1887

Abstract

Open reading frame (ORF) 67 () of the nucleopolyhedrovirus (BmNPV) is a highly conserved gene that is found in all completely sequenced baculoviruses; its function is unknown. In the present study, a -knockout virus was generated for studying the role of Bm67 in the BmNPV infection cycle. Furthermore, a -repair bacmid was constructed by transposing the native promoter-promoted ORF into the locus of the -knockout bacmid. After these recombinant bacmids were transfected into BmN cells, the -knockout bacmid caused defects in the production of infectious budded viruses. However, the -repair bacmid could rescue the defect, and budded virus titres reached wild-type levels. Quantitative real-time PCR analysis indicated that Bm67 is required for normal levels of DNA synthesis or for the stability of nascent viral DNA at the early stage. Electron microscopic analysis revealed that the formation of normal-appearing nucleocapsids is reduced in -knockout bacmid-transfected cells, and nucleocapsids are rarely found in the cytoplasm. The presence of ‘enveloped’ nucleocapsids at the nucleoplasm bilayer indicated that they are enveloped abnormally. These results indicated that Bm67 is required for the production of infectious budded viruses and for assembly of envelope and nucleocapsids.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.83398-0
2008-03-01
2019-10-17
Loading full text...

Full text loading...

/deliver/fulltext/jgv/89/3/766.html?itemId=/content/journal/jgv/10.1099/vir.0.83398-0&mimeType=html&fmt=ahah

