1887

Journal of General Virology header logo

Volume 83, Issue 8

Characterization of late gene expression factors -9 and -8 from nucleopolyhedrovirus Free

Abstract

Late gene expression factors, LEF-4, LEF-8, LEF-9 and P47 constitute the primary components of the multinucleocapsid polyhedrovirus (AcMNPV)-encoded RNA polymerase, which initiates transcription from late and very late promoters. Here, characterization of -9 and -8, which encode their corresponding counterparts, from NPV is reported. Transcription of -9 initiated at two independent sites: from a CCT sequence located at −38 nt and a TCTT sequence located at −50 nt, with respect to the +1 ATG of the open reading frame. The 3′ end of the transcript was mapped to a site 17 nt downstream of a canonical polyadenylation signal located 7 nt downstream of the first of the two tandem translational termination codons. Maximum synthesis of LEF-9 was seen from 36 h post-infection (p.i.). The transcription of -8 initiated early in infection from a TGCAAT sequence that differed in the corresponding region from its AcMNPV counterpart (GCG), with consequent elimination of the consensus early transcription start site motif (underlined). Peak levels of -8 transcripts were attained by 24 h p.i. Immunocopurification analyses suggested that there was an association between LEF-8 and LEF-9 .

  • Received:
  • Accepted:
  • Published Online:
© Microbiology Society
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-83-8-2015
2002-08-01
2024-04-27
Loading full text...

Full text loading...

/deliver/fulltext/jgv/83/8/0832015a.html?itemId=/content/journal/jgv/10.1099/0022-1317-83-8-2015&mimeType=html&fmt=ahah

References

  1. Acharya A., Gopinathan K. P. 2001; Identification of an enhancer-like element in the polyhedrin gene upstream region of Bombyx mori nucleopolyhedrovirus. Journal of General Virology 82:2811–2819
     [Google Scholar]
  2. Ahrens C. H., Russell R. L., Funk C. J., Evans J. T., Harwood S. H., Rohrmann G. F. 1997; The sequence of the Orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus genome. Virology 229:381–399
     [Google Scholar]
  3. 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
     [Google Scholar]
  4. Beniya H., Funk C. J., Rohrmann G. F., Weaver R. F. 1996; Purification of a virus-induced RNA polymerase from Autographa californica nucleopolyhedro virus-infected Spodoptera frugiperda cells that accurately initiates late and very late transcription in vitro . Virology 216:12–19
     [Google Scholar]
  5. Blissard G. W., Rohrmann G. F. 1990; Baculovirus diversity and molecular biology. Annual Review of Entomology 35:127–155
     [Google Scholar]
  6. Carstens E. B., Lu A. L., Chan H. L. B. 1993; Sequence, transcriptional mapping, and overexpression of p47, a baculovirus gene regulating late gene expression. Journal of Virology 67:2513–2520
     [Google Scholar]
  7. Carstens E. B., Chan H., Yu H., Williams G. V., Casselman R. 1994; Genetic analyses of temperature-sensitive mutations in baculovirus late expression factors. Virology 204:323–337
     [Google Scholar]
  8. Chomczynski P., Sacchi N. 1987; Single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction. Analytical Biochemistry 162:156–159
     [Google Scholar]
  9. 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]
  10. Friesen P. D., Miller L. K. 1985; Temporal regulation of baculoviral RNA: overlapping early and late transcripts. Journal of Virology 54:392–400
     [Google Scholar]
  11. Fuchs L. Y., Woods M. S., Weaver R. F. 1983; Viral transcription during AcNPV infection: a novel RNA polymerase induced in infected Spodoptera frugiperda cells. Journal of Virology 48:641–646
     [Google Scholar]
  12. Gomi S., Zhou C. E., Yih W., Majima K., Maeda S. 1997; Deletion analysis of four of eighteen late gene expression factor gene homologues of the baculovirus, BmNPV. Virology 230:35–47
     [Google Scholar]
  13. Gomi S., Majima K., Maeda S. 1999; Sequence analysis of the genome of Bombyx mori nucleopolyhedrovirus. Journal of General Virology 80:1323–1337
     [Google Scholar]
  14. Grula M. A., Buller P. L., Weaver R. F. 1981; α-amanitin-resistant viral RNA synthesis in nuclei isolated from nuclear polyhedrosis virus infected Heliothios zea larvae and Spodoptera frugiperda cells. Journal of Virology 38:916–921
     [Google Scholar]
  15. Guarino L. A., Xu B., Jin J., Dong W. 1998a; A virus-encoded RNA polymerase purified from baculovirus-infected cells. Journal of Virology 72:7985–7991
     [Google Scholar]
  16. Guarino L. A., Jin J., Dong W. 1998b; Guanylyltransferase activity of the LEF-4 subunit of baculovirus RNA polymerase. Journal of Virology 72:10003–10010
     [Google Scholar]
  17. Jin J., Guarino L. A. 2000; 3′-end formation of baculovirus late RNAs. Journal of Virology 74:8930–8937
     [Google Scholar]
  18. Jin J., Dong W., Guarino L. A. 1998; The LEF-4 subunit of baculovirus RNA polymerase has RNA 5′-triphosphatase and ATPase activities. Journal of Virology 72:10011–10019
     [Google Scholar]
  19. King L. A., Possee R. D. 1992 The Baculovirus Expression System: a Laboratory Guide London: Chapman & Hall;
     [Google Scholar]
  20. Kuzio J., Pearson M. N., Harwood S. H., Funk C. J., Evans J. T., Slavicek J. M., Rohrmann G. F. 1999; Sequence and analysis of the genome of a baculovirus pathogenic for Lymantria dispar . Virology 253:17–34
     [Google Scholar]
  21. Li L., Harwood S. H., Rohrmann G. F. 1999; Identification of additional genes that influence baculovirus late gene expression. Virology 255:9–19
     [Google Scholar]
  22. Lu A., Miller L. K. 1994; Identification of three late gene expression factors within the 33·8- to 43·4-map-unit region of Autographa californica nuclear polyhedrosis virus. Journal of Virology 68:6710–6718
     [Google Scholar]
  23. Lu A., Miller L. K. 1995; The roles of eighteen baculovirus late gene expression factor genes in transcription and DNA replication. Journal of Virology 69:975–982
     [Google Scholar]
  24. McDevitt M. A., Hart R. P., Wong W. W., Nevins J. R. 1986; Sequences capable of restoring poly(A) site function define two distinct downstream elements. EMBO Journal 5:2907–2913
     [Google Scholar]
  25. McLachlin J. R., Miller L. K. 1994; Identification and characterization of vlf-1 , a baculovirus gene involved in very late gene expression. Journal of Virology 68:7746–7756
     [Google Scholar]
  26. McLauchlan J., Gaffnay D., Whitton J. L., Clements J. B. 1985; The consensus sequence YGTGTTYY located downstream from the AATAAA signal is required for efficient formation of mRNA 3′ termini. Nucleic Acids Research 13:1347–1368
     [Google Scholar]
  27. Mikhailov V. S. 2000; Helix-destabilizing properties of the baculovirus single-stranded DNA-binding protein (LEF-3). Virology 270:180–189
     [Google Scholar]
  28. O’Reilly D. R., Miller L. K., Luckow V. E. 1992 Baculovirus Expression Vectors: a Laboratory Manual New York: W. H. Freeman;
     [Google Scholar]
  29. Palhan V. B., Gopinathan K. P. 1996; Characterization of a local isolate of Bombyx mori nuclear polyhedrosis virus. Current Science (Bangalore 70:147–153
     [Google Scholar]
  30. Palhan V. B., Sumathy S., Gopinathan K. P. 1995; Baculovirus mediated high-level expression of luciferase in silkworm cells and larvae. Biotechniques 19:97–105
     [Google Scholar]
  31. Passarelli A. L., Todd J. W., Miller L. K. 1994; A baculovirus gene involved in late gene expression predicts a large polypeptide with a conserved motif of RNA polymerases. Journal of Virology 68:4673–4678
     [Google Scholar]
  32. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  33. Sehgal D., Gopinathan K. P. 1998; Recombinant Bombyx mori nucleopolyhedrovirus harboring green fluorescent protein. Biotechniques 25:997–1004
     [Google Scholar]
  34. Sriram S., Gopinathan K. P. 1998; A potential role of late gene expression factor, lef2 , from Bombyx mori nuclear polyhedrosis virus in very late gene transcription and DNA replication. Virology 251:108–122
     [Google Scholar]
  35. Todd J. W., Passarelli A. L., Miller L. K. 1995; Eighteen baculovirus genes, including lef-11 , p35 , 39K, and p47 , support late gene expression. Journal of Virology 69:968–974
     [Google Scholar]
  36. Westwood J. A., Jones I. M., Bishop D. H. L. 1993; Analyses of alternative poly(A) signals for use in baculovirus expression vectors. Virology 195:90–99
     [Google Scholar]
  37. Yang S., Miller L. K. 1998; Control of baculovirus polyhedrin gene expression by very late factor 1. Virology 248:131–138
     [Google Scholar]
  38. Yang S., Miller L. K. 1999; Activation of baculovirus very late promoters by interaction with very late factor 1. Journal of Virology 73:3404–3409
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-83-8-2015
Loading
Characterization of late gene expression factors lef-9 and lef-8 from Bombyx mori nucleopolyhedrovirus
J. Gen. Virol. 83, 2015 (2002); https://doi.org/10.1099/0022-1317-83-8-2015
/content/journal/jgv/10.1099/0022-1317-83-8-2015
/content/journal/jgv/10.1099/0022-1317-83-8-2015
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