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. 1992 Feb;66(2):655–663. doi: 10.1128/jvi.66.2.655-663.1992

Transcription analysis of the EcoRI D region of the baculovirus Autographa californica nuclear polyhedrosis virus identifies an early 4-kilobase RNA encoding the essential p143 gene.

A Lu 1, E B Carstens 1
PMCID: PMC240764  PMID: 1731106

Abstract

We have investigated the transcriptional activity of the 60.1- to 68.3-map-unit region of the baculovirus Autographa californica nuclear polyhedrosis virus (AcMNPV). Twelve transcripts mapping to this region were expressed at various times during infection. An early 4.0-kb transcript, potentially coding for a 143-kDa peptide essential for viral DNA replication, was maximally abundant at 6 h postinfection (p.i.). Transcripts of 0.5, 1.1, 1.4, 2.1, and 3.1 kb were most abundant at 12 h p.i., while two large transcripts of 5.2 and 6.8 kb were expressed maximally at 24 h p.i. In the presence of cycloheximide, and in ts8-infected cells at the nonpermissive temperature, only the 4.0-kb RNA was expressed. Northern (RNA) blot analysis using DNA subfragments from the EcoRI D fragment as probes suggested that many of the transcripts overlapped. Strand-specific cRNA probes revealed that the majority of the RNAs were transcribed in the counterclockwise direction. S1 nuclease and primer extension analysis were used to map the 5' ends of transcripts coded within the 60.1- to 64.8-map-unit region. Mapping of the 3' ends of the 1.1-, 4.0-, 5.2-, and 6.8-kb transcripts suggested that these RNAs were all coterminal at their 3' ends. A minicistron was found between the early 4.0-kb transcription start site and the predicted ATG start codon of the p143 gene. Several similar sequence motifs were identified in the promoter regions of the p143 gene and the AcMNPV DNA polymerase gene.

