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- PDB-9xae: Cryo-EM structure of the pre-40S ribosome (Enp1-Rrp12 WT) from Ch... -

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Basic information

Entry
Database: PDB / ID: 9xae
TitleCryo-EM structure of the pre-40S ribosome (Enp1-Rrp12 WT) from Chaetomium thermophilum, state Rio2-C
Components
  • (40S ribosomal protein ...) x 21
  • (Small ribosomal subunit protein ...) x 4
  • 18S rRNA
  • Bms1-type G domain-containing protein
  • Bystin-like protein
  • Pre-rRNA-processing protein PNO1
  • Putative low-temperature viability protein
  • Putative ribosomal protein
  • Ribosomal protein s27-like protein
KeywordsRIBOSOME / 90S pre-ribosome / Rrp12 / Enp1 / ribosome biogenesis
Function / homology
Function and homology information


endonucleolytic cleavage of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / U3 snoRNA binding / snoRNA binding / preribosome, small subunit precursor / 90S preribosome / ribosomal small subunit export from nucleus / cytosolic ribosome / methyltransferase activity / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA ...endonucleolytic cleavage of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / U3 snoRNA binding / snoRNA binding / preribosome, small subunit precursor / 90S preribosome / ribosomal small subunit export from nucleus / cytosolic ribosome / methyltransferase activity / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / small-subunit processome / protein tag activity / rRNA processing / ribosomal small subunit assembly / ribosome biogenesis / ribosomal small subunit biogenesis / small ribosomal subunit / small ribosomal subunit rRNA binding / cytosolic small ribosomal subunit / methylation / cytoplasmic translation / rRNA binding / structural constituent of ribosome / ribosome / translation / ribonucleoprotein complex / GTPase activity / GTP binding / nucleolus / RNA binding / nucleus / cytoplasm / cytosol
Similarity search - Function
Low temperature viability protein Ltv1 / Low temperature viability protein / Tsr1, G-like domain / Bystin / Bystin / : / : / Eukaryotic type KH-domain (KH-domain type I) / Ribosome biogenesis protein BMS1/TSR1, C-terminal / AARP2CN ...Low temperature viability protein Ltv1 / Low temperature viability protein / Tsr1, G-like domain / Bystin / Bystin / : / : / Eukaryotic type KH-domain (KH-domain type I) / 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 S21e, conserved site / Ribosomal protein S21e signature. / : / Ribosomal protein S12e signature. / Ribosomal protein S12e / Ribosomal protein S5, eukaryotic/archaeal / Ribosomal protein S21e / Ribosomal protein S21e superfamily / Ribosomal protein S21e / Ribosomal protein S2, eukaryotic / S27a-like superfamily / Ribosomal protein S25 / S25 ribosomal protein / Ribosomal protein S8e subdomain, eukaryotes / : / Ribosomal protein S17e, conserved site / Ribosomal protein S17e signature. / Ribosomal protein S7e signature. / Ribosomal protein S27a / Ribosomal protein S27a / Ribosomal protein S27a / Ribosomal protein S2, eukaryotic/archaeal / Ribosomal protein S3Ae, conserved site / Ribosomal protein S3Ae signature. / 40S ribosomal protein S4, C-terminal domain / 40S ribosomal protein S4 C-terminus / Ribosomal protein