1887

Abstract

A recent study found that the Autographa californica multiple nucleopolyhedrovirus (AcMNPV) Ac132 is a nucleocapsid-associated protein and required for budded virion (BV) production. We therefore initiated experiments aimed at understanding how Ac132 is involved in AcMNPV infection. An 80 bp region of ac132 was replaced with a chloramphenicol resistance gene to construct vAc132KO. Transfection of vAc132KO into Sf9 cells resulted in a single-cell infection phenotype, consistent with findings reported in a previous study. Interestingly, BVs were observable in the supernatants, and the BV production in the supernatant was comparable with that present in supernatants from a WT control. These results suggest that the ac132 deletion does not affect the egress of nucleocapsids from the transfected cells to form BVs, but the BVs are non-infectious. Transfection with DNA extracted from vAc132KO BVs could establish infection in Sf9 cells, indicating that the deletion does not affect the integrity of the viral genomic DNA in non-infectious progeny BVs. To monitor the traffic of nucleocapsids without Ac132, two mCherry proteins were fused with the major capsid protein VP39 to construct vAc132KO : 2mC. Using confocal microscopy, we observed that the nucleocapsids of vAc132KO : 2mC could not enter the nucleus and instead remained docked at the nuclear membrane. This study provides a new understanding of the nuclear entry of baculoviruses.

Keyword(s): ac132 , AcMNPV , Baculovirus , NEBU and nuclear entry
Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.000602
2016-11-10
2019-10-20
Loading full text...

Full text loading...

/deliver/fulltext/jgv/97/11/3030.html?itemId=/content/journal/jgv/10.1099/jgv.0.000602&mimeType=html&fmt=ahah

