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- EMDB-24280: State E2 nucleolar 60S ribosomal intermediate - Local Map for Noc... -

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Basic information

Entry
Database: EMDB / ID: EMD-24280
TitleState E2 nucleolar 60S ribosomal intermediate - Local Map for Noc2/Noc3 region
Map dataState E2 Noc2/Noc3 locally refined map
Sample
  • Complex: Nucleolar 60S intermediate purified with tags on Ytm1 and Spb4
    • RNA: x 1 types
    • Protein or peptide: x 15 types
Keywordsribosome biogenesis / DEAD-box ATPases / methyltransferases / nucleolus / RIBOSOME
Function / homology
Function and homology information


25S rRNA (cytosine2870-C5)-methyltransferase / 27S pre-rRNA (guanosine2922-2'-O)-methyltransferase / Noc1p-Noc2p complex / rRNA (guanosine-2'-O-)-methyltransferase activity / Noc2p-Noc3p complex / nuclear division / exonucleolytic trimming to generate mature 5'-end of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA (uridine-2'-O-)-methyltransferase activity / rRNA (guanine) methyltransferase activity / rRNA (cytosine-C5-)-methyltransferase activity ...25S rRNA (cytosine2870-C5)-methyltransferase / 27S pre-rRNA (guanosine2922-2'-O)-methyltransferase / Noc1p-Noc2p complex / rRNA (guanosine-2'-O-)-methyltransferase activity / Noc2p-Noc3p complex / nuclear division / exonucleolytic trimming to generate mature 5'-end of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA (uridine-2'-O-)-methyltransferase activity / rRNA (guanine) methyltransferase activity / rRNA (cytosine-C5-)-methyltransferase activity / endonucleolytic cleavage in ITS1 upstream of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / RNA strand annealing activity / intracellular mRNA localization / RNA folding chaperone / pre-replicative complex assembly involved in nuclear cell cycle DNA replication / endonucleolytic cleavage in 5'-ETS of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / PeBoW complex / endonucleolytic cleavage to generate mature 5'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / nuclear pre-replicative complex / rRNA primary transcript binding / maturation of 5.8S rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA base methylation / rRNA methylation / protein folding chaperone complex / Major pathway of rRNA processing in the nucleolus and cytosol / SRP-dependent cotranslational protein targeting to membrane / GTP hydrolysis and joining of the 60S ribosomal subunit / preribosome, large subunit precursor / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / Formation of a pool of free 40S subunits / L13a-mediated translational silencing of Ceruloplasmin expression / ribosomal large subunit export from nucleus / DNA replication initiation / mRNA transport / ribonucleoprotein complex binding / maturation of LSU-rRNA / 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) / nuclear periphery / assembly of large subunit precursor of preribosome / ribosomal large subunit biogenesis / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / rRNA processing / ribosome biogenesis / ribosome binding / 5S rRNA binding / ribosomal large subunit assembly / large ribosomal subunit rRNA binding / cytosolic large ribosomal subunit / cytoplasmic translation / negative regulation of translation / rRNA binding / structural constituent of ribosome / ribonucleoprotein complex / cell division / mRNA binding / chromatin binding / nucleolus / mitochondrion / DNA binding / RNA binding / nucleoplasm / identical protein binding / nucleus / cytoplasm / cytosol
Similarity search - Function
Nucleolar protein 12 / Nucleolar protein 12 (25kDa) / 60S ribosomal subunit assembly/export protein Loc1 / Nucleolar complex protein 2 / Noc2p family / Nucleolar complex-associated protein 3, N-terminal / Nucleolar complex-associated protein 3 / Nucleolar complex-associated protein / Ribosome biogenesis factor, NIP7 / RNA (C5-cytosine) methyltransferase, NOP2 ...Nucleolar protein 12 / Nucleolar protein 12 (25kDa) / 60S ribosomal subunit assembly/export protein Loc1 / Nucleolar complex protein 2 / Noc2p family / Nucleolar complex-associated protein 3, N-terminal / Nucleolar complex-associated protein 3 / Nucleolar complex-associated protein / Ribosome biogenesis factor, NIP7 / RNA (C5-cytosine) methyltransferase, NOP2 / : / UPF0113, pre-PUA domain / UPF0113 PUA domain / NIP7/UPF0113, pre-PUA domain / UPF0113, PUA domain / Nop2p / Bacterial Fmu (Sun)/eukaryotic nucleolar NOL1/Nop2p, conserved site / NOL1/NOP2/sun family signature. / Ribosomal RNA methyltransferase, SPB1-like, C-terminal / Ribosomal RNA methyltransferase Spb1, domain of unknown function DUF3381 / AdoMet-dependent rRNA methyltransferase SPB1-like / Spb1 C-terminal domain / Ribosomal RNA methyltransferase Spb1, DUF3381 / Eukaryotic rRNA processing / Eukaryotic rRNA processing protein EBP2 / : / Ribosomal RNA large subunit methyltransferase E / Ribosome biogenesis protein BRX1 / BOP1, N-terminal domain / WD repeat BOP1/Erb1 / BOP1NT (NUC169) domain / BOP1NT (NUC169) domain / SAM-dependent methyltransferase RsmB/NOP2-type / RNA (C5-cytosine) methyltransferase / : / 16S rRNA methyltransferase RsmB/F / SAM-dependent MTase RsmB/NOP-type domain profile. / Putative RNA-binding Domain in PseudoUridine synthase and Archaeosine transglycosylase / CCAAT-binding factor / CBF/Mak21 family / PUA domain / PUA domain superfamily / PUA domain profile. / Ribosomal biogenesis NSA2 family / Brix domain / Brix domain / Brix domain profile. / Brix / PUA-like superfamily / Ribosomal protein L34e, conserved site / Ribosomal protein L34e signature. / Ribosomal protein L36e signature. / : / Ribosomal protein L34Ae / Ribosomal protein L34e / Ribosomal protein L7A/L8 / Ribosomal protein L36e / Ribosomal protein L36e domain superfamily / Ribosomal protein L36e / Ribosomal RNA methyltransferase, FtsJ domain / FtsJ-like methyltransferase / Ribosomal protein L15e, conserved site / Ribosomal protein L15e signature. / Ribosomal_L15e / Ribosomal protein L15e / Ribosomal protein L15e core domain superfamily / Ribosomal L15 / Ribosomal protein L7Ae conserved site / Ribosomal protein L7Ae signature. / Ribosomal protein S8e/ribosomal biogenesis NSA2 / Ribosomal protein S8e / Ribosomal protein L7Ae/L8/Nhp2 family / : / Ribosomal protein L7Ae/L30e/S12e/Gadd45 / Ribosomal protein L7Ae/L30e/S12e/Gadd45 family / 50S ribosomal protein L30e-like / WD domain, G-beta repeat / Ribosomal protein L23/L15e core domain superfamily / Armadillo-type fold / 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 / S-adenosyl-L-methionine-dependent methyltransferase superfamily
Similarity search - Domain/homology
RRP17 isoform 1 / Ribosome biogenesis protein NSA2 / Large ribosomal subunit protein eL36A / Large ribosomal subunit protein eL15A / Large ribosomal subunit protein eL8A / 27S pre-rRNA (guanosine(2922)-2'-O)-methyltransferase / rRNA-processing protein EBP2 / Nucleolar complex protein 2 / 25S rRNA (cytosine(2870)-C(5))-methyltransferase / 60S ribosomal subunit assembly/export protein LOC1 ...RRP17 isoform 1 / Ribosome biogenesis protein NSA2 / Large ribosomal subunit protein eL36A / Large ribosomal subunit protein eL15A / Large ribosomal subunit protein eL8A / 27S pre-rRNA (guanosine(2922)-2'-O)-methyltransferase / rRNA-processing protein EBP2 / Nucleolar complex protein 2 / 25S rRNA (cytosine(2870)-C(5))-methyltransferase / 60S ribosomal subunit assembly/export protein LOC1 / Large ribosomal subunit protein eL34A / Ribosome biogenesis protein ERB1 / Nucleolar complex-associated protein 3 / Ribosome biogenesis protein BRX1 / 60S ribosome subunit biogenesis protein NIP7
Similarity search - Component
Biological speciesSaccharomyces cerevisiae BY4741 (yeast)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.17 Å
AuthorsCruz VE / Sekulski K / Peddada N / Erzberger JP
Funding support United States, 3 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM135617-01 United States
Cancer Prevention and Research Institute of Texas (CPRIT)RR150074 United States
Robert A. Welch FoundationI-1897 United States
CitationJournal: Nat Struct Mol Biol / Year: 2022
Title: Sequence-specific remodeling of a topologically complex RNP substrate by Spb4.
Authors: Victor Emmanuel Cruz / Kamil Sekulski / Nagesh Peddada / Carolin Sailer / Sahana Balasubramanian / Christine S Weirich / Florian Stengel / Jan P Erzberger /
Abstract: DEAD-box ATPases are ubiquitous enzymes essential in all aspects of RNA biology. However, the limited in vitro catalytic activities described for these enzymes are at odds with their complex cellular ...DEAD-box ATPases are ubiquitous enzymes essential in all aspects of RNA biology. However, the limited in vitro catalytic activities described for these enzymes are at odds with their complex cellular roles, most notably in driving large-scale RNA remodeling steps during the assembly of ribonucleoproteins (RNPs). We describe cryo-EM structures of 60S ribosomal biogenesis intermediates that reveal how context-specific RNA unwinding by the DEAD-box ATPase Spb4 results in extensive, sequence-specific remodeling of rRNA secondary structure. Multiple cis and trans interactions stabilize Spb4 in a post-catalytic, high-energy intermediate that drives the organization of the three-way junction at the base of rRNA domain IV. This mechanism explains how limited strand separation by DEAD-box ATPases is leveraged to provide non-equilibrium directionality and ensure efficient and accurate RNP assembly.
History
DepositionJun 22, 2021-
Header (metadata) releaseNov 9, 2022-
Map releaseNov 9, 2022-
UpdateJun 5, 2024-
Current statusJun 5, 2024Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_24280.png

