+Open data
-Basic information
Entry | Database: PDB / ID: 6yj6 | ||||||
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Title | Structure of the TFIIIC subcomplex tauA | ||||||
Components |
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Keywords | TRANSCRIPTION / TFIIIC / tauA / Transcription initiation / Pol III / TFIIIB / Transcription factor | ||||||
Function / homology | Function and homology information 5S class rRNA transcription by RNA polymerase III / transcription factor TFIIIC complex / RNA Polymerase III Transcription Initiation From Type 2 Promoter / transcription initiation at RNA polymerase III promoter / phosphatase activity / transcription by RNA polymerase III / DNA binding / nucleoplasm / nucleus / cytoplasm Similarity search - Function | ||||||
Biological species | Saccharomyces cerevisiae (brewer's yeast) | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.1 Å | ||||||
Authors | Vorlaender, M.K. / Muller, C.W. | ||||||
Citation | Journal: Nat Commun / Year: 2020 Title: Structure of the TFIIIC subcomplex τA provides insights into RNA polymerase III pre-initiation complex formation. Authors: Matthias K Vorländer / Anna Jungblut / Kai Karius / Florence Baudin / Helga Grötsch / Jan Kosinski / Christoph W Müller / Abstract: Transcription factor (TF) IIIC is a conserved eukaryotic six-subunit protein complex with dual function. It serves as a general TF for most RNA polymerase (Pol) III genes by recruiting TFIIIB, but it ...Transcription factor (TF) IIIC is a conserved eukaryotic six-subunit protein complex with dual function. It serves as a general TF for most RNA polymerase (Pol) III genes by recruiting TFIIIB, but it is also involved in chromatin organization and regulation of Pol II genes through interaction with CTCF and condensin II. Here, we report the structure of the S. cerevisiae TFIIIC subcomplex τA, which contains the most conserved subunits of TFIIIC and is responsible for recruitment of TFIIIB and transcription start site (TSS) selection at Pol III genes. We show that τA binding to its promoter is auto-inhibited by a disordered acidic tail of subunit τ95. We further provide a negative-stain reconstruction of τA bound to the TFIIIB subunits Brf1 and TBP. This shows that a ruler element in τA achieves positioning of TFIIIB upstream of the TSS, and suggests remodeling of the complex during assembly of TFIIIB by TFIIIC. | ||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | Molecule: MolmilJmol/JSmol |
-Downloads & links
-Download
PDBx/mmCIF format | 6yj6.cif.gz | 586.1 KB | Display | PDBx/mmCIF format |
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PDB format | pdb6yj6.ent.gz | 475.7 KB | Display | PDB format |
PDBx/mmJSON format | 6yj6.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 6yj6_validation.pdf.gz | 1.1 MB | Display | wwPDB validaton report |
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Full document | 6yj6_full_validation.pdf.gz | 1.1 MB | Display | |
Data in XML | 6yj6_validation.xml.gz | 56.5 KB | Display | |
Data in CIF | 6yj6_validation.cif.gz | 84.9 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/yj/6yj6 ftp://data.pdbj.org/pub/pdb/validation_reports/yj/6yj6 | HTTPS FTP |
-Related structure data
Related structure data | 10817MC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
#1: Protein | Mass: 120746.969 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Saccharomyces cerevisiae (brewer's yeast) Gene: TFC4, PCF1, YGR047C / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: P33339 |
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#2: Protein | Mass: 77363.664 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Saccharomyces cerevisiae (brewer's yeast) Gene: TFC1, YBR123C, YBR0919 / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: P32367 |
#3: Protein | Mass: 49198.898 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Saccharomyces cerevisiae (brewer's yeast) Gene: TFC7, YOR110W, O3234, YOR3234w / Production host: Trichoplusia ni (cabbage looper) / References: UniProt: Q12415 |
-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: TFIIIC subcomplex tauA / Type: COMPLEX Details: Generated by co-expression of subunits in insect cells Entity ID: all / Source: RECOMBINANT |
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Molecular weight | Value: 0.245 MDa / Experimental value: NO |
Source (natural) | Organism: Saccharomyces cerevisiae (brewer's yeast) |
Source (recombinant) | Organism: Trichoplusia ni (cabbage looper) / Strain: Hi 5 cells |
Buffer solution | pH: 7.5 / Details: 20 mM HEPES, 5 mM DTT, 75 mM NaCl |
Specimen | Conc.: 1.3 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES / Details: 4 mM CHAPSO added prior to freezing |
Specimen support | Details: Pelco Easygglow instrument / Grid material: COPPER / Grid mesh size: 200 divisions/in. / Grid type: Quantifoil R2/1 |
Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 298 K / Details: Blot force 2, Blot time 4 |
-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 magnification: 130000 X / Nominal defocus max: -2000 nm / Nominal defocus min: -500 nm / Cs: 2.7 mm / C2 aperture diameter: 70 µm / Alignment procedure: ZEMLIN TABLEAU |
Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Average exposure time: 48.73 sec. / Electron dose: 1.35 e/Å2 / Detector mode: COUNTING / Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Num. of grids imaged: 1 / Num. of real images: 5824 / Details: collected 8 frames per hole using beam shift |
EM imaging optics | Energyfilter slit width: 20 eV |
Image scans | Movie frames/image: 36 / Used frames/image: 1-36 |
-Processing
EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||
Particle selection | Num. of particles selected: 702583 / Details: BoxNet2_20180918 was used for particle picking | ||||||||||||||||||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||||||||||||||
3D reconstruction | Resolution: 3.1 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 254700 / Symmetry type: POINT | ||||||||||||||||||||||||||||||||||||
Atomic model building | Protocol: AB INITIO MODEL / Space: REAL |