKHSFYKP WKEASATAVK DFKVLLILKD SDIKKVSEDD LFDQLDSEGI KVDEIICGKD L KTVYKAYE ...String:
MAKKSNSKKS TPVSTPSKEK KKVIEKKSST AIPRERVIKA VNELIKFTSK PQDENNEEGN NGKKNLLEDD EEELKKDLQL IVVNNKSFT GTSKSFKLKL LNVKHSFYKP WKEASATAVK DFKVLLILKD SDIKKVSEDD LFDQLDSEGI KVDEIICGKD L KTVYKAYE ARNAFISQFS LILADDSIVT SLPKLMGGKA YNKVETTPIS IRTHANKEFS LTTLTNNIKK VYMNQLPVKL PR GTTLNVH LGNLEWLRPE EFVDNVELIS EQLIKAYQIR SIFIKTNRSP VLPLYYNQDV LDELEAKKDK IEETHEDDMV TID GVQVHL STFNKGLMEI ANPSELGSIF SKQINNAKKR SSSELEKESS ESEAVKKAKS

UniProtKB: Proteasome-interacting protein CIC1

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Macromolecule #8: 60S ribosomal protein L13-A

MacromoleculeName: 60S ribosomal protein L13-A / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 22.604164 KDa
SequenceString: MAISKNLPIL KNHFRKHWQE RVKVHFDQAG KKVSRRNARA TRAAKIAPRP LDLLRPVVRA PTVKYNRKVR AGRGFTLAEV KAAGLTAAY ARTIGIAVDH RRQNRNQEIF DANVQRLKEY QSKIIVFPRN GKAPEAEQVL SAAATFPIAQ PATDVEARAV Q DNGESAFR ...String:
MAISKNLPIL KNHFRKHWQE RVKVHFDQAG KKVSRRNARA TRAAKIAPRP LDLLRPVVRA PTVKYNRKVR AGRGFTLAEV KAAGLTAAY ARTIGIAVDH RRQNRNQEIF DANVQRLKEY QSKIIVFPRN GKAPEAEQVL SAAATFPIAQ PATDVEARAV Q DNGESAFR TLRLARSEKK FRGIREKRAR EKAEAEAEKK K

UniProtKB: Large ribosomal subunit protein eL13A

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Macromolecule #9: 60S ribosomal protein L15-A

MacromoleculeName: 60S ribosomal protein L15-A / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 24.482357 KDa
SequenceString: MGAYKYLEEL QRKKQSDVLR FLQRVRVWEY RQKNVIHRAA RPTRPDKARR LGYKAKQGFV IYRVRVRRGN RKRPVPKGAT YGKPTNQGV NELKYQRSLR ATAEERVGRR AANLRVLNSY WVNQDSTYKY FEVILVDPQH KAIRRDARYN WICDPVHKHR E ARGLTATG ...String:
MGAYKYLEEL QRKKQSDVLR FLQRVRVWEY RQKNVIHRAA RPTRPDKARR LGYKAKQGFV IYRVRVRRGN RKRPVPKGAT YGKPTNQGV NELKYQRSLR ATAEERVGRR AANLRVLNSY WVNQDSTYKY FEVILVDPQH KAIRRDARYN WICDPVHKHR E ARGLTATG KKSRGINKGH KFNNTKAGRR KTWKRQNTLS LWRYRK

UniProtKB: Large ribosomal subunit protein eL15A

+
Macromolecule #10: Ribosome biogenesis protein BRX1

MacromoleculeName: Ribosome biogenesis protein BRX1 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 33.585414 KDa
SequenceString: MSSIYKALAG KSKDNKSEKK QGNVKQFMNK QRTLLISSRG VNYRHRHLIQ DLSGLLPHSR KEPKLDTKKD LQQLNEIAEL YNCNNVLFF EARKHQDLYL WLSKPPNGPT IKFYIQNLHT MDELNFTGNC LKGSRPVLSF DQRFESSPHY QLIKELLVHN F GVPPNARK ...String:
MSSIYKALAG KSKDNKSEKK QGNVKQFMNK QRTLLISSRG VNYRHRHLIQ DLSGLLPHSR KEPKLDTKKD LQQLNEIAEL YNCNNVLFF EARKHQDLYL WLSKPPNGPT IKFYIQNLHT MDELNFTGNC LKGSRPVLSF DQRFESSPHY QLIKELLVHN F GVPPNARK SKPFIDHVMS FSIVDDKIWV RTYEISHSTK NKEEYEDGEE DISLVEIGPR FVMTVILILE GSFGGPKIYE NK QYVSPNV VRAQIKQQAA EEAKSRAEAA VERKIKRREN VLAADPLSND ALFK