Downloads & links

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Map

FileDownload / File: emd_24280.map.gz / Format: CCP4 / Size: 282.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationState E2 Noc2/Noc3 locally refined map
Projections & slices

Image control

Size
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Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.08 Å/pix.
x 420 pix.
= 453.6 Å		1.08 Å/pix.
x 420 pix.
= 453.6 Å		1.08 Å/pix.
x 420 pix.
= 453.6 Å

Surface

Projections

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Images are generated by Spider.

Voxel sizeX=Y=Z: 1.08 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.0125
Minimum - Maximum-0.038572192 - 0.08519787
Average (Standard dev.)0.000023658962 (±0.0015540959)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions420420420
Spacing420420420
CellA=B=C: 453.6 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: State E2 Noc2/Noc3 locally refined half map 1

Fileemd_24280_half_map_1.map
AnnotationState E2 Noc2/Noc3 locally refined half map 1
Projections & Slices
AxesZYX

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Histogram (log scale)

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Half map: State E2 Noc2/Noc3 locally refined half map 2

Fileemd_24280_half_map_2.map
AnnotationState E2 Noc2/Noc3 locally refined half map 2
Projections & Slices
AxesZYX

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Sample components

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Entire : Nucleolar 60S intermediate purified with tags on Ytm1 and Spb4

EntireName: Nucleolar 60S intermediate purified with tags on Ytm1 and Spb4
Components
  • Complex: Nucleolar 60S intermediate purified with tags on Ytm1 and Spb4
    • RNA: 25S rRNA
    • Protein or peptide: 60S ribosomal subunit assembly/export protein LOC1
    • Protein or peptide: Nucleolar complex protein 2
    • Protein or peptide: Ribosome biogenesis protein BRX1
    • Protein or peptide: 60S ribosomal protein L8-A
    • Protein or peptide: Nucleolar complex-associated protein 3
    • Protein or peptide: rRNA-processing protein EBP2
    • Protein or peptide: 60S ribosomal protein L15-A
    • Protein or peptide: 60S ribosomal protein L34-A
    • Protein or peptide: 60S ribosomal protein L36-A
    • Protein or peptide: 60S ribosome subunit biogenesis protein NIP7
    • Protein or peptide: Ribosome biogenesis protein ERB1
    • Protein or peptide: 25S rRNA (cytosine(2870)-C(5))-methyltransferase
    • Protein or peptide: Ribosome biogenesis protein NSA2
    • Protein or peptide: 27S pre-rRNA (guanosine(2922)-2'-O)-methyltransferase
    • Protein or peptide: RRP17 isoform 1

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Supramolecule #1: Nucleolar 60S intermediate purified with tags on Ytm1 and Spb4

SupramoleculeName: Nucleolar 60S intermediate purified with tags on Ytm1 and Spb4
type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast)
Molecular weightTheoretical: 3.3 MDa

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Macromolecule #1: 25S rRNA

MacromoleculeName: 25S rRNA / type: rna / ID: 1 / Number of copies: 1
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 49.28523 KDa
SequenceString:
GCUGAACUUA AGCAUAAAGC GGAGUCGAGU UGUUUGCAGC UCUAAGCUAC GGUGAUCGUC GAAGGGGCGA AAGACAAGAU GGGGAAGCU CCGUUUCAAU GUUGAGCUUG ACUCUAGUUG UGGGGAGUAA AGUUACCACA UCGUGAGACA GGU

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Macromolecule #2: 60S ribosomal subunit assembly/export protein LOC1

MacromoleculeName: 60S ribosomal subunit assembly/export protein LOC1 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 23.652645 KDa
SequenceString: MAPKKPSKRQ NLRREVAPEV FQDSQARNQL ANVPHLTEKS AQRKPSKTKV KKEQSLARLY GAKKDKKGKY SEKDLNIPTL NRAIVPGVK IRRGKKGKKF IADNDTLTLN RLITTIGDKY DDIAESKLEK ARRLEEIREL KRKEIERKEA LKQDKLEEKK D EIKKKSSV ...String:
MAPKKPSKRQ NLRREVAPEV FQDSQARNQL ANVPHLTEKS AQRKPSKTKV KKEQSLARLY GAKKDKKGKY SEKDLNIPTL NRAIVPGVK IRRGKKGKKF IADNDTLTLN RLITTIGDKY DDIAESKLEK ARRLEEIREL KRKEIERKEA LKQDKLEEKK D EIKKKSSV ARTIRRKNKR DMLKSEAKAS ESKTEGRKVK KVSFAQ

UniProtKB: 60S ribosomal subunit assembly/export protein LOC1

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Macromolecule #3: Nucleolar complex protein 2

MacromoleculeName: Nucleolar complex protein 2 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 81.719289 KDa
SequenceString: MGKVSKSTKK FQSKHLKHTL DQRRKEKIQK KRIQGRRGNK TDQEKADAAG TREQQQLKKS AKEEVFKDMS VETFFEKGIE IPKENKKLK KKTTKEQSDE DSSSSEEEED MGQSMAKLAE KDPEFYKYLE ENDKDLLDFA GTNPLDGIDS QDEGEDAERN S NIEEKSEQ ...String:
MGKVSKSTKK FQSKHLKHTL DQRRKEKIQK KRIQGRRGNK TDQEKADAAG TREQQQLKKS AKEEVFKDMS VETFFEKGIE IPKENKKLK KKTTKEQSDE DSSSSEEEED MGQSMAKLAE KDPEFYKYLE ENDKDLLDFA GTNPLDGIDS QDEGEDAERN S NIEEKSEQ MELEKEKIEL SLKLVRKWKK QLHDSPSLKL LRNIISAFKV AVNLNKEENI EDYKYAITDE KAFHELMFMV LK DVPQAIQ KMAPYKIVKG ARTLPNGGNV SRVSSIVKSH AGSLLILLND ITNTETAALV LHSVNELMPY LLSYRRILKE LIK SIVGVW STTRELETQI ASFAFLINTT KEFKKSMLET TLKTTYSTFI KSCRKTNMRS MPLINFQKNS AAELFGIDEV LGYQ VGFEY IRQLAIHLRN TMNATTKKSS KINSAEAYKI VYNWQFCHSL DFWSRVLSFA CQPEKENGSE SPLRQLIYPL VQVTL GVIR LIPTPQFFPL RFYLIKSLIR LSQNSGVFIP IYPLLSEILT STAFTKAPKK SPNLAAFDFE HNIKCTQAYL NTKIYQ EGL SEQFVDLLGD YFALYCKNIA FPELVTPVII SLRRYIKTST NVKLNKRLST VVEKLNQNST FIQEKRSDVE FGPTNKS EV SRFLNDVAWN KTPLGSYVAV QREVKEEKAR LMRESMEEQD KERETEEAKL LNSLESDDDN EDVEMSDA

