Acknowledgement
We thank E. Tunkle for preparation of the manuscript and J. Choi at Dongduk University College of Pharmacy for technical assistance. This work was supported by grants from the National Research Foundation of Korea (NRF-2019R1A2C2084535, NRF-2021R1A2C3003496, NRF-2022R1A2C3004609) funded by the Korean government (MSIP), and a grant from the National Research Foundation of Korea (NRF-2020R1I1A1A01072977) funded by the Korean government (MOE).
References
- Ahn, S.Y., Kim, N.H., Lee, K., Cha, Y.H., Yang, J.H., Cha, S.Y., Cho, E.S., Lee, Y., Cha, J.S., Cho, H.S., et al. (2017a). Niclosamide is a potential therapeutic for familial adenomatosis polyposis by disrupting Axin-GSK3 interaction. Oncotarget 8, 31842-31855. https://doi.org/10.18632/oncotarget.16252
- Ahn, S.Y., Yang, J.H., Kim, N.H., Lee, K., Cha, Y.H., Yun, J.S., Kang, H.E., Lee, Y., Choi, J., Kim, H.S., et al. (2017b). Anti-helminthic niclosamide inhibits Ras-driven oncogenic transformation via activation of GSK-3. Oncotarget 8, 31856-31863. https://doi.org/10.18632/oncotarget.16255
- Alfhili, M.A., Alsughayyir, J., McCubrey, J.A., and Akula, S.M. (2020). GSK-3-associated signaling is crucial to virus infection of cells. Biochim. Biophys. Acta Mol. Cell Res. 1867, 118767.
- Al-Kuraishy, H.M., Al-Gareeb, A.I., Alzahrani, K.J., Alexiou, A., and Batiha, G.E. (2021). Niclosamide for Covid-19: bridging the gap. Mol. Biol. Rep. 48, 8195-8202. https://doi.org/10.1007/s11033-021-06770-7
- Cabantous, S., Terwilliger, T.C., and Waldo, G.S. (2005). Protein tagging and detection with engineered self-assembling fragments of green fluorescent protein. Nat. Biotechnol. 23, 102-107. https://doi.org/10.1038/nbt1044
- Carlson, C.R., Asfaha, J.B., Ghent, C.M., Howard, C.J., Hartooni, N., Safari, M., Frankel, A.D., and Morgan, D.O. (2020). Phosphoregulation of phase separation by the SARS-CoV-2 N protein suggests a biophysical basis for its dual functions. Mol. Cell 80, 1092-1103.e4. https://doi.org/10.1016/j.molcel.2020.11.025
- Chang, C.K., Hou, M.H., Chang, C.F., Hsiao, C.D., and Huang, T.H. (2014). The SARS coronavirus nucleocapsid protein--forms and functions. Antiviral Res. 103, 39-50. https://doi.org/10.1016/j.antiviral.2013.12.009
- Chen, H., Gill, A., Dove, B.K., Emmett, S.R., Kemp, C.F., Ritchie, M.A., Dee, M., and Hiscox, J.A. (2005). Mass spectroscopic characterization of the coronavirus infectious bronchitis virus nucleoprotein and elucidation of the role of phosphorylation in RNA binding by using surface plasmon resonance. J. Virol. 79, 1164-1179. https://doi.org/10.1128/JVI.79.2.1164-1179.2005
- Cohen, P. and Frame, S. (2001). The renaissance of GSK3. Nat. Rev. Mol. Cell Biol. 2, 769-776. https://doi.org/10.1038/35096075
- Cubuk, J., Alston, J.J., Incicco, J.J., Singh, S., Stuchell-Brereton, M.D., Ward, M.D., Zimmerman, M.I., Vithani, N., Griffith, D., Wagoner, J.A., et al. (2021). The SARS-CoV-2 nucleocapsid protein is dynamic, disordered, and phase separates with RNA. Nat. Commun. 12, 1936.
- Dajani, R., Fraser, E., Roe, S.M., Yeo, M., Good, V.M., Thompson, V., Dale, T.C., and Pearl, L.H. (2003). Structural basis for recruitment of glycogen synthase kinase 3beta to the axin-APC scaffold complex. EMBO J. 22, 494-501. https://doi.org/10.1093/emboj/cdg068
- de Haan, C.A. and Rottier, P.J. (2005). Molecular interactions in the assembly of coronaviruses. Adv. Virus Res. 64, 165-230. https://doi.org/10.1016/S0065-3527(05)64006-7
- Doble, B.W. and Woodgett, J.R. (2003). GSK-3: tricks of the trade for a multi-tasking kinase. J. Cell Sci. 116, 1175-1186. https://doi.org/10.1242/jcs.00384
- Fraser, E., Young, N., Dajani, R., Franca-Koh, J., Ryves, J., Williams, R.S., Yeo, M., Webster, M.T., Richardson, C., Smalley, M.J., et al. (2002). Identification of the Axin and Frat binding region of glycogen synthase kinase-3. J. Biol. Chem. 277, 2176-2185. https://doi.org/10.1074/jbc.M109462200
- Fujimuro, M., Liu, J., Zhu, J., Yokosawa, H., and Hayward, S.D. (2005). Regulation of the interaction between glycogen synthase kinase 3 and the Kaposi's sarcoma-associated herpesvirus latency-associated nuclear antigen. J. Virol. 79, 10429-10441. https://doi.org/10.1128/JVI.79.16.10429-10441.2005
- Hedgepeth, C.M., Deardorff, M.A., Rankin, K., and Klein, P.S. (1999). Regulation of glycogen synthase kinase 3beta and downstream Wnt signaling by axin. Mol. Cell. Biol. 19, 7147-7157. https://doi.org/10.1128/mcb.19.10.7147
- Howng, S.L., Hwang, C.C., Hsu, C.Y., Hsu, M.Y., Teng, C.Y., Chou, C.H., Lee, M.F., Wu, C.H., Chiou, S.J., Lieu, A.S., et al. (2010). Involvement of the residues of GSKIP, AxinGID, and FRATtide in their binding with GSK3beta to unravel a novel C-terminal scaffold-binding region. Mol. Cell. Biochem. 339, 23-33. https://doi.org/10.1007/s11010-009-0366-0
- Kaidanovich-Beilin, O. and Woodgett, J.R. (2011). GSK-3: functional insights from cell biology and animal models. Front. Mol. Neurosci. 4, 40.
- Kim, N.H., Kim, H.S., Li, X.Y., Lee, I., Choi, H.S., Kang, S.E., Cha, S.Y., Ryu, J.K., Yoon, D., Fearon, E.R., et al. (2011). A p53/miRNA-34 axis regulates Snail1- dependent cancer cell epithelial-mesenchymal transition. J. Cell Biol. 195, 417-433. https://doi.org/10.1083/jcb.201103097
- Kinoshita, E., Kinoshita-Kikuta, E., Takiyama, K., and Koike, T. (2006). Phosphate-binding tag, a new tool to visualize phosphorylated proteins. Mol. Cell. Proteomics 5, 749-757. https://doi.org/10.1074/mcp.T500024-MCP200
- Ko, M., Jeon, S., Ryu, W.S., and Kim, S. (2021). Comparative analysis of antiviral efficacy of FDA-approved drugs against SARS-CoV-2 in human lung cells. J. Med. Virol. 93, 1403-1408. https://doi.org/10.1002/jmv.26397
- Krutikov, M., Palmer, T., Tut, G., Fuller, C., Shrotri, M., Williams, H., Davies, D., Irwin-Singer, A., Robson, J., Hayward, A., et al. (2021). Incidence of SARS-CoV-2 infection according to baseline antibody status in staff and residents of 100 long-term care facilities (VIVALDI): a prospective cohort study. Lancet Healthy Longev. 2, e362-e370. https://doi.org/10.1016/S2666-7568(21)00093-3
- Lee, D.G., Kim, H.S., Lee, Y.S., Kim, S., Cha, S.Y., Ota, I., Kim, N.H., Cha, Y.H., Yang, D.H., Lee, Y., et al. (2014). Helicobacter pylori CagA promotes Snail-mediated epithelial-mesenchymal transition by reducing GSK-3 activity. Nat. Commun. 5, 4423.
