Structural variability and complexity of the giant Pithovirus sibericum particle revealed by high-voltage electron cryo-tomography and energy-filtered electron cryo-microscopy

Kenta Okamoto , Naoyuki Miyazaki (1) , Chihong Song , Filipe Maia , Hemanth Reddy , Chantal Abergel (2) , Jean-Michel Claverie (2) , Janos Hajdu , Martin Svenda (3) , Kazuyoshi Murata (1)

1 National Institutes of Natural Sciences
2 IGS - Information génomique et structurale
3 Department of Cell and Molecular Biology [Uppsala]

The Pithoviridae giant virus family exhibits the largest viral particle known so far, a prolate spheroid up to 2.5 μm in length and 0.9 μm in diameter. These particles show significant variations in size. Little is known about the structure of the intact virion due to technical limitations with conventional electron cryo-microscopy (cryo-EM) when imaging thick specimens. Here we present the intact structure of the giant Pithovirus sibericum particle at near native conditions using high-voltage electron cryo-tomography (cryo-ET) and energy-filtered cryo-EM. We detected a previously undescribed low-density outer layer covering the tegument and a periodical structuring of the fibres in the striated apical cork. Energy-filtered Zernike phase-contrast cryo-EM images show distinct substructures inside the particles, implicating an internal compartmentalisation. The density of the interior volume of Pithovirus particles is three quarters lower than that of the Mimivirus. However, it is remarkably high given that the 600 kbp Pithovirus genome is only half the size of the Mimivirus genome and is packaged in a volume up to 100 times larger. These observations suggest that the interior is densely packed with macromolecules in addition to the genomic nucleic acid.

Domaines: Virologie

Kenta Okamoto, Naoyuki Miyazaki, Chihong Song, Filipe Maia, Hemanth Reddy, et al.. Structural variability and complexity of the giant Pithovirus sibericum particle revealed by high-voltage electron cryo-tomography and energy-filtered electron cryo-microscopy. Scientific Reports, 2017, 7 (1), pp.13291. ⟨10.1038/s41598-017-13390-4⟩. ⟨hal-01785112⟩