UniProtKB: Ribosome biogenesis protein BRX1

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Macromolecule #11: 60S ribosomal protein L36-A

MacromoleculeName: 60S ribosomal protein L36-A / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 11.151259 KDa
SequenceString:
MTVKTGIAIG LNKGKKVTSM TPAPKISYKK GAASNRTKFV RSLVREIAGL SPYERRLIDL IRNSGEKRAR KVAKKRLGSF TRAKAKVEE MNNIIAASRR H

UniProtKB: Large ribosomal subunit protein eL36A

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Macromolecule #12: rRNA-processing protein EBP2

MacromoleculeName: rRNA-processing protein EBP2 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 49.842613 KDa
SequenceString: MAKGFKLKEL LSHQKEIEKA EKLENDLKKK KSQELKKEEP TIVTASNLKK LEKKEKKADV KKEVAADTEE YQSQALSKKE KRKLKKELK KMQEQDATEA QKHMSGDEDE SGDDREEEEE EEEEEEGRLD LEKLAKSDSE SEDDSESEND SEEDEDVVAK E ESEEKEEQ ...String:
MAKGFKLKEL LSHQKEIEKA EKLENDLKKK KSQELKKEEP TIVTASNLKK LEKKEKKADV KKEVAADTEE YQSQALSKKE KRKLKKELK KMQEQDATEA QKHMSGDEDE SGDDREEEEE EEEEEEGRLD LEKLAKSDSE SEDDSESEND SEEDEDVVAK E ESEEKEEQ EEEQDVPLSD VEFDSDADVV PHHKLTVNNT KAMKHALERV QLPWKKHSFQ EHQSVTSETN TDEHIKDIYD DT ERELAFY KQSLDAVLVA RDELKRLKVP FKRPLDYFAE MVKSDEHMDK IKGKLIEEAS DKKAREEARR QRQLKKFGKQ VQN ATLQKR QLEKRETLEK IKSLKNKRKH NEIDHSEFNV GVEEEVEGKR FDRGRPNGKR AAKNAKYGQG GMKRFKRKND ATSS ADVSG FSSRKMKGKT NRPGKSRRAR RF

UniProtKB: rRNA-processing protein EBP2

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Macromolecule #13: Pescadillo homolog

MacromoleculeName: Pescadillo homolog / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 69.984148 KDa
SequenceString: MRIKKKNTRG NARNFITRSQ AVRKLQVSLA DFRRLCIFKG IYPREPRNKK KANKGSTAPT TFYYAKDIQY LMHEPVLAKF REHKTFARK LTRALGRGEV SSAKRLEENR DSYTLDHIIK ERYPSFPDAI RDIDDALNML FLFSNLPSTN QVSSKIINDA Q KICNQWLA ...String:
MRIKKKNTRG NARNFITRSQ AVRKLQVSLA DFRRLCIFKG IYPREPRNKK KANKGSTAPT TFYYAKDIQY LMHEPVLAKF REHKTFARK LTRALGRGEV SSAKRLEENR DSYTLDHIIK ERYPSFPDAI RDIDDALNML FLFSNLPSTN QVSSKIINDA Q KICNQWLA YVAKERLVRK VFVSIKGVYY QANIKGEEVR WLVPFKFPEN IPSDVDFRIM LTFLEFYSTL LHFVLYKLYT DS GLIYPPK LDLKKDKIIS GLSSYILESR QEDSLLKLDP TEIEEDVKVE SLDASTLKSA LNADEANTDE TEKEEEQEKK QEK EQEKEQ NEETELDTFE DNNKNKGDIL IQPSKYDSPV ASLFSAFVFY VSREVPIDIL EFLILSCGGN VISEAAMDQI ENKK DIDMS KVTHQIVDRP VLKNKVAGRT YIQPQWIFDC INKGELVPAN KYLPGEALPP HLSPWGDAIG YDPTAPVEEG EEEES ESES ESEDQVEEED QEVVAGEEDD DDDEELQAQK ELELEAQGIK YSETSEADKD VNKSKNKKRK VDEEEEEKKL KMIMMS NKQ KKLYKKMKYS NAKKEEQAEN LKKKKKQIAK QKAKLNKLDS KK