- Lee, Y., Kim, N.H., Cho, E.S., Yang, J.H., Cha, Y.H., Kang, H.E., Yun, J.S., Cho, S.B., Lee, S.H., Paclikova, P., et al. (2018). Dishevelled has a YAP nuclear export function in a tumor suppressor context-dependent manner. Nat. Commun. 9, 2301.
- Liu, X., Verma, A., Garcia, G., Jr., Ramage, H., Lucas, A., Myers, R.L., Michaelson, J.J., Coryell, W., Kumar, A., Charney, A.W., et al. (2021). Targeting the coronavirus nucleocapsid protein through GSK-3 inhibition. Proc. Natl. Acad. Sci. U. S. A. 118, e2113401118.
- Lu, S., Ye, Q., Singh, D., Cao, Y., Diedrich, J.K., Yates, J.R., 3rd, Villa, E., Cleveland, D.W., and Corbett, K.D. (2021). The SARS-CoV-2 nucleocapsid phosphoprotein forms mutually exclusive condensates with RNA and the membrane-associated M protein. Nat. Commun. 12, 502.
- Meyer, B., Drosten, C., and Muller, M.A. (2014). Serological assays for emerging coronaviruses: challenges and pitfalls. Virus Res. 194, 175-183. https://doi.org/10.1016/j.virusres.2014.03.018
- Peng, T.Y., Lee, K.R., and Tarn, W.Y. (2008). Phosphorylation of the arginine/serine dipeptide-rich motif of the severe acute respiratory syndrome coronavirus nucleocapsid protein modulates its multimerization, translation inhibitory activity and cellular localization. FEBS J. 275, 4152-4163. https://doi.org/10.1111/j.1742-4658.2008.06564.x
- Shah, M. and Woo, H.G. (2021). Molecular perspectives of SARS-CoV-2: pathology, immune evasion, and therapeutic interventions. Mol. Cells 44, 408-421. https://doi.org/10.14348/molcells.2021.0026
- Shan, D., Johnson, J.M., Fernandes, S.C., Suib, H., Hwang, S., Wuelfing, D., Mendes, M., Holdridge, M., Burke, E.M., Beauregard, K., et al. (2021). N-protein presents early in blood, dried blood and saliva during asymptomatic and symptomatic SARS-CoV-2 infection. Nat. Commun. 12, 1931.
- Surjit, M., Liu, B., Chow, V.T., and Lal, S.K. (2006). The nucleocapsid protein of severe acute respiratory syndrome-coronavirus inhibits the activity of cyclin-cyclin-dependent kinase complex and blocks S phase progression in mammalian cells. J. Biol. Chem. 281, 10669-10681. https://doi.org/10.1074/jbc.M509233200
- Syed, A.M., Taha, T.Y., Tabata, T., Chen, I.P., Ciling, A., Khalid, M.M., Sreekumar, B., Chen, P.Y., Hayashi, J.M., Soczek, K.M., et al. (2021). Rapid assessment of SARS-CoV-2-evolved variants using virus-like particles. Science 374, 1626-1632. https://doi.org/10.1126/science.abl6184
- Tabibzadeh, A., Esghaei, M., Soltani, S., Yousefi, P., Taherizadeh, M., Safarnezhad Tameshkel, F., Golahdooz, M., Panahi, M., Ajdarkosh, H., Zamani, F., et al. (2021). Evolutionary study of COVID-19, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as an emerging coronavirus: phylogenetic analysis and literature review. Vet. Med. Sci. 7, 559-571. https://doi.org/10.1002/vms3.394
- Tan, Y.J., Goh, P.Y., Fielding, B.C., Shen, S., Chou, C.F., Fu, J.L., Leong, H.N., Leo, Y.S., Ooi, E.E., Ling, A.E., et al. (2004). Profiles of antibody responses against severe acute respiratory syndrome coronavirus recombinant proteins and their potential use as diagnostic markers. Clin. Diagn. Lab. Immunol. 11, 362-371.
