1887

Abstract

Viral infections result in cellular stress responses, which can trigger protein translation shutoff via phosphorylation of eukaryotic initiation factor 2 alpha (eIF2α). Newcastle disease virus (NDV) causes severe disease in poultry and selectively kills human tumour cells. In this report, we determined that infection of HeLa human cervical cancer cells and DF1 chicken fibroblast cells with NDV maintained protein at early infection times, 0–12 h post-infection (p.i.), and gradually inhibited global protein translation at late infection times, 12–24 h p.i. Mechanistic studies showed that translation inhibition at late infection times was accompanied by phosphorylation of eIF2α, a checkpoint of translation initiation. Meanwhile, the eIF2α kinase, PKR, was upregulated and activated by phosphorylation and another eIF2α kinase, PERK, was phosphorylated and cleaved into two fragments. Pharmacological inhibition experiments revealed that only PKR activity was required for eIF2α phosphorylation, suggesting that recognition of viral dsRNA by PKR was responsible for translation shutoff. High levels of phospho-eIF2α led to preferential translation of the transcription factor ATF4 and an increase in GADD34 expression. Functionally, GADD34, in conjunction with PP1, dephosphorylated eIF2a and restored protein translation, benefiting virus protein synthesis. However, PP1 was degraded at late infection times, functionally counteracting the upregulation of GADD34. Taken together, our data support that NDV-induced translation shutoff at late infection times was attributed to sustaining phosphorylation of eIF2α, which is mediated by continual activation of PKR and degradation of PP1.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.000426
2016-04-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/97/4/867.html?itemId=/content/journal/jgv/10.1099/jgv.0.000426&mimeType=html&fmt=ahah

References

  1. Alexander D. J. 2000; Newcastle disease and other avian paramyxoviruses. Rev Sci Tech 19:443–462[PubMed]
    [Google Scholar]
  2. Axten J. M., Medina J. R., Feng Y., Shu A., Romeril S. P., Grant S. W., Li W. H., Heerding D. A., Minthorn E. other authors 2012; Discovery of 7-methyl-5-(1-{[3-(trifluoromethyl)phenyl]acetyl}-2,3-dihydro-1H-indol-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (GSK2606414), a potent and selective first-in-class inhibitor of protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK). J Med Chem 55:7193–7207 [CrossRef]
    [Google Scholar]
  3. Brush M. H., Weiser D. C., Shenolikar S. 2003; Growth arrest and DNA damage-inducible protein GADD34 targets protein phosphatase 1 alpha to the endoplasmic reticulum and promotes dephosphorylation of the alpha subunit of eukaryotic translation initiation factor 2. Molecular and cellular biology 23:1292–1303 [CrossRef]
    [Google Scholar]
  4. Childs K., Stock N., Ross C., Andrejeva J., Hilton L., Skinner M., Randall R., Goodbourn S. 2007; mda-5, but not RIG-I, is a common target for paramyxovirus V proteins. Virology 359:190–200 [CrossRef]
    [Google Scholar]
  5. Cohen P., Klumpp S., Schelling D. L. 1989; An improved procedure for identifying and quantitating protein phosphatases in mammalian tissues. FEBS letters 250:596–600 [CrossRef]
    [Google Scholar]
  6. Cruz J. L., Sola I., Becares M., Alberca B., Plana J., Enjuanes L., Zuñiga S. 2011; Coronavirus gene 7 counteracts host defenses and modulates virus virulence. PLoS Pathog 7:e1002090 [View Article][PubMed]
    [Google Scholar]
  7. Davis M. E., Wang M. K., Rennick L. J., Full F., Gableske S., Mesman A. W., Gringhuis S. I., Geijtenbeek T. B., Duprex W. P., Gack M. U. 2014; Antagonism of the phosphatase PP1 by the measles virus V protein is required for innate immune escape of MDA5. Cell Host Microbe 16:19–30 [View Article][PubMed]
    [Google Scholar]
  8. Gale M. Jr., Katze M. G. 1998; Molecular mechanisms of interferon resistance mediated by viral-directed inhibition of PKR, the interferon-induced protein kinase. Pharmacol Ther 78:29–46 [View Article][PubMed]
    [Google Scholar]
  9. Gu M., Zhang T., lin W., Liu Z., Lai R., Xia D., Huang H., Wang X. 2014; Protein phosphatase PP1 negatively regulates the Toll-like receptor- and RIG-I-like receptor-triggered production of type I interferon by inhibiting IRF3 phosphorylation at serines 396 and 385 in macrophage. Cell Signal 26:2930–2939 [View Article][PubMed]
    [Google Scholar]
  10. Harding H. P., Calfon M., Urano F., Novoa I., Ron D. 2002; Transcriptional and translational control in the Mammalian unfolded protein response. Annu Rev Cell Dev Bi 18:575–599 [View Article][PubMed]
    [Google Scholar]
  11. Harding H. P., Novoa I., Zhang Y., Zeng H., Wek R., Schapira M., Ron D. 2000; Regulated translation initiation controls stress-induced gene expression in mammalian cells. Mol Cell 6:1099–1108 [CrossRef]
    [Google Scholar]
  12. Harding H. P., Zhang Y., Ron D. 1999; Protein translation and folding are coupled by an endoplasmic-reticulum-resident kinase. Nature 397:271–274 [View Article][PubMed]
    [Google Scholar]
  13. He B., Gross M., Roizman B. 1997; The gamma(1)34.5 protein of herpes simplex virus 1 complexes with protein phosphatase 1alpha to dephosphorylate the alpha subunit of the eukaryotic translation initiation factor 2 and preclude the shutoff of protein synthesis by double-stranded RNA-activated protein kinase. Proc Natl Acad Sci U S A 94:843–848 [View Article][PubMed]
    [Google Scholar]
  14. He B., Gross M., Roizman B. 1998; The gamma134.5 protein of herpes simplex virus 1 has the structural and functional attributes of a protein phosphatase 1 regulatory subunit and is present in a high molecular weight complex with the enzyme in infected cells. J Biol Chem 273:20737–20743 [View Article][PubMed]
    [Google Scholar]
  15. Hinnebusch A. G., Lorsch J. R. 2012; The mechanism of eukaryotic translation initiation: new insights and challenges. Cold Spring Harb Perspect Biol 4:4 [View Article][PubMed]
    [Google Scholar]
  16. Hu Y., Conway T. W. 1993; 2-Aminopurine inhibits the double-stranded RNA-dependent protein kinase both in vitro and in vivo . J Interferon Res 13:323–328 [CrossRef]
    [Google Scholar]
  17. Ishihara H., Martin B. L., Brautigan D. L., Karaki H., Ozaki H., Kato Y., Fusetani N., Watabe S., Hashimoto K., other authors. 1989; Calyculin A and okadaic acid: inhibitors of protein phosphatase activity. Biochem Bioph Res Co 159:871–877 [CrossRef]
    [Google Scholar]
  18. Jackson R. J., Hellen C. U., Pestova T. V. 2010; The mechanism of eukaryotic translation initiation and principles of its regulation. Nat Rev Mol Cell Biol 11:113–127 [View Article][PubMed]
    [Google Scholar]
  19. Kazemi S., Papadopoulou S., Li S., Su Q., Wang S., Yoshimura A., Matlashewski G., Dever T. E., Koromilas A. E. 2004; Control of alpha subunit of eukaryotic translation initiation factor 2 (eIF2 alpha) phosphorylation by the human papillomavirus type 18 E6 oncoprotein: implications for eIF2 alpha-dependent gene expression and cell death. Mol Cell Biol 24:3415–3429 [View Article][PubMed]
    [Google Scholar]
  20. Kwon Y. G., Lee S. Y., Choi Y., Greengard P., Nairn A. C. 1997; Cell cycle-dependent phosphorylation of mammalian protein phosphatase 1 by cdc2 kinase. Proc Natl Acad Sci U S A 94:2168–2173 [View Article][PubMed]
    [Google Scholar]
  21. Li H. Y., Liu H., Wang C. H., Zhang J. Y., Man J. H., Gao Y. F., Zhang P. J., Li W. H., Zhao J., other authors. 2008; Deactivation of the kinase IKK by CUEDC2 through recruitment of the phosphatase PP1. Nat Immunol 9:533–541 [View Article][PubMed]
    [Google Scholar]
  22. Ma Y., Brewer J. W., Diehl J. A., Hendershot L. M. 2002; Two distinct stress signaling pathways converge upon the CHOP promoter during the mammalian unfolded protein response. J Mol Biol 318:1351–1365 [View Article][PubMed]
    [Google Scholar]
  23. Mesman A. W., Zijlstra-Willems E. M., Kaptein T. M., de Swart R. L., Davis M. E., Ludlow M., Duprex W. P., Gack M. U., Gringhuis S. I., Geijtenbeek T. B. 2014; Measles virus suppresses RIG-I-like receptor activation in dendritic cells via DC-SIGN-mediated inhibition of PP1 phosphatases. Cell Host Microbe 16:31–42 [View Article][PubMed]
    [Google Scholar]
  24. Mulvey M., Arias C., Mohr I. 2007; Maintenance of endoplasmic reticulum (ER) homeostasis in herpes simplex virus type 1-infected cells through the association of a viral glycoprotein with PERK, a cellular ER stress sensor. J Virol 81:3377–3390 [View Article][PubMed]
    [Google Scholar]
  25. Novoa I., Zeng H., Harding H. P., Ron D. 2001; Feedback inhibition of the unfolded protein response by GADD34-mediated dephosphorylation of eIF2alpha. J Cell Biol 153:1011–1022 [View Article][PubMed]
    [Google Scholar]
  26. Pavio N., Romano P. R., Graczyk T. M., Feinstone S. M., Taylor D. R. 2003; Protein synthesis and endoplasmic reticulum stress can be modulated by the hepatitis C virus envelope protein E2 through the eukaryotic initiation factor 2alpha kinase PERK. J Virol 77:3578–3585 [View Article][PubMed]
    [Google Scholar]
  27. Proud C. G. 2005; eIF2 and the control of cell physiology. Semin Cell Dev Biol 16:3–12 [CrossRef]
    [Google Scholar]
  28. Qu L., Ji Y., Zhu X., Zheng X. 2015; hCINAP negatively regulates NF-κB signaling by recruiting the phosphatase PP1 to deactivate IKK complex. J Mol Cell Biol 7:529–542 [View Article][PubMed]
    [Google Scholar]
  29. Schmidt E. K., Clavarino G., Ceppi M., Pierre P. 2009; SUnSET, a nonradioactive method to monitor protein synthesis. Nat Methods 6:275–277 [View Article][PubMed]
    [Google Scholar]
  30. Schröder M., Kaufman R. J. 2005; The mammalian unfolded protein response. Annu Rev Biochem 74:739–789 [View Article][PubMed]
    [Google Scholar]
  31. Van Opdenbosch N., Van den Broeke C., De Regge N., Tabarés E., Favoreel H. W. 2012; The IE180 protein of pseudorabies virus suppresses phosphorylation of translation initiation factor eIF2α. J Virol 86:7235–7240 [View Article][PubMed]
    [Google Scholar]
  32. Wang X., Liao Y., Yap P. L., Png K. J., Tam J. P., Liu D. X. 2009; Inhibition of protein kinase R activation and upregulation of GADD34 expression play a synergistic role in facilitating coronavirus replication by maintaining de novo protein synthesis in virus-infected cells. J Virol 83:12462–12472 [View Article][PubMed]
    [Google Scholar]
  33. Wies E., Wang M. K., Maharaj N. P., Chen K., Zhou S., Finberg R. W., Gack M. U. 2013; Dephosphorylation of the RNA sensors RIG-I and MDA5 by the phosphatase PP1 is essential for innate immune signaling. Immunity 38:437–449 [View Article][PubMed]
    [Google Scholar]
  34. Williams B. R. 1999; PKR; a sentinel kinase for cellular stress. Oncogene 18:6112–6120 [View Article][PubMed]
    [Google Scholar]
  35. Xu L., Zhou X., Peppelenbosch M. P., Pan Q. 2015; Inhibition of hepatitis E virus replication by proteasome inhibitor is nonspecific. Arch Virol 160:435–439 [CrossRef]
    [Google Scholar]
  36. Zhang F., Moon A., Childs K., Goodbourn S., Dixon L. K. 2010a; The African swine fever virus DP71L protein recruits the protein phosphatase 1 catalytic subunit to dephosphorylate eIF2alpha and inhibits CHOP induction but is dispensable for these activities during virus infection. J Virol 84:10681–10689 [View Article][PubMed]
    [Google Scholar]
  37. Zhang S., Sun Y., Chen H., Dai Y., Zhan Y., Yu S., Qiu X., Tan L., Song C., Ding C. 2014; Activation of the PKR/eIF2α signaling cascade inhibits replication of Newcastle disease virus. Virol J 11:62 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.000426
Loading
/content/journal/jgv/10.1099/jgv.0.000426
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error