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Open data
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Basic information
| Entry | Database: PDB / ID: 9ikz | |||||||||||||||||||||
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| Title | SARS-CoV-2 E-RTC bound to pRNA-nsp9 and GDP-BeF3- | |||||||||||||||||||||
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Keywords | VIRAL PROTEIN/RNA / SARS-CoV-2 / RTC / nsp12 / nsp13 / nsp9 / GDP-BeF3- / VIRAL PROTEIN-RNA complex | |||||||||||||||||||||
| Function / homology | Function and homology informationviral genome replication / methyltransferase activity / protein guanylyltransferase activity / RNA endonuclease activity producing 3'-phosphomonoesters, hydrolytic mechanism / mRNA guanylyltransferase activity / 5'-3' RNA helicase activity / Lyases; Phosphorus-oxygen lyases / Assembly of the SARS-CoV-2 Replication-Transcription Complex (RTC) / symbiont-mediated suppression of host cytoplasmic pattern recognition receptor signaling pathway via inhibition of TBK1 activity / Maturation of replicase proteins ...viral genome replication / methyltransferase activity / protein guanylyltransferase activity / RNA endonuclease activity producing 3'-phosphomonoesters, hydrolytic mechanism / mRNA guanylyltransferase activity / 5'-3' RNA helicase activity / Lyases; Phosphorus-oxygen lyases / Assembly of the SARS-CoV-2 Replication-Transcription Complex (RTC) / symbiont-mediated suppression of host cytoplasmic pattern recognition receptor signaling pathway via inhibition of TBK1 activity / Maturation of replicase proteins / TRAF3-dependent IRF activation pathway / ISG15-specific peptidase activity / Transcription of SARS-CoV-2 sgRNAs / snRNP Assembly / Translation of Replicase and Assembly of the Replication Transcription Complex / Replication of the SARS-CoV-2 genome / Hydrolases; Acting on ester bonds; Exoribonucleases producing 5'-phosphomonoesters / double membrane vesicle viral factory outer membrane / SARS coronavirus main proteinase / host cell endoplasmic reticulum-Golgi intermediate compartment / 5'-3' DNA helicase activity / endonuclease activity / 3'-5'-RNA exonuclease activity / host cell endosome / symbiont-mediated degradation of host mRNA / mRNA guanylyltransferase / symbiont-mediated suppression of host ISG15-protein conjugation / symbiont-mediated suppression of host toll-like receptor signaling pathway / G-quadruplex RNA binding / symbiont-mediated suppression of host cytoplasmic pattern recognition receptor signaling pathway via inhibition of IRF3 activity / omega peptidase activity / methylation / mRNA (guanine-N7)-methyltransferase / methyltransferase cap1 / SARS-CoV-2 modulates host translation machinery / host cell Golgi apparatus / symbiont-mediated suppression of host NF-kappaB cascade / DNA helicase / symbiont-mediated perturbation of host ubiquitin-like protein modification / methyltransferase cap1 activity / ubiquitinyl hydrolase 1 / cysteine-type deubiquitinase activity / mRNA 5'-cap (guanine-N7-)-methyltransferase activity / Hydrolases; Acting on peptide bonds (peptidases); Cysteine endopeptidases / single-stranded RNA binding / regulation of autophagy / viral protein processing / lyase activity / host cell perinuclear region of cytoplasm / host cell endoplasmic reticulum membrane / RNA helicase / symbiont-mediated suppression of host type I interferon-mediated signaling pathway / symbiont-mediated suppression of host gene expression / copper ion binding / viral translational frameshifting / symbiont-mediated activation of host autophagy / RNA-directed RNA polymerase / cysteine-type endopeptidase activity / viral RNA genome replication / RNA-directed RNA polymerase activity / DNA-templated transcription / lipid binding / host cell nucleus / SARS-CoV-2 activates/modulates innate and adaptive immune responses / ATP hydrolysis activity / proteolysis / RNA binding / zinc ion binding / ATP binding / membrane Similarity search - Function | |||||||||||||||||||||
| Biological species | ![]() | |||||||||||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.14 Å | |||||||||||||||||||||
Authors | Yan, L.M. / Huang, Y.C. / Liu, Y.X. / Rao, Z.H. / Lou, Z.Y. | |||||||||||||||||||||
| Funding support | China, 2items
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Citation | Journal: Cell / Year: 2025Title: Response to: The mechanism for GTP-mediated RNA capping by the SARS-CoV-2 NiRAN domain remains unresolved. Authors: Yucen Huang / Liping Tan / Yixiao Liu / Hesheng Zhao / Junbo Wang / Ji Ge / Sihan Ye / Zhenyu Liu / Weiqi Lan / Bo Huang / Hongbo Zhang / Yan Gao / Liming Yan / Zihe Rao / Zhiyong Lou / ![]() Abstract: The SARS-CoV-2 polymerase NiRAN domain initiates RNA capping. Previous results showed that both GTP and GDP can be utilized by NiRAN to yield GpppA together with RNAylated nsp9 (RNA-nsp9); however, ...The SARS-CoV-2 polymerase NiRAN domain initiates RNA capping. Previous results showed that both GTP and GDP can be utilized by NiRAN to yield GpppA together with RNAylated nsp9 (RNA-nsp9); however, the G-pocket substrate selection and the working mechanism of NiRAN remain unclear. Small et