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Open data
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Basic information
Entry | Database: PDB / ID: 6kf3 | |||||||||||||||
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Title | Cryo-EM structure of Thermococcus kodakarensis RNA polymerase | |||||||||||||||
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![]() | TRANSCRIPTION / apo-RNA polymerase | |||||||||||||||
Function / homology | ![]() RNA polymerase III activity / tRNA transcription by RNA polymerase III / RNA polymerase I activity / DNA-directed RNA polymerase complex / RNA polymerase II activity / DNA-templated transcription initiation / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / protein dimerization activity ...RNA polymerase III activity / tRNA transcription by RNA polymerase III / RNA polymerase I activity / DNA-directed RNA polymerase complex / RNA polymerase II activity / DNA-templated transcription initiation / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / protein dimerization activity / nucleotide binding / DNA-templated transcription / magnesium ion binding / DNA binding / zinc ion binding / cytoplasm Similarity search - Function | |||||||||||||||
Biological species | ![]() ![]() | |||||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.9 Å | |||||||||||||||
![]() | Jun, S.-H. / Hyun, J. / Jeong, H. / Cha, J.S. / Kim, H. / Bartlett, M.S. / Cho, H.-S. / Murakami, K.S. | |||||||||||||||
Funding support | ![]() ![]()
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![]() | ![]() Title: Direct binding of TFEα opens DNA binding cleft of RNA polymerase. Authors: Sung-Hoon Jun / Jaekyung Hyun / Jeong Seok Cha / Hoyoung Kim / Michael S Bartlett / Hyun-Soo Cho / Katsuhiko S Murakami / ![]() ![]() ![]() Abstract: Opening of the DNA binding cleft of cellular RNA polymerase (RNAP) is necessary for transcription initiation but the underlying molecular mechanism is not known. Here, we report on the cryo-electron ...Opening of the DNA binding cleft of cellular RNA polymerase (RNAP) is necessary for transcription initiation but the underlying molecular mechanism is not known. Here, we report on the cryo-electron microscopy structures of the RNAP, RNAP-TFEα binary, and RNAP-TFEα-promoter DNA ternary complexes from archaea, Thermococcus kodakarensis (Tko). The structures reveal that TFEα bridges the RNAP clamp and stalk domains to open the DNA binding cleft. Positioning of promoter DNA into the cleft closes it while maintaining the TFEα interactions with the RNAP mobile modules. The structures and photo-crosslinking results also suggest that the conserved aromatic residue in the extended winged-helix domain of TFEα interacts with promoter DNA to stabilize the transcription bubble. This study provides a structural basis for the functions of TFEα and elucidates the mechanism by which the DNA binding cleft is opened during transcription initiation in the stalk-containing RNAPs, including archaeal and eukaryotic RNAPs. | |||||||||||||||
History |
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Structure visualization
Movie |
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Structure viewer | Molecule: ![]() ![]() |
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Downloads & links
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Download
PDBx/mmCIF format | ![]() | 590.8 KB | Display | ![]() |
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PDB format | ![]() | 473 KB | Display | ![]() |
PDBx/mmJSON format | ![]() | Tree view | ![]() | |
Others | ![]() |
-Validation report
Summary document | ![]() | 1 MB | Display | ![]() |
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Full document | ![]() | 1.1 MB | Display | |
Data in XML | ![]() | 91.5 KB | Display | |
Data in CIF | ![]() | 140.8 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 9960M ![]() 6kf4C ![]() 6kf9C ![]() 6plnC ![]() 6xjfC M: map data used to model this data C: citing same article ( |
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Similar structure data |
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Links
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Assembly
Deposited unit | ![]()
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1 |
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Components
