Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1987 May;61(5):1448–1456. doi: 10.1128/jvi.61.5.1448-1456.1987

Construction of a genetic switch for inducible trans-activation of gene expression in eucaryotic cells.

S L Rhode 3rd
PMCID: PMC254122  PMID: 3033274

Abstract

The cotransfection of selectable marker genes and the gene for the nonstructural proteins NS1 and NS2 of the autonomous parvovirus H-1 failed to produce cell lines that constitutively expressed NS1. A plasmid, pP38NS1cat, was constructed that expressed the NS1-NS2 gene from the H-1 P38 coat protein promoter in place of the natural P4 promoter. The P38 promoter is constitutively weak and is trans-activated by NS1. Stable cell lines were isolated that contained pP38NS1cat that was constitutively silent, but inducible with exogenous NS1 by superinfection or by treatment with sodium butyrate. The cells that were induced for this self-stimulatory genetic circuit did not remain in the culture, suggesting that expression of NS1-NS2 is cytotoxic or that the expression is not sustained. The properties of these cell lines and an example of the construction of a cell line inducible for expression of the viral coat protein gene and the bacterial gene for chloramphenicol acetyltransferase (cat) are described.

Full text

PDF
1448

Images in this article

Selected References

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

  1. Astell C. R., Gardiner E. M., Tattersall P. DNA sequence of the lymphotropic variant of minute virus of mice, MVM(i), and comparison with the DNA sequence of the fibrotropic prototype strain. J Virol. 1986 Feb;57(2):656–669. doi: 10.1128/jvi.57.2.656-669.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Astell C. R., Thomson M., Merchlinsky M., Ward D. C. The complete DNA sequence of minute virus of mice, an autonomous parvovirus. Nucleic Acids Res. 1983 Feb 25;11(4):999–1018. doi: 10.1093/nar/11.4.999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Candido E. P., Reeves R., Davie J. R. Sodium butyrate inhibits histone deacetylation in cultured cells. Cell. 1978 May;14(1):105–113. doi: 10.1016/0092-8674(78)90305-7. [DOI] [PubMed] [Google Scholar]
  4. Cotmore S. F., Sturzenbecker L. J., Tattersall P. The autonomous parvovirus MVM encodes two nonstructural proteins in addition to its capsid polypeptides. Virology. 1983 Sep;129(2):333–343. doi: 10.1016/0042-6822(83)90172-1. [DOI] [PubMed] [Google Scholar]
  5. Cotmore S. F., Tattersall P. Organization of nonstructural genes of the autonomous parvovirus minute virus of mice. J Virol. 1986 Jun;58(3):724–732. doi: 10.1128/jvi.58.3.724-732.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gorman C. M., Howard B. H., Reeves R. Expression of recombinant plasmids in mammalian cells is enhanced by sodium butyrate. Nucleic Acids Res. 1983 Nov 11;11(21):7631–7648. doi: 10.1093/nar/11.21.7631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Graham F. L., Smiley J., Russell W. C., Nairn R. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol. 1977 Jul;36(1):59–74. doi: 10.1099/0022-1317-36-1-59. [DOI] [PubMed] [Google Scholar]
  8. Jongeneel C. V., Sahli R., McMaster G. K., Hirt B. A precise map of splice junctions in the mRNAs of minute virus of mice, an autonomous parvovirus. J Virol. 1986 Sep;59(3):564–573. doi: 10.1128/jvi.59.3.564-573.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Keller J. M., Alwine J. C. Activation of the SV40 late promoter: direct effects of T antigen in the absence of viral DNA replication. Cell. 1984 Feb;36(2):381–389. doi: 10.1016/0092-8674(84)90231-9. [DOI] [PubMed] [Google Scholar]
  10. Labow M. A., Hermonat P. L., Berns K. I. Positive and negative autoregulation of the adeno-associated virus type 2 genome. J Virol. 1986 Oct;60(1):251–258. doi: 10.1128/jvi.60.1.251-258.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lebovitz R. M., Roeder R. G. Parvovirus H-1 expression: mapping of the abundant cytoplasmic transcripts and identification of promoter sites and overlapping transcription units. J Virol. 1986 May;58(2):271–280. doi: 10.1128/jvi.58.2.271-280.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Molitor T. W., Joo H. S., Collett M. S. Identification and characterization of a porcine parvovirus nonstructural polypeptide. J Virol. 1985 Sep;55(3):554–559. doi: 10.1128/jvi.55.3.554-559.