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- EMDB-13165: Mumps viral factory in a non-stressed HeLa cell in a chronic infe... -
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Open data
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Basic information
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Title | Mumps viral factory in a non-stressed HeLa cell in a chronic infection stage | |||||||||
![]() | Tomogram of a viral factory in unstressed HeLa cells chronically infected by mumps virus | |||||||||
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![]() | Cryo-ET / cryo-FIB / Helical filament / Nucleocapsid / Protein-RNA complex / Viral factories / Host-virus / VIRUS | |||||||||
Biological species | ![]() | |||||||||
Method | electron tomography / cryo EM | |||||||||
![]() | Mahamid J / Zhang X | |||||||||
Funding support | European Union, 1 items
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![]() | ![]() Title: Molecular mechanisms of stress-induced reactivation in mumps virus condensates. Authors: Xiaojie Zhang / Sindhuja Sridharan / Ievgeniia Zagoriy / Christina Eugster Oegema / Cyan Ching / Tim Pflaesterer / Herman K H Fung / Isabelle Becher / Ina Poser / Christoph W Müller / ...Authors: Xiaojie Zhang / Sindhuja Sridharan / Ievgeniia Zagoriy / Christina Eugster Oegema / Cyan Ching / Tim Pflaesterer / Herman K H Fung / Isabelle Becher / Ina Poser / Christoph W Müller / Anthony A Hyman / Mikhail M Savitski / Julia Mahamid / ![]() Abstract: Negative-stranded RNA viruses can establish long-term persistent infection in the form of large intracellular inclusions in the human host and cause chronic diseases. Here, we uncover how cellular ...Negative-stranded RNA viruses can establish long-term persistent infection in the form of large intracellular inclusions in the human host and cause chronic diseases. Here, we uncover how cellular stress disrupts the metastable host-virus equilibrium in persistent infection and induces viral replication in a culture model of mumps virus. Using a combination of cell biology, whole-cell proteomics, and cryo-electron tomography, we show that persistent viral replication factories are dynamic condensates and identify the largely disordered viral phosphoprotein as a driver of their assembly. Upon stress, increased phosphorylation of the phosphoprotein at its interaction interface with the viral polymerase coincides with the formation of a stable replication complex. By obtaining atomic models for the authentic mumps virus nucleocapsid, we elucidate a concomitant conformational change that exposes the viral genome to its replication machinery. These events constitute a stress-mediated switch within viral condensates that provide an environment to support upregulation of viral replication. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 362.8 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 12.3 KB 12.3 KB | Display Display | ![]() |
Images | ![]() | 291.1 KB | ||
Masks | ![]() | 821.3 MB | ![]() | |
Filedesc metadata | ![]() | 4.1 KB | ||
Others | ![]() | 6.9 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 661.3 KB | Display | ![]() |
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Full document | ![]() | 660.9 KB | Display | |
Data in XML | ![]() | 5.4 KB | Display | |
Data in CIF | ![]() | 7.1 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
File | ![]() | ||||||||||||||||||||
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Annotation | Tomogram of a viral factory in unstressed HeLa cells chronically infected by mumps virus | ||||||||||||||||||||
Voxel size | X=Y=Z: 13.48 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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Projections & Slices |
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Density Histograms |
-Additional map: Tomogram and nucleocapsid tracing of a viral factory...
File | emd_13165_additional_1.map | ||||||||||||
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Annotation | Tomogram and nucleocapsid tracing of a viral factory in unstressed HeLa cells chronically infected by mumps virus | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : A mumps viral factory in the cytoplasm of a HeLa cell at a chroni...
Entire | Name: A mumps viral factory in the cytoplasm of a HeLa cell at a chronic infection stage |
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Components |
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-Supramolecule #1: A mumps viral factory in the cytoplasm of a HeLa cell at a chroni...
Supramolecule | Name: A mumps viral factory in the cytoplasm of a HeLa cell at a chronic infection stage type: cell / ID: 1 / Parent: 0 |
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Source (natural) | Organism: ![]() |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | electron tomography |
Aggregation state | cell |
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Sample preparation
Buffer | pH: 7.5 |
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Grid | Model: Quantifoil / Material: GOLD / Pretreatment - Type: GLOW DISCHARGE |
Vitrification | Cryogen name: ETHANE / Instrument: LEICA EM GP |
Sectioning | Focused ion beam - Instrument: OTHER / Focused ion beam - Ion: OTHER / Focused ion beam - Voltage: 30 / Focused ion beam - Current: 0.05 / Focused ion beam - Duration: 600 / Focused ion beam - Temperature: 100 K / Focused ion beam - Initial thickness: 1000 / Focused ion beam - Final thickness: 150 Focused ion beam - Details: Rough milling was done with currents of 1 nA, gradually reduced to lower currents, down to 50 pA for the final polishing step. The value given for _em_focused_ion_beam. ...Focused ion beam - Details: Rough milling was done with currents of 1 nA, gradually reduced to lower currents, down to 50 pA for the final polishing step. The value given for _em_focused_ion_beam.instrument is Aquilos Cryo-FIB. This is not in a list of allowed values {'DB235', 'OTHER'} so OTHER is written into the XML file. |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Phase plate: VOLTA PHASE PLATE / Energy filter - Name: GIF Quantum LS / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Digitization - Dimensions - Width: 3708 pixel / Digitization - Dimensions - Height: 3838 pixel / Digitization - Frames/image: 1-8 / Number real images: 58 / Average exposure time: 1.6 sec. / Average electron dose: 2.2 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus min: 3.5 µm / Nominal magnification: 42000 |
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
Final reconstruction | Algorithm: BACK PROJECTION / Software - Name: ![]() |
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