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Yorodumi- PDB-6vyx: Escherichia coli transcription-translation complex C4 (TTC-C4) co... -
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-Basic information
Entry | Database: PDB / ID: 6vyx | ||||||
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Title | Escherichia coli transcription-translation complex C4 (TTC-C4) containing mRNA with a 21 nt long spacer, transcription factor NusG, and fMet-tRNAs at P-site and E-site | ||||||
Components |
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Keywords | Ribosome/transcription/translation / bacterial coupled transcription-translation complex / TRANSCRIPTION / Ribosome-transcription-translation complex | ||||||
Function / homology | Function and homology information DNA-templated transcription elongation / positive regulation of ribosome biogenesis / DnaA-L2 complex / DNA-directed RNA polymerase complex / negative regulation of DNA-templated DNA replication initiation / assembly of large subunit precursor of preribosome / cytosolic ribosome assembly / regulation of DNA-templated transcription elongation / transcription antitermination / regulation of cell growth ...DNA-templated transcription elongation / positive regulation of ribosome biogenesis / DnaA-L2 complex / DNA-directed RNA polymerase complex / negative regulation of DNA-templated DNA replication initiation / assembly of large subunit precursor of preribosome / cytosolic ribosome assembly / regulation of DNA-templated transcription elongation / transcription antitermination / regulation of cell growth / DNA-templated transcription termination / ribonucleoside binding / mRNA 5'-UTR binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / small ribosomal subunit rRNA binding / large ribosomal subunit / ribosome binding / 5S rRNA binding / transferase activity / small ribosomal subunit / cytosolic small ribosomal subunit / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / tRNA binding / protein dimerization activity / rRNA binding / ribosome / structural constituent of ribosome / translation / ribonucleoprotein complex / mRNA binding / DNA-templated transcription / magnesium ion binding / DNA binding / RNA binding / zinc ion binding / metal ion binding / cytosol / cytoplasm Similarity search - Function | ||||||
Biological species | Escherichia coli (E. coli) synthetic construct (others) | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 9.9 Å | ||||||
Authors | Molodtsov, V. / Wang, C. / Su, M. / Ebright, R.H. | ||||||
Funding support | United States, 1items
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Citation | Journal: Science / Year: 2020 Title: Structural basis of transcription-translation coupling. Authors: Chengyuan Wang / Vadim Molodtsov / Emre Firlar / Jason T Kaelber / Gregor Blaha / Min Su / Richard H Ebright / Abstract: In bacteria, transcription and translation are coupled processes in which the movement of RNA polymerase (RNAP)-synthesizing messenger RNA (mRNA) is coordinated with the movement of the first ...In bacteria, transcription and translation are coupled processes in which the movement of RNA polymerase (RNAP)-synthesizing messenger RNA (mRNA) is coordinated with the movement of the first ribosome-translating mRNA. Coupling is modulated by the transcription factors NusG (which is thought to bridge RNAP and the ribosome) and NusA. Here, we report cryo-electron microscopy structures of transcription-translation complexes (TTCs) containing different-length mRNA spacers between RNAP and the ribosome active-center P site. Structures of TTCs containing short spacers show a state incompatible with NusG bridging and NusA binding (TTC-A, previously termed "expressome"). Structures of TTCs containing longer spacers reveal a new state compatible with NusG bridging and NusA binding (TTC-B) and reveal how NusG bridges and NusA binds. We propose that TTC-B mediates NusG- and NusA-dependent transcription-translation coupling. | ||||||
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 | 6vyx.cif.gz | 6.6 MB | Display | PDBx/mmCIF format |
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PDB format | pdb6vyx.ent.gz | Display | PDB format | |
PDBx/mmJSON format | 6vyx.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 6vyx_validation.pdf.gz | 1.4 MB | Display | wwPDB validaton report |
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Full document | 6vyx_full_validation.pdf.gz | 1.5 MB | Display | |
Data in XML | 6vyx_validation.xml.gz | 264.3 KB | Display | |
Data in CIF | 6vyx_validation.cif.gz | 466 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/vy/6vyx ftp://data.pdbj.org/pub/pdb/validation_reports/vy/6vyx | HTTPS FTP |
-Related structure data
Related structure data | 21475MC 6vu3C 6vyqC 6vyrC 6vysC 6vytC 6vyuC 6vywC 6vyyC 6vyzC 6vz2C 6vz3C 6vz5C 6vz7C 6vzjC 6x6tC 6x7fC 6x7kC 6x9qC 6xdqC 6xdrC 6xgfC 6xiiC 6xijC 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
+50S ribosomal protein ... , 29 types, 29 molecules 01234bcefghijklmnopqrstuvwxyz
-DNA chain , 2 types, 2 molecules 56
#6: DNA chain | Mass: 8281.355 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) synthetic construct (others) |
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#7: DNA chain | Mass: 8155.232 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) synthetic construct (others) |
-RNA chain , 6 types, 7 molecules 7ABDYad
#8: RNA chain | Mass: 5196.096 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) synthetic construct (others) | ||||||||
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#9: RNA chain | Mass: 24496.617 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Escherichia coli (E. coli) / Production host: Escherichia coli (E. coli) / References: GenBank: 1848954948 #16: RNA chain | | Mass: 499690.031 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Escherichia coli (E. coli) / Production host: Escherichia coli (E. coli) / References: GenBank: 1732209707 #37: RNA chain | | Mass: 873.540 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) synthetic construct (others) #38: RNA chain | | Mass: 941635.438 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Escherichia coli (E. coli) / Production host: Escherichia coli (E. coli) / References: GenBank: 937521852 #41: RNA chain | | Mass: 38790.090 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Escherichia coli (E. coli) / Production host: Escherichia coli (E. coli) / References: GenBank: 1273279017 |
-DNA-directed RNA polymerase subunit ... , 4 types, 5 molecules AAACADAEAF
#10: Protein | Mass: 150820.875 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Escherichia coli (E. coli) / Strain: MRE600 / Gene: rpoB / Production host: Escherichia coli (E. coli) / References: UniProt: P0A8V4, DNA-directed RNA polymerase | ||||
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#12: Protein | Mass: 36558.680 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Escherichia coli (E. coli) / Strain: MRE600 / Gene: rpoA / Production host: Escherichia coli (E. coli) References: UniProt: A0A073G207, DNA-directed RNA polymerase #13: Protein | | Mass: 155398.859 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Escherichia coli (E. coli) / Strain: MRE600 / Gene: rpoC / Production host: Escherichia coli (E. coli) References: UniProt: A0A4S1NBU2, DNA-directed RNA polymerase #14: Protein | | Mass: 10249.547 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Escherichia coli (E. coli) / Strain: MRE600 / Gene: rpoZ / Production host: Escherichia coli (E. coli) / References: UniProt: P0A802, DNA-directed RNA polymerase |
-Protein , 1 types, 1 molecules AB
#11: Protein | Mass: 20560.523 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Escherichia coli (E. coli) / Gene: nusG / Production host: Escherichia coli (E. coli) / References: UniProt: V8K3D7 |
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+30S ribosomal protein ... , 21 types, 21 molecules CEFGHIJKLMNOPQRSTUVWX
-Non-polymers , 2 types, 3 molecules
#64: Chemical | ChemComp-MG / |
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#65: Chemical |
-Details
Has ligand of interest | N |
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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 |
-Sample preparation
Component | Name: Escherichia coli transcription-translation complex C4 (TTC-C4) containing mRNA with a 21 nt long spacer, transcription factor NusG, and fMet-tRNAs at P-site and E-site Type: COMPLEX / Entity ID: #1-#63 / Source: RECOMBINANT |
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Molecular weight | Experimental value: NO |
Source (natural) | Organism: Escherichia coli (E. coli) |
Source (recombinant) | Organism: Escherichia coli (E. coli) |
Buffer solution | pH: 7.6 |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Vitrification | Cryogen name: ETHANE |
-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 |
Image recording | Electron dose: 45 e/Å2 / Film or detector model: GATAN K2 SUMMIT (4k x 4k) |
-Processing
CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION |
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3D reconstruction | Resolution: 9.9 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 6233 / Symmetry type: POINT |