1887

Abstract

PIF3 is one of the six conserved infectivity factors (PIFs) of baculoviruses. In this study, PIF3 of nucleopolyhedrovirus (HearNPV) was analysed by infectivity bioassays using a series of recombinant viruses harbouring various PIF3 truncation/substitution mutants. The results demonstrated that the N-terminal region (L26–Y45) and C-terminal region (T160–Q199) are essential for HearNPV oral infectivity. In the C-terminal T160–Q199 region, there are three conserved cysteines (C162, C164 and C185). Our results showed that substitutions of C162 or C164, predicted to be involved in disulfide-bond formation, led to a severe decrease in HearNPV infectivity. Mutation of C185, predicted not to be involved in disulfide-bond formation, did not affect the infectivity. The data suggest that disulfide bonds are important for PIF3 conformation and function. Immunofluorescence assays showed that none of the mutations affected the subcellular localization of PIF3 to the nuclear ring zone region of infected cells. Western blot results showed that all mutants except C162G and C185G failed to incorporate PIF3 into occlusion-derived viruses, which resulted in impaired oral infectivity of the latter. The data provide insights for future study of PIF3 function.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.035865-0
2012-02-01
2020-11-24
Loading full text...

Full text loading...

/deliver/fulltext/jgv/93/2/374.html?itemId=/content/journal/jgv/10.1099/vir.0.035865-0&mimeType=html&fmt=ahah

