European Union, France, Germany, United Kingdom, 7 items
Organization
Grant number
Country
European Research Council (ERC)
832601
European Union
Agence Nationale de la Recherche (ANR)
ANR-16-CE11-0033-01
France
German Research Foundation (DFG)
(INST 152/772-1|152/774-1|152/775-1|152/776-1
Germany
Wellcome Trust
107806/Z/15/Z
United Kingdom
German Federal Ministry for Education and Research
05K18BHA
Germany
Alexander von Humboldt Foundation
FRA 1200789 HFST-P
Germany
Agence Nationale de la Recherche (ANR)
ANR-10-INBS-04
France
Citation
Journal: Elife / Year: 2022 Title: The giant mimivirus 1.2 Mb genome is elegantly organized into a 30-nm diameter helical protein shield. Authors: Alejandro Villalta / Alain Schmitt / Leandro F Estrozi / Emmanuelle R J Quemin / Jean-Marie Alempic / Audrey Lartigue / Vojtěch Pražák / Lucid Belmudes / Daven Vasishtan / Agathe M G ...Authors: Alejandro Villalta / Alain Schmitt / Leandro F Estrozi / Emmanuelle R J Quemin / Jean-Marie Alempic / Audrey Lartigue / Vojtěch Pražák / Lucid Belmudes / Daven Vasishtan / Agathe M G Colmant / Flora A Honoré / Yohann Couté / Kay Grünewald / Chantal Abergel / Abstract: Mimivirus is the prototype of the family of giant dsDNA viruses. Little is known about the organization of the 1.2 Mb genome inside the membrane-limited nucleoid filling the ~0.5 µm icosahedral ...Mimivirus is the prototype of the family of giant dsDNA viruses. Little is known about the organization of the 1.2 Mb genome inside the membrane-limited nucleoid filling the ~0.5 µm icosahedral capsids. Cryo-electron microscopy, cryo-electron tomography, and proteomics revealed that it is encased into a ~30-nm diameter helical protein shell surprisingly composed of two GMC-type oxidoreductases, which also form the glycosylated fibrils decorating the capsid. The genome is arranged in 5- or 6-start left-handed super-helices, with each DNA-strand lining the central channel. This luminal channel of the nucleoprotein fiber is wide enough to accommodate oxidative stress proteins and RNA polymerase subunits identified by proteomics. Such elegant supramolecular organization would represent a remarkable evolutionary strategy for packaging and protecting the genome, in a state ready for immediate transcription upon unwinding in the host cytoplasm. The parsimonious use of the same protein in two unrelated substructures of the virion is unexpected for a giant virus with thousand genes at its disposal.
pH: 7.5 / Component - Concentration: 40.0 mM / Component - Formula: (HOCH2)3CNH2 / Component - Name: Tris buffer
Vitrification
Cryogen name: ETHANE
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Electron microscopy
Microscope
FEI TITAN KRIOS
Image recording
Film or detector model: GATAN K2 QUANTUM (4k x 4k) / Average electron dose: 50.6 e/Å2
Electron beam
Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron optics
Illumination mode: OTHER / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.0 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
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Image processing
Startup model
Type of model: OTHER / Details: filtered Relion reconstruction, after subtraction.
Final reconstruction
Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.3 Å / Resolution method: FSC 0.5 CUT-OFF Details: Map obtained after focused refine of the compacted 5-start genomic fibre of Mimivirus Number images used: 95722
Initial angle assignment
Type: PROJECTION MATCHING
Final angle assignment
Type: PROJECTION MATCHING
FSC plot (resolution estimation)
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