UniProtKB: Pescadillo homolog

+
Macromolecule #14: Ribosome biogenesis protein 15

MacromoleculeName: Ribosome biogenesis protein 15 / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 25.499186 KDa
SequenceString: MVKSTSKTST KETVTKQPTE EKPIQEKEEL ALETSSSSSD EEDEKDEDEI EGLAASDDEQ SGTHKIKRLN PKKQANEKKS KDKKTLEEY SGIIYVSRLP HGFHEKELSK YFAQFGDLKE VRLARNKKTG NSRHYGFLEF VNKEDAMIAQ ESMNNYLLMG H LLQVRVLP ...String:
MVKSTSKTST KETVTKQPTE EKPIQEKEEL ALETSSSSSD EEDEKDEDEI EGLAASDDEQ SGTHKIKRLN PKKQANEKKS KDKKTLEEY SGIIYVSRLP HGFHEKELSK YFAQFGDLKE VRLARNKKTG NSRHYGFLEF VNKEDAMIAQ ESMNNYLLMG H LLQVRVLP KGAKIEKLYK YKKRVLVEKG ITKPVKQLKD NMKQKHEERI KKLAKSGIEF KW

UniProtKB: Ribosome biogenesis protein 15

+
Macromolecule #15: ATP-dependent RNA helicase HAS1

MacromoleculeName: ATP-dependent RNA helicase HAS1 / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO / EC number: RNA helicase
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 56.798348 KDa
SequenceString: MATPSNKRSR DSESTEEPVV DEKSTSKQNN AAPEGEQTTC VEKFEELKLS QPTLKAIEKM GFTTMTSVQA RTIPPLLAGR DVLGAAKTG SGKTLAFLIP AIELLHSLKF KPRNGTGIIV ITPTRELALQ IFGVARELME FHSQTFGIVI GGANRRQEAE K LMKGVNML ...String:
MATPSNKRSR DSESTEEPVV DEKSTSKQNN AAPEGEQTTC VEKFEELKLS QPTLKAIEKM GFTTMTSVQA RTIPPLLAGR DVLGAAKTG SGKTLAFLIP AIELLHSLKF KPRNGTGIIV ITPTRELALQ IFGVARELME FHSQTFGIVI GGANRRQEAE K LMKGVNML IATPGRLLDH LQNTKGFVFK NLKALIIDEA DRILEIGFED EMRQIIKILP NEDRQSMLFS ATQTTKVEDL AR ISLRPGP LFINVVPETD NSTADGLEQG YVVCDSDKRF LLLFSFLKRN QKKKIIVFLS SCNSVKYYAE LLNYIDLPVL ELH GKQKQQ KRTNTFFEFC NAERGILICT DVAARGLDIP AVDWIIQFDP PDDPRDYIHR VGRTARGTKG KGKSLMFLTP NELG FLRYL KASKVPLNEY EFPENKIANV QSQLEKLIKS NYYLHQTAKD GYRSYLQAYA SHSLKTVYQI DKLDLAKVAK SYGFP VPPK VNITIGASGK TPNTKRRKTH K

