Virologica Sinica

, Volume 34, Issue 6, pp 712–721 | Cite as

Autographa Californica Multiple Nucleopolyhedrovirus P48 (Ac103) Is Required for the Efficient Formation of Virus-Induced Intranuclear Microvesicles

  • Yan Wang
  • Qingyun Cai
  • Jiannan Chen
  • Zhihong Huang
  • Wenbi Wu
  • Meijin YuanEmail author
  • Kai Yang
Research Article


Our previous study has shown that the Autographa californica multiple nucleopolyhedrovirus (AcMNPV) p48 (ac103) gene is essential for the nuclear egress of nucleocapsids and the formation of occlusion-derived virions (ODVs). However, the exact role of p48 in the morphogenesis of ODVs remains unknown. In this study, we demonstrated that p48 was required for the efficient formation of intranuclear microvesicles. To further understand its functional role in intranuclear microvesicle formation, we characterized the distribution of the P48 protein, which was found to be associated with the nucleocapsid and envelope fractions of both budded virions and ODVs. In AcMNPV-infected cells, P48 was predominantly localized to nucleocapsids in the virogenic stroma and the nucleocapsids enveloped in ODVs, with a limited but discernible distribution in the plasma membrane, nuclear envelope, intranuclear microvesicles, and ODV envelope. Furthermore, coimmunoprecipitation assays showed that among the viral proteins required for intranuclear microvesicle formation, P48 associated with Ac93 in the absence of viral infection.


P48 Baculovirus Intranuclear microvesicle formation Protein association Ac93 



We thank Prof. Zhihong Hu (Wuhan Institute of Virology) for the generous gift of E25 polyclonal antiserum. This research was supported by the National Natural Science Foundation of China (31572056 and 31872025), the Key Project of Natural Science Foundation of Guangdong Province (2018B030311018), the National Key R&D Program of China (2017YFD0200404), and the Guangzhou Science and Technology Project (201707020003).

Author Contributions

YW and MY conceived the experiments. YW constructed recombinant viruses and conducted TEM analyses, TCID50 EPDA, BV and ODV purification, IEM analyses, and coimmunoprecipitation assays. QC and JC constructed plasmids. ZH, WW, KY and MY analyzed the results. MY is responsible for the financial support of the project. All authors reviewed the manuscript.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Animal and Human Rights Statement

This article does not contain any studies with human or animal subjects performed by any of the authors.

Supplementary material

12250_2019_147_MOESM1_ESM.pdf (346 kb)
Supplementary material 1 (PDF 347 kb)


