Journal: Microbiology Spectrum
Article Title: O-GlcNAcylation at S659 enhances SARS-CoV-2 spike protein stability and pseudoparticle packaging efficiency
doi: 10.1128/spectrum.00527-25
Figure Lengend Snippet: O-GlcNAcylation of S at S659 regulates pseudoviral packaging of SARS-CoV-2. ( A ) S-S659A does not affect the binding between S and Ace2. HEK293T cells were transfected with the plasmids expressing HA-ACE2 and GFP-S-WT, GFP-S-659A, or empty vector. The cell lysates were subject to IP. ( B ) Quantitation of ( A ). ( C ) Diagram of four-plasmid system for SARS-CoV-2pp production. The four-plasmid system for preparing pseudovirus SARS-CoV-2pp includes Luc (pLenti6) encoding luciferase, HIV Gag/pol (pLP1), HIV Rev (pLP2), and GFP-S. ( D ) S659A mutation in SARS-CoV-2 S reduced the SARS-CoV2pp package efficiency. The luciferase activity of lysates from Huh7.5.1 cells infected with SARS-CoV-2pp generated from HEK293T cells based on either S or the S-S659A was assayed. ns, no significance; ****, P < 0.0001.
Article Snippet: The human ACE2 ORF cDNA clone expression plasmid with a C-HA tag was sourced from Sino Biological (HG10108-CY).
Techniques: Binding Assay, Transfection, Expressing, Plasmid Preparation, Quantitation Assay, Luciferase, Mutagenesis, Activity Assay, Infection, Generated