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Comment
. 2021 Jun 11;6(1):231.
doi: 10.1038/s41392-021-00651-y.

Palmitoylation of SARS-CoV-2 S protein is essential for viral infectivity

Zhuanchang Wu #  1 , Zhaoying Zhang #  2 , Xin Wang #  3 , Jing Zhang #  4 , Caiyue Ren  2 , Yuming Li  5 , Lifen Gao  2   6   7 , Xiaohong Liang  2   6   7 , Peihui Wang  8 , Chunhong Ma  9   10   11   12
Affiliations
Comment

Palmitoylation of SARS-CoV-2 S protein is essential for viral infectivity

Zhuanchang Wu et al. Signal Transduct Target Ther. .
No abstract available

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Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
Palmitoylation of SARS-CoV-2 S protein contributes to membrane fusion and viral infection. a SARS-CoV-2 S protein is palmitoylated. Flag-S was transfected into HEK293T cells, 48 h later, its palmitoylation was detected by ABE assay in the presence of hydroxylamine (HAM). b ZDHHC5 and GOLGA7 interact with S protein. Flag-S, HA-ZDHHC5 and Myc-GOLGA7 constructs were cotransfected into HEK293T cells, 48 h later, protein interactions were measured by Co-IP. c ZDHHC5 and GOLGA7 contribute to S protein palmitoylation. Flag-S was coexpressed with HA-ZDHHC5 or HA-ZDHHC5/Myc-GOLGA7 in HEK293T cells for 48 h, then the palmitoylation levels of S were detected. d The palmitoyltransferases activity of ZDHHC5 is essential for regulating S protein palmitoylation. Flag-S plasmid was cotransfected with HA-ZDHHC5 or HA-ZDHHC5-C143S plasmids into HEK293T cells, 48 h later, the palmitoylation levels of S were measured by ABE assay. e The palmitoylation sites of S protein are Cys residues at C-terminus. The palmitoylation sites of S were predicted by CSS-Palm tool. The blue fonts indicated the predicated palmitoylation sites (left panel), and the palmitoylation levels of wild-type S, C-C15A and S-∆C-Palm mutants were measured by ABE assay (right panel). f Palmitoylation of S protein is required for the infectivity of SARS-CoV-2 pseudoviruses. HEK293T-ACE2 cells were infected with lentiviruses pseudotyped with S-WT or S-∆C-Palm for 72 h, viral infection rate was analysed through detecting firefly luciferase activity relative to the level (set as 100) at S-WT (n = 3). Unpaired t-test, *P < 0.05; **P < 0.01. g S-trimer formation depends on its palmitoylation. Lentiviruses pseudotyped with S-WT or S-∆C-Palm were packaged from HEK293T cells and purified through supercentrifuging under 20% sucrose cushion, then S protein expression on pseudoviruses particles was detected by western blot, HIV-1 p24 antigen as loading control (upper panel). S-WT and S-∆C-Palm were overexpressed in HEK293T for 48 h, the relative S-trimer/monomer levels were detected by western blot (lower panel). h Palmitoylation of S protein is essential for S-mediated cell–cell fusion. S/GFP and S-∆C-Palm/GFP coexpressed HEK293T cells were cocultured with Dil-labelled Huh7 cells, cell fusion was measured with flow cytometry (n = 3) and visualized by fluorescent imaging at indicated time. The scale bar indicates 50 µm. One-way ANOVA, *P < 0.05; **P < 0.01. i ZDHHC5 knockdown inhibits SARS-CoV-2 pseudovirus infection. HEK293T cells were transfected with shZDHHC5 for 24 h, Flag-S plasmid alone or together with other packing plasmids were transfected into these cells. Another 48 h later, the SARS-CoV-2 pseudoviruses were collected to infect HEK293T-ACE2 cells. S palmitoylation levels were measured by ABE assay. S-mediated cell–cell fusion and SARS-CoV-2 pseudoviruses infection rate of HEK293T-ACE2 cells were detected as in 1h and 1f (n = 3). Unpaired t-test, *P < 0.05; **P < 0.01. j 2-BP represses SARS-CoV-2 pseudoviruses infection. HEK293T cells were transfected with Flag-S alone or together with other packing plasmids for 12 h and subsequently treated with 2-BP at 25 μM for another 36 h. S protein palmitoylation levels were measured by ABE assay. S-mediated cell fusion and pseudovirus infection rate were detected as in 1h and 1f (n = 3). Unpaired t-test, *P < 0.05; **P < 0.01
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Comment on

  • Palmitoylation of NOD1 and NOD2 is required for bacterial sensing.
    Lu Y, Zheng Y, Coyaud É, Zhang C, Selvabaskaran A, Yu Y, Xu Z, Weng X, Chen JS, Meng Y, Warner N, Cheng X, Liu Y, Yao B, Hu H, Xia Z, Muise AM, Klip A, Brumell JH, Girardin SE, Ying S, Fairn GD, Raught B, Sun Q, Neculai D. Lu Y, et al. Science. 2019 Oct 25;366(6464):460-467. doi: 10.1126/science.aau6391. Epub 2019 Oct 24. Science. 2019. PMID: 31649195

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