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Yorodumi- EMDB-8282: Structure of RelA bound to ribosome in presence of A/R tRNA (Stru... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-8282 | ||||||||||||
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Title | Structure of RelA bound to ribosome in presence of A/R tRNA (Structure IV) | ||||||||||||
Map data | Ribosome*RelA structures reveal the mechanism of stringent response activation (Structure IV) | ||||||||||||
Sample |
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Keywords | ribosome / RelA | ||||||||||||
Function / homology | Function and homology information guanosine tetraphosphate metabolic process / guanosine-3',5'-bis(diphosphate) 3'-diphosphatase activity / GTP diphosphokinase / GTP diphosphokinase activity / guanosine tetraphosphate biosynthetic process / nucleobase-containing small molecule interconversion / negative regulation of cytoplasmic translational initiation / response to starvation / stringent response / ornithine decarboxylase inhibitor activity ...guanosine tetraphosphate metabolic process / guanosine-3',5'-bis(diphosphate) 3'-diphosphatase activity / GTP diphosphokinase / GTP diphosphokinase activity / guanosine tetraphosphate biosynthetic process / nucleobase-containing small molecule interconversion / negative regulation of cytoplasmic translational initiation / response to starvation / stringent response / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / positive regulation of ribosome biogenesis / negative regulation of cytoplasmic translation / four-way junction DNA binding / translational termination / DnaA-L2 complex / translation repressor activity / negative regulation of DNA-templated DNA replication initiation / negative regulation of translational initiation / regulation of mRNA stability / mRNA regulatory element binding translation repressor activity / ribosome assembly / assembly of large subunit precursor of preribosome / positive regulation of RNA splicing / transcription elongation factor complex / cytosolic ribosome assembly / regulation of DNA-templated transcription elongation / DNA endonuclease activity / response to reactive oxygen species / transcription antitermination / regulation of cell growth / translational initiation / DNA-templated transcription termination / maintenance of translational fidelity / response to radiation / mRNA 5'-UTR binding / ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / large ribosomal subunit / ribosome biogenesis / ribosome binding / kinase activity / regulation of translation / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / large ribosomal subunit rRNA binding / transferase activity / cytosolic small ribosomal subunit / ribosomal large subunit assembly / cytoplasmic translation / cytosolic large ribosomal subunit / tRNA binding / molecular adaptor activity / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / translation / response to antibiotic / negative regulation of DNA-templated transcription / mRNA binding / GTP binding / DNA binding / RNA binding / zinc ion binding / ATP binding / membrane / cytosol / cytoplasm Similarity search - Function | ||||||||||||
Biological species | Escherichia coli (strain K12) (bacteria) / Escherichia coli K-12 (bacteria) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.9 Å | ||||||||||||
Authors | Loveland AB / Bah E | ||||||||||||
Funding support | United States, 3 items
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Citation | Journal: Elife / Year: 2016 Title: Ribosome•RelA structures reveal the mechanism of stringent response activation. Authors: Anna B Loveland / Eugene Bah / Rohini Madireddy / Ying Zhang / Axel F Brilot / Nikolaus Grigorieff / Andrei A Korostelev / Abstract: Stringent response is a conserved bacterial stress response underlying virulence and antibiotic resistance. RelA/SpoT-homolog proteins synthesize transcriptional modulators (p)ppGpp, allowing ...Stringent response is a conserved bacterial stress response underlying virulence and antibiotic resistance. RelA/SpoT-homolog proteins synthesize transcriptional modulators (p)ppGpp, allowing bacteria to adapt to stress. RelA is activated during amino-acid starvation, when cognate deacyl-tRNA binds to the ribosomal A (aminoacyl-tRNA) site. We report four cryo-EM structures of E. coli RelA bound to the 70S ribosome, in the absence and presence of deacyl-tRNA accommodating in the 30S A site. The boomerang-shaped RelA with a wingspan of more than 100 Å wraps around the A/R (30S A-site/RelA-bound) tRNA. The CCA end of the A/R tRNA pins the central TGS domain against the 30S subunit, presenting the (p)ppGpp-synthetase domain near the 30S spur. The ribosome and A/R tRNA are captured in three conformations, revealing hitherto elusive states of tRNA engagement with the ribosomal decoding center. Decoding-center rearrangements are coupled with the step-wise 30S-subunit 'closure', providing insights into the dynamics of high-fidelity tRNA decoding. | ||||||||||||
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
Map data | emd_8282.map.gz | 138.2 MB | EMDB map data format | |
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Header (meta data) | emd-8282-v30.xml emd-8282.xml | 83.2 KB 83.2 KB | Display Display | EMDB header |
Images | emd_8282.png | 147.6 KB | ||
Filedesc metadata | emd-8282.cif.gz | 14.7 KB | ||
Others | emd_8282_additional.map.gz emd_8282_half_map_1.map.gz emd_8282_half_map_2.map.gz | 139.4 MB 139.5 MB 139.5 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-8282 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-8282 | HTTPS FTP |
-Validation report
Summary document | emd_8282_validation.pdf.gz | 1004.1 KB | Display | EMDB validaton report |
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Full document | emd_8282_full_validation.pdf.gz | 1003.7 KB | Display | |
Data in XML | emd_8282_validation.xml.gz | 18.2 KB | Display | |
Data in CIF | emd_8282_validation.cif.gz | 21.7 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-8282 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-8282 | HTTPS FTP |
-Related structure data
Related structure data | 5kpxMC 8279C 8280C 8281C 5kpsC 5kpvC 5kpwC C: citing same article (ref.) M: atomic model generated by this map |
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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_8282.map.gz / Format: CCP4 / Size: 421.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Ribosome*RelA structures reveal the mechanism of stringent response activation (Structure IV) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.82 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Additional map: Ribosome*RelA structures reveal the mechanism of stringent response...
File | emd_8282_additional.map | ||||||||||||
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Annotation | Ribosome*RelA structures reveal the mechanism of stringent response activation (Structure IV, additional map) | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Ribosome*RelA structures reveal the mechanism of stringent response...
File | emd_8282_half_map_1.map | ||||||||||||
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Annotation | Ribosome*RelA structures reveal the mechanism of stringent response activation (Structure IV, half map 1) | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Ribosome*RelA structures reveal the mechanism of stringent response...
File | emd_8282_half_map_2.map | ||||||||||||
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Annotation | Ribosome*RelA structures reveal the mechanism of stringent response activation (Structure IV, half map 2) | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : RelA bound to 70S ribosome in the presence of A/R tRNA (Structure IV)
+Supramolecule #1: RelA bound to 70S ribosome in the presence of A/R tRNA (Structure IV)
+Macromolecule #1: 50S ribosomal protein L2
+Macromolecule #2: 50S ribosomal protein L3
+Macromolecule #3: 50S ribosomal protein L4
+Macromolecule #4: 50S ribosomal protein L5
+Macromolecule #5: 50S ribosomal protein L6
+Macromolecule #6: 50S ribosomal protein L9
+Macromolecule #7: 50S ribosomal protein L10
+Macromolecule #8: 50S ribosomal protein L11
+Macromolecule #9: 50S ribosomal protein L13
+Macromolecule #10: 50S ribosomal protein L14
+Macromolecule #11: 50S ribosomal protein L15
+Macromolecule #12: 50S ribosomal protein L16
+Macromolecule #13: 50S ribosomal protein L17
+Macromolecule #14: 50S ribosomal protein L18
+Macromolecule #15: 50S ribosomal protein L19
+Macromolecule #16: 50S ribosomal protein L20
+Macromolecule #17: 50S ribosomal protein L21
+Macromolecule #18: 50S ribosomal protein L22
+Macromolecule #19: 50S ribosomal protein L23
+Macromolecule #20: 50S ribosomal protein L24
+Macromolecule #21: 50S ribosomal protein L25
+Macromolecule #22: 50S ribosomal protein L27
+Macromolecule #23: 50S ribosomal protein L28
+Macromolecule #24: 50S ribosomal protein L29
+Macromolecule #25: 50S ribosomal protein L30
+Macromolecule #26: 50S ribosomal protein L31
+Macromolecule #27: 50S ribosomal protein L32
+Macromolecule #28: 50S ribosomal protein L33
+Macromolecule #29: 50S ribosomal protein L34
+Macromolecule #30: 50S ribosomal protein L35
+Macromolecule #31: 50S ribosomal protein L36
+Macromolecule #32: 30S ribosomal protein S2
+Macromolecule #33: 30S ribosomal protein S3
+Macromolecule #34: 30S ribosomal protein S4
+Macromolecule #35: 30S ribosomal protein S5
+Macromolecule #36: 30S ribosomal protein S6
+Macromolecule #37: 30S ribosomal protein S7
+Macromolecule #38: 30S ribosomal protein S8
+Macromolecule #39: 30S ribosomal protein S9
+Macromolecule #40: 30S ribosomal protein S10
+Macromolecule #41: 30S ribosomal protein S11
+Macromolecule #42: 30S ribosomal protein S12
+Macromolecule #43: 30S ribosomal protein S13
+Macromolecule #44: 30S ribosomal protein S14
+Macromolecule #45: 30S ribosomal protein S15
+Macromolecule #46: 30S ribosomal protein S16
+Macromolecule #47: 30S ribosomal protein S17
+Macromolecule #48: 30S ribosomal protein S18
+Macromolecule #49: 30S ribosomal protein S19
+Macromolecule #50: 30S ribosomal protein S20
+Macromolecule #51: 30S ribosomal protein S21
+Macromolecule #59: GTP pyrophosphokinase
+Macromolecule #52: 16S ribosomal RNA
+Macromolecule #53: 23S ribosomal RNA
+Macromolecule #54: 5S ribosomal RNA
+Macromolecule #55: mRNA
+Macromolecule #56: A-site tRNAPhe
+Macromolecule #57: P-site tRNAfMet
+Macromolecule #58: E-site tRNAfMet
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.1 mg/mL |
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Buffer | pH: 7.4 |
Grid | Model: C-flat-1.2/1.3 / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 45 sec. / Pretreatment - Atmosphere: AIR |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 75 % / Chamber temperature: 295 K / Instrument: GATAN CRYOPLUNGE 3 Details: Apply 2 uL sample and blot 4 seconds before plunging into liquid ethane (GATAN CRYOPLUNGE 3).. |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Digitization - Frames/image: 1-25 / Number grids imaged: 5 / Number real images: 5763 / Average exposure time: 0.4 sec. / Average electron dose: 1.6 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 100.0 µm / Calibrated defocus max: 2.2 µm / Calibrated defocus min: 0.5 µm / Calibrated magnification: 30488 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.2 µm / Nominal defocus min: 0.5 µm / Nominal magnification: 30488 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |