1887

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Volume 102, Issue 7

Targeting cap-dependent translation to inhibit Chikungunya virus replication: selectivity of p38 MAPK inhibitors to virus-infected cells due to autophagy-mediated down regulation of phospho-ERK No Access

Abstract

The 5′ capped, message-sense RNA genome of Chikungunya virus (CHIKV) utilizes the host cell machinery for translation. Translation is regulated by eIF2 alpha at the initiation phase and by eIF4F at cap recognition. Translational suppression by eIF2 alpha phosphorylation occurs as an early event in many alphavirus infections. We observe that in CHIKV-infected HEK293 cells, this occurs as a late event, by which time the viral replication has reached an exponential phase, implying its minimal role in virus restriction. The regulation by eIF4F is mediated through the PI3K-Akt-mTOR, p38 MAPK and RAS-RAF-MEK-ERK pathways. A kinetic analysis revealed that CHIKV infection did not modulate AKT phosphorylation, but caused a significant reduction in p38 MAPK phosphorylation. It caused degradation of phospho-ERK 1/2 by increased autophagy, leaving the PI3K-Akt-mTOR and p38 MAPK pathways for pharmacological targeting. mTOR inhibition resulted in moderate reduction in viral titre, but had no effect on CHIKV E2 protein expression, indicating a minimal role of the mTOR complex in virus replication. Inhibition of p38 MAPK using SB202190 caused a significant reduction in viral titre and CHIKV E2 and nsP3 protein expression. Furthermore, inhibiting the two pathways together did not offer any synergism, indicating that inhibiting the p38 MAPK pathway alone is sufficient to cause restriction of CHIKV replication. Meanwhile, in uninfected cells the fully functional RAS-RAF-MEK-ERK pathway can circumvent the effect of p38 MAPK inhibition on cap-dependent translation. Thus, our results show that host-directed antiviral strategies targeting cellular p38 MAPK are worth exploring against Chikungunya as they could be selective against CHIKV-infected cells with minimal effects on uninfected host cells.

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  • Accepted:
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Keyword(s): autophagy , cap-dependent translation , Chikungunya , LC3 , phospho-ERK , torin and Wortmannin
Funding
This study was supported by the:
  • Department of Biotechnology, Govt. of India (Award No.BT/PR13801/MED/29/952/2015)
    • Principle Award Recipient: EaswaranSreekumar
© 2021 The Authors
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/content/journal/jgv/10.1099/jgv.0.001629
2021-07-30
2024-04-27
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References

  1. Schwartz O, Albert ML. Biology and pathogenesis of chikungunya virus. Nat Rev Microbiol 2010; 8:491–500 [View Article] [PubMed]
     [Google Scholar]
  2. Arpino C, Curatolo P, Rezza G. Chikungunya and the nervous system: what we do and do not know. Rev Med Virol 2009; 19:121–129 [View Article] [PubMed]
     [Google Scholar]
  3. Khan AH, Morita K, del Carmen Parquet M, Hasebe F, Mathenge EG et al. Complete nucleotide sequence of chikungunya virus and evidence for an internal polyadenylation site. J Gen Virol 2002; 83:3075–3084 [View Article] [PubMed]
     [Google Scholar]
  4. Solignat M, Gay B, Higgs S, Briant L, Devaux C. Replication cycle of chikungunya: a re-emerging arbovirus. Virology 2009; 393:183–197 [View Article] [PubMed]
     [Google Scholar]
