1887

Abstract

Our previous study showed that the nucleopolyhedrovirus (BmNPV) F-like protein Bm14 is intrinsically related to the production of occlusion bodies, occlusion-derived virus (ODV) embedding and virulence in infected larvae. However, the exact mechanism by which Bm14 affects primary infection remains unknown. In this report, we characterized the detailed distribution and topology of Bm14 in occlusion bodies (OBs) and ODVs, and then further investigated the functional role of Bm14 in primary infection. A combination of Western blot and immunoelectron microscopy showed that Bm14 is mainly present on the surface of ODVs within OBs, but rarely in the OB matrix. Further phase separation and topology analysis of Bm14 by selective permeabilization revealed that Bm14 is a type I integral membrane protein with an N-terminus hidden in the endoplasmic reticulum (ER) lumen and a C-terminus exposed to the cytosol. assays demonstrated that the disruption of impaired the interactions of ODV with midgut epithelia, resulting in delayed spread in larval tissues. As the essential trigger of primary infection, some infectivity factors (PIFs) were verified to interact with Bm14 via a series of coimmunoprecipitation analyses. Further partially denaturing SDS-PAGE and BN-PAGE assays clearly showed that the deletion of did not affect the formation and presence of the PIF complex. In conclusion, Bm14 functions as a type I integral membrane protein to regulate ODV attachment to the midgut epithelial cells.

Keyword(s): BmNPV , F-like , PIFs and primary infection
Funding
This study was supported by the:
  • Natural Science Foundation of Zhejiang Province (Award LZ20C170001)
    • Principle Award Recipient: Xiaofeng Wu
  • National Natural Science Foundation of China (Award 31972619)
    • Principle Award Recipient: Xiaofeng Wu
  • National Natural Science Foundation of China (CN) (Award 31772675)
    • Principle Award Recipient: Xiaofeng Wu
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2020-01-31
2024-04-20
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References

  1. Blissard G, Black B, Crook N, Keddie B, Possee R et al. Baculoviridae: taxonomic structure and properties of the family, p195-202. MHV van Regenmortel, et al; 2002
  2. Jehle JA, Blissard GW, Bonning BC, Cory JS, Herniou EA et al. On the classification and nomenclature of baculoviruses: a proposal for revision. Arch Virol 2006; 151:1257–1266 [View Article]
    [Google Scholar]
  3. Rohrmann GF. Baculovirus molecular biology; 2011
  4. Boogaard B, van Oers M, van Lent J. An advanced view on baculovirus per OS infectivity factors. Insects 2018; 9:84 [View Article]
    [Google Scholar]
  5. Slack J, Arif BM. The baculoviruses occlusion‐derived virus: virion structure and function. Advances in virus research 2006; 69:99–165
    [Google Scholar]
  6. Wang X, Liu X, Makalliwa GA, Li J, Wang H et al. Per OS infectivity factors: a complicated and evolutionarily conserved entry machinery of baculovirus. Sci China Life Sci 2017; 60:806–815 [View Article]
    [Google Scholar]
  7. Keddie B, Aponte G, Volkman L. The pathway of infection of Autographa californica nuclear polyhedrosis virus in an insect host. Science 1989; 243:1728–1730 [View Article]
    [Google Scholar]
  8. Passarelli AL. Barriers to success: how baculoviruses establish efficient systemic infections. Virology 2011; 411:383–392 [View Article]
    [Google Scholar]
  9. Lung OY, Cruz-Alvarez M, Blissard GW. Ac23, an envelope fusion protein homolog in the baculovirus Autographa californica multicapsid nucleopolyhedrovirus, is a viral pathogenicity factor. J Virol 2003; 77:328–339 [View Article]
    [Google Scholar]
  10. Xu W, Fan Y, Wang H, Feng M, Wu X. Bombyx mori nucleopolyhedrovirus F-like protein Bm14 affects the morphogenesis and production of occlusion bodies and the embedding of ODVs. Virology 2019a; 526:61–71 [View Article]
    [Google Scholar]
  11. Xu W, Wang H, Liu H, Wu X. Bombyx mori nucleopolyhedrovirus F-like protein Bm14 is a cofactor for GP64-Mediated efficient infection via forming a complex on the envelope of budded virus. Virology 2019b