al. questioned the binding of the non-hydrolyzable GTP analog GMPPNP in the G-pocket of the RTC:RNA-nsp9:GMPPNP structure (PDB: 8GWE) and proposed that the GTP-mediated RNA-capping mechanism remains unresolved. Here, we show the optimized density derived from the original data to support the modeling of GMPPNP, and we reveal why the alternative data processing method failed to obtain density results. We provide additional biochemical and structural evidence by using GTP, GDP, GMPPNP, and GDP⋅BeF as probes to clarify the GTP-mediated RNA-capping mechanism and reconcile the two currently known models by using GTP and GDP as substrates. This Matters Arising Response addresses the Small et al. (2025) Matters Arising paper, published concurrently in Cell. | |||||||||||||||||||||
| History |
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Structure visualization
| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 9ikz.cif.gz | 554.1 KB | Display | PDBx/mmCIF format |
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| PDB format | pdb9ikz.ent.gz | 439.5 KB | Display | PDB format |
| PDBx/mmJSON format | 9ikz.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Summary document | 9ikz_validation.pdf.gz | 536.4 KB | Display | wwPDB validaton report |
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| Full document | 9ikz_full_validation.pdf.gz | 615.5 KB | Display | |
| Data in XML | 9ikz_validation.xml.gz | 62.1 KB | Display | |
| Data in CIF | 9ikz_validation.cif.gz | 95.1 KB | Display | |
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/ik/9ikz ftp://data.pdbj.org/pub/pdb/validation_reports/ik/9ikz | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 60663MC M: map data used to model this data C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
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Assembly
| Deposited unit | ![]()
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Components
-Protein , 3 types, 4 molecules AEFG
| #1: Protein | Mass: 106652.852 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: rep, 1a-1b / Production host: ![]() References: UniProt: P0DTD1, RNA-directed RNA polymerase, mRNA guanylyltransferase | ||
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| #4: Protein | Mass: 65989.438 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: rep, 1a-1b / Production host: ![]() #5: Protein | | Mass: 12391.171 Da / Num. of mol.: 1 / Fragment: UNP RESIDUES 4141-4253 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: rep, 1a-1b / Production host: ![]() |
-Non-structural protein ... , 2 types, 3 molecules BDC
| #2: Protein | Mass: 20803.787 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: rep, 1a-1b / Production host: ![]() #3: Protein | | Mass: 8678.143 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Production host: ![]() |
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-RNA chain , 2 types, 2 molecules IJ
| #6: RNA chain | Mass: 8172.932 Da / Num. of mol.: 1 / Source method: obtained synthetically Source: (synth.) ![]() |
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| #7: RNA chain | Mass: 8522.079 Da / Num. of mol.: 1 / Source method: obtained synthetically Source: (synth.) ![]() |
-Non-polymers , 4 types, 11 molecules 






| #8: Chemical | ChemComp-ZN / #9: Chemical | ChemComp-GDP / | #10: Chemical | ChemComp-BEF / | #11: Chemical | ChemComp-MG / | |
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-Details
| Has ligand of interest | Y |
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| Has protein modification | Y |
-Experimental details
-Experiment
| Experiment | Method: ELECTRON MICROSCOPY |
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| EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
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Sample preparation
| Component | Name: SARS-CoV-2 E-RTC bound to pRNA-nsp9 and GDP-BeF3- / Type: COMPLEX / Entity ID: #1-#7 / Source: RECOMBINANT |
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| Source (natural) | Organism: ![]() |
| Source (recombinant) | Organism: ![]() |
| Buffer solution | pH: 7 |
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
| Vitrification | Cryogen name: ETHANE |
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Electron microscopy imaging
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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| Microscopy | Model: TFS KRIOS |
| Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
| Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 2000 nm / Nominal defocus min: 1000 nm |
| Image recording | Electron dose: 60 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
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Processing
| EM software | Name: PHENIX / Category: model refinement |
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION |
| 3D reconstruction | Resolution: 3.14 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 156590 / Symmetry type: POINT |
| Refinement | Highest resolution: 3.14 Å Stereochemistry target values: REAL-SPACE (WEIGHTED MAP SUM AT ATOM CENTERS) |
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