  5. Delang L, Li C, Tas A, Quérat G, Albulescu I et al. The viral capping enzyme nsP1: a novel target for the inhibition of chikungunya virus infection. Sci Rep 2016; 6:1–10 [View Article]
     [Google Scholar]
  6. Karpe YA, Aher PP, Lole KS. NTPase and 5′-RNA triphosphatase activities of Chikungunya virus nsP2 protein. PLoS One 2011; 6:e22336 [View Article] [PubMed]
     [Google Scholar]
  7. Gao Y, Goonawardane N, Ward J, Tuplin A, Harris M. Multiple roles of the non-structural protein 3 (nsP3) alphavirus unique domain (AUD) during Chikungunya virus genome replication and transcription. PLoS Pathog 2019; 15:e1007239 [View Article] [PubMed]
     [Google Scholar]
  8. Abraham R, Hauer D, McPherson RL, Utt A, Kirby IT et al. ADP-ribosyl–binding and hydrolase activities of the alphavirus nsP3 macrodomain are critical for initiation of virus replication. Proc Natl Acad Sci U S A 2018; 115:E10457–E10466 [View Article]
     [Google Scholar]
  9. Pietilä MK, Hellström K, Ahola T. Alphavirus polymerase and RNA replication. Virus Res 2017; 234:44–57 [View Article] [PubMed]
     [Google Scholar]
  10. Pialoux G, Gaüzère BA, Jauréguiberry S, Strobel M. Chikungunya, an epidemic arbovirosis. Lancet Infect Dis 2007; 7:319–327 [View Article] [PubMed]
     [Google Scholar]
  11. Ganesan VK, Duan B, Reid SP. Chikungunya virus: pathophysiology, mechanism, and modeling. Viruses 2017; 9:368 [View Article]
     [Google Scholar]
  12. Staples JE, Breiman RF, Powers AM. Chikungunya fever: an epidemiological review of a re-emerging infectious disease. Clin Infect Dis 2009; 49:942–948 [View Article] [PubMed]
     [Google Scholar]
  13. Gingras AC, Raught B, Sonenberg N. eIF4 initiation factors: effectors of mRNA recruitment to ribosomes and regulators of translation. Annu Rev Biochem 1999; 68:913–963 [View Article] [PubMed]
     [Google Scholar]
  14. Sonenberg N, Hinnebusch AG. Regulation of translation initiation in eukaryotes: mechanisms and biological targets. Cell 2009; 136:731–745 [View Article] [PubMed]
     [Google Scholar]
  15. Ventoso I, Sanz MA, Molina S, Berlanga JJ, Carrasco L et al. Translational resistance of late alphavirus mRNA to eIF2α phosphorylation: a strategy to overcome the antiviral effect of protein kinase PKR. Genes Dev 2006; 20:87–100 [View Article] [PubMed]
     [Google Scholar]
  16. Bhat M, Robichaud N, Hulea L, Sonenberg N, Pelletier J et al. Targeting the translation machinery in cancer. Nat Rev Drug Discov 2015; 14:261–278 [View Article] [PubMed]
     [Google Scholar]
  17. Pestova TV, Kolupaeva VG, Lomakin IB, Pilipenko EV, Shatsky IN et al. Molecular mechanisms of translation initiation in eukaryotes. Proc Natl Acad Sci U S A 2001; 98:7029–7036 [View Article] [PubMed]
     [Google Scholar]
  18. Tanabe IS, Tanabe EL, Santos EC, Martins WV, Araújo IM et al. Cellular and molecular immune response to Chikungunya virus infection. Front Cell Infect Microbiol 2018; 8:345 [View Article] [PubMed]
     [Google Scholar]
  19. Glick D, Barth S, Macleod KF. Autophagy: cellular and molecular mechanisms. J Pathol 2010; 221:3–12 [View Article] [PubMed]
     [Google Scholar]
  20. Balachandran S, Roberts PC, Brown LE, Truong H, Pattnaik AK et al. Essential role for the dsRNA-dependent protein kinase PKR in innate immunity to viral infection. Immunity 2000; 13:129–141 [View Article] [PubMed]
     [Google Scholar]
  21. Clemens MJ. Initiation factor eIF2 alpha phosphorylation in stress responses and apoptosis. Prog Mol Subcell Biol 2001; 27:57–89 [View Article] [PubMed]
