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Yorodumi- EMDB-33787: In situ structure of polymerase complex of mammalian reovirus in ... -
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Basic information
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| Title | In situ structure of polymerase complex of mammalian reovirus in virion | |||||||||
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Keywords | mammalian reovirus / cryo-em / RNA dependent RNA polymerase / transcription / VIRAL PROTEIN / VIRAL PROTEIN-RNA complex | |||||||||
| Function / homology | Function and homology informationT=2 icosahedral viral capsid / viral inner capsid / host cytoskeleton / viral outer capsid / 7-methylguanosine mRNA capping / viral genome replication / viral capsid / mRNA guanylyltransferase activity / viral nucleocapsid / host cell cytoplasm ...T=2 icosahedral viral capsid / viral inner capsid / host cytoskeleton / viral outer capsid / 7-methylguanosine mRNA capping / viral genome replication / viral capsid / mRNA guanylyltransferase activity / viral nucleocapsid / host cell cytoplasm / mRNA 5'-cap (guanine-N7-)-methyltransferase activity / RNA helicase activity / hydrolase activity / RNA-directed RNA polymerase / RNA-directed RNA polymerase activity / GTP binding / structural molecule activity / RNA binding / ATP binding Similarity search - Function | |||||||||
| Biological species | Mammalian orthoreovirus 3 | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.4 Å | |||||||||
Authors | Bao KY / Zhang XL / Li DY / Zhu P | |||||||||
| Funding support | China, 2 items
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Citation | Journal: Proc Natl Acad Sci U S A / Year: 2022Title: In situ structures of polymerase complex of mammalian reovirus illuminate RdRp activation and transcription regulation. Authors: Keyan Bao / Xueli Zhang / Dongyu Li / Wei Sun / Zhenzhao Sun / Jingfei Wang / Ping Zhu / ![]() Abstract: Mammalian reovirus (reovirus) is a multilayered, turreted member of characterized by transcription of dsRNA genome within the innermost capsid shell. Here, we present high-resolution in situ ...Mammalian reovirus (reovirus) is a multilayered, turreted member of characterized by transcription of dsRNA genome within the innermost capsid shell. Here, we present high-resolution in situ structures of reovirus transcriptase complex in an intact double-layered virion, and in the uncoated single-layered core particles in the unloaded, reloaded, pre-elongation, and elongation states, respectively, obtained by cryo-electron microscopy and sub-particle reconstructions. At the template entry of RNA-dependent RNA polymerase (RdRp), the RNA-loading region gets flexible after uncoating resulting in the unloading of terminal genomic RNA and inactivity of transcription. However, upon adding transcriptional substrates, the RNA-loading region is recovered leading the RNAs loaded again. The priming loop in RdRp was found to play a critical role in regulating transcription, which hinders the elongation of transcript in virion and triggers the rearrangement of RdRp C-terminal domain (CTD) during elongation, resulting in splitting of template-transcript hybrid and opening of transcript exit. With the integration of these structures, a transcriptional model of reovirus with five states is proposed. Our structures illuminate the RdRp activation and regulation of the multilayered turreted reovirus. | |||||||||
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_33787.map.gz | 118 MB | EMDB map data format | |
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| Header (meta data) | emd-33787-v30.xml emd-33787.xml | 24.4 KB 24.4 KB | Display Display | EMDB header |
| Images | emd_33787.png | 99.4 KB | ||
| Filedesc metadata | emd-33787.cif.gz | 8.7 KB | ||
| Others | emd_33787_half_map_1.map.gz emd_33787_half_map_2.map.gz | 116 MB 116 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-33787 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-33787 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 7yfeMC ![]() 7yedC ![]() 7yevC ![]() 7yezC ![]() 7yf0C M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_33787.map.gz / Format: CCP4 / Size: 125 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.36 Å | ||||||||||||||||||||||||||||||||||||
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Half map: #1
| File | emd_33787_half_map_1.map | ||||||||||||
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-Half map: #2
| File | emd_33787_half_map_2.map | ||||||||||||
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| Density Histograms |
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Sample components
+Entire : Mammalian orthoreovirus 3
+Supramolecule #1: Mammalian orthoreovirus 3
+Supramolecule #3: RNA
+Supramolecule #2: Mammalian orthoreovirus 3
+Macromolecule #1: RNA helicase
+Macromolecule #2: Lambda-2 protein
+Macromolecule #5: RNA-directed RNA polymerase
+Macromolecule #7: Mu-2 protein
+Macromolecule #3: RNA (5'-R(P*GP*CP*U)-3')
+Macromolecule #4: RNA (5'-R(P*AP*UP*CP*GP*U)-3')
+Macromolecule #6: RNA (5'-R(P*AP*CP*GP*AP*UP*UP*AP*GP*C)-3')
+Macromolecule #8: ZINC ION
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Buffer | pH: 8 |
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| Vitrification | Cryogen name: ETHANE |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 40.0 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 1.2 µm |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
| Startup model | Type of model: NONE |
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| Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.4 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 50031 |
| Initial angle assignment | Type: RANDOM ASSIGNMENT |
| Final angle assignment | Type: RANDOM ASSIGNMENT |
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Mammalian orthoreovirus 3
Keywords
Authors
China, 2 items
Citation













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FIELD EMISSION GUN
