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SARS-CoV-2 spike protein expression in the ACE2-expressing cell surface induces cell\u2013cell membrane fusion, thus forming syncytia. To exert its fusogenic activity, the spike protein is typically processed at a specific site (the S1\/S2 site) by cellular proteases such as furin. The C488 residue, located at the spike\u2013ACE2 interacting surface, is critical for the fusogenic and infectious roles of the SARS-CoV-2 spike protein. We have demonstrated that the C488 residue of the spike protein is involved in subcellular targeting and S1\/S2 processing. C488 mutant spike localization to the Golgi apparatus and cell surface were impaired. Consequently, the S1\/S2 processing of the spike protein, probed by anti-Ser-686-cleaved spike antibody, markedly decreased in C488 mutant spike proteins. Moreover, brefeldin-A-mediated endoplasmic-reticulum-to-Golgi traffic suppression also suppressed spike protein S1\/S2 processing. As brefeldin A treatment and C488 mutation inhibited S1\/S2 processing and syncytia formation, the C488 residue of spike protein is required for functional spike protein processing.<\/jats:p>","DOI":"10.3390\/ijms232415834","type":"journal-article","created":{"date-parts":[[2022,12,14]],"date-time":"2022-12-14T01:52:34Z","timestamp":1670982754000},"page":"15834","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["SARS-CoV-2 Spike Protein Mutation at Cysteine-488 Impairs Its Golgi Localization and Intracellular S1\/S2 Processing"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1992-1806","authenticated-orcid":false,"given":"Yuichiro","family":"Yamamoto","sequence":"first","affiliation":[{"name":"Laboratory of Molecular Targeted Therapy, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda 278-8510, Chiba, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tetsuya","family":"Inoue","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Targeted Therapy, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda 278-8510, Chiba, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miyu","family":"Inoue","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Targeted Therapy, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda 278-8510, Chiba, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mana","family":"Murae","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Targeted Therapy, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda 278-8510, Chiba, Japan"},{"name":"Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masayoshi","family":"Fukasawa","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Targeted Therapy, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda 278-8510, Chiba, Japan"},{"name":"Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4158-9208","authenticated-orcid":false,"given":"Mika K.","family":"Kaneko","sequence":"additional","affiliation":[{"name":"Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Sendai 980-8575, Miyagi, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5385-8201","authenticated-orcid":false,"given":"Yukinari","family":"Kato","sequence":"additional","affiliation":[{"name":"Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Sendai 980-8575, Miyagi, Japan"},{"name":"Department of Molecular Pharmacology, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Sendai 980-8575, Miyagi, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9080-0807","authenticated-orcid":false,"given":"Kohji","family":"Noguchi","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Targeted Therapy, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda 278-8510, Chiba, Japan"},{"name":"Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"5333","DOI":"10.1038\/s41467-021-25589-1","article-title":"Sequences in the Cytoplasmic Tail of SARS-CoV-2 Spike Facilitate Expression at the Cell Surface and Syncytia Formation","volume":"12","author":"Welch","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"779","DOI":"10.1016\/j.molcel.2020.04.022","article-title":"A Multibasic Cleavage Site in the Spike Protein of SARS-CoV-2 Is Essential for Infection of Human Lung Cells","volume":"78","author":"Hoffmann","year":"2020","journal-title":"Mol. Cell"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1016\/j.cell.2020.02.058","article-title":"Structure, Function, and Antigenicity of the SARS-CoV-2 Spike Glycoprotein","volume":"181","author":"Walls","year":"2020","journal-title":"Cell"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1038\/s41580-021-00418-x","article-title":"Mechanisms of SARS-CoV-2 Entry into Cells","volume":"23","author":"Jackson","year":"2022","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3910","DOI":"10.1038\/s41467-020-17796-z","article-title":"Morphogenesis and Cytopathic Effect of SARS-CoV-2 Infection in Human Airway Epithelial Cells","volume":"11","author":"Zhu","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Buchrieser, J., Dufloo, J., Hubert, M., Monel, B., Planas, D., Rajah, M.M., Planchais, C., Porrot, F., Guivel-Benhassine, F., and van der Werf, S. (2020). Syncytia Formation by SARS-CoV-2-infected Cells. EMBO J., 39.","DOI":"10.15252\/embj.2020106267"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Saraste, J., and Prydz, K. (2021). Assembly and Cellular Exit of Coronaviruses: Hijacking an Unconventional Secretory Pathway from the Pre-Golgi Intermediate Compartment via the Golgi Ribbon to the Extracellular Space. Cells, 10.","DOI":"10.3390\/cells10030503"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2418","DOI":"10.1128\/JVI.02146-06","article-title":"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","volume":"81","author":"McBride","year":"2007","journal-title":"J. Virol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1758","DOI":"10.1002\/1873-3468.14109","article-title":"A Weak COPI Binding Motif in the Cytoplasmic Tail of SARS-CoV-2 Spike Glycoprotein Is Necessary for Its Cleavage, Glycosylation, and Localization","volume":"595","author":"Jennings","year":"2021","journal-title":"FEBS Lett."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/j.bbrc.2022.01.106","article-title":"The Function of SARS-CoV-2 Spike Protein Is Impaired by Disulfide-Bond Disruption with Mutation at Cysteine-488 and by Thiol-Reactive N-Acetyl-Cysteine and Glutathione","volume":"597","author":"Murae","year":"2022","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Hackstadt, T., Chiramel, A.I., Hoyt, F.H., Williamson, B.N., Dooley, C.A., Beare, P.A., de Wit, E., Best, S.M., and Fischer, E.R. (2021). Disruption of the Golgi Apparatus and Contribution of the Endoplasmic Reticulum to the SARS-CoV-2 Replication Complex. Viruses, 13.","DOI":"10.3390\/v13091798"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"753","DOI":"10.1038\/nrm934","article-title":"Furin at the Cutting Edge: From Protein Traffic to Embryogenesis and Disease","volume":"3","author":"Thomas","year":"2002","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2424","DOI":"10.1002\/j.1460-2075.1995.tb07240.x","article-title":"Two Independent Targeting Signals in the Cytoplasmic Domain Determine Trans-Golgi Network Localization and Endosomal Trafficking of the Proprotein Convertase Furin","volume":"14","author":"Stroh","year":"1995","journal-title":"EMBO J."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e01626-2","DOI":"10.1128\/jvi.01626-21","article-title":"Analysis of Glycosylation and Disulfide Bonding of Wild-Type SARS-CoV-2 Spike Glycoprotein","volume":"96","author":"Zhang","year":"2022","journal-title":"J. Virol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"706","DOI":"10.1038\/s41586-022-04474-x","article-title":"Altered TMPRSS2 Usage by SARS-CoV-2 Omicron Impacts Infectivity and Fusogenicity","volume":"603","author":"Meng","year":"2022","journal-title":"Nature"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"100111","DOI":"10.1074\/jbc.RA120.016175","article-title":"The SARS-CoV-2 Envelope and Membrane Proteins Modulate Maturation and Retention of the Spike Protein, Allowing Assembly of Virus-like Particles","volume":"296","author":"Boson","year":"2021","journal-title":"J. Biol. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"eabl4895","DOI":"10.1126\/sciadv.abl4895","article-title":"SARS-CoV-2 Nucleocapsid Protein Adheres to Replication Organelles before Viral Assembly at the Golgi\/ERGIC and Lysosome-Mediated Egress","volume":"8","author":"Scherer","year":"2022","journal-title":"Sci. Adv."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"11398","DOI":"10.1016\/S0021-9258(18)67398-3","article-title":"Novel Blockade by Brefeldin A of Intracellular Transport of Secretory Proteins in Cultured Rat Hepatocytes","volume":"261","author":"Misumi","year":"1986","journal-title":"J. Biol. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1520","DOI":"10.1016\/j.cell.2020.10.039","article-title":"\u03b2-Coronaviruses Use Lysosomes for Egress Instead of the Biosynthetic Secretory Pathway","volume":"183","author":"Ghosh","year":"2020","journal-title":"Cell"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1295","DOI":"10.1016\/j.cell.2020.08.012","article-title":"Deep Mutational Scanning of SARS-CoV-2 Receptor Binding Domain Reveals Constraints on Folding and ACE2 Binding","volume":"182","author":"Starr","year":"2020","journal-title":"Cell"},{"key":"ref_21","first-page":"e21651","article-title":"Disruption of Disulfides within RBD of SARS-CoV-2 Spike Protein Prevents Fusion and Represents a Target for Viral Entry Inhibition by Registered Drugs","volume":"35","author":"Govednik","year":"2021","journal-title":"FASEB J."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Goh, G.K.-M., Dunker, A.K., Foster, J.A., and Uversky, V.N. (2022). A Study on the Nature of SARS-CoV-2 Using the Shell Disorder Models: Reproducibility, Evolution, Spread, and Attenuation. Biomolecules, 12.","DOI":"10.3390\/biom12101353"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2991","DOI":"10.3390\/v6082991","article-title":"The Coronavirus Nucleocapsid Is a Multifunctional Protein","volume":"6","author":"McBride","year":"2014","journal-title":"Viruses"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"104709","DOI":"10.1016\/j.isci.2022.104709","article-title":"Glycosylation and S-Palmitoylation Regulate SARS-CoV-2 Spike Protein Intracellular Trafficking","volume":"25","author":"Tien","year":"2022","journal-title":"iScience"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"103589","DOI":"10.1016\/j.isci.2021.103589","article-title":"Functional Evaluation of the P681H Mutation on the Proteolytic Activation of the SARS-CoV-2 Variant B.1.1.7 (Alpha) Spike","volume":"25","author":"Lubinski","year":"2022","journal-title":"iScience"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1161","DOI":"10.1038\/s41564-022-01143-7","article-title":"SARS-CoV-2 Omicron Is an Immune Escape Variant with an Altered Cell Entry Pathway","volume":"7","author":"Willett","year":"2022","journal-title":"Nat. Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"111220","DOI":"10.1016\/j.celrep.2022.111220","article-title":"SARS-CoV-2 Spike N-Terminal Domain Modulates TMPRSS2-Dependent Viral Entry and Fusogenicity","volume":"40","author":"Meng","year":"2022","journal-title":"Cell Rep."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Yamaoka, Y., Jeremiah, S.S., Funabashi, R., Miyakawa, K., Morita, T., Mihana, Y., Kato, H., and Ryo, A. (2022). Characterization and Utilization of Disulfide-Bonded SARS-CoV-2 Receptor Binding Domain of Spike Protein Synthesized by Wheat Germ Cell-Free Production System. Viruses, 14.","DOI":"10.3390\/v14071461"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1016\/j.cell.2020.09.032","article-title":"Structural and Functional Analysis of the D614G SARS-CoV-2 Spike Protein Variant","volume":"183","author":"Yurkovetskiy","year":"2020","journal-title":"Cell"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/24\/15834\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:40:31Z","timestamp":1760146831000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/24\/15834"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,13]]},"references-count":29,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2022,12]]}},"alternative-id":["ijms232415834"],"URL":"https:\/\/doi.org\/10.3390\/ijms232415834","relation":{"has-preprint":[{"id-type":"doi","id":"10.20944\/preprints202210.0245.v1","asserted-by":"object"}]},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,13]]}}}