References

  1. Braunagel, S. C. & Summers, M. D. ( 1994; ). Autographa californica nuclear polyhedrosis virus, PDV, and ECV viral envelopes and nucleocapsids: structural proteins, antigens, lipid and fatty acid profiles. Virology 202, 315–328.[CrossRef]
    [Google Scholar]
  2. 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 A 100, 9797–9802.[CrossRef]
    [Google Scholar]
  3. Clem, R. J. & Miller, L. K. ( 1993; ). Apoptosis reduces both the in vitro replication and the in vivo infectivity of a baculovirus. J Virol 67, 3730–3738.
    [Google Scholar]
  4. Croizier, G., Croizier, L., Argaud, O. & Poudevigne, D. ( 1994; ). Extension of Autographa californica nuclear polyhedrosis virus host range by interspecific replacement of a short DNA sequence in the p143 helicase gene. Proc Natl Acad Sci U S A 91, 48–52.[CrossRef]
    [Google Scholar]
  5. Federici, B. A. ( 1997; ). Baculovirus pathogenesis. In The Baculoviruses, pp. 33–60. Edited by L. K. Miller. New York: Plenum Press.
  6. Gomi, S., Zhou, C. E., Yih, W., Majima, K. & Maeda, S. ( 1997; ). Deletion analysis of four of eighteen late gene expression factor gene homologues of the baculovirus, BmNPV. Virology 230, 35–47.[CrossRef]
    [Google Scholar]
  7. Gomi, S., Majima, K. & Maeda, S. ( 1999; ). Sequence analysis of the genome of Bombyx mori nucleopolyhedrovirus. J Gen Virol 80, 1323–1337.
    [Google Scholar]
  8. Hefferon, K. L. ( 2003; ). Characterization of HCF-1, a determinant of Autographa californica multiple nucleopolyhedrovirus host specificity. Insect Mol Biol 12, 651–658.[CrossRef]
    [Google Scholar]
  9. Hefferon, K. L. & Miller, L. K. ( 2002; ). Reconstructing the replication complex of AcMNPV. Eur J Biochem 269, 6233–6240.[CrossRef]
    [Google Scholar]
  10. Herniou, E. A., Olszewski, J. A., Cory, J. S. & O'Reilly, D. R. ( 2003; ). The genome sequence and evolution of baculoviruses. Annu Rev Entomol 48, 211–234.[CrossRef]
    [Google Scholar]
  11. Ikeda, M., Reimbold, E. A. & Thiem, S. M. ( 2005; ). Functional analysis of the baculovirus host range gene, hrf-1. Virology 332, 602–613.[CrossRef]
    [Google Scholar]
  12. Iwanaga, M., Kurihara, M., Kobayashi, M. & Kang, W. ( 2002; ). Characterization of Bombyx mori nucleopolyhedrovirus orf68 gene that encodes a novel structural protein of budded virus. Virology 297, 39–47.[CrossRef]
    [Google Scholar]
  13. Iwanaga, M., Takaya, K., Katsuma, S., Ote, M., Tanaka, S., Kamita, S. G., Kang, W., Shimada, T. & Kobayashi, M. ( 2004; ). Expression profiling of baculovirus genes in permissive and nonpermissive cell lines. Biochem Biophys Res Commun 323, 599–614.[CrossRef]
    [Google Scholar]
  14. 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 Virol 151, 1257–1266.[CrossRef]
    [Google Scholar]
  15. Knudson, D. L. & Harrap, K. A. ( 1975; ). Replication of nuclear polyhedrosis virus in a continuous cell culture of Spodoptera frugiperda: microscopy study of the sequence of events of the virus infection. J Virol 17, 254–268.
    [Google Scholar]
  16. Lin, G. & Blissard, G. W. ( 2002a; ). Analysis of an Autographa californica multicapsid nucleopolyhedrovirus lef-6-null virus: LEF-6 is not essential for viral replication but appears to accelerate late gene transcription. J Virol 76, 5503–5514.[CrossRef]
    [Google Scholar]
  17. Lin, G. & Blissard, G. W. ( 2002b; ). Analysis of an Autographa californica nucleopolyhedrovirus lef-11 knockout: LEF-11 is essential for viral DNA replication. J Virol 76, 2770–2779.[CrossRef]
    [Google Scholar]
  18. Lu, A. & Miller, L. K. ( 1996; ). Species-specific effects of the hcf-1 gene on baculovirus virulence. J Virol 70, 5123–5130.
    [Google Scholar]
  19. 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 Virol 67, 4566–4579.
    [Google Scholar]
  20. Lung, O. Y., Cruz-Alvarez, M. & Blissard, G. W. ( 2003; ). Ac23, an envelope fusion protein homolog in the baculovirus Autographa californica multicapsid nucleopolyhedrovirus, is a viral pathogenicity factor. J Virol 77, 328–339.[CrossRef]
    [Google Scholar]
  21. Mainz, D., Quadt, I. & Knebel-Morsdorf, D. ( 2002; ). Nuclear IE2 structures are related to viral DNA replication sites during baculovirus infection. J Virol 76, 5198–5207.[CrossRef]
    [Google Scholar]
  22. Mikhailov, V. S. ( 2003; ). Replication of the baculovirus genome. Mol Biol (Mosk) 37, 288–299.
    [Google Scholar]
  23. Monsma, S. A., Oomens, A. G. & Blissard, G. W. ( 1996; ). The GP64 envelope fusion protein is an essential baculovirus protein required for cell-to-cell transmission of infection. J Virol 70, 4607–4616.
    [Google Scholar]
  24. Morris, T. D. & Miller, L. K. ( 1993; ). Characterization of productive and non-productive AcMNPV infection in selected insect cell lines. Virology 197, 339–348.[CrossRef]
    [Google Scholar]
  25. O'Reilly, D. R., Miller, L. K. & Luckow, V. A. ( 1994; ). Baculovirus Expression Vectors: A Laboratory Manual. New York: Oxford University Press.
  26. Okano, K., Mikhailov, V. S. & Maeda, S. ( 1999; ). Colocalization of baculovirus IE-1 and two DNA-binding proteins, DBP and LEF-3, to viral replication factories. J Virol 73, 110–119.
    [Google Scholar]
  27. Okano, K., Vanarsdall, A. L., Mikhailov, V. S. & Rohrmann, G. F. ( 2006; ). Conserved molecular systems of the Baculoviridae. Virology 344, 77–87.[CrossRef]
    [Google Scholar]
  28. Okano, K., Vanarsdall, A. L. & Rohrmann, G. F. ( 2007; ). A baculovirus alkaline nuclease knockout construct produces fragmented DNA and aberrant capsids. Virology 359, 46–54.[CrossRef]
    [Google Scholar]
  29. Olszewski, J. & Miller, L. K. ( 1997a; ). Identification and characterization of a baculovirus structural protein, VP1054, required for nucleocapsid formation. J Virol 71, 5040–5050.
    [Google Scholar]
  30. Olszewski, J. & Miller, L. K. ( 1997b; ). A role for baculovirus GP41 in budded virus production. Virology 233, 292–301.[CrossRef]
    [Google Scholar]
  31. Oomens, A. G. & Blissard, G. W. ( 1999; ). Requirement for GP64 to drive efficient budding of Autographa californica multicapsid nucleopolyhedrovirus. Virology 254, 297–314.[CrossRef]
    [Google Scholar]
  32. Pan, L., Li, Z., Gong, Y., Yu, M., Yang, K. & Pang, Y. ( 2005; ). Characterization of gp41 gene of Spodoptera litura multicapsid nucleopolyhedrovirus. Virus Res 110, 73–79.[CrossRef]
    [Google Scholar]
  33. Perera, O., Green, T. B., Stevens, S. M., Jr, White, S. & Becnel, J. J. ( 2007; ). Proteins associated with Culex nigripalpus nucleopolyhedrovirus occluded virions. J Virol 81, 4585–4590.[CrossRef]
    [Google Scholar]
  34. Vanarsdall, A. L., Okano, K. & Rohrmann, G. F. ( 2004; ). Characterization of a baculovirus with a deletion of vlf-1. Virology 326, 191–201.[CrossRef]
    [Google Scholar]
  35. Vanarsdall, A. L., Okano, K. & Rohrmann, G. F. ( 2005; ). Characterization of the replication of a baculovirus mutant lacking the DNA polymerase gene. Virology 331, 175–180.[CrossRef]
    [Google Scholar]
  36. Vanarsdall, A. L., Okano, K. & Rohrmann, G. F. ( 2006; ). Characterization of the role of very late expression factor 1 in baculovirus capsid structure and DNA processing. J Virol 80, 1724–1733.[CrossRef]
    [Google Scholar]
  37. Whitford, M. & Faulkner, P. ( 1992a; ). Nucleotide sequence and transcriptional analysis of a gene encoding gp41, a structural glycoprotein of the baculovirus Autographa californica nuclear polyhedrosis virus. J Virol 66, 4763–4768.
    [Google Scholar]
  38. Whitford, M. & Faulkner, P. ( 1992b; ). A structural polypeptide of the baculovirus Autographa californica nuclear polyhedrosis virus contains O-linked N-acetylglucosamine. J Virol 66, 3324–3329.
    [Google Scholar]
  39. Wu, W. W., Wang, J. W., Xie, F., Long, Q. X. & Wang, X. Z. ( 2003; ). Baculovirus p74 gene is a species-specific gene. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 35, 834–840. (in Chinese)
    [Google Scholar]
  40. 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 Virol 80, 11475–11485.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.83398-0
Loading
/content/journal/jgv/10.1099/vir.0.83398-0
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