1887

Journal of General Virology header logo

Volume 77, Issue 5

Baculovirus infection of larvae: expression driven by promoters of early genes pe38 and me53 in larval tissue Free

Abstract

To follow the progression of infection of multicapsid nuclear polyhedrosis virus (AcMNPV) within tissues of its larval host, we have constructed AcMNPV recombinants carrying reporter genes under the control of the early virus promoters pe38 and me53, in addition to the authentic genes. The early promoter- gene cassettes were located upstream of the very late polyhedrin gene. In infected insect cell lines, pe38 transcription is initiated at an early promoter, while me53 transcripts start from both early and late sites. Transcriptional mapping of the duplicated me53 and pe38 promoters driving expression showed that they initiated at the same start sites as in the authentic genes. Expression of by these recombinants was compared to a recombinant driving β-glucuronidase expression from the very late p10 promoter and expression from the constitutive heat shock protein 70 promoter of . After infection of larvae with the different recombinants, we followed reporter gene expression and polyhedron formation in different tissues using immunohistochemistry and electron microscopy. LacZ expression, indicative of early viral transcriptional activity, was detected in nearly all larval tissues during the course of infection. In most tissues these early events were followed by pathophysiological changes associated with late and very late gene expression. However, p10 transcription and polyhedron formation were not observed in midgut goblet cells, Malpighian tubules and salivary glands. These results suggest that expression of early virus genes, such as me53 and pe38, is not restricted to larval tissues that are permissive for AcMNPV replication.

  • Received:
  • Accepted:
  • Published Online:
© Journal of General Virology 1996
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-77-5-815
1996-05-01
2024-04-25
Loading full text...

Full text loading...

/deliver/fulltext/jgv/77/5/JV0770050815.html?itemId=/content/journal/jgv/10.1099/0022-1317-77-5-815&mimeType=html&fmt=ahah

References

  1. Becker D., Knebel-Mörsdorf D. 1993; Sequence and temporal appearance of the early transcribed baculovirus gene HE65. Journal of Virology 67:5867–5872
     [Google Scholar]
  2. Blissard G. W., Rohrmann G. F. 1990; Baculovirus diversity and molecular biology. Annual Review of Entomology 35:127–155
     [Google Scholar]
  3. Carson D. D., Guarino L. A., Summers M. D. 1988; Functional mapping of an AcNPV immediate early gene which augments expression of the IE-1 trans-activated 39K gene. Virology 162:444–451
     [Google Scholar]
  4. Dickson J. A., Friesen P. D. 1991; Identification of upstream promoter elements mediating early transcription from the 35, 000-molecular-weight protein gene of Autographa californica nuclear polyhedrosis virus. Journal of Virology 65:4006–4016
     [Google Scholar]
  5. Engelhard E. K., Kam-Morgan L. N. W., Washburn J. O., Volkman L. E. 1994; The insect tracheal system: a conduit for the systemic spread of Autographa californica M nuclear polyhedrosis virus. Proceedings of the National Academy of Sciences, USA 91:3224–3227
     [Google Scholar]
  6. Flipsen J. T. M., van Lent J. W. M., Goldbach R. W., Vlak J. M. 1993; Expression of polyhedrin and p10 in the midgut of AcMNPV-infected Spodoptera exigua larvae: an immunoelectron microscopic investigation. Journal of Invertebrate Pathology 61:17–23
     [Google Scholar]
  7. Flipsen J. T. M., Martens J. W. M., van Oers M. M., Vlak J. M., van Lent J. W. M. 1995a; Passage of Autographa californica nuclear polyhedrosis virus through the midgut epithelium of Spodoptera exigua larvae. Virology 208:328–335
     [Google Scholar]
  8. Flipsen J. T. M., Mans R. M. W., Kleefsman A. W. F., Knebel-Mörsdorf D., Vlak J. M. 1995b; Deletion of the baculovirus ecdysteroid UDP-glucosyltransferase gene induces early degeneration of Malpighian tubules in infected insects. Journal of Virology 69:4529–4532
     [Google Scholar]
  9. Gardiner G. R., Stockdale H. 1975; Two tissue culture media for production of lepidopteran cells and nuclear polyhedrosis viruses. Journal of Invertebrate Pathology 25:363–370
     [Google Scholar]
  10. Gearing K. L., Possee R. D. 1990; Functional analysis of a 603 nucleotide open reading frame upstream of the polyhedrin gene of Autographa californica nuclear polyhedrosis virus. Journal of General Virology 71:251–262
     [Google Scholar]
  11. Granados R. R., Williams K. A. 1986; In vivo infection and replication of baculoviruses. In The Biology of Baculoviruses vol 1Biological Properties and Molecular Biology pp 89–108 Edited by Granados R. R., Federici B. A. Boca Raton: CRC Press;
     [Google Scholar]
  12. Guarino L. A., Summers M. D. 1986; Functional mapping of a trans-activating gene required for expression of a baculovirus delayed-early gene. Journal of Virology 57:563–571
     [Google Scholar]
  13. Guarino L. A., Smith M. 1992; Regulation of delayed-early gene transcription by dual TATA boxes. Journal of Virology 66:3733–3739
     [Google Scholar]
  14. Hink W. F. 1970; Established insect cell line from the cabbage looper, Trichoplusia ni . Nature 226:466–467
     [Google Scholar]
  15. Keddie B. A., Aponte G. W., Volkman L. E. 1989; The pathway of infection of Autographa californica nuclear polyhedrosis virus in an insect host. Science 243:1728–1730
     [Google Scholar]
  16. Knebel-Mörsdorf D., Kremer A., Jahnel F. 1993; Baculovirus gene ME53, which contains a putative zinc finger motif, is one of the major early transcribed genes. Journal of Virology 67:753–758
     [Google Scholar]
  17. Krappa R., Knebel-Mörsdorf D. 1991; Identification of the very early transcribed baculovirus gene PE 38. Journal of Virology 65:805–812
     [Google Scholar]
  18. Krappa R., Behn-Krappa A., Jahnel F., Doerfler W., Knebel-Morsdorf D. 1992; Differential factor binding at the promoter of early baculovirus gene PE38 during viral infection: GATA motif is recognized by an insect protein. Journal of Virology 66:3494–3503
     [Google Scholar]
  19. Krappa R., Roncarati R., Knebel-Mörsdorf D. 1995; Expression of PE38 and IE2, viral members of the C3HC4 finger family, during baculovirus infection: PE38 and IE2 localize to distinct nuclear regions. Journal of Virology 69:5287–5293
     [Google Scholar]
  20. OoI B. G., Miller L. K. 1990; Transcription of the baculovirus polyhedrin gene reduces the levels of an antisense transcript initiated downstream. Journal of Virology 64:3126–3129
     [Google Scholar]
  21. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. 1988; Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
     [Google Scholar]
  22. Smith G. E., Summers M. D. 1978; Analysis of baculovirus genomes with restriction enzyme endonucleases. Virology 98:517–527
     [Google Scholar]
  23. Smith G. E., Summers M. D. 1981; Application of a novel radio-immunoassay to identify baculovirus structural proteins that share interspecies antigenic determinants. Journal of Virology 39:125–137
     [Google Scholar]
  24. Smits P. H. 1987 Nuclear polyhedrosis virus as biological control agent of Spodoptera exigua PhD thesis Wageningen Agricultural University:
     [Google Scholar]
  25. Tjia S. T., Carstens E. B., Doerfler W. 1979; Infection of Spodoptera frugiperda cells with Autographa californica nuclear polyhedrosis virus. II. The viral DNA and the kinetics of its replication. Virology 99:399–409
     [Google Scholar]
  26. van der Wilk F., van Lent J. W. M., Vlak J. M. 1987; Immunogold detection of polyhedrin, p10 and virion antigens in Autographa californica nuclear polyhedrosis virus-infected Spodoptera frugiperda cells. Journal of General Virology 68:2615–2623
     [Google Scholar]
  27. Vaughn J. L., Goodwin R. H., Tompkins G. J., McCawley P. 1977; The establishment of two cell lines from the insect Spodoptera frugiperda . In Vitro 13:213–217
     [Google Scholar]
  28. Volkman L. E., Keddie B. A. 1990; Nuclear polyhedrosis virus pathogenesis. Seminars in Virology 1:249–256
     [Google Scholar]
  29. Washburn J. O., Kirkpatrick B. A., Volkman L. E. 1995; Comparative pathogenesis of Autographa californica M nuclear polyhedrosis virus in larvae of Trichoplusia ni and Heliothis virescens . Virology 209:561–568
     [Google Scholar]
  30. Weyer U., Knight S., Possee R. D. 1990; Analysis of very late gene expression by Autographa californica nuclear polyhedrosis virus and the further development of multiple expression vectors. Journal of General Virology 71:1525–1534
     [Google Scholar]
  31. Zuidema D., Schouten A., Usmany M., Maule A. J., Belsham G. J., Roosien J., Klinge-Roode E. C., van Lent J. W. M., Vlak J. M. 1990; Expression of cauliflower mosaic virus gene I in insect cells using a novel polyhedron-based baculovirus expression vector. Journal of General Virology 71:2201–2209
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-77-5-815
Loading
Baculovirus infection of Spodoptera exigua larvae: lacZ expression driven by promoters of early genes pe38 and me53 in larval tissue
J. Gen. Virol. 77, 815 (1996); https://doi.org/10.1099/0022-1317-77-5-815
/content/journal/jgv/10.1099/0022-1317-77-5-815
/content/journal/jgv/10.1099/0022-1317-77-5-815
Loading

Data & Media loading...

Most cited 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