UniProtKB: ATP-dependent RNA helicase HAS1

+
Macromolecule #16: Ribosome biogenesis protein ERB1

MacromoleculeName: Ribosome biogenesis protein ERB1 / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 91.830609 KDa
SequenceString: MMAKNNKTTE AKMSKKRAAS EESDVEEDED KLLSVDGLID AEASESDEDD DEYESAVEEK ESSSDKEAQD DSDDDSDAEL NKLLAEEEG DGEEDYDSSE FSDDTTSLTD RLSGVKLQTI VDPNIYSKYA DGSDRIIKPE INPVYDSDDS DAETQNTIGN I PLSAYDEM ...String:
MMAKNNKTTE AKMSKKRAAS EESDVEEDED KLLSVDGLID AEASESDEDD DEYESAVEEK ESSSDKEAQD DSDDDSDAEL NKLLAEEEG DGEEDYDSSE FSDDTTSLTD RLSGVKLQTI VDPNIYSKYA DGSDRIIKPE INPVYDSDDS DAETQNTIGN I PLSAYDEM PHIGYDINGK RIMRPAKGSA LDQLLDSIEL PEGWTGLLDK NSGSSLNLTK EELELISKIQ RNEQTDDSIN PY EPLIDWF TRHEEVMPLT AVPEPKRRFV PSKNEAKRVM KIVRAIREGR IIPPKKLKEM KEKEKIENYQ YDLWGDSTET NDH VMHLRA PKLPPPTNEE SYNPPEEYLL SPEEKEAWEN TEYSERERNF IPQKYSALRK VPGYGESIRE RFERSLDLYL APRV RKNKL NIDPNSLIPE LPSPKDLRPF PIRCSTIYAG HKGKVRTLSI DPSGLWLATG SDDGTVRVWE ILTGREVYRT TLIDD EENP DYHIECIEWN PDANNGILAV AVGENIHLIV PPIFGYDIEN NGKTKIEDGF GYDTFGTVKK SNLEVNENGD GDEDGE NES AKNAVKKQVA QWNKPSQKQL EKDICITISC KKTVKKLSWH RKGDYFVTVQ PDSGNTSVLI HQVSKHLTQS PFKKSKG II MDAKFHPFKP QLFVCSQRYV RIYDLSQQIL VKKLLPGARW LSKIDIHPRG DNLIASSFDK RVLWHDLDLA STPYKTLR Y HEKAVRSVNF HKKLPLFSSA ADDGTIHVFH ATVYDDMMKN PMIVPLKKLT GHKVINSLGV LDAIWHPREA WLFSAGADN TARLWTT

UniProtKB: Ribosome biogenesis protein ERB1

+
Macromolecule #17: Ribosome biogenesis protein RLP7

MacromoleculeName: Ribosome biogenesis protein RLP7 / type: protein_or_peptide / ID: 17 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 36.621074 KDa
SequenceString: MSSTQDSKAQ TLNSNPEILL RKRRNADRTR IERQELAKKK REEQIKKKRS NKNKFVRAES IVAKTLATSR EKERIKRVSI LEDKKAKNE TQHIASGKDF ILKITEKANG AEENSVDLEE TEEEEDDGLI REKTTYDGKP ALLFIVRVRG PLAVNIPNKA F KILSLLRL ...String:
MSSTQDSKAQ TLNSNPEILL RKRRNADRTR IERQELAKKK REEQIKKKRS NKNKFVRAES IVAKTLATSR EKERIKRVSI LEDKKAKNE TQHIASGKDF ILKITEKANG AEENSVDLEE TEEEEDDGLI REKTTYDGKP ALLFIVRVRG PLAVNIPNKA F KILSLLRL VETNTGVFVK LTKNVYPLLK VIAPYVVIGK PSLSSIRSLI QKRGRIIYKG ENEAEPHEIV LNDNNIVEEQ LG DHGIICV EDIIHEIATM GESFSVCNFF LQPFKLNREV SGFGSLNRLR KIKQREAESR TRQFSNAATA PVIEVDIDSL LAK LN

UniProtKB: Ribosome biogenesis protein RLP7

+
Macromolecule #18: MAGNESIUM ION

MacromoleculeName: MAGNESIUM ION / type: ligand / ID: 18 / Number of copies: 5 / Formula: MG
Molecular weightTheoretical: 24.305 Da

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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: 7.5
GridModel: Quantifoil R3.5/1 / Material: GOLD / Mesh: 400 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: AIR
VitrificationCryogen name: ETHANE / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeTFS KRIOS
SoftwareName: SerialEM
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 51.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 1.7 µm / Nominal defocus min: 0.7000000000000001 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

CTF correctionSoftware - Name: cryoSPARC / Type: PHASE FLIPPING AND AMPLITUDE CORRECTION
Startup modelType of model: NONE
Final reconstructionResolution.type: BY AUTHOR / Resolution: 2.78 Å / Resolution method: OTHER / Software - Name: cryoSPARC / Number images used: 443499
Initial angle assignmentType: RANDOM ASSIGNMENT / Software - Name: cryoSPARC
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC

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Atomic model buiding 1

Initial modelPDB ID:

8e5t


Chain - Source name: PDB / Chain - Initial model type: experimental model
RefinementSpace: REAL / Protocol: RIGID BODY FIT
Output model

PDB-11aa:
Yeast co-transcriptional pre-60S assembly intermediate Nop12 RNP

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