-Protein , 1 types, 1 molecules A
#1: Protein | Mass: 103038.633 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: RpoA' Source: (natural) ![]() ![]() Strain: ATCC BAA-918 / JCM 12380 / KOD1 / References: UniProt: Q5JE33, DNA-directed RNA polymerase |
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-DNA-directed RNA polymerase subunit ... , 8 types, 8 molecules BCDHKLNP
#2: Protein | Mass: 127468.039 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: RpoB Source: (natural) ![]() ![]() Strain: ATCC BAA-918 / JCM 12380 / KOD1 / References: UniProt: Q5JE32, DNA-directed RNA polymerase |
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#3: Protein | Mass: 43727.410 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: RpoA'' Source: (natural) ![]() ![]() Strain: ATCC BAA-918 / JCM 12380 / KOD1 / References: UniProt: Q5JE34, DNA-directed RNA polymerase |
#4: Protein | Mass: 29429.645 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: RpoD Source: (natural) ![]() ![]() Strain: ATCC BAA-918 / JCM 12380 / KOD1 / References: UniProt: Q5JJF4, DNA-directed RNA polymerase |
#7: Protein | Mass: 9522.031 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: RpoH Source: (natural) ![]() ![]() Strain: ATCC BAA-918 / JCM 12380 / KOD1 / References: UniProt: Q5JE31, DNA-directed RNA polymerase |
#8: Protein | Mass: 6286.519 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: RpoK Source: (natural) ![]() ![]() Strain: ATCC BAA-918 / JCM 12380 / KOD1 / References: UniProt: Q5JJD0, DNA-directed RNA polymerase |
#9: Protein | Mass: 11013.504 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: RpoL Source: (natural) ![]() ![]() Strain: ATCC BAA-918 / JCM 12380 / KOD1 / References: UniProt: Q5JE88, DNA-directed RNA polymerase |
#10: Protein | Mass: 7601.975 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: RpoN Source: (natural) ![]() ![]() Strain: ATCC BAA-918 / JCM 12380 / KOD1 / References: UniProt: Q5JJC9, DNA-directed RNA polymerase |
#11: Protein/peptide | Mass: 5553.708 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Details: RpoP Source: (natural) ![]() ![]() Strain: ATCC BAA-918 / JCM 12380 / KOD1 / References: UniProt: Q5JDM8, DNA-directed RNA polymerase |
-DNA-directed RNA polymerase, subunit ... , 2 types, 2 molecules EF
#5: Protein | Mass: 21893.438 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: RpoE Source: (gene. exp.) ![]() ![]() Strain: ATCC BAA-918 / JCM 12380 / KOD1 / Gene: TK1699 / Production host: ![]() ![]() |
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#6: Protein | Mass: 14519.659 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: RpoF Source: (gene. exp.) ![]() ![]() Strain: ATCC BAA-918 / JCM 12380 / KOD1 / Gene: TK0901 / Production host: ![]() ![]() |
-Non-polymers , 2 types, 6 molecules ![](data/chem/img/MG.gif)
![](data/chem/img/ZN.gif)
![](data/chem/img/ZN.gif)
#12: Chemical | ChemComp-MG / |
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#13: Chemical | ChemComp-ZN / |
-Details
Has ligand of interest | N |
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-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: RNA polymerase / Type: COMPLEX / Entity ID: #1-#11 / Source: MULTIPLE SOURCES |
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Molecular weight | Value: 0.38 MDa |
Source (natural) | Organism: ![]() ![]() |
Source (recombinant) | Organism: ![]() ![]() |
Buffer solution | pH: 8 |
Specimen | Conc.: 0.3 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Specimen support | Grid material: COPPER / Grid mesh size: 200 divisions/in. / Grid type: Quantifoil R1.2/1.3 |
Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 90 % / Chamber temperature: 278 K |
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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: ![]() |
Electron lens | Mode: BRIGHT FIELD |
Image recording | Electron dose: 35 e/Å2 / Film or detector model: FEI FALCON II (4k x 4k) |
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Processing
EM software |
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CTF correction | Type: PHASE FLIPPING ONLY | ||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||
3D reconstruction | Resolution: 3.9 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 139242 / Algorithm: BACK PROJECTION / Num. of class averages: 1 / Symmetry type: POINT |