S19A/S15e / Ribosomal protein S19e / Ribosomal protein S19e / Ribosomal_S19e / Ribosomal protein S4e, N-terminal, conserved site / Ribosomal protein S4e signature. / Ribosomal protein S17e / Ribosomal protein S17e-like superfamily / Ribosomal S17 / Ribosomal protein S6, eukaryotic / 40S ribosomal protein S1/3, eukaryotes / 40S ribosomal protein S11, N-terminal / Ribosomal_S17 N-terminal / Ribosomal protein S7e / Ribosomal protein S7e / : / Ribosomal S24e conserved site / Ribosomal protein S24e signature. / Ribosomal protein S4e, N-terminal / RS4NT (NUC023) domain / Ribosomal protein S4, KOW domain / Ribosomal protein S4e / Ribosomal protein S4e, central region / Ribosomal protein S4e, central domain superfamily / Ribosomal family S4e / Ribosomal protein S28e conserved site / Ribosomal protein S28e signature. / Ribosomal protein S6/S6e/A/B/2, conserved site / Ribosomal protein S17, archaeal/eukaryotic / Ribosomal protein S6e signature. / Ribosomal protein S23, eukaryotic/archaeal / Ribosomal protein S8e / Ribosomal protein S27 / Ribosomal protein S24e / Ribosomal protein S27, zinc-binding domain superfamily / Ribosomal protein S24e / Ribosomal protein S27 / Ribosomal protein S28e / Ribosomal protein S28e / Ribosomal protein S3Ae / Ribosomal S3Ae family / Ribosomal S3Ae family / Ribosomal protein S5/S7, eukaryotic/archaeal / Ribosomal protein S6e / Ribosomal protein S6e / Ribosomal protein S6e / Ribosomal protein S13/S15, N-terminal / Ribosomal protein S15P / Ribosomal S13/S15 N-terminal domain / Ribosomal S13/S15 N-terminal domain / Ribosomal protein S4/S9, eukaryotic/archaeal / Ribosomal protein S8e/ribosomal biogenesis NSA2 / Ribosomal protein S8e / Ribosomal protein L7Ae/L30e/S12e/Gadd45 / Ribosomal protein L7Ae/L30e/S12e/Gadd45 family / 50S ribosomal protein L30e-like
Similarity search - Domain/homology
RNA / RNA (> 10) / RNA (> 100) / RNA (> 1000) / 40S ribosomal protein s23-like protein / 40S ribosomal protein S6 / 40S ribosomal protein S8 / Small ribosomal subunit protein uS2 / Bms1-type G domain-containing protein / 40S ribosomal protein S13-like protein ...RNA / RNA (> 10) / RNA (> 100) / RNA (> 1000) / 40S ribosomal protein s23-like protein / 40S ribosomal protein S6 / 40S ribosomal protein S8 / Small ribosomal subunit protein uS2 / Bms1-type G domain-containing protein / 40S ribosomal protein S13-like protein / 40S ribosomal protein s15-like protein / 40S ribosomal protein s9-like protein / 40S ribosomal protein S4 / Ribosomal protein s27-like protein / 40S ribosomal protein S25 / 40S ribosomal protein s5-like protein / 40S ribosomal protein S27a-like protein / Pre-rRNA-processing protein PNO1 / Bystin-like protein / Putative ribosomal protein / Small ribosomal subunit protein eS1 / 40S ribosomal protein S17-like protein / 40S ribosomal protein S7 / 40S ribosomal protein S28-like protein / Small ribosomal subunit protein uS5 / 40S ribosomal protein S16-like protein / Low-temperature viability protein / 40S ribosomal protein S11-like protein / 40S ribosomal protein S19-like protein / 40S ribosomal protein S14-like protein / 40S ribosomal protein S12 / 40S ribosomal protein S22-like protein / 40S ribosomal protein S21-like protein / Small ribosomal subunit protein eS24
Similarity search - Component