    [Google Scholar]
  12. Wang X, Shang Y, Chen C, Liu S, Chang M et al. Baculovirus Per Os Infectivity Factor Complex: Components and Assembly. J Virol 2019; 93:e02053–02018 [View Article]
    [Google Scholar]
  13. Shen Y, Wang H, Xu W, Wu X. Bombyx mori nucleopolyhedrovirus orf133 and orf134 are involved in the embedding of occlusion-derived viruses into polyhedra. J Gen Virol 2018; 99:717–729 [View Article]
    [Google Scholar]
  14. Rosas-Acosta G, Braunagel SC, Summers MD. Effects of deletion and overexpression of the Autographa californica nuclear polyhedrosis virus FP25K gene on synthesis of two occlusion-derived virus envelope proteins and their transport into virus-induced intranuclear membranes. J Virol 2001; 75:10829–10842 [View Article]
    [Google Scholar]
  15. Wei D, Wang Y, Zhang X, Hu Z, Yuan M et al. Autographa californica nucleopolyhedrovirus Ac76: a dimeric type II integral membrane protein that contains an inner nuclear membrane-sorting motif. J Virol 2014; 88:1090–1103 [View Article]
    [Google Scholar]
  16. Haas-Stapleton EJ, Washburn JO, Volkman LE. P74 mediates specific binding of Autographa californica M nucleopolyhedrovirus occlusion-derived virus to primary cellular targets in the midgut epithelia of Heliothis virescens larvae. J Virol 2004; 78:6786–6791 [View Article]
    [Google Scholar]
  17. Nussbaum O, Loyter A. Quantitative determination of virus-membrane fusion events fusion of influenza virions with plasma membranes and membranes of endocytic vesicles in living cultured cells. FEBS Lett 1987; 221:61–67 [View Article]
    [Google Scholar]
  18. Ke P, Oers MM, Van Z, Lent H, Van JWM et al. Baculovirus per OS infectivity factors form a complex on the surface of occlusion-derived virus. Journal of Virology 2010; 84:9497–9504
    [Google Scholar]
  19. Wang X, Chen C, Zhang N, Li J, Deng F et al. The group I alphabaculovirus-specific protein, AC5, is a novel component of the occlusion body but is not associated with ODVs or the PIF complex. Journal of General Virology 2018; 99:585–595 [View Article]
    [Google Scholar]
  20. Guo Z-J, Yu M-H, Dong X-Y, Wang W-L, Tian T et al. Protein composition analysis of polyhedra matrix of Bombyx mori nucleopolyhedrovirus (BmNPV) showed powerful capacity of polyhedra to encapsulate foreign proteins. Sci Rep 2017; 7:8768 [View Article]
    [Google Scholar]
  21. Braunagel SC, Williamson ST, Saksena S, Zhong Z, Russell WK et al. Trafficking of ODV-E66 is mediated via a sorting motif and other viral proteins: facilitated trafficking to the inner nuclear membrane. Proc Natl Acad Sci U S A 2004; 101:8372–8377 [View Article]
    [Google Scholar]
  22. Gang L, Marcel W, Hualin W, Vlak JM, Zhihong H. Function, oligomerization and N-linked glycosylation of the Helicoverpa armigera single nucleopolyhedrovirus envelope fusion protein. Journal of General Virology 2006; 87:839
    [Google Scholar]
  23. Shao S, Hegde RS. Membrane protein insertion at the endoplasmic reticulum. Annu Rev Cell Dev Biol 2011; 27:2556 [View Article]
    [Google Scholar]
  24. Haas-Stapleton EJ, Washburn JO, Volkman LE. Spodoptera frugiperda resistance to oral infection by Autographa californica multiple nucleopolyhedrovirus linked to aberrant occlusion-derived virus binding in the midgut. Journal of General Virology 2005; 86:1349–1355 [View Article]
    [Google Scholar]
  25. Gowda MK, Peter A, Naika MB, Niranjana P, Shamprasad P. Identification of in-vitro red fluorescent protein with Antipathogenic activity from the midgut of the silkworm (Bombyx mori L.). Protein & Peptide Letters 2018; 25:
    [Google Scholar]
  26. Faulkner P, Wilson JA, Kuzio J, Williams GV. Analysis of p74, a PDV envelope protein of Autographa californica nucleopolyhedrovirus required for occlusion body infectivity in vivo. Journal of General Virology 1997; 78:3091–3100 [View Article]
    [Google Scholar]
  27. Kuzio J, Jaques R, Faulkner P. Identification of p74, a gene essential for virulence of baculovirus occlusion bodies. Virology 1989; 173:759–763 [View Article]
    [Google Scholar]
