Coronavirus S protein-induced fusion is blocked prior to hemifusion by Abl kinase inhibitors Free

Abstract

Enveloped viruses gain entry into host cells by fusing with cellular membranes, a step that is required for virus replication. Coronaviruses, including the severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV) and infectious bronchitis virus (IBV), fuse at the plasma membrane or use receptor-mediated endocytosis and fuse with endosomes, depending on the cell or tissue type. The virus spike (S) protein mediates fusion with the host cell membrane. We have shown previously that an Abelson (Abl) kinase inhibitor, imatinib, significantly reduces SARS-CoV and MERS-CoV viral titres and prevents endosomal entry by HIV SARS S and MERS S pseudotyped virions. SARS-CoV and MERS-CoV are classified as BSL-3 viruses, which makes experimentation into the cellular mechanisms involved in infection more challenging. Here, we use IBV, a BSL-2 virus, as a model for studying the role of Abl kinase activity during coronavirus infection. We found that imatinib and two specific Abl kinase inhibitors, GNF2 and GNF5, reduce IBV titres by blocking the first round of virus infection. Additionally, all three drugs prevented IBV S-induced syncytia formation prior to the hemifusion step. Our results indicate that membrane fusion (both virus–cell and cell–cell) is blocked in the presence of Abl kinase inhibitors. Studying the effects of Abl kinase inhibitors on IBV will be useful in identifying the host cell pathways required for coronavirus infection. This will provide an insight into possible therapeutic targets to treat infections by current as well as newly emerging coronaviruses.

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2018-05-01
2024-03-29
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References

  1. Mingo RM, Simmons JA, Shoemaker CJ, Nelson EA, Schornberg KL et al. Ebola virus and severe acute respiratory syndrome coronavirus display late cell entry kinetics: evidence that transport to NPC1+ endolysosomes is a rate-defining step. J Virol 2015; 89:2931–2943 [View Article][PubMed]
    [Google Scholar]
  2. Millet JK, Whittaker GR. Host cell entry of Middle East respiratory syndrome coronavirus after two-step, furin-mediated activation of the spike protein. Proc Natl Acad Sci USA 2014; 111:15214–15219 [View Article][PubMed]
    [Google Scholar]
  3. Burkard C, Verheije MH, Wicht O, van Kasteren SI, van Kuppeveld FJ et al. Coronavirus cell entry occurs through the endo-/lysosomal pathway in a proteolysis-dependent manner. PLoS Pathog 2014; 10:e1004502 [View Article][PubMed]
    [Google Scholar]
  4. Millet JK, Whittaker GR. Host cell proteases: critical determinants of coronavirus tropism and pathogenesis. Virus Res 2015; 202:120–134 [View Article][PubMed]
    [Google Scholar]
  5. Dyall J, Coleman CM, Hart BJ, Venkataraman T, Holbrook MR et al. Repurposing of clinically developed drugs for treatment of Middle East respiratory syndrome coronavirus infection. Antimicrob Agents Chemother 2014; 58:4885–4893 [View Article][PubMed]
    [Google Scholar]
  6. Gaki GS, Papavassiliou AG. Oxidative stress-induced signaling pathways implicated in the pathogenesis of Parkinson's disease. Neuromolecular Med 2014; 16:217–230 [View Article][PubMed]
    [Google Scholar]
  7. Colicelli J. ABL tyrosine kinases: evolution of function, regulation, and specificity. Sci Signal 2010; 3:re6 [View Article][PubMed]
    [Google Scholar]
  8. Cheng WH, von Kobbe C, Opresko PL, Fields KM, Ren J et al. Werner syndrome protein phosphorylation by abl tyrosine kinase regulates its activity and distribution. Mol Cell Biol 2003; 23:6385–6395 [View Article][PubMed]
    [Google Scholar]
  9. Rogers EM, Spracklen AJ, Bilancia CG, Sumigray KD, Allred SC et al. Abelson kinase acts as a robust, multifunctional scaffold in regulating embryonic morphogenesis. Mol Biol Cell 2016; 27:2613–2631 [View Article][PubMed]
    [Google Scholar]
  10. Khatri A, Wang J, Pendergast AM. Multifunctional Abl kinases in health and disease. J Cell Sci 2016; 129:9–16 [View Article][PubMed]
    [Google Scholar]
  11. Buchdunger E, Zimmermann J, Mett H, Meyer T, Müller M et al. Inhibition of the Abl protein-tyrosine kinase in vitro and in vivo by a 2-phenylaminopyrimidine derivative. Cancer Res 1996; 56:100–104[PubMed]
    [Google Scholar]
  12. Rowley JD. Letter: a new consistent chromosomal abnormality in chronic myelogenous leukaemia identified by quinacrine fluorescence and Giemsa staining. Nature 1973; 243:290–293 [View Article][PubMed]
    [Google Scholar]
  13. Heisterkamp N, Stephenson JR, Groffen J, Hansen PF, de Klein A et al. Localization of the c-ab1 oncogene adjacent to a translocation break point in chronic myelocytic leukaemia. Nature 1983; 306:239–242 [View Article][PubMed]
    [Google Scholar]
  14. Bartram CR, de Klein A, Hagemeijer A, van Agthoven T, Geurts van Kessel A et al. Translocation of c-ab1 oncogene correlates with the presence of a Philadelphia chromosome in chronic myelocytic leukaemia. Nature 1983; 306:277–280 [View Article][PubMed]
    [Google Scholar]
  15. Coleman CM, Sisk JM, Mingo RM, Nelson EA, White JM et al. Abelson kinase inhibitors are potent inhibitors of severe acute respiratory syndrome coronavirus and middle east respiratory syndrome coronavirus fusion. J Virol 2016; 90:8924–8933 [View Article][PubMed]
    [Google Scholar]
  16. Coyne CB, Bergelson JM. Virus-induced Abl and Fyn kinase signals permit coxsackievirus entry through epithelial tight junctions. Cell 2006; 124:119–131 [View Article][PubMed]
    [Google Scholar]
  17. Newsome TP, Weisswange I, Frischknecht F, Way M. Abl collaborates with Src family kinases to stimulate actin-based motility of vaccinia virus. Cell Microbiol 2006; 8:233–241 [View Article][PubMed]
    [Google Scholar]
  18. Harmon B, Campbell N, Ratner L. Role of Abl kinase and the Wave2 signaling complex in HIV-1 entry at a post-hemifusion step. PLoS Pathog 2010; 6:e1000956 [View Article][PubMed]
    [Google Scholar]
  19. Thomas A, Mariani-Floderer C, López-Huertas MR, Gros N, Hamard-Péron E et al. Involvement of the Rac1-IRSp53-Wave2-Arp2/3 signaling pathway in HIV-1 gag particle release in CD4 T cells. J Virol 2015; 89:8162–8181 [View Article][PubMed]
    [Google Scholar]
  20. Reeves PM, Smith SK, Olson VA, Thorne SH, Bornmann W et al. Variola and monkeypox viruses utilize conserved mechanisms of virion motility and release that depend on abl and SRC family tyrosine kinases. J Virol 2011; 85:21–31 [View Article][PubMed]
    [Google Scholar]
  21. García M, Cooper A, Shi W, Bornmann W, Carrion R et al. Productive replication of Ebola virus is regulated by the c-Abl1 tyrosine kinase. Sci Transl Med 2012; 4:123ra24 [View Article][PubMed]
    [Google Scholar]
  22. Kouznetsova J, Sun W, Martínez-Romero C, Tawa G, Shinn P et al. Identification of 53 compounds that block Ebola virus-like particle entry via a repurposing screen of approved drugs. Emerg Microbes Infect 2014; 3:e84 [View Article][PubMed]
    [Google Scholar]
  23. Yamauchi S, Takeuchi K, Chihara K, Sun X, Honjoh C et al. Hepatitis C virus particle assembly involves phosphorylation of NS5A by the c-Abl tyrosine kinase. J Biol Chem 2015; 290:21857–21864 [View Article][PubMed]
    [Google Scholar]
  24. Cluett EB, Kuismanen E, Machamer CE. Heterogeneous distribution of the unusual phospholipid semilysobisphosphatidic acid through the Golgi complex. Mol Biol Cell 1997; 8:2233–2240 [View Article][PubMed]
    [Google Scholar]
  25. Hogue BMC. Coronavirus Structural Proteins and Virus Assembly Washington, DC: ASM Press; 2008 [Crossref]
    [Google Scholar]
  26. Schindler T, Bornmann W, Pellicena P, Miller WT, Clarkson B et al. Structural mechanism for STI-571 inhibition of abelson tyrosine kinase. Science 2000; 289:1938–1942 [View Article][PubMed]
    [Google Scholar]
  27. Nagar B, Bornmann WG, Pellicena P, Schindler T, Veach DR et al. Crystal structures of the kinase domain of c-Abl in complex with the small molecule inhibitors PD173955 and imatinib (STI-571). Cancer Res 2002; 62:4236–4243[PubMed]
    [Google Scholar]
  28. Zhang J, Adrián FJ, Jahnke W, Cowan-Jacob SW, Li AG, Ag L et al. Targeting Bcr-Abl by combining allosteric with ATP-binding-site inhibitors. Nature 2010; 463:501–506 [View Article][PubMed]
    [Google Scholar]
  29. Greuber EK, Smith-Pearson P, Wang J, Pendergast AM. Role of ABL family kinases in cancer: from leukaemia to solid tumours. Nat Rev Cancer 2013; 13:559–571 [View Article][PubMed]
    [Google Scholar]
  30. Reddy EP, Aggarwal AK. The ins and outs of bcr-abl inhibition. Genes Cancer 2012; 3:447–454 [View Article][PubMed]
    [Google Scholar]
  31. Stertz S, Reichelt M, Spiegel M, Kuri T, Martínez-Sobrido L et al. The intracellular sites of early replication and budding of SARS-coronavirus. Virology 2007; 361:304–315 [View Article][PubMed]
    [Google Scholar]
  32. Lontok E, Corse E, Machamer CE. Intracellular targeting signals contribute to localization of coronavirus spike proteins near the virus assembly site. J Virol 2004; 78:5913–5922 [View Article][PubMed]
    [Google Scholar]
  33. Bradley WD, Koleske AJ. Regulation of cell migration and morphogenesis by Abl-family kinases: emerging mechanisms and physiological contexts. J Cell Sci 2009; 122:3441–3454 [View Article][PubMed]
    [Google Scholar]
  34. Huang Y, Comiskey EO, Dupree RS, Li S, Koleske AJ et al. The c-Abl tyrosine kinase regulates actin remodeling at the immune synapse. Blood 2008; 112:111–119 [View Article][PubMed]
    [Google Scholar]
  35. Gu JJ, Lavau CP, Pugacheva E, Soderblom EJ, Moseley MA et al. Abl family kinases modulate T cell-mediated inflammation and chemokine-induced migration through the adaptor HEF1 and the GTPase Rap1. Sci Signal 2012; 5:ra51 [View Article][PubMed]
    [Google Scholar]
  36. Koleske AJ, Gifford AM, Scott ML, Nee M, Bronson RT et al. Essential roles for the Abl and Arg tyrosine kinases in neurulation. Neuron 1998; 21:1259–1272 [View Article][PubMed]
    [Google Scholar]
  37. Smith-Pearson PS, Greuber EK, Yogalingam G, Pendergast AM. Abl kinases are required for invadopodia formation and chemokine-induced invasion. J Biol Chem 2010; 285:40201–40211 [View Article][PubMed]
    [Google Scholar]
  38. Mader CC, Oser M, Magalhaes MA, Bravo-Cordero JJ, Condeelis J et al. An EGFR-Src-Arg-cortactin pathway mediates functional maturation of invadopodia and breast cancer cell invasion. Cancer Res 2011; 71:1730–1741 [View Article][PubMed]
    [Google Scholar]
  39. Lapetina S, Mader CC, Machida K, Mayer BJ, Koleske AJ. Arg interacts with cortactin to promote adhesion-dependent cell edge protrusion. J Cell Biol 2009; 185:503–519 [View Article][PubMed]
    [Google Scholar]
  40. MacGrath SM, Koleske AJ. Arg/Abl2 modulates the affinity and stoichiometry of binding of cortactin to F-actin. Biochemistry 2012; 51:6644–6653 [View Article][PubMed]
    [Google Scholar]
  41. Lin YC, Yeckel MF, Koleske AJ. Abl2/Arg controls dendritic spine and dendrite arbor stability via distinct cytoskeletal control pathways. J Neurosci 2013; 33:1846–1857 [View Article][PubMed]
    [Google Scholar]
  42. Courtemanche N, Gifford SM, Simpson MA, Pollard TD, Koleske AJ. Abl2/Abl-related gene stabilizes actin filaments, stimulates actin branching by actin-related protein 2/3 complex, and promotes actin filament severing by cofilin. J Biol Chem 2015; 290:4038–4046 [View Article][PubMed]
    [Google Scholar]
  43. Woodring PJ, Hunter T, Wang JY. Inhibition of c-Abl tyrosine kinase activity by filamentous actin. J Biol Chem 2001; 276:27104–27110 [View Article][PubMed]
    [Google Scholar]
  44. Hernández SE, Krishnaswami M, Miller AL, Koleske AJ. How do Abl family kinases regulate cell shape and movement?. Trends Cell Biol 2004; 14:36–44 [View Article][PubMed]
    [Google Scholar]
  45. Selbach M, Backert S. Cortactin: an Achilles' heel of the actin cytoskeleton targeted by pathogens. Trends Microbiol 2005; 13:181–189 [View Article][PubMed]
    [Google Scholar]
  46. Taylor MP, Koyuncu OO, Enquist LW. Subversion of the actin cytoskeleton during viral infection. Nat Rev Microbiol 2011; 9:427–439 [View Article][PubMed]
    [Google Scholar]
  47. Wessler S, Backert S. Abl family of tyrosine kinases and microbial pathogenesis. Int Rev Cell Mol Biol 2011; 286:271–300 [View Article][PubMed]
    [Google Scholar]
  48. Levaot N, Simoncic PD, Dimitriou ID, Scotter A, La Rose J et al. 3BP2-deficient mice are osteoporotic with impaired osteoblast and osteoclast functions. J Clin Invest 2011; 121:3244–3257 [View Article][PubMed]
    [Google Scholar]
  49. Hindi SM, Tajrishi MM, Kumar A. Signaling mechanisms in mammalian myoblast fusion. Sci Signal 2013; 6:re2 [View Article][PubMed]
    [Google Scholar]
  50. Kim JH, Jin P, Duan R, Chen EH. Mechanisms of myoblast fusion during muscle development. Curr Opin Genet Dev 2015; 32:162–170 [View Article][PubMed]
    [Google Scholar]
  51. Shilagardi K, Li S, Luo F, Marikar F, Duan R et al. Actin-propelled invasive membrane protrusions promote fusogenic protein engagement during cell-cell fusion. Science 2013; 340:359–363 [View Article][PubMed]
    [Google Scholar]
  52. Georgess D, Machuca-Gayet I, Blangy A, Jurdic P. Podosome organization drives osteoclast-mediated bone resorption. Cell Adh Migr 2014; 8:192–204 [View Article][PubMed]
    [Google Scholar]
  53. Youn S, Collisson EW, Machamer CE. Contribution of trafficking signals in the cytoplasmic tail of the infectious bronchitis virus spike protein to virus infection. J Virol 2005; 79:13209–13217 [View Article][PubMed]
    [Google Scholar]
  54. McBride CE, Li J, Machamer CE. 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. J Virol 2007; 81:2418–2428 [View Article][PubMed]
    [Google Scholar]
  55. McBride CE, Machamer CE. A single tyrosine in the severe acute respiratory syndrome coronavirus membrane protein cytoplasmic tail is important for efficient interaction with spike protein. J Virol 2010; 84:1891–1901 [View Article][PubMed]
    [Google Scholar]
  56. Wickramasinghe IN, de Vries RP, Gröne A, de Haan CA, Verheije MH. Binding of avian coronavirus spike proteins to host factors reflects virus tropism and pathogenicity. J Virol 2011; 85:8903–8912 [View Article][PubMed]
    [Google Scholar]
  57. Promkuntod N, Wickramasinghe IN, de Vrieze G, Gröne A, Verheije MH. Contributions of the S2 spike ectodomain to attachment and host range of infectious bronchitis virus. Virus Res 2013; 177:127–137 [View Article][PubMed]
    [Google Scholar]
  58. Wang L, Parr RL, King DJ, Collisson EW. A highly conserved epitope on the spike protein of infectious bronchitis virus. Arch Virol 1995; 140:2201–2213 [View Article][PubMed]
    [Google Scholar]
  59. Simmons G, Reeves JD, Rennekamp AJ, Amberg SM, Piefer AJ et al. Characterization of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) spike glycoprotein-mediated viral entry. Proc Natl Acad Sci USA 2004; 101:4240–4245 [View Article][PubMed]
    [Google Scholar]
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