{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T16:30:43Z","timestamp":1785429043653,"version":"3.56.0"},"reference-count":76,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2026,4,21]],"date-time":"2026-04-21T00:00:00Z","timestamp":1776729600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,4,21]],"date-time":"2026-04-21T00:00:00Z","timestamp":1776729600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Virol"],"published-print":{"date-parts":[[2026,4,21]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>The spike (S) protein of SARS-CoV-2 is extensively glycosylated, with N-glycosylation sites remaining highly conserved during viral evolution. While inhibiting N-glycosylation has been shown to significantly suppress SARS-CoV-2 infection, the underlying molecular mechanisms remain incompletely characterized. Here, we identify that two N-glycosylation sites, N61 and N343, are critical for spike maturation. We demonstrate that asparagine-to-aspartic acid substitutions (N to D) at these sites lead to endoplasmic reticulum (ER) retention of the S protein, with consequent abrogation of S1\/S2 cleavage and near-complete elimination of viral infectivity. IP-MS analysis further reveals that the COPI complex, which facilitates retrograde Golgi-to-ER transport, is a key participant in this ER retention process. Additionally, inhibition of COPI effectively restores the plasma membrane localization of N61D- and N343D-mutated S proteins and enhanced viral infectivity. More importantly, a specific inhibitor has been developed that effectively blocks the ER-to-Golgi trafficking of the S protein, thereby broadly abolishing viral infectivity across SARS-CoV-2 variants. Overall, our study reveals the unique roles of N-glycosylation in the regulation of S protein maturation, providing a potential mechanistic target for antiviral drug development.<\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>N-glycosylation of the spike protein is critical for SARS-CoV-2. While most studies have focused on the effects on spike-ACE2 binding and neutralizing antibody recognition, few studies have reported how N-glycosylation regulates S protein maturation, with the underlying molecular mechanisms remaining poorly understood. Here, we demonstrate that N-glycosylation at N61\/ N343 contributes to spike ER-to-Golgi trafficking. Specifically, defects in S protein\u2019s N-glycosylation (including mutations at N61 or N343, N-glycosylation inhibitors treatment, and MOGS depletion) result in ER retention through COPI-mediated retrograde Golgi-to-ER transport, and thus, the S proteins are not effectively cleaved by furin in the Golgi. This impairment of S protein maturation leads to a significant reduction in viral infectivity, which highlights the key role of N-glycosylation at residues N61 and N343 in SARS-CoV-2 life cycle. Overall, our findings uncover the molecular mechanism by which N-glycosylation controls SARS-CoV-2 spike intracellular trafficking, offering novel insights for anti-SARS-CoV-2 strategies.<\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/jvi.00270-26","type":"journal-article","created":{"date-parts":[[2026,3,30]],"date-time":"2026-03-30T13:25:36Z","timestamp":1774877136000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Disruption of spike protein N-glycosylation induces its endoplasmic reticulum retention and attenuates SARS-CoV-2 infectivity"],"prefix":"10.1128","volume":"100","author":[{"given":"Weili","family":"Kong","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01zntxs11","id-type":"ROR","asserted-by":"publisher"}],"name":"Key Laboratory of Medical Molecular Virology (MOE\/NHC\/CAMS), Shanghai Frontiers Science Center of Pathogenic 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