Skip to main content
SpringerLink
Log in

Grass carp reovirus NS26 interacts with cellular lipopolysaccharide-induced tumor necrosis factor-alpha factor, LITAF

  • Published:
Virus Genes Aims and scope Submit manuscript

Abstract

The nonstructural protein NS26 of grass carp reovirus (GCRV) is encoded by the 11th genomic dsRNA segment, homolog of which is not found in orthoreoviruses. The role of NS26 in GCRV pathogenesis is still unclear. Previously, grass carp LITAF/SIMPLE protein was identified as a putative binding partner for NS26 in a yeast two-hybrid screen. Here, we further characterized the association between NS26 and LITAF using in vivo and in vitro protein interaction assays. Soluble GST-NS26 and His6-LITAF were expressed and purified from E. coli; recombinant NS26 tagged with myc and LITAF tagged with GFP were expressed in Ctenopharyngon idellus kidney cells (CIK) by transient transfection experiments. A GST pulldown assay demonstrated that GST-tagged NS26 efficiently bound to His6-LITAF. Co-immunoprecipitation assays demonstrated that GCRV NS26 reciprocally precipitated endogenous LITAF in CIK cells. Double-immunofluorescent analyses revealed myc-NS26 colocalized with GFP-LITAF in CIK cells. Taken together, the current in vitro and in vivo data demonstrated the interaction between cellular LITAF and GCRV NS26.

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.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. S. Blindheim, A. Nylund, K. Watanabe, H. Plarre, B. Erstad, S. Nylund, A new aquareovirus causing high mortality in farmed Atlantic halibut fry in Norway. Arch. Virol. 160(1), 91–102 (2015)

    Article  CAS  PubMed  Google Scholar 

  2. J.M. Carballeda, S. Chimeno Zoth, E. Gómez, J.M. Gravisaco, A. Berinstein, Activation of the immune response against infectious bursal disease virus after intramuscular inoculation of an intermediate strain. Immunobiology 216(9), 1028–1033 (2011)

    Article  CAS  PubMed  Google Scholar 

  3. L. Cheng, Q. Fang, S. Shah, I.C. Atanasov, Z.H. Zhou, Subnanometer-resolution structures of the grass carp reovirus core and virion. J. Mol. Biol. 382(1), 213–222 (2008)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. H.E. Eaton, A. Ferreira Lacerda, G. Desrochers, J. Metcalf, A. Angers, C.R. Brunetti, Cellular LITAF interacts with frog virus 3 75L protein and alters its subcellular localization. J. Virol. 87(2), 716–723 (2013)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Y. Fan, S. Rao, L. Zeng, J. Ma, Y. Zhou, J. Xu, H. Zhang, Identification and genomic characterization of a novel fish reovirus, Hubei grass carp disease reovirus, isolated in 2009 in China. J. Gen. Virol. 94(Pt 10), 2266–2277 (2013)

    Article  CAS  PubMed  Google Scholar 

  6. Q. Fang, S. Shah, Y. Liang, Z.H. Zhou, 3D reconstruction and capsid protein characterization of grass carp reovirus. Sci. China C (Life Sci) 48(6), 593–600 (2005)

    Article  CAS  Google Scholar 

  7. H. Guo, Q. Chen, L. Yan, J. Zhang, S. Yan, F. Zhang, Q. Fang, Identification of a functional motif in the AqRV NS26 protein required for enhancing the fusogenic activity of FAST protein NS16. J. Gen. Virol. 96(Pt 5), 1080–1085 (2015)

    Article  CAS  PubMed  Google Scholar 

  8. H. Guo, X. Sun, L. Yan, L. Shao, Q. Fang, The NS16 protein of aquareovirus-C is a fusion-associated small transmembrane (FAST) protein, and its activity can be enhanced by the nonstructural protein NS26. Virus Res. 171(1), 129–137 (2013)

    Article  CAS  PubMed  Google Scholar 

  9. Y. He, Y. Jiang, L. Lu, Serodiagnosis of grass carp reovirus infection in grass carp Ctenopharyngodon idella by a novel Western blot technique. J. Virol. Methods 194(1–2), 14–20 (2013)

    Article  CAS  PubMed  Google Scholar 

  10. Y.H. Hong, H.S. Lillehoj, S.H. Lee, D. Park, E.P. Lillehoj, Molecular cloning and characterization of chicken lipopolysaccharide-induced TNF-alpha factor (LITAF). Dev. Comp. Immunol. 30(10), 919–929 (2006)

    Article  CAS  PubMed  Google Scholar 

  11. X. Huang, Y. Huang, J. Gong, Y. Yan, Q. Qin, Identification and characterization of a putative lipopolysaccharide-induced TNF-alpha factor (LITAF) homolog from Singapore grouper iridovirus. Biochem. Biophys. Res. Commun. 373(1), 140–145 (2008)

    Article  CAS  PubMed  Google Scholar 

  12. S.M. Lee, L.S. Chin, L. Li, Charcot-Marie-Tooth disease-linked protein SIMPLE functions with the ESCRT machinery in endosomal trafficking. J. Cell Biol. 199(5), 799–816 (2012)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. S.M. Lee, J.A. Olzmann, L.S. Qin, L. Li, Mutations associated with Charcot-Marine-Tooth disease cause SIMPLE protein mislocation and degradation by the proteasome and aggresome-autophagy pathways. J. Cell Sci. 124(Pt19), 3319–3331 (2011)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. L. Lu, H. Xu, Y. He, J. Li, Protection of grass carp, Ctenopharyngon idellus (Valenciennes), through oral administration of a subunit vaccine against reovirus. J. Fish Dis. 34(12), 939–942 (2011)

    Article  CAS  PubMed  Google Scholar 

  15. C. Mavian, A. López-Bueno, M.P. FernándezSomalo, A. Alcamí, A. Alejo, Complete genome sequence of the European sheatfish virus. J. Virol. 86(11), 6365–6366 (2012)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. J.C. Merrill, J. You, C. Constable, S.E. Leeman, S. Amar, Whole-body deletion of LPS-induced TNF-α factor (LITAF) markedly improves experimental endotoxic shock and inflammatory arthritis. Proc. Natl. Acad. Sci. USA 108(52), 21247–21252 (2011)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. F. Mohd Jaafar, A.E. Goodwin, M. Belhouchet, G. Merry, Q. Fang, J.F. Cantaloube, P. Biagini, P. de Micco, P.P. Mertens, H. Attoui, Complete characterisation of the American grass carp reovirus genome (genus Aquareovirus: family Reoviridae) reveals an evolutionary link between aquareoviruses and coltiviruses. Virology 373(2), 310–321 (2008)

    Article  CAS  PubMed  Google Scholar 

  18. C. Pei, F. Ke, Z.Y. Chen, Q.Y. Zhang, Complete genome sequence and comparative analysis of grass carp reovirus strain 109 (GCReV-109) with other grass carp reovirus strains reveals no significant correlation with regional distribution. Arch. Virol. 159(9), 2435–2440 (2014)

    Article  CAS  PubMed  Google Scholar 

  19. X. Tang, D.L. Marciano, S.E. Leeman, S. Amar, LPS induces the interaction of a transcription factor, LPS-induced TNF-alpha factor, and STAT6(B) with effection on multiple cytokines. Proc. Natl. Acad. Sci. USA 102(14), 5132–5137 (2005)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. X. Tang, D. Metzger, S. Leeman, S. Amar, LPS-induced TNF-alpha factor (LITAF)-deficient mice express reduced LPS-induced cytokine: evidence for LITAF-dependent LPS signaling pathways. Proc. Natl. Acad. Sci. USA 103(37), 13777–13782 (2006)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. H. Wang, X. Shen, D. Xu, L. Lu, Lipopolysaccharide-induced TNF-α factor in grass carp (Ctenopharyngodon idella): evidence for its involvement in antiviral innate immunity. Fish Shellfish Immunol. 34(2), 538–545 (2013)

    Article  CAS  PubMed  Google Scholar 

  22. Q. Wang, W. Zeng, C. Liu, C. Zhang, Y. Wang, C. Shi, S. Wu, Complete genome sequence of a reovirus isolated from grass carp, indicating different genotypes of GCRV in China. J. Virol. 86(22), 12466 (2012)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. T. Wang, J. Li, L. Lu, Quantitative in vivo and in vitro characterization of co-infection by two genetically distant grass carp reoviruses. J. Gen. Virol. 94(Pt 6), 1301–1309 (2013)

    Article  CAS  PubMed  Google Scholar 

  24. X. Ye, Y.Y. Tian, G.C. Deng, Y.Y. Chi, X.Y. Jiang, Complete genomic sequence of a reovirus isolated from grass carp in China. Virus Res. 163(1), 275–283 (2012)

    Article  CAS  PubMed  Google Scholar 

  25. X. Zhang, L. Jin, Q. Fang, W.H. Hui, Z.H. Zhou, 3.3 Å cryo-EM structure of a non enveloped virus reveals a priming mechanism for cell entry. Cell 141(3), 472–482 (2010)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Y. Zhang, H. Wang, Y. Li, D. Xu, L. Lu, Molecular cloning and expression analysis of the Ajuba gene of grass carp (Ctenopharyngodon idella) involved in cellular response to viral infection. Dev. Comp. Immunol. 48(1), 164–170 (2015)

    Article  CAS  PubMed  Google Scholar 

  27. H. Zhu, S. Guariglia, R.Y. Yu, W. Li, D. Brancho, H. Peinado, D. Lyden, J. Salzer, C. Bennett, C.W. Chow, Mutation of SIMPLE in Charcot-Marie-Tooth 1C alters production of exosomes. Mol. Biol. Cell 24(11), 1619–1637 (2013)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Aquatic Pathogen Collection Center, MOA Key Laboratory of Freshwater Fishery Germplasm Resources, Shanghai Ocean University, Shanghai, 201306, China

    Jianfei Lu, Hao Wang, Yanan Zhang, Yan Li & Liqun Lu

Authors
  1. Jianfei Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hao Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yanan Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yan Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Liqun Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Liqun Lu.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All the applicable international and national guidelines for the care and use of animals were followed.

Funding

This study was funded by the National Natural Science Foundation of China (No. 31372561), the Funding program for outstanding dissertations of Shanghai Ocean University (A1-0209-14-0902-5).

Additional information

Edited by Zhen F. Fu.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lu, J., Wang, H., Zhang, Y. et al. Grass carp reovirus NS26 interacts with cellular lipopolysaccharide-induced tumor necrosis factor-alpha factor, LITAF. Virus Genes 52, 789–796 (2016). https://doi.org/10.1007/s11262-016-1370-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11262-016-1370-6

Keywords

Search

Navigation