  1. Ayres MD, Howard SC, Kuzio J, Lopez-Ferber M, Possee RD (1994) The complete DNA sequence of Autographa californica nuclear polyhedrosis virus. Virology 202:586–605PubMedCrossRefPubMedCentralGoogle Scholar
  2. Blissard GW, Wenz JR (1992) Baculovirus gp64 envelope glycoprotein is sufficient to mediate pH-dependent membrane fusion. J Virol 66:6829–6835PubMedPubMedCentralGoogle Scholar
  3. Blissard GW, Theilmann DA (2018) Baculovirus entry and egress from insect cells. Annu Rev Virol 5:113–139PubMedCrossRefPubMedCentralGoogle Scholar
  4. Braunagel SC, Summers MD (1994) Autographa californica nuclear polyhedrosis virus, PDV, and ECV viral envelopes and nucleocapsids: structural proteins, antigens, lipid and fatty acid profiles. Virology 202:315–328PubMedCrossRefPubMedCentralGoogle Scholar
  5. Braunagel SC, Summers MD (2007) Molecular biology of the baculovirus occlusion-derived virus envelope. Curr Drug Targets 8:1084–1095PubMedCrossRefPubMedCentralGoogle Scholar
  6. Braunagel SC, Russell WK, Rosas-Acosta G, Russell DH, Summers MD (2003) Determination of the protein composition of the occlusion-derived virus of Autographa californica nucleopolyhedrovirus. Proc Natl Acad Sci USA 100:9797–9802PubMedCrossRefPubMedCentralGoogle Scholar
  7. Braunagel SC, Williamson ST, Saksena S, Zhong Z, Russell WK, Russell DH, Summers MD (2004) 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 USA 101:8372–8377PubMedCrossRefPubMedCentralGoogle Scholar
  8. Braunagel SC, Cox V, Summers MD (2009) Baculovirus data suggest a common but multifaceted pathway for sorting proteins to the inner nuclear membrane. J Virol 83:1280–1288PubMedCrossRefPubMedCentralGoogle Scholar
  9. Cai Y, Long Z, Qiu J, Yuan M, Li G, Yang K (2012) An ac34 deletion mutant of Autographa californica nucleopolyhedrovirus exhibits delayed late gene expression and a lack of virulence in vivo. J Virol 86:10432–10443PubMedPubMedCentralCrossRefGoogle Scholar
  10. Chen L, Hu X, Xiang X, Yu S, Yang R, Wu X (2012) Autographa californica multiple nucleopolyhedrovirus odv-e25 (Ac94) is required for budded virus infectivity and occlusion-derived virus formation. Arch Virol 157:617–625PubMedCrossRefPubMedCentralGoogle Scholar
  11. Chen Y, Zhong S, Fei Z, Hashimoto Y, Xiang J, Zhang S, Blissard GW (2013) The transcriptome of the baculovirus Autographa californica multiple nucleopolyhedrovirus in Trichoplusia ni cells. J Virol 87:6391–6405PubMedPubMedCentralCrossRefGoogle Scholar
  12. Garavaglia M, Miele S, Iserte J, Belaich M, Ghiringhelli P (2012) The ac53, ac78, ac101, and ac103 genes are newly discovered core genes in the family Baculoviridae. J Virol 86:12069–12079PubMedPubMedCentralCrossRefGoogle Scholar
  13. Ge JQ, Gao GH, Xu YP, Zhang CX (2011) Characterization of a late gene, ORF75 from Bombyx mori nucleopolyhedrovirus. Mol Biol Rep 38:2141–2149PubMedCrossRefPubMedCentralGoogle Scholar
  14. Guan Z, Zhong L, Li C, Wu W, Yuan M, Yang K (2016) The Autographa californica multiple nucleopolyhedrovirus ac54 gene is crucial for localization of the major capsid protein VP39 at the site of nucleocapsid assembly. J Virol 90:4115–4126PubMedPubMedCentralCrossRefGoogle Scholar
  15. Guo Y, Yue Q, Gao J, Wang Z, Chen YR, Blissard GW, Liu TX, Li Z (2017) Roles of cellular NSF protein in entry and nuclear egress of budded virions of Autographa californica multiple nucleopolyhedrovirus. J Virol 91:e01111–e01117PubMedPubMedCentralGoogle Scholar
  16. Harrison RL, Herniou EA, Jehle JA, Theilmann DA, Burand JP, Becnel JJ, Krell PJ, van Oers MM, Mowery JD, Bauchan GR, ICTV Report C (2018) ICTV virus taxonomy profile: baculoviridae. J Gen Virol 99:1185–1186PubMedCrossRefPubMedCentralGoogle Scholar
  17. Hellberg T, Passvogel L, Schulz KS, Klupp BG, Mettenleiter TC (2016) Nuclear egress of Herpesviruses: the prototypic vesicular nucleocytoplasmic transport. Adv Virus Res 94:81–140PubMedCrossRefPubMedCentralGoogle Scholar
  18. Hodgson JJ, Arif BM, Krell PJ (2007) Reprogramming the chiA expression profile of Autographa californica multiple nucleopolyhedrovirus. J Gen Virol 88:2479–2487PubMedCrossRefPubMedCentralGoogle Scholar
