Archives of Virology

, Volume 163, Issue 10, pp 2719–2725 | Cite as

Inhibition of expression of BmNPV cg30 by bmo-miRNA-390 is a host response to baculovirus invasion

  • Lequn Kang
  • Minglei Wang
  • Xueliang Cao
  • Shunming Tang
  • Dingguo Xia
  • Xingjia Shen
  • Qiaoling Zhao
Original Article


Bombyx mori larvae exhibit in vivo defensive reactions immediately after invasion by a virus. One of these defense systems is to express appropriate microRNAs (miRNAs) to respond to the infection. A novel Bombyx mori–encoded miRNA, bmo-miR-390, was identified previously by high-throughput sequencing. Based on bioinformatic predictions, the Bombyx mori nuclear polyhedrosis virus cg30 gene (BmNPV-cg30) is one of the target genes of bmo-miR-390. In this study, expression vectors with an enhanced green fluorescence protein (EGFP) or a luciferase (luc) reporter gene together with bm-miR-390 or the cg30 3’ UTR were constructed and used to co-transfect BmN cells. Using a dual luciferase reporter (DLR) assay, we found that bmo-miR-390 significantly downregulates the expression of BmNPV-cg30 (P < 0.05) in vitro. Moreover, artificially synthesized bmo-miR-390 mimics enhanced the regulatory effect of bmo-miR-390, while an inhibitor eliminated the inhibitory effect. These results show for the first time that bmo-miR-390 can effectively downregulate the expression of BmNPV-cg30 in BmNPV-infected BmN cells.


Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


This work was supported by the National Natural Science Foundation of China (Grant nos. 31372378 and 31372377).


  1. 1.
    Asgari S (2011) Role of microRNAs in insect host–microorganism interactions. Front Physiol 2:48CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Ayres MD, Howard SC, Kuzio J, Lopez-Ferber M, Possee RD (1994) The complete DNA sequence of Autographa californica nuclear polyhedrosis virus. Virology 202:586–605CrossRefPubMedGoogle Scholar
  3. 3.
    Cao X, Huang Y, Xia D, Qiu Z, Shen X, Guo X, Zhao Q (2015) BmNPV-miR-415 up-regulates the expression of TOR2 via Bmo-miR-5738. Saudi J Biol Sci 24:1614–1619CrossRefPubMedCentralPubMedGoogle Scholar
  4. 4.
    Chen A, Xia D, Qiu Z, Gao P, Tang S, Shen X, Zhu F, Zhao Q (2013) Expression of a vitelline membrane protein, BmVMP23, is repressed by bmo-miR-1a-3p in silkworm, Bombyx mori. FEBS Lett 587:970–975CrossRefPubMedGoogle Scholar
  5. 5.
    Chen C, Ridzon DA, Broomer AJ, Zhou Z, Lee DH, Nguyen JT, Barbisin M, Xu NL, Mahuvakar VR, Andersen MR (2005) Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res 33:e179CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Granados RR, Lawler KA (1981) In vivo pathway of Autographa californica baculovirus invasion and infection. Virology 108:297–308CrossRefPubMedGoogle Scholar
  7. 7.
    Grey F, Meyers H, White EA, Spector DH, Nelson J (2007) A human cytomegalovirus-encoded microRNA regulates expression of multiple viral genes involved in replication. PLoS Pathog 3:e163CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Hatfield S, Ruohola-Baker H (2008) MicroRNA and stem cell function. Cell Tissue Res 331:57–66CrossRefPubMedGoogle Scholar
  9. 9.
    He J, Zhang JF, Yi C, Lv Q, Xie WD, Li JN, Wan G, Cui K, Kung HF, Yang J, Yang BB, Zhang Y (2010) miRNA-mediated functional changes through co-regulating function related genes. PLoS One 5:e13558CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Ho BC, Yang PC, Yu SL (2016) MicroRNA and pathogenesis of enterovirus infection. Viruses. CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Horton HM, Burand JP (1993) Saturable attachment sites for polyhedron-derived baculovirus on insect cells and evidence for entry via direct membrane fusion. J Virol 67:1860–1868PubMedPubMedCentralGoogle Scholar
  12. 12.
    Hussain M, Frentiu FD, Moreira LA, O’Neill SL, Asgari S (2011) Wolbachia uses host microRNAs to manipulate host gene expression and facilitate colonization of the dengue vector Aedes aegypti. Proc Natl Acad Sci USA 108:9250–9255CrossRefPubMedGoogle Scholar
  13. 13.
    Imai N, Matsuda N, Tanaka K, Nakano A, Matsumoto S, Kang W (2003) Ubiquitin ligase activities of Bombyx mori nucleopolyhedrovirus RING finger proteins. J Virol 77:923–930CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Ishihara G, Shimada T, Katsuma S (2013) Functional characterization of Bombyx mori nucleopolyhedrovirus CG30 protein. Virus Res 174:52–59CrossRefPubMedGoogle Scholar
  15. 15.
    Keddie BA, Aponte GW, Volkman LE (1989) The pathway of infection of Autographa californica nuclear polyhedrosis virus in an insect host. Science 243:1728–1730CrossRefPubMedGoogle Scholar
  16. 16.
    Lakshmipathy U, Love B, Goff LA, Jornsten R, Graichen R, Hart RP, Chesnut JD (2007) MicroRNA expression pattern of undifferentiated and differentiated human embryonic stem cells. Stem Cells Dev 16:1003–1016CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Lemaitre B, Hoffmann J (2007) The host defense of Drosophila melanogaster. Annu Rev Immunol 25:697–743CrossRefPubMedGoogle Scholar
  18. 18.
    Lewis BP, Burge CB, Bartel DP (2005) Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 120:15–20CrossRefPubMedGoogle Scholar
  19. 19.
    Lu M, Iatrou K (1996) The genes encoding the P39 and CG30 proteins of Bombyx mori nuclear polyhedrosis virus. J Gen Virol 77(Pt 12):3135–3143CrossRefPubMedGoogle Scholar
  20. 20.
    Lucas KJ, Zhao B, Liu S, Raikhel AS (2015) Regulation of physiological processes by microRNAs in insects. Curr Opin Insect Sci 11:1–7CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Morin RD, O’Connor MD, Griffith M, Kuchenbauer F, Delaney A, Prabhu AL, Zhao Y, McDonald H, Zeng T, Hirst M, Eaves CJ, Marra MA (2008) Application of massively parallel sequencing to microRNA profiling and discovery in human embryonic stem cells. Genome Res 18:610–621CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Nair V, Zavolan M (2006) Virus-encoded microRNAs: novel regulators of gene expression. Trends Microbiol 14:169–175CrossRefPubMedGoogle Scholar
  23. 23.
    Passarelli AL, Miller LK (1994) In vivo and in vitro analyses of recombinant baculoviruses lacking a functional cg30 gene. J Virol 68:1186–1190PubMedPubMedCentralGoogle Scholar