Biological speciesThermochaetoides thermophila (fungus)
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3 Å
AuthorsLau, B. / Li, Y. / Zhu, J. / Fischer, P. / Hong, X. / Yuan, R. / Beckmann, R. / Hurt, E. / Cheng, J.
Funding support1items
OrganizationGrant numberCountry
Not funded
CitationJournal: Cell Rep / Year: 2026
Title: Nucleoplasmic checkpoint of the 40S ribosomal decoding center maturation.
Authors: Benjamin Lau / Yi Li / Jingyi Zhu / Xianwen Ye / Paulina Fischer / Xiaying Hong / Rui Yuan / Roland Beckmann / Ed Hurt / Jingdong Cheng /
Abstract: The decoding center (DC) is a key ribosomal structure for accurate translation, assembled in a multi-step process that starts on nucleolar pre-ribosomes and ends in the cytoplasm. While late ...The decoding center (DC) is a key ribosomal structure for accurate translation, assembled in a multi-step process that starts on nucleolar pre-ribosomes and ends in the cytoplasm. While late cytoplasmic steps and their checkpoint mechanisms are well characterized, the regulation of early nucleoplasmic DC assembly is unclear. Here, we show that the essential assembly factor Rrp12 plays a central coordinating role. Using Chaetomium thermophilum and cryo-electron microscopy analyses of fifteen pre-40S intermediates, we demonstrate that Rrp12 C terminus truncation: (1) inhibits release of the Utp14-Dhr1 pair, (2) displaces Tsr1, (3) promotes premature stabilization of h28, and (4) prevents h44 formation. These defects impair final 18S rRNA processing and prematurely activate the quality control kinase Rio1. Our results reveal a nucleoplasmic checkpoint during DC formation and establish Rrp12 as a critical regulator ensuring accurate assembly and orderly ribosome maturation.
History
DepositionOct 22, 2025Deposition site: PDBJ / Processing site: PDBC
Revision 1.0Jun 24, 2026Provider: repository / Type: Initial release
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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
D: Small ribosomal subunit protein uS2
E: Small ribosomal subunit protein eS1
F: Small ribosomal subunit protein uS5
H: 40S ribosomal protein S4
I: 40S ribosomal protein s5-like protein
J: 40S ribosomal protein S6
K: 40S ribosomal protein S7
L: 40S ribosomal protein S8
M: 40S ribosomal protein s9-like protein
O: 40S ribosomal protein S11-like protein
P: 40S ribosomal protein S12
Q: 40S ribosomal protein S13-like protein
R: 40S ribosomal protein S14-like protein
S: 40S ribosomal protein s15-like protein
T: 40S ribosomal protein S16-like protein
U: 40S ribosomal protein S17-like protein
V: Putative ribosomal protein
W: 40S ribosomal protein S19-like protein
Y: 40S ribosomal protein S21-like protein
Z: 40S ribosomal protein S22-like protein
a: 40S ribosomal protein s23-like protein
b: Small ribosomal subunit protein eS24
c: 40S ribosomal protein S25
e: Ribosomal protein s27-like protein
f: 40S ribosomal protein S28-like protein
h: 40S ribosomal protein S30
i: 40S ribosomal protein S27a-like protein
A: 18S rRNA
2: Bms1-type G domain-containing protein
3: Pre-rRNA-processing protein PNO1
4: Bystin-like protein
5: Putative low-temperature viability protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)1,306,13435
Polymers1,306,02032
Non-polymers1143
Water724
1