     [Google Scholar]
  22. Kudchodkar SB, Levine B. Viruses and autophagy. Rev Med Virol 2009; 19:359–378 [View Article] [PubMed]
     [Google Scholar]
  23. Levine B. Eating oneself and uninvited guests: autophagy-related pathways in cellular defense. Cell 2005; 120:159–162 [View Article] [PubMed]
     [Google Scholar]
  24. Matsumoto H, Miyazaki S, Matsuyama S, Takeda M, Kawano M et al. Selection of autophagy or apoptosis in cells exposed to ER-stress depends on ATF4 expression pattern with or without CHOP expression. Biol Open 2013; 2:1084–1090 [View Article] [PubMed]
     [Google Scholar]
  25. Krejbich-Trotot P, Gay B, Li-Pat-Yuen G, Hoarau JJ, Jaffar-Bandjee MC et al. Chikungunya triggers an autophagic process which promotes viral replication. Virol J 2011; 8:432 [View Article] [PubMed]
     [Google Scholar]
  26. Rathore AP, M-L N, Vasudevan SG. Differential unfolded protein response during Chikungunya and Sindbis virus infection: CHIKV nsP4 suppresses eIF2α phosphorylation. Virol J 2013; 10:36 [View Article] [PubMed]
     [Google Scholar]
  27. Thaa B, Biasiotto R, Eng K, Neuvonen M, Götte B et al. Differential phosphatidylinositol-3-kinase-Akt-mTOR activation by Semliki Forest and chikungunya viruses is dependent on nsP3 and connected to replication complex internalization. J Virol 2015; 89:11420–11437 [View Article] [PubMed]
     [Google Scholar]
  28. Nayak TK, Mamidi P, Sahoo SS, Kumar PS, Mahish C et al. P38 and Jnk mitogen-activated protein kinases interact with chikungunya virus non-structural protein-2 and regulate TNF induction during viral infection in macrophages. Front Immunol 2019; 10:786 [View Article] [PubMed]
     [Google Scholar]
  29. Sreekumar E, Issac A, Nair S, Hariharan R, Janki M et al. Genetic characterization of 2006–2008 isolates of Chikungunya virus from Kerala, South India, by whole genome sequence analysis. Virus Genes 2010; 40:14–27 [View Article] [PubMed]
     [Google Scholar]
  30. Abraham R, Mudaliar P, Jaleel A, Srikanth J, Sreekumar E. High throughput proteomic analysis and a comparative review identify the nuclear chaperone, Nucleophosmin among the common set of proteins modulated in Chikungunya virus infection. J Proteomics 2015; 120:126–141 [View Article] [PubMed]
     [Google Scholar]
  31. Van Huizen E, McInerney GM. Activation of the PI3K-AKT pathway by old world Alphaviruses. Cells 2020; 9:970 [View Article]
     [Google Scholar]
  32. Martinez-Lopez N, Athonvarangkul D, Mishall P, Sahu S, Singh R. Autophagy proteins regulate ERK phosphorylation. Nat Commun 2013; 4:1–14 [View Article]
     [Google Scholar]
  33. Joubert PE, Werneke SW, de la Calle C, Guivel-Benhassine F, Giodini A et al. Chikungunya virus-induced autophagy delays caspase-dependent cell death. J Exp Med 2012; 209:1029–1047 [View Article] [PubMed]
     [Google Scholar]
  34. Judith D, Couderc T, Lecuit M. Chikungunya virus-induced autophagy and apoptosis. Okeoma C. eds In Chikungunya Virus Springer.Cham: 2016 pp 149–159
     [Google Scholar]
  35. Abraham R, Mudaliar P, Padmanabhan A, Sreekumar E. Induction of cytopathogenicity in human glioblastoma cells by chikungunya virus. PLoS One 2013; 8:e75854 [View Article] [PubMed]
     [Google Scholar]
  36. Hart PD, Young MR. Ammonium chloride, an inhibitor of phagosome-lysosome fusion in macrophages, concurrently induces phagosome-endosome fusion, and opens a novel pathway: studies of a pathogenic mycobacterium and a nonpathogenic yeast. J Exp Med 1991; 174:881–889 [View Article] [PubMed]