References

  1. Alfonso V., Maroniche G. A., Reca S. R., López M. G., del Vas M., Taboga O..( 2012;). AcMNPV core gene ac109 is required for budded virion transport to the nucleus and for occlusion of viral progeny. . PLoS One7:e46146. [CrossRef][PubMed]
    [Google Scholar]
  2. Au S., Panté N..( 2012;). Nuclear transport of baculovirus: revealing the nuclear pore complex passage. . J Struct Biol177:90–98. [CrossRef][PubMed]
    [Google Scholar]
  3. Ayres M. D., Howard S. C., Kuzio J., Lopez-Ferber M., Possee R. D..( 1994;). The complete DNA sequence of Autographa californica nuclear polyhedrosis virus. . Virology202:586–605. [CrossRef][PubMed]
    [Google Scholar]
  4. Braunagel S. C., Russell W. K., Rosas-Acosta G., Russell D. H., Summers M. D..( 2003;). Determination of the protein composition of the occlusion-derived virus of Autographa californica nucleopolyhedrovirus. . Proc Natl Acad Sci U S A100:9797–9802. [CrossRef][PubMed]
    [Google Scholar]
  5. 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 Virol86:10432–10443. [CrossRef][PubMed]
    [Google Scholar]
  6. Chiu J., March P. E., Lee R., Tillett D..( 2004;). Site-directed, Ligase-Independent Mutagenesis (SLIM): a single-tube methodology approaching 100 % efficiency in 4 h. . Nucleic Acids Res32:e174. [CrossRef][PubMed]
    [Google Scholar]
  7. Cox M. M..( 2012;). Recombinant protein vaccines produced in insect cells. . Vaccine30:1759–1766. [CrossRef][PubMed]
    [Google Scholar]
  8. Fang M., Nie Y., Theilmann D. A..( 2009;). Deletion of the AcMNPV core gene ac109 results in budded virions that are non-infectious. . Virology389:66–74. [CrossRef][PubMed]
    [Google Scholar]
  9. Hou D., Zhang L., Deng F., Fang W., Wang R., Liu X., Guo L., Rayner S., Chen X. et al.( 2013;). Comparative proteomics reveal fundamental structural and functional differences between the two progeny phenotypes of a baculovirus. . J Virol87:829–839. [CrossRef][PubMed]
    [Google Scholar]
  10. Jehle J. A., Blissard G. W., Bonning B. C., Cory J. S., Herniou E. A., Rohrmann G. F., Theilmann D. A., Thiem S. M., Vlak J. M..( 2006;). On the classification and nomenclature of baculoviruses: a proposal for revision. . Arch Virol151:1257–1266. [CrossRef][PubMed]
    [Google Scholar]
  11. Lanier L. M., Volkman L. E..( 1998;). Actin binding and nucleation by Autographa california M nucleopolyhedrovirus. . Virology243:167–177. [CrossRef][PubMed]
    [Google Scholar]
  12. Larkin M. A., Blackshields G., Brown N. P., Chenna R., McGettigan P. A., McWilliam H., Valentin F., Wallace I. M., Wilm A. et al.( 2007;). clustal w and clustal x version 2.0. . Bioinformatics23:2947–2948. [CrossRef][PubMed]
    [Google Scholar]
  13. Lehiy C. J., Wu W., Berretta M. F., Passarelli A. L..( 2013;). Autographa californica M nucleopolyhedrovirus open reading frame 109 affects infectious budded virus production and nucleocapsid envelopment in the nucleus of cells. . Virology435:442–452. [CrossRef][PubMed]
    [Google Scholar]
  14. Letunic I., Doerks T., Bork P..( 2009;). smart 6: recent updates and new developments. . Nucleic Acids Res37:D229–232. [CrossRef][PubMed]
    [Google Scholar]
  15. Li L., Li Z., Chen W., Pang Y..( 2007;). Cloning, expression of Autographa californica nucleopolyhedro virus vp39 gene in Escherichia coli and preparation of its antibody. . Biotechnology17:3–5.
    [Google Scholar]
  16. Luckow V. A., Lee S. C., Barry G. F., Olins P. O..( 1993;). Efficient generation of infectious recombinant baculoviruses by site-specific transposon-mediated insertion of foreign genes into a baculovirus genome propagated in Escherichia coli. . J Virol67:4566–4579.[PubMed]
    [Google Scholar]
  17. Maghodia A. B., Jarvis D. L., Geisler C..( 2014;). Complete genome sequence of the Autographa californica multiple nucleopolyhedrovirus strain E2. . Genome Announc2:e0120201214. [CrossRef][PubMed]
    [Google Scholar]
  18. Marek M., Merten O. W., Galibert L., Vlak J. M., van Oers M. M..( 2011;). Baculovirus VP80 protein and the F-actin cytoskeleton interact and connect the viral replication factory with the nuclear periphery. . J Virol85:5350–5362. [CrossRef][PubMed]
    [Google Scholar]
  19. Nicholas K. B., Nicholas H. B., Deerfield D. W..( 1997;). Genedoc: analysis and visualization of genetic variation. . EMBNEW News4:14.
    [Google Scholar]
  20. O'Reilly D. R., Miller L., Luckow V. A..( 1992;). Baculovirus Expression Vectors: A Laboratory Manual. New York:: W.H. Freeman;.
    [Google Scholar]
  21. Ohkawa T., Volkman L. E., Welch M. D..( 2010;). Actin-based motility drives baculovirus transit to the nucleus and cell surface. . J Cell Biol190:187–195. [CrossRef][PubMed]
    [Google Scholar]
  22. Ono C., Kamagata T., Taka H., Sahara K., Asano S., Bando H..( 2012;). Phenotypic grouping of 141 BmNPVs lacking viral gene sequences. . Virus Res165:197–206. [CrossRef][PubMed]
    [Google Scholar]
  23. Pfuhl M., Winder S. J., Castiglione Morelli M. A., Labeit S., Pastore A..( 1996;). Correlation between conformational and binding properties of nebulin repeats. . J Mol Biol257:367–384. [CrossRef][PubMed]
    [Google Scholar]
  24. Rivera-Gonzalez G. C., Swift S. L., Dussupt V., Georgopoulos L. J., Maitland N. J..( 2011;). Baculoviruses as gene therapy vectors for human prostate cancer. . J Invertebr Pathol107:S59–70. [CrossRef][PubMed]
    [Google Scholar]
  25. Rohrmann G. F..( 2013;). Baculovirus Molecular Biology. Bethesda, MD:: US National Library of Medicine, National Center for Biotechnology Information;.
    [Google Scholar]
  26. Summers M. D..( 1971;). Electron microscopic observations on granulosis virus entry, uncoating and replication processes during infection of the midgut cells of Trichoplusia ni. . J Ultrastruct Res35:606–625. [CrossRef][PubMed]
    [Google Scholar]
  27. Vaughn J. L., Goodwin R. H., Tompkins G. J., McCawley P..( 1977;). The establishment of two cell lines from the insect Spodoptera frugiperda (Lepidoptera; Noctuidae). . In Vitro13:213–217. [CrossRef][PubMed]
    [Google Scholar]
  28. Wang R., Deng F., Hou D., Zhao Y., Guo L., Wang H., Hu Z..( 2010;). Proteomics of the Autographa californica nucleopolyhedrovirus budded virions. . J Virol84:7233–7242. [CrossRef][PubMed]
    [Google Scholar]
  29. Wang Y., Choi J. Y., Roh J. Y., Liu Q., Tao X. Y., Park J. B., Kim J. S., Je Y. H..( 2011;). Genomic sequence analysis of granulovirus isolated from the tobacco cutworm, Spodoptera litura. . PLoS One6:e28163. [CrossRef][PubMed]
    [Google Scholar]
  30. Wei D., Wang Y., Zhang X., Hu Z., Yuan M., Yang K..( 2013;). Autographa californica nucleopolyhedrovirus Ac76: a dimeric type II integral membrane protein that contains an inner nuclear membrane-sorting motif. . J Virol88:1090–1103. [CrossRef][PubMed]
    [Google Scholar]
  31. 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 38K gene. . J Virol80:11475–11485. [CrossRef][PubMed]
    [Google Scholar]
  32. Wu W., Liang H., Kan J., Liu C., Yuan M., Liang C., Yang K., Pang Y..( 2008;). Autographa californica multiple nucleopolyhedrovirus 38K is a novel nucleocapsid protein that interacts with VP1054, VP39, VP80, and itself. . J Virol82:12356–12364. [CrossRef][PubMed]
    [Google Scholar]
  33. Yang M., Wang S., Yue X. L., Li L. L..( 2014;). Autographa californica multiple nucleopolyhedrovirus orf132 encodes a nucleocapsid-associated protein required for budded-virus and multiply enveloped occlusion-derived virus production. . J Virol88:12586–12598. [CrossRef][PubMed]
    [Google Scholar]
  34. 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 Virol85:11664–11674. [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.000602
Loading
/content/journal/jgv/10.1099/jgv.0.000602
Loading

Data & Media loading...

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error