Skip to main content

Advertisement

SpringerLink
Log in

Betanodavirus up-regulates chaperone GRP78 via ER stress: roles of GRP78 in viral replication and host mitochondria-mediated cell death

  • Original Paper
  • Published:
Apoptosis Aims and scope Submit manuscript

Abstract

Whether viral pathogens that induce ER stress responses benefit the host or the virus remains controversial. In this study we show that betanodavirus induced ER stress responses up-regulate GRP78, which regulates the viral replication and host cellular mitochondrial-mediated cell death. Betanodavirus (redspotted grouper nervous necrosis virus, RGNNV) infection resulted in the following increased ER stress responses in fish GF-1 grouper fin cells: (1) IRE-1 and ATF-6 sensors at 48 h post-infection (p.i.) that up-regulated chaperone protein GRP78; (2) activation of caspase-12; and (3) PERK phosphorylation and down-regulation of Bcl-2. Analyses of GRP78 functions during viral replication using either loss-of-function or gain-of-function approaches showed that GRP78 over-expression also enhanced viral replication and induced cell death. Then, we found that zfGRP78 localization gradually increased in mitochondria after RGNNV infection by EGFP tagging approach. Furthermore, zfGRP78 can interact with viral RNA-dependent RNA polymerase (RdRp) by using immunofluorescent and immunoprecipitation assays. Finally, we found that blocking GRP78-mediated ER signals can reduce the viral death factors protein α and protein B2 expression and decrease the Bcl-2 down-regulation mediated mitochondria-dependent cell death, which also enhances host cellular viability. Taken together, our results suggest that RGNNV infection and expression can trigger ER stress responses, which up-regulate the chaperone GRP78 at early replication stage. Then, GRP78 can interact with RdRp that may enhance the viral replication for increasing viral death factors’ expressions at middle-late replication stage, which can enhance mitochondrial-mediated cell death pathway and viral spreading. These results may provide new insights into the mechanism of ER stress-mediated cell death in RNA viruses.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. Ball LA, Johnson KL (1999) Reverse genetics of nodaviruses. Adv Virus Res 53:229–244

    Article  PubMed  CAS  Google Scholar 

  2. Munday BL, Kwang J, Moody N (2002) Betanodavirus infections of teleost fish: a review. J Fish Dis 25:127–142

    Article  Google Scholar 

  3. Schneemann A, Reddy V, Johson JE (1998) The structural and function of nodavirus particles: a paradigm for understanding chemical biology. Adv Virus Res 50:381–446

    Article  PubMed  CAS  Google Scholar 

  4. Toffolo V, Negrisolo E, Maltese C, Bovo G, Belvedere P, Colombo L, Valle LD (2006) Phylogeny of betanodaviruses and molecular evolution of their RNA polymerase and coat proteins. Mol Phylogenet Evol 43:298–308

    Article  PubMed  Google Scholar 

  5. Mori K, Nakai T, Muroga K, Arimoto M, Mushiake K, Furusawa I (1992) Properties of a new virus belonging to nodaviridae found in larval striped jack (Pseudocaranx dentex) with nervous necrosis. Virology 187:368–371

    Article  PubMed  CAS  Google Scholar 

  6. Guo YX, Wei T, Dallmann K, Kwang J (2003) Induction of caspase-dependent apoptosis by betanodaviruses GGNNV and demonstration of protein alpha as an apoptosis inducer. Virology 308:74–82

    Article  PubMed  CAS  Google Scholar 

  7. Wu HC, Chiu CS, Wu JL, Gong HY, Chen MC, Lu MW, Hong JR (2008) Zebrafish anti-apoptotic protein zfBcl-xL can block betanodavirus protein alpha-induced mitochondria-mediated secondary necrosis cell death. Fish Shellfish Immunol 24:436–449

    Article  PubMed  CAS  Google Scholar 

  8. Chen LJ, Su YC, Hong JR (2009) Betanodavirus non-structural protein B1: a novel anti-necrotic death factor that modulates cell death in early replication cycle in fish cells. Virology 385:444–454

    Article  PubMed  CAS  Google Scholar 

  9. Fenner BJ, Thiagarajan R, Chua HK, Kwang J (2006) Betanodavirus B2 is an RNA interference antagonist that facilitates intracellular viral RNA accumulation. J Virol 80:85–94

    Article  PubMed  CAS  Google Scholar 

  10. Iwamoto T, Mise K, Takeda A, Okinaka Y, Mori K, Arimoto M, Okuno T, Nakai T (2005) Characterization of Striped jack nervous necrosis virus subgenomic RNA3 and biological activities of its encoded protein B2. J Gen Virol 86:2807–2816