- Tay, M.Z., Poh, C.M., Renia, L., MacAry, P.A., and Ng, L.F.P. (2020). The trinity of COVID-19: immunity, inflammation and intervention. Nat. Rev. Immunol. 20, 363-374. https://doi.org/10.1038/s41577-020-0311-8
- Wang, H.Y., Juo, L.I., Lin, Y.T., Hsiao, M., Lin, J.T., Tsai, C.H., Tzeng, Y.H., Chuang, Y.C., Chang, N.S., Yang, C.N., et al. (2012). WW domain-containing oxidoreductase promotes neuronal differentiation via negative regulation of glycogen synthase kinase 3β. Cell Death Differ. 19, 1049-1059. https://doi.org/10.1038/cdd.2011.188
- Wu, C.H., Chen, P.J., and Yeh, S.H. (2014). Nucleocapsid phosphorylation and RNA helicase DDX1 recruitment enables coronavirus transition from discontinuous to continuous transcription. Cell Host Microbe 16, 462-472. https://doi.org/10.1016/j.chom.2014.09.009
- Wu, C.H., Yeh, S.H., Tsay, Y.G., Shieh, Y.H., Kao, C.L., Chen, Y.S., Wang, S.H., Kuo, T.J., Chen, D.S., and Chen, P.J. (2009). Glycogen synthase kinase-3 regulates the phosphorylation of severe acute respiratory syndrome coronavirus nucleocapsid protein and viral replication. J. Biol. Chem. 284, 5229-5239. https://doi.org/10.1074/jbc.M805747200
- Wu, C.J., Jan, J.T., Chen, C.M., Hsieh, H.P., Hwang, D.R., Liu, H.W., Liu, C.Y., Huang, H.W., Chen, S.C., Hong, C.F., et al. (2004). Inhibition of severe acute respiratory syndrome coronavirus replication by niclosamide. Antimicrob. Agents Chemother. 48, 2693-2696. https://doi.org/10.1128/AAC.48.7.2693-2696.2004
- Yamamoto, H., Kishida, S., Kishida, M., Ikeda, S., Takada, S., and Kikuchi, A. (1999). Phosphorylation of axin, a Wnt signal negative regulator, by glycogen synthase kinase-3beta regulates its stability. J. Biol. Chem. 274, 10681-10684. https://doi.org/10.1074/jbc.274.16.10681
- Yook, J.I., Li, X.Y., Ota, I., Fearon, E.R., and Weiss, S.J. (2005). Wnt-dependent regulation of the E-cadherin repressor snail. J. Biol. Chem. 280, 11740-11748. https://doi.org/10.1074/jbc.M413878200
- Yook, J.I., Li, X.Y., Ota, I., Hu, C., Kim, H.S., Kim, N.H., Cha, S.Y., Ryu, J.K., Choi, Y.J., Kim, J., et al. (2006). A Wnt-Axin2-GSK3beta cascade regulates Snail1 activity in breast cancer cells. Nat. Cell Biol. 8, 1398-1406. https://doi.org/10.1038/ncb1508
- Zhou, P., Yang, X.L., Wang, X.G., Hu, B., Zhang, L., Zhang, W., Si, H.R., Zhu, Y., Li, B., Huang, C.L., et al. (2020). A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature 579, 270-273. https://doi.org/10.1038/s41586-020-2012-7