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. 1993 Jun;67(6):3481–3488. doi: 10.1128/jvi.67.6.3481-3488.1993

Identification and characterization of lef-1, a baculovirus gene involved in late and very late gene expression.

A L Passarelli 1, L K Miller 1
PMCID: PMC237694  PMID: 8497062

Abstract

An Autographa californica nuclear polyhedrosis virus (AcMNPV) gene required in transient expression assays for late and very late viral gene expression was identified, sequenced, and transcriptionally mapped. This gene, designated late expression factor 1 (lef-1), was located between 7.4 and 8.7 map units of the AcMNPV physical map. It was identified by cotransfecting Spodoptera frugiperda cultured cells with a collection of overlapping cloned DNA fragments covering the entire AcMNPV genome and a reporter gene controlled by an early, late, or very late AcMNPV promoter. Omission of the DNA fragment containing lef-1 curtailed most late and very late gene expression but not early gene expression. lef-1 was found to be an early gene transcribed as a 1.8-kb RNA in the presence of the protein synthesis inhibitor cycloheximide. The C terminus of the predicted polypeptide product, LEF-1, contained a sequence motif characteristic of nucleoside triphosphate-binding sites.

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Selected References

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  1. 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]
  2. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. 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]
  6. 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]
  7. 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]
  8. Grula M. A., Buller P. L., Weaver R. F. alpha-Amanitin-Resistant Viral RNA Synthesis in Nuclei Isolated from Nuclear Polyhedrosis Virus-Infected Heliothis zea Larvae and Spodoptera frugiperda Cells. J Virol. 1981 Jun;38(3):916–921. doi: 10.1128/jvi.38.3.916-921.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. 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]
  12. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  13. Hoopes R. R., Jr, Rohrmann G. F. In vitro transcription of baculovirus immediate early genes: accurate mRNA initiation by nuclear extracts from both insect and human cells. Proc Natl Acad Sci U S A. 1991 May 15;88(10):4513–4517. doi: 10.1073/pnas.88.10.4513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Huh N. E., Weaver R. F. Identifying the RNA polymerases that synthesize specific transcripts of the Autographa californica nuclear polyhedrosis virus. J Gen Virol. 1990 Jan;71(Pt 1):195–201. doi: 10.1099/0022-1317-71-1-195. [DOI] [PubMed] [Google Scholar]
  15. Hunter T. A thousand and one protein kinases. Cell. 1987 Sep 11;50(6):823–829. doi: 10.1016/0092-8674(87)90509-5. [DOI] [PubMed] [Google Scholar]
  16. Lee H. H., Miller L. K. Isolation of genotypic variants of Autographa californica nuclear polyhedrosis virus. J Virol. 1978 Sep;27(3):754–767. doi: 10.1128/jvi.27.3.754-767.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. O'Reilly D. R., Miller L. K. A baculovirus blocks insect molting by producing ecdysteroid UDP-glucosyl transferase. Science. 1989 Sep 8;245(4922):1110–1112. doi: 10.1126/science.2505387. [DOI] [PubMed] [Google Scholar]
  19. O'Reilly D. R., Miller L. K. Regulation of expression of a baculovirus ecdysteroid UDPglucosyltransferase gene. J Virol. 1990 Mar;64(3):1321–1328. doi: 10.1128/jvi.64.3.1321-1328.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. O'Reilly D. R., Passarelli A. L., Goldman I. F., Miller L. K. Characterization of the DA26 gene in a hypervariable region of the Autographa californica nuclear polyhedrosis virus genome. J Gen Virol. 1990 May;71(Pt 5):1029–1037. doi: 10.1099/0022-1317-71-5-1029. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Ooi B. G., Miller L. K. The influence of antisense RNA on transcriptional mapping of the 5' terminus of a baculovirus RNA. J Gen Virol. 1991 Mar;72(Pt 3):527–534. doi: 10.1099/0022-1317-72-3-527. [DOI] [PubMed] [Google Scholar]
  23. Ooi B. G., Rankin C., Miller L. K. Downstream sequences augment transcription from the essential initiation site of a baculovirus polyhedrin gene. J Mol Biol. 1989 Dec 20;210(4):721–736. doi: 10.1016/0022-2836(89)90105-8. [DOI] [PubMed] [Google Scholar]
  24. Passarelli A. L., Miller L. K. Three baculovirus genes involved in late and very late gene expression: ie-1, ie-n, and lef-2. J Virol. 1993 Apr;67(4):2149–2158. doi: 10.1128/jvi.67.4.2149-2158.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Possee R. D., Sun T. P., Howard S. C., Ayres M. D., Hill-Perkins M., Gearing K. L. Nucleotide sequence of the Autographa californica nuclear polyhedrosis 9.4 kbp EcoRI-I and -R (polyhedrin gene) region. Virology. 1991 Nov;185(1):229–241. doi: 10.1016/0042-6822(91)90770-c. [DOI] [PubMed] [Google Scholar]
  26. Rankin C., Ooi B. G., Miller L. K. Eight base pairs encompassing the transcriptional start point are the major determinant for baculovirus polyhedrin gene expression. Gene. 1988 Oct 15;70(1):39–49. doi: 10.1016/0378-1119(88)90102-3. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. 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]
  29. Theilmann D. A., Stewart S. Identification and characterization of the IE-1 gene of Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus. Virology. 1991 Feb;180(2):492–508. doi: 10.1016/0042-6822(91)90063-h. [DOI] [PubMed] [Google Scholar]
  30. Thiem S. M., Miller L. K. Differential gene expression mediated by late, very late and hybrid baculovirus promoters. Gene. 1990 Jul 2;91(1):87–94. doi: 10.1016/0378-1119(90)90166-o. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. 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]
  33. 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]
  34. Weaver R. F., Weissmann C. Mapping of RNA by a modification of the Berk-Sharp procedure: the 5' termini of 15 S beta-globin mRNA precursor and mature 10 s beta-globin mRNA have identical map coordinates. Nucleic Acids Res. 1979 Nov 10;7(5):1175–1193. doi: 10.1093/nar/7.5.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]

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