+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-11422 | ||||||||||||||||||
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Title | E. coli 70S-RNAP expressome complex in uncoupled state 1 | ||||||||||||||||||
Map data | Uncoupled E. coli expressome with intervening mRNA length 38 nucleotides, state 1 of 6: composite map. | ||||||||||||||||||
Sample |
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Function / homology | Function and homology information RNA polymerase complex / submerged biofilm formation / cellular response to cell envelope stress / cytosolic DNA-directed RNA polymerase complex / regulation of DNA-templated transcription initiation / bacterial-type flagellum assembly / bacterial-type flagellum-dependent cell motility / transcriptional attenuation / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity ...RNA polymerase complex / submerged biofilm formation / cellular response to cell envelope stress / cytosolic DNA-directed RNA polymerase complex / regulation of DNA-templated transcription initiation / bacterial-type flagellum assembly / bacterial-type flagellum-dependent cell motility / transcriptional attenuation / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / positive regulation of ribosome biogenesis / nitrate assimilation / negative regulation of cytoplasmic translation / DnaA-L2 complex / translation repressor activity / negative regulation of DNA-templated DNA replication initiation / ribosome assembly / mRNA regulatory element binding translation repressor activity / response to reactive oxygen species / assembly of large subunit precursor of preribosome / transcription elongation factor complex / DNA-directed RNA polymerase complex / cytosolic ribosome assembly / regulation of DNA-templated transcription elongation / transcription antitermination / cell motility / regulation of cell growth / DNA-templated transcription initiation / DNA-templated transcription termination / : / response to radiation / ribonucleoside binding / mRNA 5'-UTR binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / ribosomal small subunit assembly / cytosolic small ribosomal subunit / large ribosomal subunit rRNA binding / ribosome binding / large ribosomal subunit / 5S rRNA binding / ribosomal large subunit assembly / cytosolic large ribosomal subunit / small ribosomal subunit / cytoplasmic translation / response to heat / transferase activity / protein-containing complex assembly / intracellular iron ion homeostasis / tRNA binding / negative regulation of translation / protein dimerization activity / ribosome / rRNA binding / hydrolase activity / structural constituent of ribosome / translation / ribonucleoprotein complex / response to antibiotic / mRNA binding / negative regulation of DNA-templated transcription / DNA-templated transcription / magnesium ion binding / DNA binding / RNA binding / zinc ion binding / membrane / metal ion binding / cytosol / cytoplasm Similarity search - Function | ||||||||||||||||||
Biological species | Escherichia coli (E. coli) / synthetic construct (others) | ||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.0 Å | ||||||||||||||||||
Authors | Webster MW / Takacs M / Weixlbaumer A | ||||||||||||||||||
Funding support | France, 5 items
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Citation | Journal: Science / Year: 2020 Title: Structural basis of transcription-translation coupling and collision in bacteria. Authors: Michael William Webster / Maria Takacs / Chengjin Zhu / Vita Vidmar / Ayesha Eduljee / Mo'men Abdelkareem / Albert Weixlbaumer / Abstract: Prokaryotic messenger RNAs (mRNAs) are translated as they are transcribed. The lead ribosome potentially contacts RNA polymerase (RNAP) and forms a supramolecular complex known as the expressome. The ...Prokaryotic messenger RNAs (mRNAs) are translated as they are transcribed. The lead ribosome potentially contacts RNA polymerase (RNAP) and forms a supramolecular complex known as the expressome. The basis of expressome assembly and its consequences for transcription and translation are poorly understood. Here, we present a series of structures representing uncoupled, coupled, and collided expressome states determined by cryo-electron microscopy. A bridge between the ribosome and RNAP can be formed by the transcription factor NusG, which stabilizes an otherwise-variable interaction interface. Shortening of the intervening mRNA causes a substantial rearrangement that aligns the ribosome entrance channel to the RNAP exit channel. In this collided complex, NusG linkage is no longer possible. These structures reveal mechanisms of coordination between transcription and translation and provide a framework for future study. | ||||||||||||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
-Related structure data
Related structure data | 6ztoMC 6ztjC 6ztlC 6ztmC 6ztnC 6ztpC 6zu1C M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_11422.map.gz / Format: CCP4 / Size: 536.4 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Uncoupled E. coli expressome with intervening mRNA length 38 nucleotides, state 1 of 6: composite map. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.052 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Additional map: Uncoupled E. coli expressome with intervening mRNA length...
+Half map: Uncoupled E. coli expressome with intervening mRNA length...
+Half map: Uncoupled E. coli expressome with intervening mRNA length...
-Sample components
+Entire : Uncoupled E. coli expressome, state 1
+Supramolecule #1: Uncoupled E. coli expressome, state 1
+Supramolecule #2: 70S ribosome within the uncoupled E. coli expressome, state 1
+Supramolecule #3: DNA-directed RNA polymerase within the uncoupled E. coli expresso...
+Supramolecule #4: synthetic mRNA and template DNA
+Macromolecule #1: 16S ribosomal RNA
+Macromolecule #22: mRNA
+Macromolecule #23: tRNA(fmet) P-site
+Macromolecule #24: Phe-NH-tRNA(Phe) A-site
+Macromolecule #25: 23S ribosomal RNA
+Macromolecule #26: 5S ribosomal RNA
+Macromolecule #2: 30S ribosomal protein S2
+Macromolecule #3: 30S ribosomal protein S3
+Macromolecule #4: 30S ribosomal protein S4
+Macromolecule #5: 30S ribosomal protein S5
+Macromolecule #6: 30S ribosomal protein S6
+Macromolecule #7: 30S ribosomal protein S7
+Macromolecule #8: 30S ribosomal protein S8
+Macromolecule #9: 30S ribosomal protein S9
+Macromolecule #10: 30S ribosomal protein S10
+Macromolecule #11: 30S ribosomal protein S11
+Macromolecule #12: 30S ribosomal protein S12
+Macromolecule #13: 30S ribosomal protein S13
+Macromolecule #14: 30S ribosomal protein S14
+Macromolecule #15: 30S ribosomal protein S15
+Macromolecule #16: 30S ribosomal protein S16
+Macromolecule #17: 30S ribosomal protein S17
+Macromolecule #18: 30S ribosomal protein S18
+Macromolecule #19: 30S ribosomal protein S19
+Macromolecule #20: 30S ribosomal protein S20
+Macromolecule #21: 30S ribosomal protein S21
+Macromolecule #27: 50S ribosomal protein L2
+Macromolecule #28: 50S ribosomal protein L3
+Macromolecule #29: 50S ribosomal protein L4
+Macromolecule #30: 50S ribosomal protein L5
+Macromolecule #31: 50S ribosomal protein L6
+Macromolecule #32: 50S ribosomal protein L9
+Macromolecule #33: 50S ribosomal protein L13
+Macromolecule #34: 50S ribosomal protein L14
+Macromolecule #35: 50S ribosomal protein L15
+Macromolecule #36: 50S ribosomal protein L16
+Macromolecule #37: 50S ribosomal protein L17
+Macromolecule #38: 50S ribosomal protein L18
+Macromolecule #39: 50S ribosomal protein L19
+Macromolecule #40: 50S ribosomal protein L20
+Macromolecule #41: 50S ribosomal protein L21
+Macromolecule #42: 50S ribosomal protein L22
+Macromolecule #43: 50S ribosomal protein L23
+Macromolecule #44: 50S ribosomal protein L24
+Macromolecule #45: 50S ribosomal protein L25
+Macromolecule #46: 50S ribosomal protein L27
+Macromolecule #47: 50S ribosomal protein L28
+Macromolecule #48: 50S ribosomal protein L29
+Macromolecule #49: 50S ribosomal protein L30
+Macromolecule #50: 50S ribosomal protein L32
+Macromolecule #51: 50S ribosomal protein L33
+Macromolecule #52: 50S ribosomal protein L34
+Macromolecule #53: 50S ribosomal protein L35
+Macromolecule #54: 50S ribosomal protein L36
+Macromolecule #55: 50S ribosomal protein L31
+Macromolecule #58: DNA-directed RNA polymerase subunit alpha
+Macromolecule #59: DNA-directed RNA polymerase subunit beta
+Macromolecule #60: DNA-directed RNA polymerase subunit beta'
+Macromolecule #61: DNA-directed RNA polymerase subunit omega
+Macromolecule #56: Non-template DNA strand
+Macromolecule #57: Template DNA strand
+Macromolecule #62: MAGNESIUM ION
+Macromolecule #63: PHENYLALANINE
+Macromolecule #64: ZINC ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 8 Component:
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Grid | Model: Quantifoil R2/2 / Material: COPPER / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Atmosphere: AIR | |||||||||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 283 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 3.5 µm / Nominal defocus min: 0.7 µm |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 42.0 e/Å2 |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |