1887

Abstract

Human astroviruses (HAstVs), non-enveloped RNA viruses with positive-sense RNA genomes, are an important cause of acute gastroenteritis in young children, although the processes that produce infectious virions are not clearly defined. To track the viral replication complex (RC) upon HAstV1 infection, the subcellular distribution of double-stranded (ds) RNA and of ORF1b, a viral RNA polymerase, was examined by immunocytochemistry. Foci that were positive for dsRNA and for ORF1b were co-localized, and both foci were also co-localized with resident proteins of the endoplasmic reticulum (ER). Focusing on the association between the HAstV RC and ER, we examined the expression of unfolded protein response (UPR) markers and found that targets of eukaryotic translation initiation factor 2α (eIF2α)-activating transcription factor 4 (ATF4), including CCAAT/enhancer-binding protein homologous protein (CHOP), a proapoptotic transcription factor, were upregulated at the late phase in HAstV-infected cells. Consistently, eIF2α was phosphorylated at the late phase of HAstV infection. The formation of foci resembling stress granules, another known downstream response to eIF2α phosphorylation, was also observed at the same period. Phosphorylation of eIF2α was attenuated in protein kinase R (PKR)-knockdown cells, suggesting that, unlike the canonical ER stress response, PKR was involved in eIF2α phosphorylation in response to HAstV infection. Studies have indicated that immature HAstV capsid protein is processed by caspases, and caspase cleavage is integral to particle release. Inhibition of CHOP upregulation reduced caspase activation and the release of HAstV RNA from cells during HAstV infection. Our results suggest that the eIF2α–ATF4–CHOP pathway participates in HAstV propagation.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.001250
2019-03-26
2019-10-15
Loading full text...

Full text loading...

References

  1. Bosch A, Pintó RM, Guix S. Human astroviruses. Clin Microbiol Rev 2014;27:1048–1074 [CrossRef][PubMed]
    [Google Scholar]
  2. Cortez V, Meliopoulos VA, Karlsson EA, Hargest V, Johnson C et al. Astrovirus Biology and Pathogenesis. Annu Rev Virol 2017;4:327–348 [CrossRef][PubMed]
    [Google Scholar]
  3. Walter JE, Mitchell DK. Astrovirus infection in children. Curr Opin Infect Dis 2003;16:247–253 [CrossRef][PubMed]
    [Google Scholar]
  4. Fuentes C, Bosch A, Pintó RM, Guix S. Identification of human astrovirus genome-linked protein (VPg) essential for virus infectivity. J Virol 2012;86:10070–10078 [CrossRef][PubMed]
    [Google Scholar]
  5. Jiang B, Monroe SS, Koonin EV, Stine SE, Glass RI. RNA sequence of astrovirus: distinctive genomic organization and a putative retrovirus-like ribosomal frameshifting signal that directs the viral replicase synthesis. Proc Natl Acad Sci USA 1993;90:10539–10543 [CrossRef][PubMed]
    [Google Scholar]
  6. Marczinke B, Bloys AJ, Brown TD, Willcocks MM, Carter MJ et al. The human astrovirus RNA-dependent RNA polymerase coding region is expressed by ribosomal frameshifting. J Virol 1994;68:5588–5595[PubMed]
    [Google Scholar]
  7. Geigenmüller U, Chew T, Ginzton N, Matsui SM. Processing of nonstructural protein 1a of human astrovirus. J Virol 2002;76:2003–2008 [CrossRef][PubMed]
    [Google Scholar]
  8. Banos-Lara MR, Méndez E. Role of individual caspases induced by astrovirus on the processing of its structural protein and its release from the cell through a non-lytic mechanism. Virology 2010;401:322–332 [CrossRef][PubMed]
    [Google Scholar]
  9. Méndez E, Salas-Ocampo E, Arias CF. Caspases mediate processing of the capsid precursor and cell release of human astroviruses. J Virol 2004;78:8601–8608 [CrossRef][PubMed]
    [Google Scholar]
  10. Ron D, Walter P. Signal integration in the endoplasmic reticulum unfolded protein response. Nat Rev Mol Cell Biol 2007;8:519–529 [CrossRef][PubMed]
    [Google Scholar]
  11. Walter P, Ron D. The unfolded protein response: from stress pathway to homeostatic regulation. Science 2011;334:1081–1086 [CrossRef][PubMed]
    [Google Scholar]
  12. Harding HP, Novoa I, Zhang Y, Zeng H, Wek R et al. Regulated translation initiation controls stress-induced gene expression in mammalian cells. Mol Cell 2000;6:1099–1108 [CrossRef][PubMed]
    [Google Scholar]
  13. Yoshida H, Matsui T, Hosokawa N, Kaufman RJ, Nagata K et al. A time-dependent phase shift in the mammalian unfolded protein response. Dev Cell 2003;4:265–271 [CrossRef][PubMed]
    [Google Scholar]
  14. Jheng JR, Ho JY, Horng JT. ER stress, autophagy, and RNA viruses. Front Microbiol 2014;5:388 [CrossRef][PubMed]
    [Google Scholar]
  15. Zhang L, Wang A. Virus-induced ER stress and the unfolded protein response. Front Plant Sci 2012;3:293 [CrossRef][PubMed]
    [Google Scholar]
  16. Pakos-Zebrucka K, Koryga I, Mnich K, Ljujic M, Samali A et al. The integrated stress response. EMBO Rep 2016;17:1374–1395 [CrossRef][PubMed]
    [Google Scholar]
  17. Donnelly N, Gorman AM, Gupta S, Samali A. The eIF2α kinases: their structures and functions. Cell Mol Life Sci 2013;70:3493–3511 [CrossRef][PubMed]
    [Google Scholar]
  18. Nover L, Scharf KD, Neumann D. Cytoplasmic heat shock granules are formed from precursor particles and are associated with a specific set of mRNAs. Mol Cell Biol 1989;9:1298–1308 [CrossRef][PubMed]
    [Google Scholar]
  19. Protter DSW, Parker R. Principles and properties of stress granules. Trends Cell Biol 2016;26:668–679 [CrossRef][PubMed]
    [Google Scholar]
  20. Anderson P, Kedersha N. Stress granules: the Tao of RNA triage. Trends Biochem Sci 2008;33:141–150 [CrossRef][PubMed]
    [Google Scholar]
  21. Reineke LC, Lloyd RE. Diversion of stress granules and P-bodies during viral infection. Virology 2013;436:255–267 [CrossRef][PubMed]
    [Google Scholar]
  22. Tsai WC, Lloyd RE. Cytoplasmic RNA granules and viral infection. Annu Rev Virol 2014;1:147–170 [CrossRef][PubMed]
    [Google Scholar]
  23. Weber F, Wagner V, Rasmussen SB, Hartmann R, Paludan SR. Double-stranded RNA is produced by positive-strand RNA viruses and DNA viruses but not in detectable amounts by negative-strand RNA viruses. J Virol 2006;80:5059–5064 [CrossRef][PubMed]
    [Google Scholar]
  24. Moser LA, Schultz-Cherry S. Suppression of astrovirus replication by an ERK1/2 inhibitor. J Virol 2008;82:7475–7482 [CrossRef][PubMed]
    [Google Scholar]
  25. Guix S, Caballero S, Bosch A, Pintó RM. C-terminal nsP1a protein of human astrovirus colocalizes with the endoplasmic reticulum and viral RNA. J Virol 2004;78:13627–13636 [CrossRef][PubMed]
    [Google Scholar]
  26. Sekine Y, Zyryanova A, Crespillo-Casado A, Fischer PM, Harding HP et al. Stress responses. Mutations in a translation initiation factor identify the target of a memory-enhancing compound. Science 2015;348:1027–1030 [CrossRef][PubMed]
    [Google Scholar]
  27. Zhang K, Daigle JG, Cunningham KM, Coyne AN, Ruan K et al. Stress granule assembly disrupts nucleocytoplasmic transport. Cell 2018;173:958–971 [CrossRef][PubMed]
    [Google Scholar]
  28. Williams BR. PKR; a sentinel kinase for cellular stress. Oncogene 1999;18:6112–6120 [CrossRef][PubMed]
    [Google Scholar]
  29. García MA, Gil J, Ventoso I, Guerra S, Domingo E et al. Impact of protein kinase PKR in cell biology: from antiviral to antiproliferative action. Microbiol Mol Biol Rev 2006;70:1032–1060 [CrossRef][PubMed]
    [Google Scholar]
  30. Guix S, Pérez-Bosque A, Miró L, Moretó M, Bosch A et al. Type I interferon response is delayed in human astrovirus infections. PLoS One 2015;10:e0123087 [CrossRef][PubMed]
    [Google Scholar]
  31. Guix S, Bosch A, Ribes E, Dora Martínez L, Pintó RM. Apoptosis in astrovirus-infected CaCo-2 cells. Virology 2004;319:249–261 [CrossRef][PubMed]
    [Google Scholar]
  32. Puthalakath H, O'Reilly LA, Gunn P, Lee L, Kelly PN et al. ER stress triggers apoptosis by activating BH3-only protein Bim. Cell 2007;129:1337–1349 [CrossRef][PubMed]
    [Google Scholar]
  33. Cazanave SC, Elmi NA, Akazawa Y, Bronk SF, Mott JL et al. CHOP and AP-1 cooperatively mediate PUMA expression during lipoapoptosis. Am J Physiol Gastrointest Liver Physiol 2010;299:G236–G243 [CrossRef][PubMed]
    [Google Scholar]
  34. Jang SY, Jeong WH, Kim MS, Lee YM, Lee JI et al. Detection of replicating negative-sense RNAs in CaCo-2 cells infected with human astrovirus. Arch Virol 2010;155:1383–1389 [CrossRef][PubMed]
    [Google Scholar]
  35. Kumar M, Carmichael GG. Antisense RNA: function and fate of duplex RNA in cells of higher eukaryotes. Microbiol Mol Biol Rev 1998;62:1415–1434[PubMed]
    [Google Scholar]
  36. Murillo A, vera-Estrella R, Barkla BJ, Méndez E, Arias CF. Identification of host cell factors associated with astrovirus replication in CaCo-2 cells. J Virol 2015;89:10359–10370 [CrossRef][PubMed]
    [Google Scholar]
  37. Fuentes C, Guix S, Bosch A, Pintó RM. The C-terminal nsP1a protein of human astrovirus is a phosphoprotein that interacts with the viral polymerase. J Virol 2011;85:4470–4479 [CrossRef][PubMed]
    [Google Scholar]
  38. Méndez E, Aguirre-Crespo G, Zavala G, Arias CF. Association of the astrovirus structural protein VP90 with membranes plays a role in virus morphogenesis. J Virol 2007;81:10649–10658 [CrossRef][PubMed]
    [Google Scholar]
  39. Emara MM, Brinton MA. Interaction of TIA-1/TIAR with West Nile and dengue virus products in infected cells interferes with stress granule formation and processing body assembly. Proc Natl Acad Sci USA 2007;104:9041–9046 [CrossRef][PubMed]
    [Google Scholar]
  40. Nikolic J, Civas A, Lama Z, Lagaudrière-Gesbert C, Blondel D. Rabies virus infection induces the formation of stress granules closely connected to the viral factories. PLoS Pathog 2016;12:e1005942 [CrossRef][PubMed]
    [Google Scholar]
  41. Anderson P, Kedersha N. RNA granules: post-transcriptional and epigenetic modulators of gene expression. Nat Rev Mol Cell Biol 2009;10:430–436 [CrossRef][PubMed]
    [Google Scholar]
  42. Onomoto K, Jogi M, Yoo JS, Narita R, Morimoto S et al. Critical role of an antiviral stress granule containing RIG-I and PKR in viral detection and innate immunity. PLoS One 2012;7:e43031 [CrossRef][PubMed]
    [Google Scholar]
  43. Oh SW, Onomoto K, Wakimoto M, Onoguchi K, Ishidate F et al. Leader-containing uncapped viral transcript activates RIG-I in antiviral stress granules. PLoS Pathog 2016;12:e1005444 [CrossRef][PubMed]
    [Google Scholar]
  44. Yoneyama M, Jogi M, Onomoto K. Regulation of antiviral innate immune signaling by stress-induced RNA granules. J Biochem 2016;159:279–286 [CrossRef][PubMed]
    [Google Scholar]
  45. Taylor RC, Cullen SP, Martin SJ. Apoptosis: controlled demolition at the cellular level. Nat Rev Mol Cell Biol 2008;9:231–241 [CrossRef][PubMed]
    [Google Scholar]
  46. Martinon F, Tschopp J. Inflammatory caspases and inflammasomes: master switches of inflammation. Cell Death Differ 2007;14:10–22 [CrossRef][PubMed]
    [Google Scholar]
  47. Koci MD, Moser LA, Kelley LA, Larsen D, Brown CC et al. Astrovirus induces diarrhea in the absence of inflammation and cell death. J Virol 2003;77:11798–11808 [CrossRef][PubMed]
    [Google Scholar]
  48. Tange S, Zhou Y, Nagakui-Noguchi Y, Imai T, Nakanishi A. Initiation of human astrovirus type 1 infection was blocked by inhibitors of phosphoinositide 3-kinase. Virol J 2013;10:153 [CrossRef][PubMed]
    [Google Scholar]
  49. Dunn KW, Kamocka MM, McDonald JH. A practical guide to evaluating colocalization in biological microscopy. Am J Physiol Cell Physiol 2011;300:C723–C742 [CrossRef][PubMed]
    [Google Scholar]
  50. Manders EMM, Verbeek FJ, Aten JA. Measurement of co-localization of objects in dual-colour confocal images. J Microsc 1993;169:375–382 [CrossRef]
    [Google Scholar]
  51. O'Donnell TB, Hyde JL, Mintern JD, MacKenzie JM. Mouse norovirus infection promotes autophagy induction to facilitate replication but prevents final autophagosome maturation. Virology 2016;492:130–139 [CrossRef][PubMed]
    [Google Scholar]
  52. Doerflinger SY, Cortese M, Romero-Brey I, Menne Z, Tubiana T et al. Membrane alterations induced by nonstructural proteins of human norovirus. PLoS Pathog 2017;13:e1006705 [CrossRef][PubMed]
    [Google Scholar]
  53. Oslowski CM, Urano F. Measuring ER stress and the unfolded protein response using mammalian tissue culture system. Methods Enzymol 2011;490:71–92 [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.001250
Loading
/content/journal/jgv/10.1099/jgv.0.001250
Loading

Data & Media loading...

Supplements

Supplementary File 1

PDF

Supplementary File 2

Supplementary File 3

Supplementary File 4

Most Cited This Month

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