1887

Abstract

Expression of and RNA and enzyme activity in wild-type (AcMNPV) was compared with that of recombinant AcMNPV viruses reprogrammed for expression of the endogenous . To establish a baseline for our recombinant AcMNPV studies, we compared, for the first time, the temporal expression profiles of both AcMNPV transcription and translation simultaneously. The rate of intracellular chitinase accumulation during AcMNPV infection followed the same pattern observed for transcription but was delayed by about 6 h. Replacement of 21 nucleotides containing the native late and promoters with a selectable cassette was sufficient to eliminate expression of both and . Viruses were generated that express from either the late or very late promoters of AcMNPV, replacing the native promoter. There was a marked difference in the temporal transcription profiles from the native, and promoters, resulting in respective specific activities of chitinase at 48 h p.i. of 62, 160 and 219 mU (mg lysate total protein). Based on temporal analysis of transcription by Northern blot, AcMNPV was transcribed from 9 h p.i. in Sf21 cells. However, expression of RNA or enzyme from a reconstructed promoter in the -reprogrammed viruses was not detected at 48 h of virus replication. Reprogramming for increased chitinase (and putatively cathepsin) expression with native baculovirus promoters might provide a means for designing environmentally benign biological insecticides.

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2007-09-01
2019-12-13
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References

  1. 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]
  2. Black, B. C., Brennan, L. A., Dierks, P. M. & Gard, I. E. ( 1997; ). Commercialization of baculovirus pesticides. In The Baculoviruses, vol. 1, pp. 341–381. Edited by L. K. Miller. New York: Plenum Press.
  3. Bowtell, D. & Sambrook, J. (editors) ( 2003; ). DNA Microarrays: a Molecular Cloning Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.
  4. Cheng, X., Krell, P. & Arif, B. ( 2001; ). P34.8 (GP37) is not essential for baculovirus replication. J Gen Virol 82, 299–305.
    [Google Scholar]
  5. Daimon, T., Katsuma, S. & Shimada, T. ( 2007; ). Mutational analysis of active site residues of chitinase from Bombyx mori nucleopolyhedrovirus. Virus Res 124, 168–175.[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. Gopalakrishnan, K., Muthukrishnan, S. & Kramer, K. J. ( 1995; ). Baculovirus-mediated expression of a Manduca sexta chitinase gene: properties of the recombinant protein. Insect Biochem Mol Biol 25, 255–265.[CrossRef]
    [Google Scholar]
  8. Harrison, R. L. & Bonning, B. C. ( 2000; ). Use of scorpion neurotoxins to improve the insecticidal activity of Rachiplusia ou multicapsid nucleopolyhedrovirus. Biol Control 17, 191–201.[CrossRef]
    [Google Scholar]
  9. Harrison, R. L. & Bonning, B. C. ( 2001; ). Use of proteases to improve the insecticidal activity of baculoviruses. Biol Control 20, 199–209.[CrossRef]
    [Google Scholar]
  10. Hawtin, R. E., Arnold, K., Ayres, M. D., Zanotto, P. M., Howard, S. C., Gooday, G. W., Chappell, L. H., Kitts, P. A., King, L. A. & Possee, R. D. ( 1995; ). Identification and preliminary characterization of a chitinase gene in the Autographa californica nuclear polyhedrosis virus genome. Virology 212, 673–685.[CrossRef]
    [Google Scholar]
  11. Hawtin, R. E., Zarkowska, T., Arnold, K., Thomas, C. J., Gooday, G. W., King, L. A., Kuzio, J. A. & Possee, R. D. ( 1997; ). Liquefaction of Autographa californica nucleopolyhedrovirus-infected insects is dependent on the integrity of virus-encoded chitinase and cathepsin genes. Virology 238, 243–253.[CrossRef]
    [Google Scholar]
  12. Hill, J. E., Kuzio, J. & Faulkner, P. ( 1995; ). Identification and characterization of the v-cath gene of the baculovirus, CfMNPV. Biochim Biophys Acta 1264, 275–278.[CrossRef]
    [Google Scholar]
  13. Hom, L. G. & Volkman, L. E. ( 2000; ). Autographa californica M nucleopolyhedrovirus chiA is required for processing of V-CATH. Virology 277, 178–183.[CrossRef]
    [Google Scholar]
  14. Hom, L. G., Ohkawa, T., Trudeau, D. & Volkman, L. E. ( 2002; ). Autographa californica M nucleopolyhedrovirus ProV-CATH is activated during infected cell death. Virology 296, 212–218.[CrossRef]
    [Google Scholar]
  15. Inceoglu, A. B., Kamita, S. G. & Hammock, B. D. ( 2006; ). Genetically modified baculoviruses: a historical overview and future outlook. Adv Virus Res 68, 323–360.
    [Google Scholar]
  16. Jarvis, D. L. & Garcia, A., Jr ( 1994; ). Biosynthesis and processing of the Autographa californica nuclear polyhedrosis virus gp64 protein. Virology 205, 300–313.[CrossRef]
    [Google Scholar]
  17. Lubbert, H. & Doerfler, W. ( 1984; ). Transcription of overlapping sets of RNAs from the genome of Autographa californica nuclear polyhedrosis virus: a novel method for mapping RNAs. J Virol 52, 255–265.
    [Google Scholar]
  18. Maeda, S. ( 1989; ). Increased insecticidal effect by a recombinant baculovirus carrying a synthetic diuretic hormone gene. Biochem Biophys Res Commun 165, 1177–1183.[CrossRef]
    [Google Scholar]
  19. McCreath, K. J. & Gooday, G. W. G. ( 1992; ). A rapid and sensitive microassay for determination of chitinolytic activity. J Microbiol Methods 14, 229–237.[CrossRef]
    [Google Scholar]
  20. Moscardi, F. ( 1999; ). Assessment of the application of baculoviruses for control of Lepidoptera. Annu Rev Entomol 44, 257–289.[CrossRef]
    [Google Scholar]
  21. Ohkawa, T., Majima, K. & Maeda, S. ( 1994; ). A cysteine protease encoded by the baculovirus Bombyx mori nuclear polyhedrosis virus. J Virol 68, 6619–6625.
    [Google Scholar]
  22. Ooi, B. G., Rankin, C. & Miller, L. K. ( 1989; ). Downstream sequences augment transcription from the essential initiation site of a baculovirus polyhedrin gene. J Mol Biol 210, 721–736.[CrossRef]
    [Google Scholar]
  23. O'Reilly, D. R., Miller, L. K. & Luckow, V. A. ( 1992; ). Baculovirus Expression Vectors: a Laboratory Manual. New York: W. H. Freeman.
  24. Possee, R. D. & Howard, S. C. ( 1987; ). Analysis of the polyhedrin gene promoter of the Autographa californica nuclear polyhedrosis virus. Nucleic Acids Res 15, 10233–10248.[CrossRef]
    [Google Scholar]
  25. Sambrook, J. & Russell, D. W. ( 2001; ). Molecular Cloning: a Laboratory Manual, 3rd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.
  26. Saville, G. P., Thomas, C. J., Possee, R. D. & King, L. A. ( 2002; ). Partial redistribution of the Autographa californica nucleopolyhedrovirus chitinase in virus-infected cells accompanies mutation of the carboxy-terminal KDEL ER-retention motif. J Gen Virol 83, 685–694.
    [Google Scholar]
  27. Saville, G. P., Patmanidi, A. L., Possee, R. D. & King, L. A. ( 2004; ). Deletion of the Autographa californica nucleopolyhedrovirus chitinase KDEL motif and in vitro and in vivo analysis of the modified virus. J Gen Virol 85, 821–831.[CrossRef]
    [Google Scholar]
  28. Slack, J. M., Kuzio, J. & Faulkner, P. ( 1995; ). Characterization of v-cath, a cathepsin L-like proteinase expressed by the baculovirus Autographa californica multiple nuclear polyhedrosis virus. J Gen Virol 76, 1091–1098.[CrossRef]
    [Google Scholar]
  29. Slack, J. M., Ribeiro, B. M. & Lobo de Souza, M. ( 2004; ). The gp64 locus of Anticarsia gemmatalis multicapsid nucleopolyhedrovirus contains a 3′ repair exonuclease homologue and lacks v-cath and ChiA genes. J Gen Virol 85, 211–219.[CrossRef]
    [Google Scholar]
  30. Thomas, C. J., Brown, H. L., Hawes, C. R., Lee, B. Y., Min, M.-K., King, L. A. & Possee, R. D. ( 1998; ). Localization of a baculovirus-induced chitinase in the insect cell endoplasmic reticulum. J Virol 72, 10207–10212.
    [Google Scholar]
  31. Thomas, C. J., Gooday, G. W., King, L. A. & Possee, R. D. ( 2000; ). Mutagenesis of the active site coding region of the Autographa californica nucleopolyhedrovirus chiA gene. J Gen Virol 81, 1403–1411.
    [Google Scholar]
  32. 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 Vitro 13, 213–217.[CrossRef]
    [Google Scholar]
  33. Whitford, M., Stewart, S., Kuzio, J. & Faulkner, P. ( 1989; ). Identification and sequence analysis of a gene encoding gp67, an abundant envelope glycoprotein of the baculovirus Autographa californica nuclear polyhedrosis virus. J Virol 63, 1393–1399.
    [Google Scholar]
  34. Wilson, M. E., Mainprize, T. H., Friesen, P. D. & Miller, L. K. ( 1987; ). Location, transcription, and sequence of a baculovirus gene encoding a small arginine-rich polypeptide. J Virol 61, 661–666.
    [Google Scholar]
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