    Article  PubMed  CAS  Google Scholar 

  11. Chen SP, Wu JL, Su YC, Hong JR (2007) Anti-Bcl-2 family members, zfBcl-x(L) and zfMcl-1a, prevent cytochrome c release from cells undergoing betanodavirus-induced secondary necrotic cell death. Apoptosis 12:1043–1060

    Article  PubMed  CAS  Google Scholar 

  12. Su HL, Liao CL, Lin YL (2002) Japanese encephalitis virus infection initiates endoplasmic reticulum stress and an unfolded protein response. J Virol 76:4162–4171

    Article  PubMed  CAS  Google Scholar 

  13. Harding HP, Calfon M, Urano F, Novoa I, Ron D (2002) Transcriptional and translational control in the Mammalian unfolded protein response. Annu Rev Cell Dev Biol 18:575–599

    Article  PubMed  CAS  Google Scholar 

  14. Kaufman RJ, Scheuner D, Schroder M, Shen X, Lee K, Liu CY, Arnold SM (2002) The unfolded protein response in nutrient sensing and differentiation. Nat Rev Mol Cell Biol 3:411–421

    Article  PubMed  CAS  Google Scholar 

  15. Li XD, Lankinen H, Putkuri N, Vapalahti O, Vaheri A (2005) Tula hantavirus triggers pro-apoptotic signals of ER stress in Vero E6 cells. Virology 333:180–189

    Article  PubMed  CAS  Google Scholar 

  16. Rao RV, Bredesen DE (2004) Misfolded proteins, endoplasmic reticulum stress and neurodegeneration. Curr Opin Cell Biol 16:653–662

    Article  PubMed  CAS  Google Scholar 

  17. Williams BL, Lipkin WI (2006) Endoplasmic reticulum stress and neurodegeneration in rats neonatally infected with borna disease virus. J Virol 80:8613–8626

    Article  PubMed  CAS  Google Scholar 

  18. Chen X, Shen J, Prywes R (2002) The luminal domain of ATF6 senses endoplasmic reticulum (ER) stress and causes translocation of ATF6 from the ER to the Golgi. J Biol Chem 277:13045–13052

    Article  PubMed  CAS  Google Scholar 

  19. Lee K, Tirasophon W, Shen X, Prywes M, Okada R, Yoshida RH, Mori K, Kaufman RJ (2002) IRE1-mediated unconventional mRNA splicing and S2P-mediated ATF6 cleavage merge to regulate XBP1 in signaling the unfolded protein response. Genes Dev 16:452–466

    Article  PubMed  CAS  Google Scholar 

  20. Szegezdi E, Logue SE, Gorman AM, Samali A (2006) Mediators of endoplasmic reticulum stress-induced apoptosis. EMBO Rep 7:880–885

    Article  PubMed  CAS  Google Scholar 

  21. Baltzis D, Qu LK, Papadopoulou S, Blais JD, Bell JC, Sonenberg N, Koromilas AE (2004) Resistance to vesicular stomatitis virus infection requires a functional cross talk between the eukaryotic translation initiation factor 2 alpha kinases PERK and PKR. J Virol 78:12747–12761

    Article  PubMed  CAS  Google Scholar 

  22. Netherton CL, Parsley JC, Wileman T (2004) African swine fever virus inhibits induction of the stress-induced proapoptotic transcription factor CHOP/GADD153. J Virol 78:10825–10838

    Article  PubMed  CAS  Google Scholar 

  23. He B (2006) Viruses, endoplasmic reticulum stress, and interferon responses. Cell Death Differ 13:393–403

    Article  PubMed  CAS  Google Scholar 

  24. Dimcheff DE, Faasse MA, McAtee FJ, Portis JL (2004) Endoplasmic reticulum (ER) stress induced by a neurovirulent mouse retrovirus is associated with prolonged BiP binding and retention of a viral protein in the ER. J Biol Chem 279:33782–33790

    Article  PubMed  CAS  Google Scholar 

  25. Tardif KD, Mori K, Siddiqui A (2002) Hepatitis C virus subgenomic replicons induce endoplasmic reticulum stress activating an intracellular signaling pathway. J Virol 76:7453–7459

    Article  PubMed  CAS  Google Scholar 

  26. Bitko V, Barik S (2001) An endoplasmic reticulum-specific stress-activated caspase (caspase-12) is implicated in the apoptosis of A549 epithelial cells by respiratory syncytial virus. J Cell Biochem 80:441–454

    Article  PubMed  CAS  Google Scholar 

  27. Farrow SN, Brown R (1996) New members of the Bcl-2 family and their protein partners. Curr Opin Genet Dev 6:45–49

    Article  PubMed  CAS  Google Scholar 

  28. Oltvai ZN, Milliman CL, Korsmeyer SJ (1993) Bcl-2 hetrodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 74:609–619

