A single C-terminal residue controls SARS-CoV-2 spike trafficking and incorporation into VLPs

Debajit Dey; Enya Qing; Yanan He; Yihong Chen; Benjamin Jennings; Whitaker Cohn; Suruchi Singh; Lokesh Gakhar; Nicholas J Schnicker; Brian G Pierce; Julian P Whitelegge; Balraj Doray; John Orban; Tom Gallagher; S Saif Hasan

doi:10.1038/s41467-023-44076-3

A single C-terminal residue controls SARS-CoV-2 spike trafficking and incorporation into VLPs

Nat Commun. 2023 Dec 15;14(1):8358. doi: 10.1038/s41467-023-44076-3.

Authors

Debajit Dey¹, Enya Qing², Yanan He³, Yihong Chen³, Benjamin Jennings⁴, Whitaker Cohn⁵, Suruchi Singh¹, Lokesh Gakhar^{6

7

8}, Nicholas J Schnicker^{7

9}, Brian G Pierce^{3

10}, Julian P Whitelegge^{5

11

12}, Balraj Doray⁴, John Orban^{3

13}, Tom Gallagher², S Saif Hasan^{14

15

16}

Affiliations

¹ Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, MD, 21201, USA.
² Department of Microbiology and Immunology, Loyola University Chicago, Maywood, IL, 60153, USA.
³ University of Maryland Institute for Bioscience and Biotechnology Research, Rockville, MD, 20850, USA.
⁴ Department of Internal Medicine, Hematology Division, Washington University School of Medicine, St. Louis, MO, 63110, USA.
⁵ Pasarow Mass Spectrometry Laboratory, The Jane and Terry Semel Institute for Neuroscience and Human Behavior, David Geffen School of Medicine, University of California, Los Angeles, CA, 90095, USA.
⁶ Department of Biochemistry and Molecular Biology, Carver College of Medicine, University of Iowa, Iowa City, IA, 52242, USA.
⁷ Protein and Crystallography Facility, Carver College of Medicine, University of Iowa, Iowa City, IA, 52242, USA.
⁸ PAQ Therapeutics, Burlington, MA, 01803, USA.
⁹ Department of Molecular Physiology and Biophysics, Carver College of Medicine, University of Iowa, Iowa City, IA, 52242, USA.
¹⁰ Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD, 20742, USA.
¹¹ Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
¹² Jonsson Comprehensive Cancer Center, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
¹³ Department of Chemistry and Biochemistry, University of Maryland, College Park, MD, 20742, USA.
¹⁴ Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, MD, 21201, USA. sshasan@som.umaryland.edu.
¹⁵ University of Maryland Marlene and Stewart Greenebaum Cancer Center, University of Maryland Medical Center, Baltimore, MD, 21201, USA. sshasan@som.umaryland.edu.
¹⁶ Center for Biomolecular Therapeutics, University of Maryland School of Medicine, Rockville, MD, 20850, USA. sshasan@som.umaryland.edu.

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 cells. However, the relevance and modulatory mechanism of this bidirectional trafficking are unclear. Here, using structure-function analyses, we show that S incorporation into virus-like particles (VLP) and VLP 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.

MeSH terms

COVID-19* / metabolism
Golgi Apparatus / metabolism
Humans
SARS-CoV-2* / genetics
Spike Glycoprotein, Coronavirus / genetics
Spike Glycoprotein, Coronavirus / metabolism

Substances

Spike Glycoprotein, Coronavirus
spike protein, SARS-CoV-2

Abstract

MeSH terms

Substances

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