References

  1. Braunagel S. C., Summers M. D. 2007; Molecular biology of the baculovirus occlusion-derived virus envelope. Curr Drug Targets 8:1084–1095 [CrossRef][PubMed]
    [Google Scholar]
  2. Braunagel S. C., Williamson S. T., Saksena S., Zhong Z., Russell W. K., Russell D. H., Summers M. D. 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 U S A 101:8372–8377 [CrossRef][PubMed]
    [Google Scholar]
  3. Braunagel S. C., Cox V., Summers M. D. 2009; Baculovirus data suggest a common but multifaceted pathway for sorting proteins to the inner nuclear membrane. J Virol 83:1280–1288 [CrossRef][PubMed]
    [Google Scholar]
  4. Chen X., IJkel W. F. J., Tarchini R., Sun X., Sandbrink H., Wang H., Peters S., Zuidema D., Lankhorst R. K.other authors 2001; The sequence of the Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus genome. J Gen Virol 82:241–257[PubMed]
    [Google Scholar]
  5. Fang M. G., Nie Y. C., Wang Q., Deng F., Wang R. R., Wang H. Z., Wang H. L., Vlak J. M., Chen X. W., Hu Z. H. 2006; Open reading frame 132 of Helicoverpa armigera nucleopolyhedrovirus encodes a functional per os infectivity factor (PIF-2). J Gen Virol 87:2563–2569 [CrossRef][PubMed]
    [Google Scholar]
  6. Fang M., Nie Y., Harris S., Erlandson M. A., Theilmann D. A. 2009; Autographa californica multiple nucleopolyhedrovirus core gene ac96 encodes a per os infectivity factor (PIF-4). J Virol 83:12569–12578 [CrossRef][PubMed]
    [Google Scholar]
  7. Faulkner P., Kuzio J., Williams G. V., Wilson J. A. 1997; Analysis of p74, a PDV envelope protein of Autographa californica nucleopolyhedrovirus required for occlusion body infectivity in vivo. J Gen Virol 78:3091–3100[PubMed]
    [Google Scholar]
  8. Haas-Stapleton E. J., Washburn J. O., Volkman L. E. 2004; 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 78:6786–6791 [CrossRef][PubMed]
    [Google Scholar]
  9. Harrison R. L., Sparks W. O., Bonning B. C. 2010; Autographa californica multiple nucleopolyhedrovirus ODV-E56 envelope protein is required for oral infectivity and can be substituted functionally by Rachiplusia ou multiple nucleopolyhedrovirus ODV-E56. J Gen Virol 91:1173–1182 [CrossRef][PubMed]
    [Google Scholar]
  10. Ho S. N., Hunt H. D., Horton R. M., Pullen J. K., Pease L. R. 1989; Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77:51–59 [CrossRef][PubMed]
    [Google Scholar]
  11. Horton H. M., Burand J. P. 1993; Saturable attachment sites for polyhedron-derived baculovirus on insect cells and evidence for entry via direct membrane fusion. J Virol 67:1860–1868[PubMed]
    [Google Scholar]
  12. Hughes P. R., van Beek N. A. M., Wood H. A. 1986; A modified droplet feeding method for rapid assay of Bacillus thuringiensis and baculoviruses in noctuid larvae. J Invertebr Pathol 48:187–192 [CrossRef]
    [Google Scholar]
  13. Kikhno I., Gutiérrez S., Croizier L., Croizier G., Ferber M. L. 2002; Characterization of pif, a gene required for the per os infectivity of Spodoptera littoralis nucleopolyhedrovirus. J Gen Virol 83:3013–3022[PubMed]
    [Google Scholar]
  14. King L. A., Possee R. D. 1992 The Baculovirus Expression System: a Laboratory Guide New York, NY: Chapman & Hall; [CrossRef]
    [Google Scholar]
  15. Kuzio J., Jaques R., Faulkner P. 1989; Identification of p74, a gene essential for virulence of baculovirus occlusion bodies. Virology 173:759–763 [CrossRef][PubMed]
    [Google Scholar]
  16. Li X., Song J., Jiang T., Liang C., Chen X. 2007; The N-terminal hydrophobic sequence of Autographa californica nucleopolyhedrovirus PIF-3 is essential for oral infection. Arch Virol 152:1851–1858 [CrossRef][PubMed]
    [Google Scholar]
  17. Mclntosh A. H., Ignoffo C. M. 1983; Characterization of five cell lines established from species of Heliothis. Appl Entomol Zool (Jpn) 18:262–269
    [Google Scholar]
  18. Ohkawa T., Washburn J. O., Sitapara R., Sid E., Volkman L. E. 2005; 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 79:15258–15264 [CrossRef][PubMed]
    [Google Scholar]
  19. Peng K., van Oers M. M., Hu Z., van Lent J. W. M., Vlak J. M. 2010; Baculovirus per os infectivity factors form a complex on the surface of occlusion-derived virus. J Virol 84:9497–9504 [CrossRef][PubMed]
    [Google Scholar]
  20. Pijlman G. P., Pruijssers A. J. P., Vlak J. M. 2003; Identification of pif-2, a third conserved baculovirus gene required for per os infection of insects. J Gen Virol 84:2041–2049 [CrossRef][PubMed]
    [Google Scholar]
  21. Robertson J. L., Preisler H. K. 1992 Pesticide Bioassays with Arthropods Baton Rouge, LA: CRC Press;
    [Google Scholar]
  22. Song J., Wang R., Deng F., Wang H., Hu Z. 2008; Functional studies of per os infectivity factors of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus. J Gen Virol 89:2331–2338 [CrossRef][PubMed]
    [Google Scholar]
  23. Sparks W. O., Harrison R. L., Bonning B. C. 2011; 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 409:69–76 [CrossRef][PubMed]
    [Google Scholar]
  24. Sun X., Zhang G., Zhang Z., Hu Z. H., Vlak J. M., Arif B. M. 1998; In vivo cloning of Helicoverpa armigera single nucleocapsid nuclear polyhedrosis virus genotypes. Virol Sin 13:83–88
    [Google Scholar]
  25. Wang H., Deng F., Pijlman G. P., Chen X., Sun X., Vlak J. M., Hu Z. 2003; Cloning of biologically active genomes from a Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus isolate by using a bacterial artificial chromosome. Virus Res 97:57–63 [CrossRef][PubMed]
    [Google Scholar]
  26. Wang Y., Bininda-Emonds O. R., van Oers M. M., Vlak J. M., Jehle J. A. 2011; The genome of Oryctes rhinoceros nudivirus provides novel insight into the evolution of nuclear arthropod-specific large circular double-stranded DNA viruses. Virus Genes 42:444–456 [CrossRef][PubMed]
    [Google Scholar]
  27. Yao L., Zhou W., Xu H., Zheng Y., Qi Y. 2004; The Heliothis armigera single nucleocapsid nucleopolyhedrovirus envelope protein P74 is required for infection of the host midgut. Virus Res 104:111–121 [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.035865-0
Loading
/content/journal/jgv/10.1099/vir.0.035865-0
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

Most cited this month Most Cited RSS feed

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