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Open data
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Basic information
| Entry | Database: PDB / ID: 9gc3 | ||||||
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| Title | yeast TFIIIC TauB subcomplex bound to a tRNA gene | ||||||
 Components | 
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 Keywords | DNA BINDING PROTEIN / Base-specific contacts / B-box / tRNA gene | ||||||
| Function / homology |  Function and homology information5S class rRNA transcription by RNA polymerase III / transcription factor TFIIIC complex / RNA polymerase III general transcription initiation factor activity / RNA Polymerase III Transcription Initiation From Type 2 Promoter / transcription initiation at RNA polymerase III promoter / transcription by RNA polymerase III / protein localization to chromatin / mitochondrion / DNA binding / nucleoplasm / nucleus Similarity search - Function  | ||||||
| Biological species | ![]()  | ||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.46 Å | ||||||
 Authors | Seifert-Davila, W. / Girbig, M. / Hauptmann, L. / Hoffmann, T. / Eustermann, S. / Mueller, C. / Chaban, A. / Duss, O. / Baudin, F. | ||||||
| Funding support | 1items 
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 Citation |  Journal: Nucleic Acids Res / Year: 2025Title: Structural and kinetic insights into tRNA promoter engagement by yeast general transcription factor TFIIIC. Authors: Wolfram Seifert-Dávila / Anastasiia Chaban / Florence Baudin / Mathias Girbig / Luis Hauptmann / Thomas Hoffmann / Olivier Duss / Sebastian Eustermann / Christoph W Müller / ![]() Abstract: Transcription of transfer RNA (tRNA) genes by RNA polymerase (Pol) III requires the general transcription factor IIIC (TFIIIC), which recognizes intragenic A-box and B-box DNA motifs of type II ...Transcription of transfer RNA (tRNA) genes by RNA polymerase (Pol) III requires the general transcription factor IIIC (TFIIIC), which recognizes intragenic A-box and B-box DNA motifs of type II gene promoters. However, the underlying mechanism has remained elusive, in part due to missing structural information for A-box recognition. In this study, we use single-particle cryogenic electron microscopy (cryo-EM) and single-molecule fluorescence resonance energy transfer (smFRET) to reveal structural and real-time kinetic insights into how the 520-kDa yeast TFIIIC complex engages A-box and B-box DNA motifs in the context of a tRNA gene promoter. Cryo-EM structures of τA and τB subcomplexes bound to the A-box and B-box were obtained at 3.7 and 2.5 Å resolution, respectively, while cryo-EM single-particle mapping determined the specific distance and relative orientation of the τA and τB subcomplexes revealing a fully engaged state of TFIIIC. smFRET experiments show that overall recruitment and residence times of TFIIIC on a tRNA gene are primarily governed by B-box recognition, while footprinting experiments suggest a key role of τA and the A-box in TFIIIB and Pol III recruitment following TFIIIC recognition of type II promoters.  | ||||||
| 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 |  9gc3.cif.gz | 409.5 KB | Display |  PDBx/mmCIF format | 
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| PDB format |  pdb9gc3.ent.gz | 310.9 KB | Display |  PDB format | 
| PDBx/mmJSON format |  9gc3.json.gz | Tree view |  PDBx/mmJSON format | |
| Others |  Other downloads | 
-Validation report
| Summary document |  9gc3_validation.pdf.gz | 1.1 MB | Display |  wwPDB validaton report | 
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| Full document |  9gc3_full_validation.pdf.gz | 1.2 MB | Display | |
| Data in XML |  9gc3_validation.xml.gz | 52.4 KB | Display | |
| Data in CIF |  9gc3_validation.cif.gz | 81.3 KB | Display | |
| Arichive directory |  https://data.pdbj.org/pub/pdb/validation_reports/gc/9gc3 ftp://data.pdbj.org/pub/pdb/validation_reports/gc/9gc3 | HTTPS FTP  | 
-Related structure data
| Related structure data | ![]() 51228MC ![]() 9gckC 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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| 1 | 
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Components
| #1: Protein |   Mass: 136602.500 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: TFC3, TSV115, YAL001C, FUN24 / Production host: ![]()  | 
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| #2: Protein |   Mass: 76484.070 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: TFC6, YDR362C / Production host: ![]()  | 
| #3: Protein |   Mass: 67755.172 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Gene: TFC8, YPL007C / Production host: ![]()  | 
| #4: DNA chain |   Mass: 12358.934 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Production host: ![]()  | 
| #5: DNA chain |   Mass: 12265.904 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() Production host: ![]()  | 
| 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: Yeast TFIIIC TauB monomer bound to a tRNA gene / Type: COMPLEX / Entity ID: all / Source: RECOMBINANT | 
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| Molecular weight | Experimental value: NO | 
| Source (natural) | Organism: ![]()  | 
| Source (recombinant) | Organism: ![]()  | 
| Buffer solution | pH: 8 | 
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | 
| Vitrification | Instrument: FEI VITROBOT MARK IV / 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: FEI TITAN KRIOS | 
| Electron gun | Electron source:  FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM | 
| Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 1700 nm / Nominal defocus min: 700 nm | 
| Image recording | Electron dose: 39.6 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) Details: dataset1: 39.6 dataset2: 43.6 dataset3: 43.2 dataset4: 43.2 dataset5: 44.4  | 
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Processing
| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| 3D reconstruction | Resolution: 2.46 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 570437 / Symmetry type: POINT | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Refinement | Resolution: 2.46→2.46 Å / Cor.coef. Fo:Fc: 0.943  / SU B: 5.693  / SU ML: 0.118  / ESU R: 0.153  Stereochemistry target values: MAXIMUM LIKELIHOOD WITH PHASES Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS 
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| Solvent computation | Solvent model: PARAMETERS FOR MASK CACLULATION | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Displacement parameters | Biso mean: 96.053 Å2 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Refinement step | Cycle: 1 / Total: 15049 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Refine LS restraints | 
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FIELD EMISSION GUN