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. 1991 Feb;65(2):805–812. doi: 10.1128/jvi.65.2.805-812.1991

Identification of the very early transcribed baculovirus gene PE-38.

R Krappa 1, D Knebel-Mörsdorf 1
PMCID: PMC239820  PMID: 1987375

Abstract

We have started to identify early viral RNAs that are transcribed at 1 h after inoculation to investigate the mechanism involved in the regulation of early gene expression of Autographa californica nuclear polyhedrosis virus (AcNPV). Cloned viral DNA fragments were hybridized to Northern (RNA) blots of polyadenylated RNA isolated from Spodoptera frugiperda cells at 1, 2, and 6 h postinfection to localize very early transcripts. Subsequently we prepared a cDNA library of polyadenylated RNA transcribed at 1 h after inoculation to analyze the cDNA clones corresponding to the major early RNAs. We identified a gene located upstream of the immediate-early gene IE-N extending in the opposite direction. Because of the very early expression during AcNPV infection and the transient expression in uninfected cells, we conclude that we found an immediate-early gene, designated PE-38. The determination of the nucleotide sequence of PE-38 revealed one open reading frame potentially encoding a gene product of 38 kDa. Results of in vitro translation experiments suggest that a PE-38-specific polypeptide of approximately 38 kDa can be expressed. We have evidence from computer analyses that the predicted amino acid sequence includes two putative DNA-binding motifs, a zinc finger, and a leucine zipper.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berg J. M. Potential metal-binding domains in nucleic acid binding proteins. Science. 1986 Apr 25;232(4749):485–487. doi: 10.1126/science.2421409. [DOI] [PubMed] [Google Scholar]
  3. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  4. Bond B. J., Davidson N. The Drosophila melanogaster actin 5C gene uses two transcription initiation sites and three polyadenylation sites to express multiple mRNA species. Mol Cell Biol. 1986 Jun;6(6):2080–2088. doi: 10.1128/mcb.6.6.2080. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Busch S. J., Sassone-Corsi P. Dimers, leucine zippers and DNA-binding domains. Trends Genet. 1990 Feb;6(2):36–40. doi: 10.1016/0168-9525(90)90071-d. [DOI] [PubMed] [Google Scholar]
  6. Carson D. D., Guarino L. A., Summers M. D. Functional mapping of an AcNPV immediately early gene which augments expression of the IE-1 trans-activated 39K gene. Virology. 1988 Feb;162(2):444–451. doi: 10.1016/0042-6822(88)90485-0. [DOI] [PubMed] [Google Scholar]
  7. Chisholm G. E., Henner D. J. Multiple early transcripts and splicing of the Autographa californica nuclear polyhedrosis virus IE-1 gene. J Virol. 1988 Sep;62(9):3193–3200. doi: 10.1128/jvi.62.9.3193-3200.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cochran M. A., Faulkner P. Location of Homologous DNA Sequences Interspersed at Five Regions in the Baculovirus AcMNPV Genome. J Virol. 1983 Mar;45(3):961–970. doi: 10.1128/jvi.45.3.961-970.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Crawford A. M., Miller L. K. Characterization of an early gene accelerating expression of late genes of the baculovirus Autographa californica nuclear polyhedrosis virus. J Virol. 1988 Aug;62(8):2773–2781. doi: 10.1128/jvi.62.8.2773-2781.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Doerfler W. Expression of the Autographa californica nuclear polyhedrosis virus genome in insect cells: homologous viral and heterologous vertebrate genes--the baculovirus vector system. Curr Top Microbiol Immunol. 1986;131:51–68. doi: 10.1007/978-3-642-71589-1_4. [DOI] [PubMed] [Google Scholar]