  • Idetical with deposited unit
  • defined by author
  • Evidence: electron microscopy, not applicable
TypeNameSymmetry operationNumber
identity operation1_5551

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Components

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Small ribosomal subunit protein ... , 4 types, 4 molecules DEFb

#1: Protein Small ribosomal subunit protein uS2


Mass: 31158.996 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0RYF9
#2: Protein Small ribosomal subunit protein eS1


Mass: 29245.158 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0S7T8
#3: Protein Small ribosomal subunit protein uS5 / 40S ribosomal protein S2


Mass: 28556.115 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0SAV4
#22: Protein Small ribosomal subunit protein eS24 / 40S ribosomal protein S24


Mass: 15535.286 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: P0CU28

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40S ribosomal protein ... , 21 types, 21 molecules HIJKLMOPQRSTUWYZacfhi

#4: Protein 40S ribosomal protein S4


Mass: 29800.764 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0S1A6
#5: Protein 40S ribosomal protein s5-like protein


Mass: 23679.225 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0S1Z0
#6: Protein 40S ribosomal protein S6


Mass: 27490.139 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0RY43
#7: Protein 40S ribosomal protein S7


Mass: 23084.650 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0S8C4
#8: Protein 40S ribosomal protein S8


Mass: 23102.416 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0RY45
#9: Protein 40S ribosomal protein s9-like protein


Mass: 22029.836 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0S0Z4
#10: Protein 40S ribosomal protein S11-like protein


Mass: 18698.182 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0SF00
#11: Protein 40S ribosomal protein S12


Mass: 15961.293 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0SHH9
#12: Protein 40S ribosomal protein S13-like protein


Mass: 16912.891 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0RZM9
#13: Protein 40S ribosomal protein S14-like protein


Mass: 16071.464 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0SFL1
#14: Protein 40S ribosomal protein s15-like protein


Mass: 17601.604 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0S084
#15: Protein 40S ribosomal protein S16-like protein


Mass: 15962.771 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0SBR7
#16: Protein 40S ribosomal protein S17-like protein


Mass: 16720.240 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0S871
#18: Protein 40S ribosomal protein S19-like protein


Mass: 17151.332 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0SFK8
#19: Protein 40S ribosomal protein S21-like protein


Mass: 10961.475 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0SI21
#20: Protein 40S ribosomal protein S22-like protein


Mass: 14905.471 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0SHI0
#21: Protein 40S ribosomal protein s23-like protein


Mass: 15934.653 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0RY17
#23: Protein 40S ribosomal protein S25


Mass: 10932.943 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0S1Y9
#25: Protein 40S ribosomal protein S28-like protein


Mass: 7741.980 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0S9M9
#26: Protein 40S ribosomal protein S30


Mass: 6917.273 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus)
#27: Protein 40S ribosomal protein S27a-like protein


Mass: 17731.760 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0S2X5

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Protein , 6 types, 6 molecules Ve2345

#17: Protein Putative ribosomal protein


Mass: 17826.707 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0S6J7
#24: Protein Ribosomal protein s27-like protein


Mass: 8923.572 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0S1S4
#29: Protein Bms1-type G domain-containing protein


Mass: 93766.930 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0RZ98
#30: Protein Pre-rRNA-processing protein PNO1


Mass: 28715.047 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0S3T2
#31: Protein Bystin-like protein


Mass: 54319.605 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0S5J4
#32: Protein Putative low-temperature viability protein


Mass: 49784.793 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus) / References: UniProt: G0SE19

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RNA chain , 1 types, 1 molecules A

#28: RNA chain 18S rRNA


Mass: 578795.562 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Thermochaetoides thermophila (fungus)

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Non-polymers , 3 types, 7 molecules

#33: Chemical ChemComp-MG / MAGNESIUM ION


Mass: 24.305 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: Mg
#34: Chemical ChemComp-ZN / ZINC ION


Mass: 65.409 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: Zn
#35: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 4 / Source method: isolated from a natural source / Formula: H2O

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Details

Has ligand of interestN
Has protein modificationN

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Experimental details

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Experiment

ExperimentMethod: ELECTRON MICROSCOPY
EM experimentAggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction

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Sample preparation

ComponentName: pre-40S ribosome, state Rio2-C / Type: RIBOSOME / Entity ID: #1-#32 / Source: NATURAL
Source (natural)Organism: Thermochaetoides thermophila (fungus)
Buffer solutionpH: 7.4
SpecimenEmbedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
VitrificationCryogen name: ETHANE

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
MicroscopyModel: TFS KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM
Electron lensMode: BRIGHT FIELD / Nominal defocus max: 2500 nm / Nominal defocus min: 1000 nm
Image recordingElectron dose: 50 e/Å2 / Film or detector model: FEI FALCON IV (4k x 4k)

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Processing

EM software
IDNameCategory
1Gautomatchparticle selection
4RELIONCTF correction
7Cootmodel fitting
10RELIONinitial Euler assignment
11RELIONfinal Euler assignment
13RELION3D reconstruction
CTF correctionDetails: Relion / Type: NONE
3D reconstructionResolution: 3 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 85384 / Symmetry type: POINT
Atomic model buildingPDB-ID: 6G18
Accession code: 6G18 / Source name: PDB / Type: experimental model

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