Deletion of a nucleopolyhedrovirus gene encoding a virion structural protein (ORF107) increases the budded virion titre and reduces infectivity Free

Abstract

The open reading frame of single nucleocapsid nucleopolyhedrovirus (HearNPV) encodes a putative protein of 51 kDa with homologues in a few group II NPVs and a granulovirus. was transcribed as polyadenylated transcripts in infected HzAM1 insect cells. The transcripts were initiated at two distinct locations, one upstream of (superoxide dismutase gene, ) and the second upstream of . By Western blot analysis, two forms of the HA107 protein were detected in infected cells, a major polypeptide of 48 kDa and a minor one of 51 kDa. Western blot and immunoelectron microscopy analyses further showed that the HA107 protein was associated with the nucleocapsids of both budded virions (BVs) and occlusion-derived virions. A knockout virus expressing enhanced green fluorescent protein and polyhedrin was constructed using bacmid technology. A one-step virus growth curve indicated that the BV titre of the knockout virus was significantly higher than that of the parental virus and a repair virus. Bioassays indicated that the knockout virus was able to infect third-instar larvae; however, its median lethal dose (LD) was significantly higher than those of the parental virus and repair virus. These data indicate that encodes a non-essential structural protein of HearNPV virions and that deletion of this gene increases the BV titre and LD of the occluded virus.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.83363-0
2007-12-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/88/12/3307.html?itemId=/content/journal/jgv/10.1099/vir.0.83363-0&mimeType=html&fmt=ahah

References

  1. Appel R. D., Baroch A., Hochstrasser D. F. 1994; A new generation of information retrieval tools for biologists: the example of the ExPASy WWW server. Trends Biochem Sci 19:258–260 [CrossRef]
    [Google Scholar]
  2. Bulach D. M., Kumar C. A., Zaia A., Liang B., Tribe D. E. 1999; Group II nucleopolyhedrovirus subgroups revealed by phylogenetic analysis of polyhedrin and DNA polymerase gene sequences. J Invertebr Pathol 73:59–73 [CrossRef]
    [Google Scholar]
  3. 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
    [Google Scholar]
  4. Chen X., Zhang W.-J., Wong J., Chun G., Lu A., McCutchen B. F., Presnail J. K., Herrmann R., Dolan M. other authors 2002; Comparative analysis of the complete genome sequences of Helicoverpa zea and Helicoverpa armigera single-nucleocapsid nucleopolyhedroviruses. J Gen Virol 83:673–684
    [Google Scholar]
  5. Datsenko K. A., Wendell B. 2000; One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci U S A 97:6640–6645 [CrossRef]
    [Google Scholar]
  6. Friesen P. D., Miller L. K. 1985; Temporal regulation of baculovirus RNA: overlapping early and late transcripts. J Virol 54:392–400
    [Google Scholar]
  7. Funk C. J., Braunagel S. C., Rohrmann G. F. 1997; Baculovirus structure. In The Baculoviruses pp 7–32 Edited by Miller L. K. New York: Plenum Press;
    [Google Scholar]
  8. Guan K. L., Dixon J. E. 1991; Eukaryotic proteins expressed in Escherichia coli : an improved thrombin cleavage and purification procedure of fusion proteins with glutathione S -transferase. Anal Biochem 192:262–267 [CrossRef]
    [Google Scholar]
  9. 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]
  10. Horton P., Nakai K. 1997; Better prediction of protein cellular localization sites with the k nearest neighbors classifier. Proc Int Conf Intell Syst Mol Biol 5:147–152
    [Google Scholar]
  11. Hou S., Chen X., Wang H., Tao M., Hu Z. 2002; Efficient method to generate homologous recombinant baculovirus genomes in E. coli . Biotechniques32783–788
    [Google Scholar]
  12. IJkel W. F., van Strien E. A., Heldens J. G., Broer R., Zuidema D., Goldbach R. W., Vlak J. M. 1999; Sequence and organization of the Spodoptera exigua multicapsid nucleopolyhedrovirus genome. J Gen Virol 80:3289–3304
    [Google Scholar]
  13. Jakubowska A. K., Peters S. A., Ziemnicka J., Vlak J. M., van Oers M. M. 2006; Genome sequence of an enhancin gene-rich nucleopolyhedrovirus (NPV) from Agrotis segetum : collinearity with Spodoptera exigua multiple NPV. J Gen Virol 87:537–551 [CrossRef]
    [Google Scholar]
  14. Jensen L. J., Gupta R., Staerfeldt H. H., Brunak S. 2003; Prediction of human protein function according to gene ontology categories. Bioinformatics 19:635–642 [CrossRef]
    [Google Scholar]
  15. Long G., Chen X., Peters D., Vlak J. M., Hu Z. 2003; Open reading frame 122 of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus encodes a novel structural protein of occlusion-derived virions. J Gen Virol 84:115–121 [CrossRef]
    [Google Scholar]
  16. McIntosh A. H., Christian P. D., Grasela J. J. 1999; The establishment of heliothine cell lines and their susceptibility to two baculovirus. In Vitro Cell Dev Biol 35:94–97
    [Google Scholar]
  17. Pang Y., Yu J., Wang L., Hu X., Bao W., Li G., Chen C., Han H., Hu S., Yang H. 2001; Sequence analysis of the Spodoptera litura multicapsid nucleopolyhedrovirus genome. Virology 287:391–404 [CrossRef]
    [Google Scholar]
  18. Shizuya H., Birren B., Kim U. J., Mancino V., Slepak T., Tachiiri Y., Simon M. 1992; Cloning and stable maintenance of 300-kilobase-pair fragments of human DNA in Escherichia coli using an F-factor-based vector. Proc Natl Acad Sci U S A 89:8794–8797 [CrossRef]
    [Google Scholar]
  19. Snedecor G. W., Cochran W. G. 1989 Statistical Methods , 8th edn. Ames: Iowa State University Press;
    [Google Scholar]
  20. 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]
  21. Theilmann D. A., Blissard G. W., Bonning B., Jehle J. A., O'Reilly D. R., Rohrmann G. F., Thiem S., Vlak J. M. 2004; Baculoviridae . In VIIIth Report of the International Committee on Taxonomy of Viruses pp 177–185 Edited by Fauquet C. M., Mayo M. A., Maniloff J., Desselberger U., Ball L. A. San Diego: Academic Press;
    [Google Scholar]
  22. Tsai J. M., Wang H. C., Leu J. H., Wang A. H., Zhuang Y., Walker P. J., Kou G. H., Lo C. F. 2006; Identification of the nucleocapsid, tegument, and envelope proteins of the shrimp white spot syndrome virus virion. J Virol 80:3021–3029 [CrossRef]
    [Google Scholar]
  23. Volkman L. E., Summers M. D. 1977; Autographa californica nuclear polyhedrosis virus: comparative infectivity of the occluded, alkali-liberated, and nonoccluded forms. J Invertebr Pathol 30:102–103 [CrossRef]
    [Google Scholar]
  24. 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]
    [Google Scholar]
  25. Zanotto P. M., Kessing B. D., Maruniak J. E. 1993; Phylogenetic interrelationships among baculoviruses: evolutionary rates and host associations. J Invertebr Pathol 62:147–164 [CrossRef]
    [Google Scholar]
  26. Zhang G. Y. 1989; Commercial viral insecticide- Heliothis armigera viral insecticide in China. IPM Pract 11:13
    [Google Scholar]
  27. Zhang C. X., Ma X. C., Guo Z. J. 2005; Comparison of the complete genome sequence between C1 and G4 isolates of the Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus. Virology 333:190–199 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.83363-0
Loading
/content/journal/jgv/10.1099/vir.0.83363-0
Loading

Data & Media loading...

Most cited Most Cited RSS feed