    Article  PubMed  CAS  Google Scholar 

  29. Zamzami N, Kroemer G (2001) The mitochondrion in apoptosis: how Pandora’s box opens. Nat Rev Mol Cell Biol 2:67–71

    Article  PubMed  CAS  Google Scholar 

  30. Oyadomari S, Mori M (2004) Roles of CHOP/GADD153 in endoplasmic reticulum stress. Cell Death Differ 11:381–389

    Article  PubMed  CAS  Google Scholar 

  31. Chen SP, Yang HL, Her GM, Lin HY, Jeng MF, Wu JL, Hong JR (2006) Betanodavirus induces phosphatidylserine exposure and loss of mitochondrial membrane potential in secondary necrotic cells, both of which are blocked by bongkrekic acid. Virology 347:379–391

    Article  PubMed  CAS  Google Scholar 

  32. Wu HC, Wu JL, Chu HL, Su YC, Hong JR (2010) RGNNV induces mitochondria-mediated cell death via newly synthesized protein dependent pathway in fish cells. Fish Shellfish Immunol 29:457–463

    Google Scholar 

  33. Su YC, Wu JL, Hong JR (2009) Betanodavirus non-structural protein B2: a novel necrotic death factor that induces mitochondria-mediated cell death in fish cells. Virology 385:143–154

    Article  PubMed  CAS  Google Scholar 

  34. Nicholson B, Dunn J (1974) Homologous viral interference in trout and Atlantic salmon cell cultures infected with infectious pancreatic necrosis virus. J Virol 14:180–182

    PubMed  CAS  Google Scholar 

  35. Yang GH, Li S, Pestka JJ (2000) Down-regulation of the endoplasmic reticulum chaperone GRP78/BiP by vomitoxin (Deoxynivalenol). Toxicol Appl Pharmacol 162:207–217

    Article  PubMed  CAS  Google Scholar 

  36. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  PubMed  CAS  Google Scholar 

  37. Kain SR, Mai K, Sinai P (1994) Human multiple tissue western blots: a new immunological tool for the analysis of tissue-specific protein expression. Biotechniques 17:982–987

    PubMed  CAS  Google Scholar 

  38. Hong JR, Wu JL (2002) Induction of apoptotic death in cells via Bad gene expression by infectious pancreatic necrosis virus infection. Cell Death Differ 9:113–124

    Article  PubMed  CAS  Google Scholar 

  39. Chen PC, Wu JL, Her GM, Hong JR (2010) Aquatic birnavirus induces necrotic cell death via mitochondria-mediated caspases pathway that inhibited by bongkrekic acid. Fish Shellfish Immunol 28:344–353

    Article  PubMed  CAS  Google Scholar 

  40. Mullen PD, Brand RJ, Parlette GN (1975) Evaluation of dye exclusion and colony inhibition techniques for detection of polyoma-specific, cell-mediated immunity. J Natl Cancer Inst 54:229–231

    PubMed  CAS  Google Scholar 

  41. Mezeth KB, Nylund S, Henrisksen H, Patel S, Nerland AH, Szilvay AM (2007) RNA-dependent RNA polymerase from Atlantic halibut nodavirus contains two signals for localization to the mitochondria. Virus Res 130:43–52

    Article  PubMed  CAS  Google Scholar 

  42. Miller DJ, Ahlquist P (2002) Flock house virus RNA polymerase is a transmembrane protein with amino-terminal sequences sufficient for mitochondrial localization and membrane insertion. J Virol 76:9856–9867

    Article  PubMed  CAS  Google Scholar 

  43. Priscilla M, Wynsberghe V, Ahlquist P (2009) 5′ cis elements direct nodavirus RNA1 recruitment to mitochondrial sites of replication complex formation. J Virol 83:2976–2988

    Article  Google Scholar 

  44. Bovo G, Nishizawa T, Maltese C, Borghesan F, Mutinelli F, Montesi F, De Mas S (1999) Viral encephalopathy and retinopathy of farmed marine fish species in Italy. Virus Res 63:143–146

    Article  PubMed  CAS  Google Scholar 

  45. Mazzarella RA, Marcus N, Haugejorden SM, Balcarek JM, Baldassare JJ, Roy B, Li LJ, Lee AS, Green M (1994) Erp61 is GRP58, a stress-inducible luminal endoplasmic reticulum protein, but is devoid of phosphatidylinositide-specific phospholipase C activity. Arch Biochem Biophys 308:454–460

