1887

Abstract

We have located, cloned, sequenced and characterized the ecdysteroid UDP-glucosyltransferase gene () gene from the baculovirus multinucleocapsid nuclear polyhedrosis virus.(LdMNPV), which is specific for the gypsy moth (). The gene from the related baculovirus multinucleocapsid nuclear polyhedrosis virus (AcMNPV) disrupts the hormonal balance of the host larva by galactosylating ecdysone, which prevents moulting. The location of the LdMNPV gene, determined by hybridization analysis using a cloned coding segment of the AcMNPV gene, was mapped to between 79·1 and 80·2 map units on the viral genome. This region contains an open reading frame of 1464 nucleotides capable of encoding a 55K polypeptide. This predicted protein exhibits a 42% amino acid identity with the AcMNPV polypeptide. Transcripts of the gene were analysed by Northern blot and primer extension. The gene is transcribed from approximately 12 to 48 h, and maximally at about 16 h post-infection. Transcription occurred in the presence of aphidicolin, a viral DNA synthesis inhibitor, but not in the presence of cycloheximide, a protein synthesis inhibitor. Therefore the LdMNPV gene is classified as a delayed early gene. The gene is transcribed in a clockwise direction with respect to the circular map, and transcription initiates at a single site. Comparisons between the two baculoviral proteins and mammalian UDP-glucuronosyltransferases reveal areas which are conserved between the mammalian and baculoviral genes, as well as areas that are only conserved in the viral proteins. The LdMNPV protein sequence appears to include a signal peptide, which would allow the protein to be secreted into the haemolymph.

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1994-04-01
2024-04-24
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References

  1. Blissard G. W., Rohrmann G. F. 1990; Baculovirus diversity and molecular biology. Annual Review of Entomology 35:127–155
    [Google Scholar]
  2. Bjornson R. M., Rohrmann G. F. 1992a; Nucleotide sequence of the polyhedron envelope protein gene region of the Lymantria dispar nuclear polyhedrosis virus. Journal of General Virology 73:1499–1504
    [Google Scholar]
  3. Bjornson R. M., Rohrmann G. F. 1992b; Nucleotide sequence of the p39-capsid gene region of the Lymantria dispar nuclear polyhedrosis virus. Journal of General Virology 73:1505–1508
    [Google Scholar]
  4. Bjornson R. M., Glockbr B., Rohrmann G. F. 1992; Characterization of the nucleotide sequence of the Lymantria dispar nuclear polyhedrosis virus DNA polymerase gene region. Journal of General Virology 73:3177–3183
    [Google Scholar]
  5. Burand J. P., Park E. J. 1992; Effect of nuclear polyhedrosis virus infection on the development and pupation of gypsy moth larvae. Journal of Invertebrate Pathology 60:171–175
    [Google Scholar]
  6. Burchell B., Coughtrie M. W. H. 1989; UDP-glucuronosyl transferases. Pharmacology and Therapeutics 43:261–289
    [Google Scholar]
  7. Crawford A. M., Miller L. K. 1988; Characterization of an early gene accelerating expression of late genes of the baculovirus Autographa californica nuclear polyhedrosis virus. Journal of Virology 62:2773–2781
    [Google Scholar]
  8. Friesen P. D., Miller L. K. 1985; Temporal regulation of baculovirus RNA: overlapping early and late transcripts. Journal of Virology 54:392–400
    [Google Scholar]
  9. Fiarrap K. A., Payne C. C. 1979; The structural properties and identification of insect viruses. Advances in Virus Research 25:273–355
    [Google Scholar]
  10. Hohn B., Collins J. 1980; A small cosmid for efficient cloning of large DNA fragments. Gene 11:291–298
    [Google Scholar]
  11. Hough J. A., Pimentel D. 1978; Influence of host foliage on development, survival, and fecundity of the gypsy moth. Environmental Entomology 7:97–102
    [Google Scholar]
  12. Kimura T., Owens I. S. 1987; Mouse UDP glucuronosyl transferase: cDNA and complete amino acid sequence and regulation. European Journal of Biochemistry 168:515–521
    [Google Scholar]
  13. Kozak M. 1986; Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell 44:283–292
    [Google Scholar]
  14. Mahmoudi M., Lin V. K. 1989; Comparison of two different hybridization systems in Northern transfer analysis. BioTechniques 7:331–333
    [Google Scholar]
  15. O’Reilly D. R., Miller L. K. 1989; A baculovirus blocks insect molting by producing ecdysteroid UDP-glucosyl transferase. Science 245:1110–1112
    [Google Scholar]
  16. O’Reilly D. R., Miller L. K. 1990; Regulation of expression of a baculovirus ecdysteroid UDP-glucosyl transferase gene. Journal of Virology 64:1321–1328
    [Google Scholar]
  17. O’Reilly D. R., Miller L. K. 1991; Improvement of a baculovirus pesticide by deletion of the egt gene. Bio/Technology 9:1086–1089
    [Google Scholar]
  18. O’Reilly D. R., Brown M. R., Miller L. K. 1992; Alteration of ecdysteroid metabolism due to baculovirus infection of the fall army worm Spodoptera frugiperda: host ecdysteroids are conjugated with galactose. Insect Biochemistry and Molecular Biology 22:313–320
    [Google Scholar]
  19. Rankin C., Ooi B. G., Miller L. K. 1988; Eight base pairs encompassing the transcriptional start point are the major determinant for baculovirus polyhedrin gene expression. Gene 70:39–49
    [Google Scholar]
  20. Ritter J. K., Sheen Y. Y., Owens I. S. 1990; Cloning and expression of human liver UDP-glucuronosyltransferase in COS-1 cells. Journal of Biological Chemistry 265:7900–7906
    [Google Scholar]
  21. Sato H., Koiwai O., Tanabe K., Kashiwamata S. 1990; Isolation and sequencing of rat liver bilirubin UDP-glucuronosyl-transferase cDNA: possible alternate splicing of a common primary transcript. Biochemical and Biophysical Research Communications 169:260–264
    [Google Scholar]
  22. Slavicek J. M. 1991; Temporal analysis and spatial mapping of Lymantria dispar nuclear polyhedrosis virus transcripts and in vitro translation polypeptides. Virus Research 20:223–236
    [Google Scholar]
  23. Slavicek J. M., Podgewaite J. D., Lanner-Herrera C. 1992; Properties of two Lymantria dispar nuclear polyhedrosis virus isolates obtained from the microbial pesticide Gypchek. Journal oj Invertebrate Pathology 59:142–148
    [Google Scholar]
  24. Smith I. R. L., Van Beek N. A. M., Podgewaite J. D., Wood H. A. 1988; Physical map and polyhedrin gene sequence of Lymantria dispar nuclear polyhedrosis virus. Gene 71:97–105
    [Google Scholar]
  25. Thiem S. M., Miller L. K. 1989; A baculovirus gene with a novel transcription pattern encodes a polypeptide with a zinc finger and a leucine zipper. Journal of Virology 63:4489–4497
    [Google Scholar]
  26. Vlak J. M., Smith G. E. 1982; Orientation of the genome of Autographa californica nuclear polyhedrosis virus: a proposal. Journal of Virology 41:1118–1121
    [Google Scholar]
  27. Von Heijne G. 1986; A new method for predicting signal sequence cleavage sites. Nucleic Acids Research 14:4683–4690
    [Google Scholar]
  28. Wilbur W. J., Lipman D. J. 1983; Rapid similarity searches of nucleic acid and protein databanks. Proceedings of the National Academy of Sciences U.S.A: 80726–730
    [Google Scholar]
  29. Wood H. A., Granados R. R. 1991; Genetically engineered baculoviruses as agents for pest control. Annual Review of Microbiology 45:69–87
    [Google Scholar]
  30. Wu J., Miller L. K. 1989; Sequence, transcription and translation of a late gene of the Autographa californica nuclear polyhedrosis virus encoding a 34-8K polypeptide. Journal of General Virology 70:2449–2459
    [Google Scholar]
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