Chikungunya virus assembly and budding visualized in situ using cryogenic electron tomography

Chikungunya virus (CHIKV) is a representative alphavirus causing debilitating arthritogenic disease in humans. Alphavirus particles assemble into two icosahedral layers: the glycoprotein spike shell embedded in a lipid envelope and the inner nucleocapsid (NC) core. In contrast to matrix-driven assembly of some enveloped viruses, the assembly/budding process of two-layered icosahedral particles remains poorly understood. Here we used cryogenic electron tomography (cryo-ET) to capture snapshots of the CHIKV assembly in infected human cells. Subvolume classification of the snapshots revealed 12 intermediates representing different stages of assembly at the plasma membrane. Further subtomogram average structures ranging from subnanometre to nanometre resolutions show that immature non-icosahedral NCs function as rough scaffolds to trigger icosahedral assembly of the spike lattice, which in turn progressively transforms the underlying NCs into icosahedral cores during budding. Further, analysis of CHIKV-infected cells treated with budding-inhibiting antibodies revealed wider spaces between spikes than in icosahedral spike lattice, suggesting that spacing spikes apart to prevent their lateral interactions prevents the plasma membrane from bending around the NC, thus blocking virus budding. These findings provide the molecular mechanisms for alphavirus assembly and antibody-mediated budding inhibition that provide valuable insights for the development of broad therapeutics targeting the assembly of icosahedral enveloped viruses. Cryogenic electron tomography analysis of Chikungunya virus particle assembly reveals 12 intermediate structural stages during virus assembly/budding at the plasma membrane and shows that non-icosahedral nucleocapsid proteins serve as scaffold to induce icosahedral assembly of the glycoprotein spike lattice. Structural analysis also shows that budding-inhibiting antibodies act by interfering with lateral spike interactions.

Automated summary

Cryogenic electron tomography analysis reveals the assembly process of Chikungunya virus in infected human cells, with non-icosahedral nucleocapsid proteins serving as a scaffold for the assembly of the glycoprotein spike lattice.