Snapshots from Cryo-ET of active SARS-CoV-2 virions

Fukuhara, H.; Dokainish, H. M.; Kita, S.; Tabata, K.; Takasu, A.; Huiskonen, J. T.; Anraku, Y.; Senda, T.; Stuart, D. I.; Sasaki, M.; Orba, Y.; Suzuki, Y.; Sawa, H.; Maenaka, K.

doi:10.1101/2023.10.10.561643

bioRxiv · 10.1101/2023.10.10.561643

Snapshots from Cryo-ET of active SARS-CoV-2 virions

Fukuhara, H.·Dokainish, H. M.·Kita, S.·Tabata, K.·Takasu, A.·Huiskonen, J. T.·Anraku, Y.·Senda, T.·Stuart, D. I.·Sasaki, M.·Orba, Y.·Suzuki, Y.·Sawa, H.·Maenaka, K.

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Abstract

Understanding the molecular properties of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is crucial for tackling future outbreaks. Current structural knowledge of the trimeric spike protein relies on truncated recombinant proteins and/or inactivated full-length forms, which may suffer from overstabilization. Here, we apply cryo-electron tomography (cryo-ET) at a Biosafety Level 3 facility to study the virus structure in its native, active state. The virus particles exhibit variable shapes and sizes with diffusible spikes, with the majority in typical prefusion conformations. Notably, we identified unprecedented, a transient open-trimer prefusion states, revealing a hidden flexibility with opened S1 conformation. Subtomogram averaging of the prefusion spikes indicates a loosely packed trimeric architecture that may facilitate the formation of open-trimer state. A cryo-EM map of recombinant Omicron BA.2.75 spike protein further confirms the presence of this loosely packed trimer as a minor conformational state. The observed dynamics uncover conserved cryptic regions that can be targeted for broadly effective vaccines. Structural analysis of active viruses profoundly impacts our understanding of the overlooked fusion mechanism and vaccine, antibody/drug design.

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Fukuhara, H., Dokainish, H. M., Kita, S., Tabata, K., Takasu, A., Huiskonen, J. T., Anraku, Y., Senda, T., Stuart, D. I., Sasaki, M., Orba, Y., Suzuki, Y., Sawa, H., Maenaka, K.. 2023-10-11. Snapshots from Cryo-ET of active SARS-CoV-2 virions. https://doi.org/10.1101/2023.10.10.561643

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