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  1. Archer T. K., Tam S. P., Deeley R. G. Kinetics of estrogen-dependent modulation of apolipoprotein A-I synthesis in human hepatoma cells. J Biol Chem. 1986 Apr 15;261(11):5067–5074. [PubMed] [Google Scholar]
  2. 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]
  3. Blissard G. W., Rohrmann G. F. Location, sequence, transcriptional mapping, and temporal expression of the gp64 envelope glycoprotein gene of the Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus. Virology. 1989 Jun;170(2):537–555. doi: 10.1016/0042-6822(89)90445-5. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. 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]
  9. 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]
  10. 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]
  11. 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]
  12. 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]
  13. 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]
  14. Gordon J. D., Carstens E. B. Phenotypic characterization and physical mapping of a temperature-sensitive mutant of Autographa californica nuclear polyhedrosis virus defective in DNA synthesis. Virology. 1984 Oct 15;138(1):69–81. doi: 10.1016/0042-6822(84)90148-x. [DOI] [PubMed] [Google Scholar]
  15. Guarino L. A., Smith M. W. Nucleotide sequence and characterization of the 39K gene region of Autographa californica nuclear polyhedrosis virus. Virology. 1990 Nov;179(1):1–8. doi: 10.1016/0042-6822(90)90266-t. [DOI] [PubMed] [Google Scholar]
  16. Guarino L. A., Summers M. D. Functional mapping of Autographa california nuclear polyhedrosis virus genes required for late gene expression. J Virol. 1988 Feb;62(2):463–471. doi: 10.1128/jvi.62.2.463-471.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Guarino L. A., Summers M. D. Nucleotide sequence and temporal expression of a baculovirus regulatory gene. J Virol. 1987 Jul;61(7):2091–2099. doi: 10.1128/jvi.61.7.2091-2099.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kozak M. Bifunctional messenger RNAs in eukaryotes. Cell. 1986 Nov 21;47(4):481–483. doi: 10.1016/0092-8674(86)90609-4. [DOI] [PubMed] [Google Scholar]
  20. Kozak M. Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. Nucleic Acids Res. 1984 Jan 25;12(2):857–872. doi: 10.1093/nar/12.2.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Krappa R., Knebel-Mörsdorf D. Identification of the very early transcribed baculovirus gene PE-38. J Virol. 1991 Feb;65(2):805–812. doi: 10.1128/jvi.65.2.805-812.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Liu A., Qin J. C., Rankin C., Hardin S. E., Weaver R. F. Nucleotide sequence of a portion of the Autographa californica nuclear polyhedrosis virus genome containing the EcoRI site-rich region (hr5) and an open reading frame just 5' of the p10 gene. J Gen Virol. 1986 Nov;67(Pt 11):2565–2570. doi: 10.1099/0022-1317-67-11-2565. [DOI] [PubMed] [Google Scholar]
  23. Lu A., Carstens E. B. Nucleotide sequence of a gene essential for viral DNA replication in the baculovirus Autographa californica nuclear polyhedrosis virus. Virology. 1991 Mar;181(1):336–347. doi: 10.1016/0042-6822(91)90500-b. [DOI] [PubMed] [Google Scholar]
  24. Lübbert H., Doerfler W. Transcription of overlapping sets of RNAs from the genome of Autographa californica nuclear polyhedrosis virus: a novel method for mapping RNAs. J Virol. 1984 Oct;52(1):255–265. doi: 10.1128/jvi.52.1.255-265.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. McMaster G. K., Carmichael G. G. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835–4838. doi: 10.1073/pnas.74.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Miller L. K., Jewell J. E., Browne D. Baculovirus induction of a DNA polymerase. J Virol. 1981 Oct;40(1):305–308. doi: 10.1128/jvi.40.1.305-308.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Müller R., Pearson M. N., Russell R. L., Rohrmann G. F. A capsid-associated protein of the multicapsid nuclear polyhedrosis virus of Orgyia pseudotsugata: genetic location, sequence, transcriptional mapping, and immunocytochemical characterization. Virology. 1990 May;176(1):133–144. doi: 10.1016/0042-6822(90)90238-m. [DOI] [PubMed] [Google Scholar]
  29. Nissen M. S., Friesen P. D. Molecular analysis of the transcriptional regulatory region of an early baculovirus gene. J Virol. 1989 Feb;63(2):493–503. doi: 10.1128/jvi.63.2.493-503.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Partington S., Yu H., Lu A., Carstens E. B. Isolation of temperature sensitive mutants of Autographa californica nuclear polyhedrosis virus: phenotype characterization of baculovirus mutants defective in very late gene expression. Virology. 1990 Mar;175(1):91–102. doi: 10.1016/0042-6822(90)90189-x. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. 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]
  34. Russell R. L., Rohrmann G. F. The p6.5 gene region of a nuclear polyhedrosis virus of Orgyia pseudotsugata: DNA sequence and transcriptional analysis of four late genes. J Gen Virol. 1990 Mar;71(Pt 3):551–560. doi: 10.1099/0022-1317-71-3-551. [DOI] [PubMed] [Google Scholar]
  35. Thiem S. M., Miller L. K. Identification, sequence, and transcriptional mapping of the major capsid protein gene of the baculovirus Autographa californica nuclear polyhedrosis virus. J Virol. 1989 May;63(5):2008–2018. doi: 10.1128/jvi.63.5.2008-2018.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Tomalski M. D., Wu J. G., Miller L. K. The location, sequence, transcription, and regulation of a baculovirus DNA polymerase gene. Virology. 1988 Dec;167(2):591–600. [PubMed] [Google Scholar]
  37. Wilson M. E., Mainprize T. H., Friesen P. D., Miller L. K. Location, transcription, and sequence of a baculovirus gene encoding a small arginine-rich polypeptide. J Virol. 1987 Mar;61(3):661–666. doi: 10.1128/jvi.61.3.661-666.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wu J. G., Miller L. K. Sequence, transcription and translation of a late gene of the Autographa californica nuclear polyhedrosis virus encoding a 34.8K polypeptide. J Gen Virol. 1989 Sep;70(Pt 9):2449–2459. doi: 10.1099/0022-1317-70-9-2449. [DOI] [PubMed] [Google Scholar]

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