     [Google Scholar]
  37. Mauthe M, Orhon I, Rocchi C, Zhou X, Luhr M et al. Chloroquine inhibits autophagic flux by decreasing autophagosome-lysosome fusion. Autophagy 2018; 14:1435–1455 [View Article] [PubMed]
     [Google Scholar]
  38. Yang J, Guo Z, Liu X, Liu Q, Wu M et al. Cytotoxicity evaluation of chloroquine and hydroxychloroquine in multiple cell lines and tissues by dynamic imaging system and physiologically based pharmacokinetic model. Front Pharmacol 2020; 11:574720 [View Article] [PubMed]
     [Google Scholar]
  39. Li L, Friedrichsen HJ, Andrews S. A TFEB nuclear export signal integrates amino acid supply and glucose availability. Nat Commun 2018; 9:2685 [View Article] [PubMed]
     [Google Scholar]
  40. Ferby IM, Waga I, Sakanaka C, Kume K, Shimizu T. Wortmannin inhibits mitogen-activated protein kinase activation induced by platelet-activating factor in guinea pig neutrophils. J Biol Chem 1994; 269:30485–30488 [View Article] [PubMed]
     [Google Scholar]
  41. ŞA D, Yerlikaya A, Zeren S, Bayhan Z, Okur E et al. Differential effects of p38 MAP kinase inhibitors SB203580 and SB202190 on growth and migration of human MDA-MB-231 cancer cell line. Cytotechnology 2017; 69:711–724 [View Article] [PubMed]
     [Google Scholar]
  42. Teng TS, Kam YW, Tan JJ, Ng LF. Host response to Chikungunya virus and perspectives for immune-based therapies. Future Virology 2011; 6:975–984 [View Article]
     [Google Scholar]
  43. Proud CG. eIF2 and the control of cell physiology. Semin Cell Dev Biol 2005; 16:3–12 [View Article] [PubMed]
     [Google Scholar]
  44. Sanz MA, González Almela E, Carrasco L. Translation of sindbis subgenomic mRNA is independent of eIF2, eIF2A and eIF2D. Sci Rep 2017; 7:43876 [View Article] [PubMed]
     [Google Scholar]
  45. Toribio R, Díaz-López I, Boskovic J, Ventoso I. An RNA trapping mechanism in Alphavirus mRNA promotes ribosome stalling and translation initiation. Nucleic Acids Res 2016; 44:4368–4380 [View Article] [PubMed]
     [Google Scholar]
  46. McInerney GM, Kedersha NL, Kaufman RJ, Anderson P, Liljestrom P. Importance of eIF2α phosphorylation and stress granule assembly in alphavirus translation regulation. Mol Biol Cell 2005; 16:3753–3763 [View Article] [PubMed]
     [Google Scholar]
  47. Walsh D, Mohr I. Viral subversion of the host protein synthesis machinery. Nat Rev Microbiol 2011; 9:860–875 [View Article] [PubMed]
     [Google Scholar]
  48. Gross JD, Moerke NJ, von der Haar T, Lugovskoy AA, Sachs AB et al. Ribosome loading onto the mRNA cap is driven by conformational coupling between eIF4G and eIF4E. Cell 2003; 115:739–750 [View Article] [PubMed]
     [Google Scholar]
  49. Shveygert M, Kaiser C, Bradrick SS, Gromeier M. Regulation of eukaryotic initiation factor 4E (eIF4E) phosphorylation by mitogen-activated protein kinase occurs through modulation of Mnk1-eIF4G interaction. Mol Cell Biol 2010; 30:5160–5167 [View Article] [PubMed]
     [Google Scholar]
  50. Cargnello M, Roux PP. Activation and function of the MAPKs and their substrates, the MAPK-activated protein kinases. Microbiol Mol Biol Rev 2011; 75:50–83 [View Article] [PubMed]
     [Google Scholar]
  51. Bose SK, Shrivastava S, Meyer K, Ray RB, Ray R. Hepatitis C virus activates the mTOR/S6K1 signaling pathway in inhibiting IRS-1 function for insulin resistance. J Virol 2012; 86:6315–6322 [View Article] [PubMed]
     [Google Scholar]
