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A single C-terminal residue controls SARS-CoV-2 spike trafficking and virion assembly

Dey, D.; Qing, E.; He, Y.; Chen, Y.; Jennings, B.; Cohn, W.; Singh, S.; Gakhar, L.; Schnicker, N. J.; Pierce, B. G.; Whitelegge, J. P.; Doray, B.; Orban, J. P.; Gallagher, T.; Hasan, S. S.

2023-03-10 biophysics
10.1101/2023.03.09.531992 bioRxiv
Show abstract

The spike (S) protein of SARS-CoV-2 is delivered to the virion assembly site in the ER-Golgi Intermediate Compartment (ERGIC) from both the ER and cis-Golgi in infected cells1-3. However, the relevance and modulatory mechanism of this bidirectional trafficking are unclear. Here, using structure-function analyses, we show that S incorporation into virions and viral fusogenicity are determined by coatomer-dependent S delivery from the cis-Golgi and restricted by S-coatomer dissociation. Although S mimicry of the host coatomer-binding dibasic motif ensures retrograde trafficking to the ERGIC, avoidance of the host-like C-terminal acidic residue is critical for S-coatomer dissociation and therefore incorporation into virions or export for cell-cell fusion. Because this C-terminal residue is the key determinant of SARS-CoV-2 assembly and fusogenicity, our work provides a framework for the export of S protein encoded in genetic vaccines for surface display and immune activation.

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