  11. Erlandson M. A., Gordon J., Carstens E. B. Size and map locations of early transcription products on the Autographa californica nuclear polyhedrosis virus genome. Virology. 1985 Apr 15;142(1):12–23. doi: 10.1016/0042-6822(85)90418-0. [DOI] [PubMed] [Google Scholar]
  12. Esche H., Schilling R., Doerfler W. In vitro translation of adenovirus type 12-specific mRNA isolated from infected and transformed cells. J Virol. 1979 Apr;30(1):21–31. doi: 10.1128/jvi.30.1.21-31.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Evans R. M., Hollenberg S. M. Zinc fingers: gilt by association. Cell. 1988 Jan 15;52(1):1–3. doi: 10.1016/0092-8674(88)90522-3. [DOI] [PubMed] [Google Scholar]
  14. Faulkner P., Carstens E. B. An overview of the structure and replication of baculoviruses. Curr Top Microbiol Immunol. 1986;131:1–19. doi: 10.1007/978-3-642-71589-1_1. [DOI] [PubMed] [Google Scholar]
  15. Friesen P. D., Miller L. K. Divergent transcription of early 35- and 94-kilodalton protein genes encoded by the HindIII K genome fragment of the baculovirus Autographa californica nuclear polyhedrosis virus. J Virol. 1987 Jul;61(7):2264–2272. doi: 10.1128/jvi.61.7.2264-2272.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Friesen P. D., Miller L. K. Temporal regulation of baculovirus RNA: overlapping early and late transcripts. J Virol. 1985 May;54(2):392–400. doi: 10.1128/jvi.54.2.392-400.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Friesen P. D., Miller L. K. The regulation of baculovirus gene expression. Curr Top Microbiol Immunol. 1986;131:31–49. doi: 10.1007/978-3-642-71589-1_3. [DOI] [PubMed] [Google Scholar]
  18. Fuchs L. Y., Woods M. S., Weaver R. F. Viral Transcription During Autographa californica Nuclear Polyhedrosis Virus Infection: a Novel RNA Polymerase Induced in Infected Spodoptera frugiperda Cells. J Virol. 1983 Dec;48(3):641–646. doi: 10.1128/jvi.48.3.641-646.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Guarino L. A., Summers M. D. Functional mapping of a trans-activating gene required for expression of a baculovirus delayed-early gene. J Virol. 1986 Feb;57(2):563–571. doi: 10.1128/jvi.57.2.563-571.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Guarino L. A., Summers M. D. Interspersed Homologous DNA of Autographa californica Nuclear Polyhedrosis Virus Enhances Delayed-Early Gene Expression. J Virol. 1986 Oct;60(1):215–223. doi: 10.1128/jvi.60.1.215-223.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  23. Landschulz W. H., Johnson P. F., McKnight S. L. The leucine zipper: a hypothetical structure common to a new class of DNA binding proteins. Science. 1988 Jun 24;240(4860):1759–1764. doi: 10.1126/science.3289117. [DOI] [PubMed] [Google Scholar]
  24. Lübbert H., Doerfler W. Mapping of Early and Late Transcripts Encoded by the Autographa californica Nuclear Polyhedrosis Virus Genome: Is Viral RNA Spliced? J Virol. 1984 May;50(2):497–506. doi: 10.1128/jvi.50.2.497-506.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ma J., Ptashne M. Deletion analysis of GAL4 defines two transcriptional activating segments. Cell. 1987 Mar 13;48(5):847–853. doi: 10.1016/0092-8674(87)90081-x. [DOI] [PubMed] [Google Scholar]
  26. Mainprize T. H., Lee K., Miller L. K. Variation in the temporal expression of overlapping baculovirus transcripts. Virus Res. 1986 Oct;6(1):85–99. doi: 10.1016/0168-1702(86)90059-6. [DOI] [PubMed] [Google Scholar]
  27. Miller J., McLachlan A. D., Klug A. Repetitive zinc-binding domains in the protein transcription factor IIIA from Xenopus oocytes. EMBO J. 1985 Jun;4(6):1609–1614. doi: 10.1002/j.1460-2075.1985.tb03825.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Morris C. E., Klement J. F., McAllister W. T. Cloning and expression of the bacteriophage T3 RNA polymerase gene. Gene. 1986;41(2-3):193–200. doi: 10.1016/0378-1119(86)90098-3. [DOI] [PubMed] [Google Scholar]
  29. Nielsen D. A., Shapiro D. J. Preparation of capped RNA transcripts using T7 RNA polymerase. Nucleic Acids Res. 1986 Jul 25;14(14):5936–5936. doi: 10.1093/nar/14.14.5936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. O'Reilly D. R., Crawford A. M., Miller L. K. Viral proliferating cell nuclear antigen. Nature. 1989 Feb 16;337(6208):606–606. doi: 10.1038/337606a0. [DOI] [PubMed] [Google Scholar]
  31. Oellig C., Happ B., Müller T., Doerfler W. Overlapping sets of viral RNAs reflect the array of polypeptides in the EcoRI J and N fragments (map positions 81.2 to 85.0) of the Autographa californica nuclear polyhedrosis virus genome. J Virol. 1987 Oct;61(10):3048–3057. doi: 10.1128/jvi.61.10.3048-3057.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ooi B. G., Miller L. K. Regulation of host RNA levels during baculovirus infection. Virology. 1988 Oct;166(2):515–523. doi: 10.1016/0042-6822(88)90522-3. [DOI] [PubMed] [Google Scholar]
  33. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rankin C., Ladin B. F., Weaver R. F. Physical mapping of temporally regulated, overlapping transcripts in the region of the 10K protein gene in Autographa californica nuclear polyhedrosis virus. J Virol. 1986 Jan;57(1):18–27. doi: 10.1128/jvi.57.1.18-27.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rice W. C., Miller L. K. Baculovirus transcription in the presence of inhibitors and in nonpermissive Drosophila cells. Virus Res. 1986 Nov;6(2):155–172. doi: 10.1016/0168-1702(86)90047-x. [DOI] [PubMed] [Google Scholar]
  36. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  37. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Scott M. R., Westphal K. H., Rigby P. W. Activation of mouse genes in transformed cells. Cell. 1983 Sep;34(2):557–567. doi: 10.1016/0092-8674(83)90388-4. [DOI] [PubMed] [Google Scholar]
  39. Short J. M., Fernandez J. M., Sorge J. A., Huse W. D. Lambda ZAP: a bacteriophage lambda expression vector with in vivo excision properties. Nucleic Acids Res. 1988 Aug 11;16(15):7583–7600. doi: 10.1093/nar/16.15.7583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  41. Struhl K. Promoters, activator proteins, and the mechanism of transcriptional initiation in yeast. Cell. 1987 May 8;49(3):295–297. doi: 10.1016/0092-8674(87)90277-7. [DOI] [PubMed] [Google Scholar]
  42. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  43. Thiem S. M., Miller L. K. A baculovirus gene with a novel transcription pattern encodes a polypeptide with a zinc finger and a leucine zipper. J Virol. 1989 Nov;63(11):4489–4497. doi: 10.1128/jvi.63.11.4489-4497.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Tjia S. T., Carstens E. B., Doerfler W. Infection of Spodoptera frugiperda cells with Autographa californica nuclear polyhedrosis virus II. The viral DNA and the kinetics of its replication. Virology. 1979 Dec;99(2):399–409. doi: 10.1016/0042-6822(79)90018-7. [DOI] [PubMed] [Google Scholar]
  45. Vaughn J. L., Goodwin R. H., Tompkins G. J., McCawley P. The establishment of two cell lines from the insect Spodoptera frugiperda (Lepidoptera; Noctuidae). In Vitro. 1977 Apr;13(4):213–217. doi: 10.1007/BF02615077. [DOI] [PubMed] [Google Scholar]
  46. Vlak J. M., Smith G. E. Orientation of the Genome of Autographa californica Nuclear Polyhedrosis Virus: a Proposal. J Virol. 1982 Mar;41(3):1118–1121. doi: 10.1128/jvi.41.3.1118-1121.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]

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