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- EMDB-51352: Pre-release fusidic acid-locked Escherichia coli 70S ribosome wit... -
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Open data
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Basic information
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Title | Pre-release fusidic acid-locked Escherichia coli 70S ribosome with Staphylococus aureus EF-G and FusB (FusB-EF-G-70S) | |||||||||||||||
![]() | Local filtered map | |||||||||||||||
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![]() | RIBOSOME / fusidic acid / EF-G / antibiotic | |||||||||||||||
Function / homology | ![]() ribosome disassembly / negative regulation of cytoplasmic translational initiation / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / translation elongation factor activity / transcriptional attenuation / positive regulation of ribosome biogenesis ...ribosome disassembly / negative regulation of cytoplasmic translational initiation / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / translation elongation factor activity / transcriptional attenuation / positive regulation of ribosome biogenesis / endoribonuclease inhibitor activity / RNA-binding transcription regulator activity / negative regulation of cytoplasmic translation / four-way junction DNA binding / DnaA-L2 complex / translation repressor activity / negative regulation of translational initiation / regulation of mRNA stability / negative regulation of DNA-templated DNA replication initiation / mRNA regulatory element binding translation repressor activity / assembly of large subunit precursor of preribosome / positive regulation of RNA splicing / ribosome assembly / transcription elongation factor complex / regulation of DNA-templated transcription elongation / cytosolic ribosome assembly / response to reactive oxygen species / DNA endonuclease activity / transcription antitermination / translational initiation / regulation of cell growth / DNA-templated transcription termination / response to radiation / maintenance of translational fidelity / mRNA 5'-UTR binding / ribosome biogenesis / regulation of translation / large ribosomal subunit / transferase activity / ribosome binding / ribosomal small subunit biogenesis / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / ribosomal large subunit assembly / cytosolic small ribosomal subunit / large ribosomal subunit rRNA binding / small ribosomal subunit rRNA binding / cytosolic large ribosomal subunit / cytoplasmic translation / tRNA binding / negative regulation of translation / rRNA binding / ribosome / structural constituent of ribosome / translation / response to antibiotic / negative regulation of DNA-templated transcription / GTPase activity / mRNA binding / GTP binding / DNA binding / RNA binding / zinc ion binding / metal ion binding / membrane / cytosol / cytoplasm Similarity search - Function | |||||||||||||||
Biological species | ![]() ![]() ![]() ![]() | |||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.79 Å | |||||||||||||||
![]() | Gonzalez-Lopez A / Selmer M | |||||||||||||||
Funding support | ![]()
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![]() | ![]() Title: Structural mechanism of FusB-mediated rescue from fusidic acid inhibition of protein synthesis. Authors: Adrián González-López / Xueliang Ge / Daniel S D Larsson / Carina Sihlbom Wallem / Suparna Sanyal / Maria Selmer / ![]() Abstract: The antibiotic resistance protein FusB rescues protein synthesis from inhibition by fusidic acid (FA), which locks elongation factor G (EF-G) to the ribosome after GTP hydrolysis. Here, we present ...The antibiotic resistance protein FusB rescues protein synthesis from inhibition by fusidic acid (FA), which locks elongation factor G (EF-G) to the ribosome after GTP hydrolysis. Here, we present time-resolved single-particle cryo-EM structures explaining the mechanism of FusB-mediated rescue. FusB binds to the FA-trapped EF-G on the ribosome, causing large-scale conformational changes of EF-G that break interactions with the ribosome, tRNA, and mRNA. This leads to dissociation of EF-G from the ribosome, followed by FA release. We also observe two independent binding sites of FusB on the classical-state ribosome, overlapping with the binding site of EF-G to each of the ribosomal subunits, yet not inhibiting tRNA delivery. The affinity of FusB to the ribosome and the concentration of FusB in S. aureus during FusB-mediated resistance support that direct binding of FusB to ribosomes could occur in the cell. Our results reveal an intricate resistance mechanism involving specific interactions of FusB with both EF-G and the ribosome, and a non-canonical release pathway of EF-G. | |||||||||||||||
History |
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Structure visualization
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Downloads & links
-EMDB archive
Map data | ![]() | 93.1 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 92.7 KB 92.7 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 22.4 KB | Display | ![]() |
Images | ![]() | 99 KB | ||
Masks | ![]() | 824 MB | ![]() | |
Filedesc metadata | ![]() | 16.9 KB | ||
Others | ![]() ![]() ![]() | 409.7 MB 764.7 MB 764.7 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 1.2 MB | Display | ![]() |
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Full document | ![]() | 1.2 MB | Display | |
Data in XML | ![]() | 29.7 KB | Display | |
Data in CIF | ![]() | 39.1 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 9ghcMC ![]() 9ghaC ![]() 9ghbC ![]() 9ghdC ![]() 9gheC ![]() 9ghfC ![]() 9ghgC ![]() 9ghhC M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Local filtered map | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.648 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
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-Additional map: Unsharpened map
File | emd_51352_additional_1.map | ||||||||||||
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-Half map: Half map A
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Annotation | Half map A | ||||||||||||
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-Half map: Half map B
File | emd_51352_half_map_2.map | ||||||||||||
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Annotation | Half map B | ||||||||||||
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Sample components
+Entire : Staphylococcus aureus EF-G on 70S ribosomes
+Supramolecule #1: Staphylococcus aureus EF-G on 70S ribosomes
+Supramolecule #2: Staphylococcus aureus FusB-EF-G
+Supramolecule #3: mRNA
+Supramolecule #4: 70S ribosome
+Macromolecule #1: 50S ribosomal protein L33
+Macromolecule #2: 50S ribosomal protein L34
+Macromolecule #3: 50S ribosomal protein L35
+Macromolecule #4: 50S ribosomal protein L36
+Macromolecule #5: 50S ribosomal protein L31
+Macromolecule #8: 30S ribosomal protein S2
+Macromolecule #9: 30S ribosomal protein S3
+Macromolecule #10: 30S ribosomal protein S4
+Macromolecule #11: 30S ribosomal protein S5
+Macromolecule #12: 30S ribosomal protein S6
+Macromolecule #13: 30S ribosomal protein S7
+Macromolecule #14: 30S ribosomal protein S8
+Macromolecule #15: 30S ribosomal protein S9
+Macromolecule #16: 30S ribosomal protein S10
+Macromolecule #17: Small ribosomal subunit protein uS11
+Macromolecule #18: 30S ribosomal protein S12
+Macromolecule #19: 30S ribosomal protein S13
+Macromolecule #20: 30S ribosomal protein S14
+Macromolecule #21: 30S ribosomal protein S15
+Macromolecule #22: 30S ribosomal protein S16
+Macromolecule #23: 30S ribosomal protein S17
+Macromolecule #24: 30S ribosomal protein S18
+Macromolecule #25: 30S ribosomal protein S19
+Macromolecule #26: 30S ribosomal protein S20
+Macromolecule #27: 30S ribosomal protein S21
+Macromolecule #28: Far1
+Macromolecule #29: Elongation factor G
+Macromolecule #33: 50S ribosomal protein L2
+Macromolecule #34: 50S ribosomal protein L3
+Macromolecule #35: 50S ribosomal protein L4
+Macromolecule #36: 50S ribosomal protein L5
+Macromolecule #37: 50S ribosomal protein L6
+Macromolecule #38: 50S ribosomal protein L9
+Macromolecule #39: 50S ribosomal protein L13
+Macromolecule #40: 50S ribosomal protein L14
+Macromolecule #41: Large ribosomal subunit protein uL15
+Macromolecule #42: 50S ribosomal protein L16
+Macromolecule #43: 50S ribosomal protein L17
+Macromolecule #44: 50S ribosomal protein L18
+Macromolecule #45: 50S ribosomal protein L19
+Macromolecule #46: 50S ribosomal protein L20
+Macromolecule #47: 50S ribosomal protein L21
+Macromolecule #48: 50S ribosomal protein L22
+Macromolecule #49: 50S ribosomal protein L23
+Macromolecule #50: 50S ribosomal protein L24
+Macromolecule #51: 50S ribosomal protein L25
+Macromolecule #52: 50S ribosomal protein L27
+Macromolecule #53: 50S ribosomal protein L28
+Macromolecule #54: Large ribosomal subunit protein uL29
+Macromolecule #55: 50S ribosomal protein L30
+Macromolecule #56: 50S ribosomal protein L32
+Macromolecule #6: mRNA
+Macromolecule #7: 16S rRNA
+Macromolecule #30: tRNA
+Macromolecule #31: 23S rRNA
+Macromolecule #32: 5S rRNA
+Macromolecule #57: ZINC ION
+Macromolecule #58: POTASSIUM ION
+Macromolecule #59: MAGNESIUM ION
+Macromolecule #60: GUANOSINE-5'-DIPHOSPHATE
+Macromolecule #61: FUSIDIC ACID
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.5 Component:
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Grid | Model: Quantifoil R2/1 / Material: COPPER / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 2 | ||||||||||||||||||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | TFS KRIOS |
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Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Number grids imaged: 1 / Number real images: 24479 / Average exposure time: 1.0 sec. / Average electron dose: 40.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | C2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 1.0 µm / Nominal defocus min: 0.7000000000000001 µm / Nominal magnification: 130000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Initial model |
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Refinement | Space: REAL | |||||||||
Output model | ![]() PDB-9ghc: |