UniProtKB: Nucleolar complex protein 2

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Macromolecule #4: Ribosome biogenesis protein BRX1

MacromoleculeName: Ribosome biogenesis protein BRX1 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
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 #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 BY4741 (yeast) / Strain: BY4741
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 complex-associated protein 3

MacromoleculeName: Nucleolar complex-associated protein 3 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 75.689008 KDa
SequenceString: MAKRNRSQFR IQERTAKKRK HEDSLLEGNV FQNAPEDMDE NTIYSAKGSS WDEEEQDYEM VPRKNRSDTS NLVEGLPIKV NGKVERKLH KAQEKPKDDD EEDEDSNDSS EDDEGPNEEQ EAEAKEDEPD TEEKILQLKE DIADLVTKVM EEPEENTAAL G RLCKMVES ...String:
MAKRNRSQFR IQERTAKKRK HEDSLLEGNV FQNAPEDMDE NTIYSAKGSS WDEEEQDYEM VPRKNRSDTS NLVEGLPIKV NGKVERKLH KAQEKPKDDD EEDEDSNDSS EDDEGPNEEQ EAEAKEDEPD TEEKILQLKE DIADLVTKVM EEPEENTAAL G RLCKMVES KNPNTCKFSM LALVPVFKSI IPGYRIRPLT ETEKKEKVSK EVSKLRNFEQ ALVYNYKNYV GRLQSLSKTP SN AAPIQVS LGILATQAAK ELISTASHFN FRTDIFTLLL RRICKPRIST DPTSIQIIQT FETLLNEDEE GSISFEILRI FNK ILKTRN FNIEESVLNM LLSLDVLHDY DPNTKLKGNV SAPKLKKKDR VHLSKKQRKA RKEMQQIEEE MRNAEQAVSA EERE RNQSE ILKIVFTIYL NILKNNAKTL IGSVLEGLTK FGNMANFDLL GDFLEVMKEL ISDTEFDNLS SAEVRKALLC IVSAF SLIS NTQYMKVNVD LSKFVDGLYA LLPYICLDAD IELSYRSLRL ADPLNNEIIK PSVNVSTKAE LLLKALDHVF FRSKSG TKE RATAFTKRLY MCISHTPEKT SIAILKFIDK LMNRYPEISG LYSSEDRIGN GHFIMEADNP SRSNPEAATL WDNALLE KH YCPVVTKGLR SLSSRSKECS K

UniProtKB: Nucleolar complex-associated protein 3

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Macromolecule #7: rRNA-processing protein EBP2

MacromoleculeName: rRNA-processing protein EBP2 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
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 #8: 60S ribosomal protein L15-A

MacromoleculeName: 60S ribosomal protein L15-A / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
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

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Macromolecule #9: 60S ribosomal protein L34-A

MacromoleculeName: 60S ribosomal protein L34-A / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 13.673196 KDa
SequenceString:
MAQRVTFRRR NPYNTRSNKI KVVKTPGGIL RAQHVKKLAT RPKCGDCGSA LQGISTLRPR QYATVSKTHK TVSRAYGGSR CANCVKERI IRAFLIEEQK IVKKVVKEQT EAAKKSEKKA KK

UniProtKB: Large ribosomal subunit protein eL34A

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Macromolecule #10: 60S ribosomal protein L36-A

MacromoleculeName: 60S ribosomal protein L36-A / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
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 #11: 60S ribosome subunit biogenesis protein NIP7

MacromoleculeName: 60S ribosome subunit biogenesis protein NIP7 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 20.411699 KDa
SequenceString:
MRQLTEEETK VVFEKLAGYI GRNISFLVDN KELPHVFRLQ KDRVYYVPDH VAKLATSVAR PNLMSLGICL GKFTKTGKFR LHITSLTVL AKHAKYKIWI KPNGEMPFLY GNHVLKAHVG KMSDDIPEHA GVIVFAMNDV PLGFGVSAKS TSESRNMQPT G IVAFRQAD IGEYLRDEDT LFT

UniProtKB: 60S ribosome subunit biogenesis protein NIP7

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Macromolecule #12: Ribosome biogenesis protein ERB1

MacromoleculeName: Ribosome biogenesis protein ERB1 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
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 #13: 25S rRNA (cytosine(2870)-C(5))-methyltransferase

MacromoleculeName: 25S rRNA (cytosine(2870)-C(5))-methyltransferase / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO / EC number: 25S rRNA (cytosine2870-C5)-methyltransferase
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 69.912164 KDa
SequenceString: MGSRRHKNKQ AAPPTLEEFQ ARKEKKANRK LEKGKRPSTT QGDEVSDRKK KKSKPFKKSR KEEEEVVEED KDLPEVDLEE LSKARKSLF DDEEDDDEAG LVDEELKDEF DLEQEYDYDE DEDNDAHPIF SDDDDEADLE ELNAQNMEAL SKKLDEEEAE E AEEAEMEL ...String:
MGSRRHKNKQ AAPPTLEEFQ ARKEKKANRK LEKGKRPSTT QGDEVSDRKK KKSKPFKKSR KEEEEVVEED KDLPEVDLEE LSKARKSLF DDEEDDDEAG LVDEELKDEF DLEQEYDYDE DEDNDAHPIF SDDDDEADLE ELNAQNMEAL SKKLDEEEAE E AEEAEMEL VEAENMQPRA DILPTEEQEE MMAQETPNLT STRTRMIEIV KVLENFKTLG AEGRSRGEYV DRLLKDICEY FG YTPFLAE KLFNLFSPAE AMEFFEANEI ARPITIRTNT LKTRRRDLAQ TLVNRGVNLQ PIGSWTKVGL QIFDSQVPIG ATP EYLAGH YILQAASSFL PVIALDPHEN ERILDMAAAP GGKTTYISAM MKNTGCVFAN DANKSRTKSL IANIHRLGCT NTIV CNYDA REFPKVIGGF DRILLDAPCS GTGVIGKDQS VKVSRTEKDF IQIPHLQKQL LLSAIDSVDC NSKHGGVIVY STCSV AVEE DEAVIDYALR KRPNVKLVDT GLAIGKEAFT SYRGKKFHPS VKLARRYYPH TYNVDGFFVA KFQKIGPSSF DDNQAS AKE KETAARKEAL EEGIIHSDFA TFEDEEDDKY IEKSVKNNLL KKGVNPKAKR PSNEK

UniProtKB: 25S rRNA (cytosine(2870)-C(5))-methyltransferase

+
Macromolecule #14: Ribosome biogenesis protein NSA2

MacromoleculeName: Ribosome biogenesis protein NSA2 / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 29.786783 KDa
SequenceString: MPQNDYIERH IKQHGKRLDH EERKRKREAR ESHKISERAQ KLTGWKGKQF AKKRYAEKVS MRKKIKAHEQ SKVKGSSKPL DTDGDALPT YLLDREQNNT AKAISSSIKQ KRLEKADKFS VPLPKVRGIS EEEMFKVIKT GKSRSKSWKR MITKHTFVGE G FTRRPVKM ...String:
MPQNDYIERH IKQHGKRLDH EERKRKREAR ESHKISERAQ KLTGWKGKQF AKKRYAEKVS MRKKIKAHEQ SKVKGSSKPL DTDGDALPT YLLDREQNNT AKAISSSIKQ KRLEKADKFS VPLPKVRGIS EEEMFKVIKT GKSRSKSWKR MITKHTFVGE G FTRRPVKM ERIIRPSALR QKKANVTHPE LGVTVFLPIL AVKKNPQSPM YTQLGVLTKG TIIEVNVSEL GMVTAGGKVV WG KYAQVTN EPDRDGCVNA VLLV

UniProtKB: Ribosome biogenesis protein NSA2

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Macromolecule #15: 27S pre-rRNA (guanosine(2922)-2'-O)-methyltransferase

MacromoleculeName: 27S pre-rRNA (guanosine(2922)-2'-O)-methyltransferase / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
EC number: 27S pre-rRNA (guanosine2922-2'-O)-methyltransferase
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 96.656172 KDa
SequenceString: MGKTQKKNSK GRLDRYYYLA KEKGYRARSS FKIIQINEKY GHFLEKSKVV IDLCAAPGSW CQVASKLCPV NSLIIGVDIV PMKPMPNVI TFQSDITTED CRSKLRGYMK TWKADTVLHD GAPNVGLGWV QDAFTQSQLT LQALKLAVEN LVVNGTFVTK I FRSKDYNK ...String:
MGKTQKKNSK GRLDRYYYLA KEKGYRARSS FKIIQINEKY GHFLEKSKVV IDLCAAPGSW CQVASKLCPV NSLIIGVDIV PMKPMPNVI TFQSDITTED CRSKLRGYMK TWKADTVLHD GAPNVGLGWV QDAFTQSQLT LQALKLAVEN LVVNGTFVTK I FRSKDYNK LIWVFQQLFE KVEATKPPAS RNVSAEIFVV CKGFKAPKRL DPRLLDPKEV FEELPDGQQN MESKIYNPEK KV RKRQGYE EGDNLLYHET SILDFVRTED PISMLGEMNK FTIDENDHEW KILKKLKQTT DEFRSCIEDL KVLGKKDFKM ILR WRKIAR EILGIEVKDD AKTEIEVVPL TEEEQIEKDL QGLQEKQRLN VKRERRRKNE MKQKELQRMQ MNMITPTDIG IEAA SLGKE SLFNLKTAEK TGILNDLAKG KKRMIFTDDE LAKDNDIYID ENIMIKDKDS AADADDLESE LNAMYSDYKT RRSER DAKF RAKQARGGDN EEEWTGFNEG SLEKKEEEGK DYIEDNDDEG VEGDSDDDEA ITNLISKLKG QEGDHKLSSK ARMIFN DPI FNNVEPDLPV NTVNDGIMSS ESVGDISKLN KKRKHEEMHQ KQDEADSSDE SSSDDSDFEI VANDNASEEF DSDYDSE EE KNQTKKEKHS RDIDIATVEA MTLAHQLALG QKNKHDLVDE GFNRYTFRDT ENLPDWFLED EKEHSKINKP ITKEAAMA I KEKIKAMNAR PIKKVAEAKA RKRMRAVARL EKIKKKAGLI NDDSDKTEKD KAEEISRLMR KVTKKPKTKP KVTLVVASG RNKGLAGRPK GVKGKYKMVD GVMKNEQRAL RRIAKKHHKK K

UniProtKB: 27S pre-rRNA (guanosine(2922)-2'-O)-methyltransferase

+
Macromolecule #16: RRP17 isoform 1

MacromoleculeName: RRP17 isoform 1 / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae BY4741 (yeast) / Strain: BY4741
Molecular weightTheoretical: 1.082234 KDa
SequenceString:
YLTKNERR

UniProtKB: RRP17 isoform 1

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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

Concentration0.45 mg/mL
BufferpH: 8
Component:
ConcentrationFormulaName
50.0 mMCH2[CH2NHC9CH20H03]2Bis-Tris-Propane
150.0 mMNaClSodium Chloride
10.0 mMMgCl2Magnesium Chloride
1.0 mMC9H15O6PHClTCEP
0.01 % (w/v)NP-40
GridModel: Quantifoil R2/2 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 0.3 / Pretreatment - Type: GLOW DISCHARGE
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 1 / Number real images: 4523 / Average exposure time: 0.05 sec. / Average electron dose: 1.2 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.2 µm / Nominal defocus min: 0.9 µm / Nominal magnification: 81000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Particle selectionNumber selected: 825096
Startup modelType of model: EMDB MAP
EMDB ID:

3891

Final reconstructionNumber classes used: 1 / Applied symmetry - Point group: C1 (asymmetric) / Algorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 3.17 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION / Number images used: 198000
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION
Final 3D classificationNumber classes: 4 / Avg.num./class: 118000
FSC plot (resolution estimation)

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Atomic model buiding 1

Initial modelPDB ID:

6elz


Chain - Source name: PDB / Chain - Initial model type: experimental model
RefinementSpace: REAL / Protocol: AB INITIO MODEL
Output model

PDB-7r6k:
State E2 nucleolar 60S ribosomal intermediate - Model for Noc2/Noc3 region

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