    Article  PubMed  Google Scholar 

  46. Pahl HL (1999) Signal transduction from the endoplasmic reticulum to the cell nucleus. Physiol Rev 79:683–701

    PubMed  CAS  Google Scholar 

  47. Buchkovich NJ, Maquire TG, Yu Y, Paton AW, Paton JC, Alwine JC (2007) Human cytomegalovirus specifically controls the levels of the endoplasmic reticulum chaperone BiP/GRP78, which is required for virion assembly. J Virol 82:31–39

    Article  PubMed  Google Scholar 

  48. Davis RJ (2000) Signal transduction by the JNK group of MAP kinases. Cell 103:239–252

    Article  PubMed  CAS  Google Scholar 

  49. Lamkanfi M, Kalai M, Vandenabeele P (2004) Caspase-12: an overview. Cell Death Differ 11:365–368

    Article  PubMed  CAS  Google Scholar 

  50. Yoneda T, Imaizumi K, Oono K, Yui D, Gomi F, Katayama T, Tohyama M (2001) Activation of caspase-12, an endoplastic reticulum (ER) resident caspase, through tumor necrosis factor receptor-associated factor 2-dependent mechanism in response to the ER stress. J Biol Chem 276:13935–13940

    PubMed  CAS  Google Scholar 

  51. Rutkowski DT, Kaufman RJ (2004) A trip to the ER: coping with stress. Trends Cell Biol 14:20–28

    Article  PubMed  CAS  Google Scholar 

  52. Dufresne PJ, Thivierge K, Cotton S, Beauchemin C, Ide C, Ubalijoro E, Laliberte JF, Fortin MG (2008) Heat shock 70 protein interaction with Turnip mosaic virus RNA-dependent RNA polymerase within virus-induced membrane vesicles. Virology 374:217–227

    Article  PubMed  CAS  Google Scholar 

  53. Miller DJ, Schwartz MD, Ahlquist P (2001) Flock house virus RNA replicates on outer mitochondrial membranes in Drosophila cells. J Virol 75:11664–11676

    Article  PubMed  CAS  Google Scholar 

  54. Weeks SA, Shield WP, Sahi C, Craig EA, Rospert S, Miller DJ (2010) A targeted analysis of cellular chaperones reveals contrasting roles for heat shock protein 70 in flock house virus RNA replication. J Virol 84:330–339

    Article  PubMed  CAS  Google Scholar 

  55. Germain M, Mathai JP, Shore GC (2002) BH-3-only BIK functions at the endoplasmic reticulum to stimulate cytochrome c release from mitochondria. J Biol Chem 277:18053–18060

    Article  PubMed  CAS  Google Scholar 

  56. Scorrano L, Oakes SA, Opferman JT, Cheng EH, Sorcinelli MD, Pozzan T, Korsmeyer SJ (2003) BAX and BAK regulation of endoplasmic reticulum Ca2+: a control point for apoptosis. Science 300:135–139

    Article  PubMed  CAS  Google Scholar 

  57. Wei MC, Zong WX, Cheng EH, Lindsten T, Panoutsakopoulou V, Ross AJ, Roth KA, MacGregor GR, Thompson CB, Korsmeyer SJ (2001) Proapoptotic BAX and BAK: a requisite gateway to mitochondrial dysfunction and death. Science 292:727–730

    Article  PubMed  CAS  Google Scholar 

  58. Kim I, Xu W, Reed JC (2008) Cell death and endoplasmic reticulum stress: disease relevance and therapeutic opportunities. Nat Rev Drug Discov 7:1013–1030

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

The authors are grateful to Dr. S. C. Chi (Institute of Zoology and Development of Life Science, Taiwan, ROC) for providing the grouper fin cell line GF-1. This work was supported by grant NSC 98-2313-B-006-004-MY3 from the National Science Council, Taiwan, Republic of China, awarded to Dr. Jainn-Ruey Hong.

Author information

Authors and Affiliations

  1. Laboratory of Molecular Virology and Biotechnology, Institute of Biotechnology, National Cheng Kung University, Tainan, 701, Taiwan, ROC

    Yu-Chin Su & Jiann-Ruey Hong

  2. Laboratory of Marine Molecular Biology and Biotechnology, Institute of Cellular and Organismic Biology, Academia Sinica, Nankang, Taipei, 115, Taiwan, ROC

    Jen-Leih Wu

Authors
  1. Yu-Chin Su
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jen-Leih Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jiann-Ruey Hong
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jiann-Ruey Hong.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Su, YC., Wu, JL. & Hong, JR. Betanodavirus up-regulates chaperone GRP78 via ER stress: roles of GRP78 in viral replication and host mitochondria-mediated cell death. Apoptosis 16, 272–287 (2011). https://doi.org/10.1007/s10495-010-0565-x

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10495-010-0565-x

Keywords

Search

Navigation