Baculovirus DNA helicases are essential for replication and are determinants of host range. Helicases of multicapsid nucleopolyhedrovirus (AcMNPV) and granulovirus (TnGV) differ markedly, although both viruses replicate efficiently in the cabbage looper, .. To determine whether the TnGV helicase (P137) could support replication of AcMNPV in . cells or larvae, the native AcMNPV helicase gene () was disrupted and substituted with . P137 did not support replication when synthesized by the P143-deficient AcMNPV. Moreover, P137 did not inhibit AcMNPV replication when co-synthesized in the presence of the AcMNPV P143. These results suggest that although TnGV and AcMNPV replicate efficiently in ., specific protein–protein or protein–DNA interactions between baculoviral helicases and viral-specific factors which form the replicase complex are required for virus replication. A novel and rapid method for disrupting AcMNPV genes in . using the commercial Bac-to-Bac AcMNPV baculovirus expression vector is described.


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  1. Ahrens, C. H. & Rohrmann, G. F. (1996). The DNA polymerase and helicase genes of a baculovirus of Orgyia pseudotsugata. Journal of General Virology 77, 825-837.[CrossRef] [Google Scholar]
  2. Argaud, O., Croizier, l., Ferber-Lopez, M. & Croizier, G. (1998). Two key mutations in the host-range specificity domain of the p143 gene of Autographa californica nucleopolyhedrovirus are required to kill Bombyxmori larvae. Journal of General Virology 79, 931-935. [Google Scholar]
  3. Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A. & Struhl, K. (1994).Current Protocols in Molecular Biology, vol. 1. New York: John Wiley and Sons.
  4. 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. Virology 202, 586-605.[CrossRef] [Google Scholar]
  5. Bideshi, D. K. & Federici, B. A. (2000). DNA-independent ATPase activity of the Trichoplusia ni granulovirus DNA helicase. Journal of General Virology 81, 1601-1604. [Google Scholar]
  6. Bideshi, D. K., Hice, R. H., Ge, B. & Federici, B. A. (1998). Molecular characterization and expression of the Trichoplusia ni granulovirus helicase gene. Journal of General Virology 79, 1309-1319. [Google Scholar]
  7. Bruckner, R. C., Crute, J. J., Dodson, M. S. & Lehman, I. R. (1991). The herpes simplex virus 1 origin binding protein, a DNA helicase. Journal of Biological Chemistry 266, 2669-2674. [Google Scholar]
  8. Chartier, C., Degryse, E., Gantzer, M., Dieterle, A., Pavirani, A. & Mehtali, M. (1996). Efficient generation of recombinant adenovirus vectors by homologous recombination in Escherichia coli. Journal of Virology 70, 4805-4810. [Google Scholar]
  9. 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. Proceedings of the National Academy Sciences, USA 91, 48-52.[CrossRef] [Google Scholar]
  10. Dodson, M. S. & Lehman, I. R. (1991). Association of DNA helicase and primase activities with a subassembly of the herpes simplex virus 1 helicase–primase composed of UL5 and UL52 gene products. Proceedings of the National Academy of Sciences, USA 88, 1105-1109.[CrossRef] [Google Scholar]
  11. Dodson, M. S., Crute, J. J., Bruckner, R. C. & Lehman, I. R. (1989). Overexpression and assembly of the herpes simplex virus type 1 helicase–primase in insect cells. Journal of Biological Chemistry 264, 20835-20838. [Google Scholar]
  12. Federici, B. A. (1993). Viral pathobiology in relation to insect control. In Parasites and Pathogens of Insects, pp. 81-101. Edited by N. Beckage, S. N. Thompson & B. A. Federici. San Diego: Academic Press.
  13. Gelernter, W. D. & Federici, B. A. (1986). Continuous cell line from Spodoptera exigua (Lepidoptera: Noctuidae) that supports replication of nuclear polyhedrosis viruses from Spodoptera exigua and Autographacalifornica. Journal of Invertebrate Pathology 48, 199-207.[CrossRef] [Google Scholar]
  14. Hanahan, D. (1983). Studies on the transformation of Escherichia coli with plasmids. Journal of Molecular Biology 166, 557-580.[CrossRef] [Google Scholar]
  15. Heldens, J. G. M., Liu, Y., Zuidema, D., Goldbach, R. W. & Vlak, J. (1997). Characterization of a putative Spodoptera exigua multicapsid nucleopolyhedrovirus helicase gene. Journal of General Virology 78, 3101-3114. [Google Scholar]
  16. Kamita, G. S. & Maeda, S. (1993). Inhibition of Bombyx mori nuclear polyhedrosis virus (NPV) replication by the putative DNA helicase of Autographa californica NPV. Journal of Virology 67, 6239-6245. [Google Scholar]
  17. Kamita, S. G. & Maeda, S. (1997). Sequencing of the putative DNA helicase-encoding gene of the Bombyx mori nuclear polyhedrosis virus and fine-mapping of a region involved in host range expansion. Gene 190, 173-179.[CrossRef] [Google Scholar]
  18. Kool, M., Voncken, J. W., van Lier, F. L. J., Tramper, J. & Vlak, J. M. (1993). Identification of seven putative origins of Autographa californica multiple nucleocapsid nuclear polyhedrosis virus DNA replication. Journal of General Virology 74, 2661-2668.[CrossRef] [Google Scholar]
  19. Kool, M., Ahrens, C., Goldbach, R. W., Rohrmann, G. F. & Vlak, J. M. (1994). Identification of genes involved in DNA replication of the Autographa californica baculovirus. Proceedings of the National Academy of Sciences, USA 91, 11212-11216.[CrossRef] [Google Scholar]
  20. Kool, M., Ahrens, H. C., Vlak, J. M. & Rohrmann, G. F. (1995). Replication of baculovirus DNA. Journal of General Virology 76, 2103-2118.[CrossRef] [Google Scholar]
  21. Leisy, D. J., Rasmussen, C., Him, H.-T. & Rohrmann, G. F. (1995). The Autographa californica nuclear polyhedrosis virus homologous region 1a: identical sequences are essential for DNA replication activity and transcriptional enhancer function. Virology 208, 742-752.[CrossRef] [Google Scholar]
  22. Liptak, L. M., Uprichard, S. L. & Knipe, D. M. (1996). Functional order of assembly of herpes simplex virus DNA replication proteins into prereplicative site structures. Journal of Virology 70, 1759-1767. [Google Scholar]
  23. Lu, A. & Carstens, E. B. (1991). Nucleotide sequence of a gene essential for viral DNA replication in the baculovirus Autographa californica nuclear polyhedrosis virus. Virology 181, 336-347.[CrossRef] [Google Scholar]
  24. Lu, A. & Miller, L. K. (1995). The roles of eighteen baculovirus late expression factor genes in transcription and DNA replication. Journal of Virology 69, 975-982. [Google Scholar]
  25. Maeda, S., Kamita, S. G. & Kondo, A. (1993). Host range expansion of Autographa californica nuclear polyhedrosis virus (NPV) following recombination of a 0·6-kilobase-pair DNA fragment originating from Bombyx mori NPV. Journal of Virology 67, 6234-6238. [Google Scholar]
  26. Martignoni, M. E. & Iwai, P. J. (1986).A Catalogue of Viral Diseases of Insects, Mites and Ticks, 4th edn. USDA Forest Service PNW-195.
  27. O’Reilly, D. R., Miller, L. K. & Luckow, V. A. (1994).Baculovirus Expression Vectors: A Laboratory Manual, pp. 132–134. New York: W. H. Freeman.
  28. Pearson, M. N. & Rohrmann, G. F. (1995).Lymantria dispar nuclear polyhedrosis virus homologous regions: characterization of their ability to function as replication origins. Journal of Virology 69, 213-221. [Google Scholar]
  29. Sanger, F., Nicklen, S. & Coulson, A. R. (1977). Nucleotide sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, USA 74, 5463-5467.[CrossRef] [Google Scholar]
  30. Shorey, H. H. & Hale, R. L. (1965). Mass-rearing of the larvae of nine noctuid species on a simple artificial medium. Journal of Economic Entomology 58, 522-524.[CrossRef] [Google Scholar]
  31. Volkman, L. E., Blissard, G. W., Friesen, P., Keddie, B. A., Possee, R. & Theilmann, D. A. (1995). Family Baculoviridae. In Virus Taxonomy. Sixth Report of the International Committee on Taxonomy of Viruses, pp. 104-113. Edited by F. A. Murphy, C. M. Fauquet, D. H. L. Bishop, S. A. Ghabrial, A. W. Jarvis, G. P. Martelli, M. A. Mayo & M. D. Summers. Vienna & New York: Springer-Verlag.

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