  52. Diehl N, Schaal H. Make yourself at home: viral hijacking of the PI3K/Akt signaling pathway. Viruses 2013; 5:3192–3212 [View Article] [PubMed]
     [Google Scholar]
  53. George A, Panda S, Kudmulwar D, Chhatbar SP, Nayak SC et al. Hepatitis C virus NS5A binds to the mRNA cap-binding eukaryotic translation initiation 4F (eIF4F) complex and up-regulates host translation initiation machinery through eIF4E-binding protein 1 inactivation. J Biol Chem 2012; 287:5042–5058 [View Article] [PubMed]
     [Google Scholar]
  54. Soares JA, Leite FG, Andrade LG, Torres AA, De Sousa LP et al. Activation of the PI3K/Akt pathway early during vaccinia and cowpox virus infections is required for both host survival and viral replication. J Virol 2009; 83:6883–6899 [View Article] [PubMed]
     [Google Scholar]
  55. Altman AM, Mahmud J, Nikolovska-Coleska Z, Chan G. HCMV modulation of cellular PI3K/AKT/mTOR signaling: New opportunities for therapeutic intervention. Antiviral Res 2019; 163:82–90 [View Article] [PubMed]
     [Google Scholar]
  56. Zhang L, Wu J, Ling MT, Zhao KN, Zhao KN. The role of the PI3K/Akt/mTOR signalling pathway in human cancers induced by infection with human papillomaviruses. Mol Cancer 2015; 14:87 [View Article] [PubMed]
     [Google Scholar]
  57. Kuss-Duerkop SK, Wang J, Mena I, White K, Metreveli G et al. Influenza virus differentially activates mTORC1 and mTORC2 signaling to maximize late stage replication. PLoS Pathog 2017; 13:e1006635 [View Article] [PubMed]
     [Google Scholar]
  58. Pleschka S, Wolff T, Ehrhardt C, Hobom G, Planz O et al. Influenza virus propagation is impaired by inhibition of the Raf/MEK/ERK signalling cascade. Nat Cell Biol 2001; 3:301–305 [View Article] [PubMed]
     [Google Scholar]
  59. Zampieri CA, Fortin JF, Nolan GP, Nabel GJ. The ERK mitogen-activated protein kinase pathway contributes to Ebola virus glycoprotein-induced cytotoxicity. J Virol 2007; 81:1230–1240 [View Article] [PubMed]
     [Google Scholar]
  60. Pei R, Zhang X, Xu S, Meng Z, Roggendorf M et al. Regulation of hepatitis C virus replication and gene expression by the MAPK-ERK pathway. Virol Sin 2012; 27:278–285 [View Article] [PubMed]
     [Google Scholar]
  61. Albarnaz JD, De Oliveira LC, Torres AA, Palhares RM, Casteluber MC et al. MEK/ERK activation plays a decisive role in yellow fever virus replication: implication as an antiviral therapeutic target. Antiviral Res 2014; 111:82–92 [View Article] [PubMed]
     [Google Scholar]
  62. Jheng JR, Ho J-Y, Horng JT. ER stress, autophagy, and RNA viruses. Front Microbiol 2014; 5:388 [View Article] [PubMed]
     [Google Scholar]
  63. Xu C, Zhang W, Liu S, Wu W, Qin M et al. Activation of the SphK1/ERK/p-ERK pathway promotes autophagy in colon cancer cells. Oncol lett 2018; 15:9719–9724 [View Article] [PubMed]
     [Google Scholar]
  64. Joubert PE, Stapleford K, Guivel-Benhassine F, Vignuzzi M, Schwartz O et al. Inhibition of mTORC1 enhances the translation of chikungunya proteins via the activation of the MnK/eIF4E pathway. PLoS Pathog 2015; 11:e1005091 [View Article]
     [Google Scholar]
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Targeting cap-dependent translation to inhibit Chikungunya virus replication: selectivity of p38 MAPK inhibitors to virus-infected cells due to autophagy-mediated down regulation of phospho-ERK
J. Gen. Virol. 102, 001629 (2021); https://doi.org/10.1099/jgv.0.001629
/content/journal/jgv/10.1099/jgv.0.001629
/content/journal/jgv/10.1099/jgv.0.001629
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