  28. Kikhno I, Gutiérrez S, Croizier L, Croizier G, Ferber ML. Characterization of PIF, a gene required for the per OS infectivity of Spodoptera littoralis nucleopolyhedrovirus. J Gen Virol 2002; 83:3013–3022 [View Article]
    [Google Scholar]
  29. Pijlman GP, Pruijssers AJ, Vlak JM. Identification of pif-2, a third conserved baculovirus gene required for per OS infection of insects. Journal of General Virology 2003; 84:2041–2049 [View Article]
    [Google Scholar]
  30. Ohkawa T, Washburn JO, Sitapara R, Sid E, Volkman LE. Specific binding of Autographa californica M nucleopolyhedrovirus Occlusion-Derived virus to midgut cells of Heliothis virescens larvae is mediated by products of PIF genes Ac119 and Ac022 but not by Ac115. J Virol 2005; 79:15258–15264 [View Article]
    [Google Scholar]
  31. Fang M, Nie Y, Harris S, Erlandson MA, Theilmann DA. Autographa californica multiple nucleopolyhedrovirus core gene ac96 encodes a per OS infectivity factor (pif-4). J Virol 2009; 83:12569–12578 [View Article]
    [Google Scholar]
  32. Sparks WO, Harrison RL, Bonning BC. Autographa californica multiple nucleopolyhedrovirus ODV-E56 is a per OS infectivity factor, but is not essential for binding and fusion of occlusion-derived virus to the host midgut. Virology 2011; 409:69–76 [View Article]
    [Google Scholar]
  33. Nie Y, Fang M, Erlandson MA, Theilmann DA. Analysis of the Autographa californica multiple nucleopolyhedrovirus overlapping gene pair lef3 and ac68 reveals that AC68 is a per OS infectivity factor and that LEF3 is critical, but not essential, for virus replication. J Virol 2012; 86:3985–3994 [View Article]
    [Google Scholar]
  34. Jianta Liu, Zhu L, Zhang S, Deng Z, Huang Z et al. The Autographa californica multiple nucleopolyhedrovirus ac110 gene encodes a new per OS infectivity factor. Virus Res 2016; 221:30–37 [View Article]
    [Google Scholar]
  35. Javed MA, Biswas S, Willis LG, Harris S, Pritchard C et al. Autographa californica Multiple Nucleopolyhedrovirus AC83 is a Per Os Infectivity Factor (PIF) Protein Required for Occlusion-Derived Virus (ODV) and Budded Virus Nucleocapsid Assembly as well as Assembly of the PIF Complex in ODV Envelopes. J Virol 2017; 91:e02115–02116 [View Article]
    [Google Scholar]
  36. Zhu S, Wang W, Wang Y, Yuan M, Yang K. The baculovirus core gene ac83 is required for nucleocapsid assembly and per OS infectivity of Autographa californica nucleopolyhedrovirus. J Virol 2013; 87:10573–10586 [View Article]
    [Google Scholar]
  37. Peng K, van Lent JW, Boeren S, Fang M, Theilmann DA et al. Characterization of novel components of the baculovirus per OS infectivity factor (PIF) complex. Journal of virology 201206801–06811
    [Google Scholar]
  38. Boogaard B, van Lent JWM, Theilmann DA, Erlandson MA, van Oers MM. Baculoviruses require an intact ODV entry-complex to resist proteolytic degradation of per OS infectivity factors by co-occluded proteases from the larval host. J Gen Virol 2017; 98:3101–3110 [View Article]
    [Google Scholar]
  39. Javed MA, Biswas S, Willis LG, Harris S, Pritchard C et al. Acmnpv AC83 is a PIF protein required for ODV and bv nucleocapsid assembly as well as assembly of the PIF complex in ODV envelopes. Journal of Virology 201602115–02116
    [Google Scholar]
  40. O'Brochta DA, Atkinson PW. Recent developments in transgenic insect technology. Parasitology Today 1997; 13:99–104 [View Article]
    [Google Scholar]
  41. Perera O, Green TB, Stevens SM, White S, Becnel JJ. Proteins associated with Culex nigripalpus nucleopolyhedrovirus occluded virions. J Virol 2007; 81:4585–4590 [View Article]
    [Google Scholar]
  42. Tang Q, Li G, Yao Q, Chen L, Lv P et al. Bm91 is an envelope component of ODV but is dispensable for the propagation of Bombyx mori nucleopolyhedrovirus. J Invertebr Pathol 2013; 113:70–77 [View Article]
    [Google Scholar]
  43. Xiang X, Shen Y, Yang R, Chen L, Hu X et al. Bombyx mori nucleopolyhedrovirus BmP95 plays an essential role in budded virus production and nucleocapsid assembly. Journal of General Virology 2013; 94:1669–1679 [View Article]
    [Google Scholar]
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