  24. 24.
    Rahman MM, Gopinathan KP (2004) Systemic and in vitro infection process of Bombyx mori nucleopolyhedrovirus. Virus Res 101:109–118CrossRefPubMedGoogle Scholar
  25. 25.
    Scaria V, Hariharan M, Maiti S, Pillai B, Brahmachari SK (2006) Host–virus interaction: a new role for microRNAs. Retrovirology 3:68CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Schratt GM, Tuebing F, Nigh EA, Kane CG, Sabatini ME, Kiebler M, Greenberg ME (2006) A brain-specific microRNA regulates dendritic spine development. Nature 439:283–289CrossRefPubMedGoogle Scholar
  27. 27.
    Serrano A, Pijlman GP, Vlak JM, Munoz D, Williams T, Caballero P (2015) Identification of Spodoptera exigua nucleopolyhedrovirus genes involved in pathogenicity and virulence. J Invertebr Pathol 126:43–50CrossRefPubMedGoogle Scholar
  28. 28.
    Shi X, Ran Z, Li S, Yin J, Zhong J (2016) The effect of microRNA bantam on baculovirus AcMNPV infection in vitro and in vivo. Viruses. CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Singh CP, Singh J, Nagaraju J (2012) A baculovirus-encoded microRNA (miRNA) suppresses its host miRNA biogenesis by regulating the exportin-5 cofactor Ran. J Virol 86:7867–7879CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Singh CP, Singh J, Nagaraju J (2014) Bmnpv-miR-3 facilitates BmNPV infection by modulating the expression of viral P6.9 and other late genes in Bombyx mori. Insect Biochem Mol Biol 49:59–69CrossRefPubMedGoogle Scholar
  31. 31.
    Slonchak A, Hussain M, Torres S, Asgari S, Khromykh AA (2014) Expression of mosquito microRNA Aae-miR-2940-5p is downregulated in response to West Nile virus infection to restrict viral replication. J Virol 88:8457–8467CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Veerappa AM, Murthy MN, Vishweswaraiah S, Lingaiah K, Suresh RV, Nachappa SA, Prashali N, Yadav SN, Srikanta MA, Manjegowda DS, Seshachalam KB, Ramachandra NB (2014) Copy number variations burden on miRNA genes reveals layers of complexities involved in the regulation of pathways and phenotypic expression. PLoS One 9:e90391CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Wu Y, Wu Y, Hui T, Wu H, Wu Y, Wang W (2013) Reaper homologue IBM1 in silkworm Bombyx mori induces apoptosis upon baculovirus infection. FEBS Lett 587:600–606CrossRefPubMedGoogle Scholar
  34. 34.
    Wu YL, Wu CP, Liu CY, Hsu PW, Wu EC, Chao YC (2011) A non-coding RNA of insect HzNV-1 virus establishes latent viral infection through microRNA. Sci Rep 1:60CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Yang N, Coukos G, Zhang L (2008) MicroRNA epigenetic alterations in human cancer: one step forward in diagnosis and treatment. Int J Cancer 122:963–968CrossRefPubMedGoogle Scholar
  36. 36.
    Yeung ML, Bennasser Y, Jeang KT (2007) miRNAs in the biology of cancers and viral infections. Curr Med Chem 14:191–197CrossRefPubMedGoogle Scholar
  37. 37.
    Yu J, Ryan DG, Getsios S, Oliveira-Fernandes M, Fatima A, Lavker RM (2008) MicroRNA-184 antagonizes microRNA-205 to maintain SHIP2 levels in epithelia. Proc Natl Acad Sci USA 105:19300–19305CrossRefPubMedGoogle Scholar
  38. 38.
    Zhang J, He Q, Zhang CD, Chen XY, Chen XM, Dong ZQ, Li N, Kuang XX, Cao MY, Lu C, Pan MH (2014) Inhibition of BmNPV replication in silkworm cells using inducible and regulated artificial microRNA precursors targeting the essential viral gene lef-11. Antiviral Res 104:143–152CrossRefPubMedGoogle Scholar
  39. 39.
    Zhang L, Xu Y, Jin X, Wang Z, Wu Y, Zhao D, Chen G, Li D, Wang X, Cao H, Xie Y, Liang Z (2015) A circulating miRNA signature as a diagnostic biomarker for non-invasive early detection of breast cancer. Breast cancer Res Treat 154:423–434CrossRefPubMedGoogle Scholar
  40. 40.
    Zhang MJ, Cheng RL, Lou YH, Ye WL, Zhang T, Fan XY, Fan HW, Zhang CX (2012) Disruption of Bombyx mori nucleopolyhedrovirus ORF71 (Bm71) results in inefficient budded virus production and decreased virulence in host larvae. Virus Genes 45:161–168CrossRefPubMedGoogle Scholar
  41. 41.
    Zhu S, Wu H, Wu F, Nie D, Sheng S, Mo YY (2008) MicroRNA-21 targets tumor suppressor genes in invasion and metastasis. Cell Res 18:350–359CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  • Lequn Kang
    • 1
  • Minglei Wang
    • 1
  • Xueliang Cao
    • 1
  • Shunming Tang
    • 1
  • Dingguo Xia
    • 1
  • Xingjia Shen
    • 1
  • Qiaoling Zhao
    • 1
  1. 1.Jiangsu University of Science and TechnologyZhenjiangChina

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