Sequences in the Cytoplasmic Tail of SARS-CoV-2 Spike Facilitate Expression at the Cell Surface and Syncytia Formation

Overview

Journal Nat Commun

Specialty Biology

Date 2021 Sep 10

PMID 34504087

Citations 64

Authors

Jerome Cattin-Ortola

Lawrence G Welch

Sarah L Maslen

Guido Papa

Leo C James

Sean Munro

Affiliations

Soon will be listed here.

Abstract

The Spike (S) protein of SARS-CoV-2 binds ACE2 to direct fusion with host cells. S comprises a large external domain, a transmembrane domain, and a short cytoplasmic tail. Understanding the intracellular trafficking of S is relevant to SARS-CoV-2 infection, and to vaccines expressing full-length S from mRNA or adenovirus vectors. Here we report a proteomic screen for cellular factors that interact with the cytoplasmic tail of S. We confirm interactions with the COPI and COPII vesicle coats, ERM family actin regulators, and the WIPI3 autophagy component. The COPII binding site promotes exit from the endoplasmic reticulum, and although binding to COPI should retain S in the early Golgi where viral budding occurs, there is a suboptimal histidine residue in the recognition motif. As a result, S leaks to the surface where it accumulates and can direct the formation of multinucleate syncytia. Thus, the trafficking signals in the tail of S indicate that syncytia play a role in the SARS-CoV-2 lifecycle.

Citing Articles

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References

Alsaadi E, Jones I . Membrane binding proteins of coronaviruses. Future Virol. 2020; 14(4):275-286. PMC: 7079996. DOI: 10.2217/fvl-2018-0144. View

Ujike M, Taguchi F . Incorporation of spike and membrane glycoproteins into coronavirus virions. Viruses. 2015; 7(4):1700-25. PMC: 4411675. DOI: 10.3390/v7041700. View

Li F . Structure, Function, and Evolution of Coronavirus Spike Proteins. Annu Rev Virol. 2016; 3(1):237-261. PMC: 5457962. DOI: 10.1146/annurev-virology-110615-042301. View

Neuman B, Buchmeier M . Supramolecular Architecture of the Coronavirus Particle. Adv Virus Res. 2016; 96:1-27. PMC: 7112365. DOI: 10.1016/bs.aivir.2016.08.005. View

Cao Y, Yang R, Lee I, Zhang W, Sun J, Wang W . Characterization of the SARS-CoV-2 E Protein: Sequence, Structure, Viroporin, and Inhibitors. Protein Sci. 2021; 30(6):1114-1130. PMC: 8138525. DOI: 10.1002/pro.4075. View

Forni D, Cagliani R, Clerici M, Sironi M . Molecular Evolution of Human Coronavirus Genomes. Trends Microbiol. 2016; 25(1):35-48. PMC: 7111218. DOI: 10.1016/j.tim.2016.09.001. View

Nal B, Chan C, Kien F, Siu L, Tse J, Chu K . Differential maturation and subcellular localization of severe acute respiratory syndrome coronavirus surface proteins S, M and E. J Gen Virol. 2005; 86(Pt 5):1423-1434. DOI: 10.1099/vir.0.80671-0. View

Krijnse-Locker J, Ericsson M, Rottier P, Griffiths G . Characterization of the budding compartment of mouse hepatitis virus: evidence that transport from the RER to the Golgi complex requires only one vesicular transport step. J Cell Biol. 1994; 124(1-2):55-70. PMC: 2119890. DOI: 10.1083/jcb.124.1.55. View

Tooze J, Tooze S, Fuller S . Sorting of progeny coronavirus from condensed secretory proteins at the exit from the trans-Golgi network of AtT20 cells. J Cell Biol. 1987; 105(3):1215-26. PMC: 2114808. DOI: 10.1083/jcb.105.3.1215. View

10.

Klein S, Cortese M, Winter S, Wachsmuth-Melm M, Neufeldt C, Cerikan B . SARS-CoV-2 structure and replication characterized by in situ cryo-electron tomography. Nat Commun. 2020; 11(1):5885. PMC: 7676268. DOI: 10.1038/s41467-020-19619-7. View

11.

Hoffmann M, Kleine-Weber H, Schroeder S, Kruger N, Herrler T, Erichsen S . SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor. Cell. 2020; 181(2):271-280.e8. PMC: 7102627. DOI: 10.1016/j.cell.2020.02.052. View

12.

Bao L, Deng W, Huang B, Gao H, Liu J, Ren L . The pathogenicity of SARS-CoV-2 in hACE2 transgenic mice. Nature. 2020; 583(7818):830-833. DOI: 10.1038/s41586-020-2312-y. View

13.

Shang J, Ye G, Shi K, Wan Y, Luo C, Aihara H . Structural basis of receptor recognition by SARS-CoV-2. Nature. 2020; 581(7807):221-224. PMC: 7328981. DOI: 10.1038/s41586-020-2179-y. View

14.

Yan R, Zhang Y, Li Y, Xia L, Guo Y, Zhou Q . Structural basis for the recognition of SARS-CoV-2 by full-length human ACE2. Science. 2020; 367(6485):1444-1448. PMC: 7164635. DOI: 10.1126/science.abb2762. View

15.

Tseng C, Tseng J, Perrone L, Worthy M, Popov V, Peters C . Apical entry and release of severe acute respiratory syndrome-associated coronavirus in polarized Calu-3 lung epithelial cells. J Virol. 2005; 79(15):9470-9. PMC: 1181546. DOI: 10.1128/JVI.79.15.9470-9479.2005. View

16.

Zhu N, Wang W, Liu Z, Liang C, Wang W, Ye F . Morphogenesis and cytopathic effect of SARS-CoV-2 infection in human airway epithelial cells. Nat Commun. 2020; 11(1):3910. PMC: 7413383. DOI: 10.1038/s41467-020-17796-z. View

17.

Buchrieser J, Dufloo J, Hubert M, Monel B, Planas D, Rajah M . Syncytia formation by SARS-CoV-2-infected cells. EMBO J. 2020; 39(23):e106267. PMC: 7646020. DOI: 10.15252/embj.2020106267. View

18.

Tian S, Hu W, Niu L, Liu H, Xu H, Xiao S . Pulmonary Pathology of Early-Phase 2019 Novel Coronavirus (COVID-19) Pneumonia in Two Patients With Lung Cancer. J Thorac Oncol. 2020; 15(5):700-704. PMC: 7128866. DOI: 10.1016/j.jtho.2020.02.010. View

19.

McBride C, Li J, Machamer C . The cytoplasmic tail of the severe acute respiratory syndrome coronavirus spike protein contains a novel endoplasmic reticulum retrieval signal that binds COPI and promotes interaction with membrane protein. J Virol. 2006; 81(5):2418-28. PMC: 1865919. DOI: 10.1128/JVI.02146-06. View

20.

Boson B, Legros V, Zhou B, Siret E, Mathieu C, Cosset F . The SARS-CoV-2 envelope and membrane proteins modulate maturation and retention of the spike protein, allowing assembly of virus-like particles. J Biol Chem. 2020; 296:100111. PMC: 7833635. DOI: 10.1074/jbc.RA120.016175. View