  19. Hong T, Summers MD, Braunagel SC (1997) N-terminal sequences from Autographa californica nuclear polyhedrosis virus envelope proteins ODV-E66 and ODV-E25 are sufficient to direct reporter proteins to the nuclear envelope, intranuclear microvesicles and the envelope of occlusion derived virus. Proc Natl Acad Sci USA 94:4050–4055PubMedCrossRefPubMedCentralGoogle Scholar
  20. Hou D, Zhang L, Deng F, Fang W, Wang R, Liu X, Guo L, Rayner S, Chen X, Wang H, Hu Z (2013) Comparative proteomics reveal fundamental structural and functional differences between the two progeny phenotypes of a baculovirus. J Virol 87:829–839PubMedPubMedCentralCrossRefGoogle Scholar
  21. Hu Z, Yuan M, Wu W, Liu C, Yang K, Pang Y (2010) Autographa californica multiple nucleopolyhedrovirus ac76 is involved in intranuclear microvesicle formation. J Virol 84:7437–7447PubMedPubMedCentralCrossRefGoogle Scholar
  22. Jehle JA, Blissard GW, Bonning BC, Cory JS, Herniou EA, Rohrmann GF, Theilmann DA, Thiem SM, Vlak JM (2006) On the classification and nomenclature of baculoviruses: a proposal for revision. Arch Virol 151:1257–1266PubMedCrossRefPubMedCentralGoogle Scholar
  23. Ke J, Wang J, Deng R, Wang X (2008) Autographa californica multiple nucleopolyhedrovirus ac66 is required for the efficient egress of nucleocapsids from the nucleus, general synthesis of preoccluded virions and occlusion body formation. Virology 374:421–431PubMedCrossRefPubMedCentralGoogle Scholar
  24. Kozak M (1987) An analysis of 5′-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res 15:8125–8148PubMedPubMedCentralCrossRefGoogle Scholar
  25. Kozak M (1991) An analysis of vertebrate mRNA sequences: intimations of translational control. J Cell Biol 115:887–903PubMedCrossRefPubMedCentralGoogle Scholar
  26. Lehiy CJ, Wu W, Berretta MF, Passarelli AL (2013) Autographa californica M nucleopolyhedrovirus open reading frame 109 affects infectious budded virus production and nucleocapsid envelopment in the nucleus of cells. Virology 435:442–452PubMedCrossRefPubMedCentralGoogle Scholar
  27. Li GH, Pang Y, Chen QJ, Su ZJ, Wen XZ (2002) Studies on the artificial diet for beet armyworm Spodoptera exigua. Chin J Biol Control 18:132–134Google Scholar
  28. Li Y, Wang JW, Deng RQ, Zhang QF, Yang K, Wang XZ (2005a) vlf-1 deletion brought AcMNPV to defect in nucleocapsid formation. Virus Genes 31:275–284PubMedCrossRefPubMedCentralGoogle Scholar
  29. Li Z, Li C, Pan L, Yu M, Yang K, Pang Y (2005b) Characterization of p24 gene of Spodoptera litura multicapsid nucleopolyhedrovirus. Virus Genes 30:349–356PubMedCrossRefPubMedCentralGoogle Scholar
  30. Li L, Li Z, Chen W, Pang Y (2007) Cloning, expression of Autographa californica nucleopolyhedrovirus vp39 gene in Escherichia coli and preparation of its antibody. Biotechnology 17:3–5Google Scholar
  31. Marek M, Merten OW, Galibert L, Vlak JM, van Oers MM (2011) Baculovirus VP80 protein and the F-actin cytoskeleton interact and connect the viral replication factory with the nuclear periphery. J Virol 85:5350–5362PubMedPubMedCentralCrossRefGoogle Scholar
  32. McCarthy CB, Dai X, Donly C, Theilmann DA (2008) Autographa californica multiple nucleopolyhedrovirus ac142, a core gene that is essential for BV production and ODV envelopment. Virology 372:325–339PubMedCrossRefPubMedCentralGoogle Scholar
  33. O’Reilly DR, Miller LK, Luckow VA (1992) Baculovirus expression vectors: a laboratory manual. Oxford University Press, New YorkGoogle Scholar
  34. Rohrmann GF (2013) Baculovirus molecular biology, 3rd edn. National Center for Biotechnology Information, BethesdaGoogle Scholar
  35. Russell RL, Rohrmann GF (1990) The p6.5 gene region of a nuclear polyhedrosis virus of Orgyia pseudotsugata: DNA sequence and transcriptional analysis of four late genes. J Gen Virol 71:551–560PubMedCrossRefPubMedCentralGoogle Scholar
  36. Shi A, Hu Z, Zuo Y, Wang Y, Wu W, Yuan M, Yang K (2018) Autographa californica multiple nucleopolyhedrovirus ac75 is required for the nuclear egress of nucleocapsids and intranuclear microvesicle formation. J Virol 92:e01509–e01517PubMedPubMedCentralCrossRefGoogle Scholar
