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Yorodumi- PDB-9eld: Full-length and internally HIS-tagged yeast E3 ubiquitin ligase T... -
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Open data
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Basic information
| Entry | Database: PDB / ID: 9eld | |||||||||
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| Title | Full-length and internally HIS-tagged yeast E3 ubiquitin ligase Tom1p in an open-conformation | |||||||||
Components | E3 ubiquitin-protein ligase TOM1 | |||||||||
Keywords | TRANSFERASE / cell cycle / stress response / solenoid / E3 ubiquitin ligase / RNA / GENE REGULATION | |||||||||
| Function / homology | Function and homology informationendonucleolytic cleavage in 5'-ETS of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage to generate mature 5'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / HECT-type E3 ubiquitin transferase / nucleocytoplasmic transport / cleavage in ITS2 between 5.8S rRNA and LSU-rRNA of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / nucleus organization / regulation of cell size / Antigen processing: Ubiquitination & Proteasome degradation / mRNA transport / 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) ...endonucleolytic cleavage in 5'-ETS of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage to generate mature 5'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / HECT-type E3 ubiquitin transferase / nucleocytoplasmic transport / cleavage in ITS2 between 5.8S rRNA and LSU-rRNA of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / nucleus organization / regulation of cell size / Antigen processing: Ubiquitination & Proteasome degradation / mRNA transport / 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) / Neutrophil degranulation / ubiquitin-protein transferase activity / ubiquitin protein ligase activity / mitotic cell cycle / ubiquitin-dependent protein catabolic process / protein ubiquitination / nucleolus / nucleus / cytoplasm Similarity search - Function | |||||||||
| Biological species | ![]() | |||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 7.1 Å | |||||||||
Authors | Madrigal, J.M. | |||||||||
| Funding support | United States, 2items
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Citation | Journal: Elife / Year: 2024Title: Tom1p ubiquitin ligase structure, interaction with Spt6p, and function in maintaining normal transcript levels and the stability of chromatin in promoters Authors: Madrigal, J. / Schubert, H.L. / Sdano, M.A. / McCullough, L. / Connell, Z. / Formosa, T. / Hill, C.P. | |||||||||
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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 | 9eld.cif.gz | 908.1 KB | Display | PDBx/mmCIF format |
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| PDB format | pdb9eld.ent.gz | Display | PDB format | |
| PDBx/mmJSON format | 9eld.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Summary document | 9eld_validation.pdf.gz | 1.3 MB | Display | wwPDB validaton report |
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| Full document | 9eld_full_validation.pdf.gz | 1.5 MB | Display | |
| Data in XML | 9eld_validation.xml.gz | 94.6 KB | Display | |
| Data in CIF | 9eld_validation.cif.gz | 142.2 KB | Display | |
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/el/9eld ftp://data.pdbj.org/pub/pdb/validation_reports/el/9eld | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 48145MC ![]() 9egkC C: citing same article ( M: map data used to model this data |
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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
| #1: Protein | Mass: 376959.844 Da / Num. of mol.: 1 / Mutation: Internal tag insertion between 1074-1075 Source method: isolated from a genetically manipulated source Details: Internal tag insertion between 1074-1075 of the native sequence: SGGGGGGSRHHHHHHHHHHHH Source: (gene. exp.) ![]() Strain: 10293-12XHIS / Gene: TOM1, SSR2, YDR457W, D8035.1 / Production host: ![]() References: UniProt: Q03280, HECT-type E3 ubiquitin transferase |
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| Has protein modification | N |
-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: Full-length and internally HIS-tagged yeast E3 ubiquitin ligase Tom1p Type: COMPLEX / Entity ID: all / Source: RECOMBINANT |
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| Molecular weight | Value: 0.377 MDa / Experimental value: YES |
| Source (natural) | Organism: ![]() |
| Source (recombinant) | Organism: ![]() |
| Buffer solution | pH: 7.5 Details: 5% glycerol, 50 mM TrisCl pH 7.5, 150 mM NaCl, 2 mM DTT |
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES Details: Specimens were prepared on Quantifoil R 2/2 Cu300 grids after glow discharge for 1 minute at 25 mA. 2.5 uL of sample was applied to grids and blotted for 2.5 seconds before vitrification by ...Details: Specimens were prepared on Quantifoil R 2/2 Cu300 grids after glow discharge for 1 minute at 25 mA. 2.5 uL of sample was applied to grids and blotted for 2.5 seconds before vitrification by plunging into liquid ethane. |
| Specimen support | Grid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R2/2 |
| 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: -800 nm |
| Image recording | Electron dose: 47 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
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Processing
| EM software | Name: PHENIX / Version: 1.20_4459 / Category: model refinement | ||||||||||||||||||||||||
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| CTF correction | Type: PHASE FLIPPING ONLY | ||||||||||||||||||||||||
| 3D reconstruction | Resolution: 7.1 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 16098 / Num. of class averages: 1 / Symmetry type: POINT | ||||||||||||||||||||||||
| Refinement | Highest resolution: 7.1 Å Stereochemistry target values: REAL-SPACE (WEIGHTED MAP SUM AT ATOM CENTERS) | ||||||||||||||||||||||||
| Refine LS restraints |
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United States, 2items
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FIELD EMISSION GUN