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mulligan R. C., Berg P. Selection for animal cells that express the Escherichia coli gene coding for xanthine-guanine phosphoribosyltransferase. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2072–2076. doi: 10.1073/pnas.78.4.2072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nelson D. A., Ferris R. C., Zhang D. E., Ferenz C. R. The beta-globin domain in immature chicken erythrocytes: enhanced solubility is coincident with histone hyperacetylation. Nucleic Acids Res. 1986 Feb 25;14(4):1667–1682. doi: 10.1093/nar/14.4.1667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nevins J. R. Mechanism of activation of early viral transcription by the adenovirus E1A gene product. Cell. 1981 Oct;26(2 Pt 2):213–220. doi: 10.1016/0092-8674(81)90304-4. [DOI] [PubMed] [Google Scholar]
  16. Ostrowski M. C., Huang A. L., Kessel M., Wolford R. G., Hager G. L. Modulation of enhancer activity by the hormone responsive regulatory element from mouse mammary tumor virus. EMBO J. 1984 Aug;3(8):1891–1899. doi: 10.1002/j.1460-2075.1984.tb02064.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Paradiso P. R. Identification of multiple forms of the noncapsid parvovirus protein NCVP1 in H-1 parvovirus-infected cells. J Virol. 1984 Oct;52(1):82–87. doi: 10.1128/jvi.52.1.82-87.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Paradiso P. R., Williams K. R., Costantino R. L. Mapping of the amino terminus of the H-1 parvovirus major capsid protein. J Virol. 1984 Oct;52(1):77–81. doi: 10.1128/jvi.52.1.77-81.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pintel D., Merchlinsky M. J., Ward D. C. Expression of minute virus of mice structural proteins in murine cell lines transformed by bovine papillomavirus-minute virus of mice plasmid chimera. J Virol. 1984 Nov;52(2):320–327. doi: 10.1128/jvi.52.2.320-327.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Reeves R., Gorman C. M., Howard B. Minichromosome assembly of non-integrated plasmid DNA transfected into mammalian cells. Nucleic Acids Res. 1985 May 24;13(10):3599–3615. doi: 10.1093/nar/13.10.3599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rhode S. L., 3rd, Paradiso P. R. Parvovirus genome: nucleotide sequence of H-1 and mapping of its genes by hybrid-arrested translation. J Virol. 1983 Jan;45(1):173–184. doi: 10.1128/jvi.45.1.173-184.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rhode S. L., 3rd Replication process of the parvovirus H-1 V. Isolation and characterization of temperature-sensitive H-1 mutants. J Virol. 1976 Feb;17(2):659–667. doi: 10.1128/jvi.17.2.659-667.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rhode S. L., 3rd trans-Activation of parvovirus P38 promoter by the 76K noncapsid protein. J Virol. 1985 Sep;55(3):886–889. doi: 10.1128/jvi.55.3.886-889.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. SHEIN H. M., ENDERS J. F. Multiplication and cytopathogenicity of Simian vacuolating virus 40 in cultures of human tissues. Proc Soc Exp Biol Med. 1962 Mar;109:495–500. doi: 10.3181/00379727-109-27246. [DOI] [PubMed] [Google Scholar]
  25. Sahli R., McMaster G. K., Hirt B. DNA sequence comparison between two tissue-specific variants of the autonomous parvovirus, minute virus of mice. Nucleic Acids Res. 1985 May 24;13(10):3617–3633. doi: 10.1093/nar/13.10.3617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sealy L., Chalkley R. The effect of sodium butyrate on histone modification. Cell. 1978 May;14(1):115–121. doi: 10.1016/0092-8674(78)90306-9. [DOI] [PubMed] [Google Scholar]
  27. Singh N., Cerutti P. Poly ADP-ribosylation of histones in tumor promoter phorbol-12-myristate-13-acetate treated mouse embryo fibroblasts C3H10T1/2. Biochem Biophys Res Commun. 1985 Oct 30;132(2):811–819. doi: 10.1016/0006-291x(85)91204-5. [DOI] [PubMed] [Google Scholar]
  28. Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
  29. Topp W. C. Normal rat cell lines deficient in nuclear thymidine kinase. Virology. 1981 Aug;113(1):408–411. doi: 10.1016/0042-6822(81)90168-9. [DOI] [PubMed] [Google Scholar]
  30. Tratschin J. D., Tal J., Carter B. J. Negative and positive regulation in trans of gene expression from adeno-associated virus vectors in mammalian cells by a viral rep gene product. Mol Cell Biol. 1986 Aug;6(8):2884–2894. doi: 10.1128/mcb.6.8.2884. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES