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- EMDB-64713: Cryo- EM structure of 75S ribosome with P- tRNA from Entamoeba hi... -
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Open data
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Basic information
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Title | Cryo- EM structure of 75S ribosome with P- tRNA from Entamoeba histolytica | |||||||||
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![]() | 75S Ribosome / P-tRNA / Entamoeba histolytica / Ribosome | |||||||||
Function / homology | ![]() inositol hexakisphosphate binding / negative regulation of translational frameshifting / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage in ITS1 to separate SSU-rRNA from 5.8S rRNA and LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / protein-RNA complex assembly / maturation of LSU-rRNA / translation regulator activity / rescue of stalled ribosome / protein kinase C binding / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) ...inositol hexakisphosphate binding / negative regulation of translational frameshifting / endonucleolytic cleavage to generate mature 3'-end of SSU-rRNA from (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / endonucleolytic cleavage in ITS1 to separate SSU-rRNA from 5.8S rRNA and LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / protein-RNA complex assembly / maturation of LSU-rRNA / translation regulator activity / rescue of stalled ribosome / protein kinase C binding / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / ribosomal large subunit biogenesis / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / small-subunit processome / maintenance of translational fidelity / rRNA processing / ribosome biogenesis / ribosome binding / ribosomal small subunit biogenesis / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / small ribosomal subunit rRNA binding / ribosomal large subunit assembly / large ribosomal subunit rRNA binding / cytosolic small ribosomal subunit / cytosolic large ribosomal subunit / cytoplasmic translation / negative regulation of translation / rRNA binding / structural constituent of ribosome / ribosome / translation / ribonucleoprotein complex / mRNA binding / nucleolus / endoplasmic reticulum / RNA binding / zinc ion binding / nucleus / cytosol Similarity search - Function | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.0 Å | |||||||||
![]() | Sharma S / Mishra S / Gourinath S / Kaushal PS | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Cryo-EM structure of ribosome from pathogenic protozoa Entamoeba histolytica reveals unique features of its architecture. Authors: Shivani Sharma / Shalini Mishra / Samudrala Gourinath / Prem S Kaushal / ![]() Abstract: Entamoeba histolytica, an anaerobic protozoan parasite, is the causative agent of amoebiasis, bloody diarrhea, and liver abscesses in humans. Amoebiasis is more predominant in tropical areas with ...Entamoeba histolytica, an anaerobic protozoan parasite, is the causative agent of amoebiasis, bloody diarrhea, and liver abscesses in humans. Amoebiasis is more predominant in tropical areas with poor sanitation conditions, and it remains the fourth leading cause of death due to a protozoan infection. E. histolytica life cycle spans between an infective 'cyst stage' and an active disease-causing 'trophozoite stage'. We have isolated ribosomes from the trophozoite stage of E. histolytica. Here, we report single particle cryo-EM structures of the 53S ribosome large subunit (LSU), and 75S associated ribosomes, with P-tRNA, A/P and P/E tRNAs, and with paromomycin antibiotic, at 2.8 Å to 3.4 Å resolution. The E. histolytica possesses a reduced ribosome with a conserved core, and the periphery evolved with species-specific unique features. The most notable features are the presence of the rRNA triple helix near the peptide exit tunnel, the expansion segment H88ES102 near the exit site on LSU, and unique insertions in r-proteins. Furthermore, the 75S ribosome paromomycin complex structure provides the atomic details of its interactions. These structures provide insights into the evolutionary adaptation of the E. histolytica translational machinery and may be explored further for amoebicidal therapeutic intervention. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 215 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 100.3 KB 100.3 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 14.2 KB | Display | ![]() |
Images | ![]() | 169.8 KB | ||
Filedesc metadata | ![]() | 19.2 KB | ||
Others | ![]() ![]() | 193.7 MB 193.8 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 1012.5 KB | Display | ![]() |
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Full document | ![]() | 1012.1 KB | Display | |
Data in XML | ![]() | 22.1 KB | Display | |
Data in CIF | ![]() | 28.9 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 9v21MC ![]() 9v1iC ![]() 9v1jC ![]() 9v1kC ![]() 9v26C ![]() 9v27C ![]() 9v28C ![]() 9v29C 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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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.07 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: #2
File | emd_64713_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_64713_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
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Sample components
+Entire : Cryo-EM structure of 75S ribosome with P-tRNA from Entamoeba hist...
+Supramolecule #1: Cryo-EM structure of 75S ribosome with P-tRNA from Entamoeba hist...
+Macromolecule #1: 25S rRNA
+Macromolecule #2: 5.8S
+Macromolecule #3: 5S rRNA
+Macromolecule #51: P-tRNA
+Macromolecule #52: mRNA
+Macromolecule #53: 17S rRNA
+Macromolecule #4: Large ribosomal subunit protein uL2
+Macromolecule #5: 60S ribosomal protein L3, putative
+Macromolecule #6: Large ribosomal subunit protein uL4
+Macromolecule #7: 60S ribosomal protein L5, putative
+Macromolecule #8: Large ribosomal subunit protein eL6
+Macromolecule #9: 60S ribosomal protein L7, putative
+Macromolecule #10: 60S ribosomal protein L7a
+Macromolecule #11: 60S ribosomal protein L9, putative
+Macromolecule #12: Ribosomal protein L10, putative
+Macromolecule #13: 60S ribosomal protein L11, putative
+Macromolecule #14: 60S ribosomal protein L13, putative
+Macromolecule #15: 60S ribosomal protein L13, putative
+Macromolecule #16: 60S ribosomal protein L14, putative
+Macromolecule #17: Ribosomal protein L15
+Macromolecule #18: 60S ribosomal protein L17, putative
+Macromolecule #19: 60S ribosomal protein L18, putative
+Macromolecule #20: 60S ribosomal protein L18a
+Macromolecule #21: Ribosomal protein L19
+Macromolecule #22: 60S ribosomal protein L21, putative
+Macromolecule #23: Large ribosomal subunit protein eL22
+Macromolecule #24: 60S ribosomal protein L23, putative
+Macromolecule #25: Ribosomal protein L23A, putative
+Macromolecule #26: 60S ribosomal protein L24, putative
+Macromolecule #27: 60S ribosomal protein L26, putative
+Macromolecule #28: 60S ribosomal protein L27, putative
+Macromolecule #29: Large ribosomal subunit protein uL15A
+Macromolecule #30: 60S ribosomal protein L29
+Macromolecule #31: 60S ribosomal protein L30, putative
+Macromolecule #32: 60S ribosomal protein L31, putative
+Macromolecule #33: 60S ribosomal protein L32, putative
+Macromolecule #34: 60S ribosomal protein L34, putative
+Macromolecule #35: uL29
+Macromolecule #36: 60S ribosomal protein L35a, putative
+Macromolecule #37: 60S ribosomal protein L36, putative
+Macromolecule #38: 60S ribosomal protein L37-A, putative
+Macromolecule #39: 60S ribosomal protein L37A, putative
+Macromolecule #40: 60S ribosomal protein L38, putative
+Macromolecule #41: Ribosomal protein L39, putative
+Macromolecule #42: 60S ribosomal protein L40, putative
+Macromolecule #43: 60S ribosomal protein L44, putative
+Macromolecule #44: nascent polypeptide (Unk)
+Macromolecule #45: 40S ribosomal protein S25
+Macromolecule #46: 40S ribosomal protein S26
+Macromolecule #47: Small ribosomal subunit protein eS27
+Macromolecule #48: 40S ribosomal protein S28, putative
+Macromolecule #49: Ribosomal protein S29, putative
+Macromolecule #50: Guanine nucleotide-binding protein subunit beta 2-like 1, putative
+Macromolecule #54: Small ribosomal subunit protein uS2
+Macromolecule #55: Small ribosomal subunit protein uS5
+Macromolecule #56: 40S ribosomal protein S3
+Macromolecule #57: Small ribosomal subunit protein eS1
+Macromolecule #58: 40S ribosomal protein S4, putative
+Macromolecule #59: Small ribosomal subunit protein uS7
+Macromolecule #60: 40S ribosomal protein S6
+Macromolecule #61: 40S ribosomal protein S7
+Macromolecule #62: 40S ribosomal protein S8
+Macromolecule #63: Small ribosomal subunit protein uS4
+Macromolecule #64: 40S ribosomal protein S10, putative
+Macromolecule #65: 40S ribosomal protein S11, putative
+Macromolecule #66: 40S ribosomal protein S12, putative
+Macromolecule #67: 40S ribosomal protein S13, putative
+Macromolecule #68: Ribosomal protein S14, putative
+Macromolecule #69: 40S ribosomal protein S15, putative
+Macromolecule #70: 40S ribosomal protein S15a, putative
+Macromolecule #71: 40S ribosomal protein S16, putative
+Macromolecule #72: 40S ribosomal protein S17, putative
+Macromolecule #73: Small ribosomal subunit protein uS13
+Macromolecule #74: Small ribosomal subunit protein eS19
+Macromolecule #75: Small ribosomal subunit protein uS10
+Macromolecule #76: 40S ribosomal protein S21
+Macromolecule #77: 40S ribosomal protein S23, putative
+Macromolecule #78: 40S ribosomal protein S24, putative
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 1 mg/mL |
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Buffer | pH: 7.4 Details: 20nM HEPES-sodium salt buffer pH 7.4, 100mM Potassium acetate, 10mM Magnesium acetate, 10mm Ammonium acetate, 3mM DTT |
Grid | Model: Quantifoil R1.2/1.3 / Material: GOLD / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 50 sec. |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 290 K / Instrument: FEI VITROBOT MARK I |
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Electron microscopy
Microscope | TFS KRIOS |
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Image recording | Film or detector model: FEI FALCON III (4k x 4k) / Detector mode: INTEGRATING / Number real images: 11840 / Average exposure time: 2.0 sec. / Average electron dose: 1.106 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.8 µm / Nominal magnification: 75000 |
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 | PDB ID: Chain - Source name: PDB / Chain - Initial model type: experimental model |
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Details | phenix.real_space_refinement |
Refinement | Space: REAL / Protocol: FLEXIBLE FIT / Overall B value: 51.95 |
Output model | ![]() PDB-9v21: |