  37. Slack J, Arif BM (2007) The baculoviruses occlusion-derived virus: virion structure and function. Adv Virus Res 69:99–165PubMedCrossRefPubMedCentralGoogle Scholar
  38. Tao XY, Choi JY, Kim WJ, An SB, Liu Q, Kim SE, Lee SH, Kim JH, Woo SD, Jin BR, Je YH (2015) Autographa californica multiple nucleopolyhedrovirus ORF11 is essential for budded-virus production and occlusion-derived-virus envelopment. J Virol 89:373–383PubMedCrossRefGoogle Scholar
  39. van Oers MM, Vlak JM (2007) Baculovirus genomics. Curr Drug Targets 8:1051–1068PubMedCrossRefPubMedCentralGoogle Scholar
  40. Vaughn JL, Goodwin RH, Tompkins GJ, McCawley P (1977) The establishment of two cell lines from the insect spodoptera frugiperda (lepidoptera; noctuidae). Vitro Cell Dev-Pl 13:213–217CrossRefGoogle Scholar
  41. Wang R, Deng F, Hou D, Zhao Y, Guo L, Wang H, Hu Z (2010) Proteomics of the Autographa californica nucleopolyhedrovirus budded virions. J Virol 84:7233–7242PubMedPubMedCentralCrossRefGoogle Scholar
  42. Wei D, Wang Y, Zhang X, Hu Z, Yuan M, Yang K (2014) Autographa californica nucleopolyhedrovirus Ac76: a dimeric type II integral membrane protein that contains an inner nuclear membrane-sorting motif. J Virol 88:1090–1103PubMedPubMedCentralCrossRefGoogle Scholar
  43. Williams GV, Faulkner P (1997) Cytological changes and viral morphogenesis during baculovirus infection. In: Miller LK (ed) The baculoviruses. Springer, Boston, pp 61–107CrossRefGoogle Scholar
  44. Wolgamot GM, Gross CH, Russell RL, Rohrmann GF (1993) Immunocytochemical characterization of p24, a baculovirus capsid-associated protein. J Gen Virol 74:103–107PubMedCrossRefPubMedCentralGoogle Scholar
  45. Wu W, Passarelli AL (2010) Autographa californica multiple nucleopolyhedrovirus Ac92 (ORF92, P33) is required for budded virus production and multiply enveloped occlusion-derived virus formation. J Virol 84:12351–12361PubMedPubMedCentralCrossRefGoogle Scholar
  46. Wu W, Lin T, Pan L, Yu M, Li Z, Pang Y, Yang K (2006) Autographa californica multiple nucleopolyhedrovirus nucleocapsid assembly is interrupted upon deletion of the 38 K gene. J Virol 80:11475–11485PubMedPubMedCentralCrossRefGoogle Scholar
  47. Wu W, Liang H, Kan J, Liu C, Yuan M, Liang C, Yang K, Pang Y (2008) Autographa californica multiple nucleopolyhedrovirus 38 K is a novel nucleocapsid protein that interacts with VP1054, VP39, VP80, and itself. J Virol 82:12356–12364PubMedPubMedCentralCrossRefGoogle Scholar
  48. Yang M, Wang S, Yue XL, Li LL (2014) Autographa californica multiple nucleopolyhedrovirus orf132 encodes a nucleocapsid-associated protein required for budded-virus and multiply enveloped occlusion-derived virus production. J Virol 88:12586–12598PubMedPubMedCentralCrossRefGoogle Scholar
  49. Yuan M, Wu W, Liu C, Wang Y, Hu Z, Yang K, Pang Y (2008) A highly conserved baculovirus gene p48 (ac103) is essential for BV production and ODV envelopment. Virology 379:87–96PubMedCrossRefPubMedCentralGoogle Scholar
  50. Yuan M, Huang Z, Hu Z, Yang K, Pang Y (2010) Expression of baculovirus AcMNPV p48 gene in insect cell. Wei Sheng Wu Xue Bao 50:465–471PubMedPubMedCentralGoogle Scholar
  51. Yuan M, Huang Z, Wei D, Hu Z, Yang K, Pang Y (2011) Identification of Autographa californica nucleopolyhedrovirus ac93 as a core gene and its requirement for intranuclear microvesicle formation and nuclear egress of nucleocapsids. J Virol 85:11664–11674PubMedPubMedCentralCrossRefGoogle Scholar
  52. Yue Q, Yu Q, Yang Q, Xu Y, Guo Y, Blissard GW, Li Z (2018) Distinct roles of cellular ESCRT-I and ESCRT-III proteins in efficient entry and egress of budded virions of Autographa californica multiple nucleopolyhedrovirus. J Virol 92:e01636-17PubMedCrossRefPubMedCentralGoogle Scholar
  53. Zhou Y, Yao Q, Shen H, Xia H, Lin F, Chen K (2010) Characterization of Bombyx mori nucleopolyhedrovirus ORF109 that encodes a 25-kDa structural protein of the occlusion-derived virion. Curr Microbiol 61:451–457PubMedCrossRefPubMedCentralGoogle Scholar

Copyright information

© Wuhan Institute of Virology, CAS 2019

Authors and Affiliations

  1. 1.State Key Laboratory of BiocontrolSun Yat-sen UniversityGuangzhouChina

Personalised recommendations