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PMID: 2821493 Published · ppublish English Journal Article

Optimizing gene expression in BPV-transformed cells: effects of cell type on enhancer/promoter interaction.

Nucleic acids research ·Vol. 15 ·No. 17 ·1987-09-11 ·Pages 7137-53

Hurwitz DR, Hodges R, Drohan W, Sarver N

Abstract

We have compared several combinations of enhancers and promoters in expressing the chloramphenicol acetyl transferase gene in transient assays, in mouse C127, the most widely used host cell for the bovine papilloma virus (BPV) expression vector. Of the various combinations tested, the unit comprised of the SV40 enhancer and adenovirus type 2 major late promoter (MLP) was the most active in BPV transformed C127 cells. We further demonstrate that untransformed and BPV transformed C127 cells respond differently to the various enhancer/promoter combinations tested. Moving the SV40 enhancer closer to the cap site of a complete MLP (from -414 to -107) reduced potentiation to less than half in BPV transformed cells. The level of potentiation with enhancer at either site was similar in human HeLa cells. In BPV transformed C127 cells, the SV40 enhancer and the MLP (at the -414 site) supports 4-5 fold greater levels of expression than the murine sarcoma virus (MSV) enhancer/mouse metallothionein (MT) promoter which has previously been extremely effective in BPV vectors. Our findings provide a basis for the improvement of the BPV vector system in supporting increased levels of expression of proteins of important therapeutic application.

MeSH Terms
Acetyltransferases/genetics Animals Base Sequence Bovine papillomavirus 1/genetics Cell Line Cell Transformation, Neoplastic Chloramphenicol O-Acetyltransferase Cytomegalovirus/genetics Enhancer Elements, Genetic Genes Genes, Viral Genetic Vectors Papillomaviridae/genetics Plasmids Promoter Regions, Genetic Simian virus 40/genetics Transcription, Genetic Transfection
Chemicals
Acetyltransferases Chloramphenicol O-Acetyltransferase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Hurwitz D R
Rorer Biotechnology Inc., Molecular Biology Development Division, Springfield, VA 22151.
Hodges R
Drohan W
Sarver N
References (49)
49 references, click to expand
  1. Expression and regulation of a human metallothionein gene carried on an autonomously replicating shuttle vector.
    Proc Natl Acad Sci U S A. 1983 Jul;80(13):4040-4 PMID: 6306660
  2. Bovine papilloma virus deoxyribonucleic acid: a novel eucaryotic cloning vector.
    Mol Cell Biol. 1981 Jun;1(6):486-96 PMID: 6100967
  3. A lymphocyte-specific cellular enhancer is located downstream of the joining region in immunoglobulin heavy chain genes.
    Cell. 1983 Jul;33(3):729-40 PMID: 6409418
  4. Enhanced production of hepatitis B surface antigen in NIH 3T3 mouse fibroblasts by using extrachromosomally replicating bovine papillomavirus vector.
    Mol Cell Biol. 1983 Jun;3(6):1032-9 PMID: 6308420
  5. Identification of two distinct regulatory regions adjacent to the human beta-interferon gene.
    Cell. 1983 Oct;34(3):865-79 PMID: 6313211
  6. Secretion of the hepatitis B virus surface antigen from mouse cells using an extra-chromosomal eucaryotic vector.
    EMBO J. 1983;2(5):669-73 PMID: 6641713
  7. Glucocorticoid regulation of transcription at an amplified, episomal promoter.
    Mol Cell Biol. 1983 Nov;3(11):2045-57 PMID: 6318079
  8. Stimulation of in vitro transcription from heterologous promoters by the simian virus 40 enhancer.
    Proc Natl Acad Sci U S A. 1984 Jan;81(2):308-12 PMID: 6320179
  9. Expression of dihydrofolate reductase, and of the adjacent EIb region, in an Ad5-dihydrofolate reductase recombinant virus.
    Nucleic Acids Res. 1984 Feb 24;12(4):1925-41 PMID: 6199745
  10. Analysis of gene expression using simian virus 40 vectors.
    Anal Biochem. 1983 Nov;135(1):1-15 PMID: 6322609
  11. Adenovirus tripartite leader sequence enhances translation of mRNAs late after infection.
    Proc Natl Acad Sci U S A. 1984 Jun;81(12):3655-9 PMID: 6587381
  12. Short and long range activation by the SV40 enhancer.
    Nucleic Acids Res. 1984 Jul 25;12(14):5589-608 PMID: 6087293
  13. Constitutive production of human interferons by mouse cells with bovine papillomavirus as a vector.
    Proc Natl Acad Sci U S A. 1984 Aug;81(16):5086-90 PMID: 6089176
  14. Promoter dependence of enhancer activity.
    Mol Cell Biol. 1984 Aug;4(8):1664-8 PMID: 6092930
  15. Synthesis and glycosylation of the common alpha subunit of human glycoprotein hormones in mouse cells.
    Proc Natl Acad Sci U S A. 1984 Nov;81(21):6701-5 PMID: 6208554
  16. Stimulation of in vitro transcription by the upstream element of the adenovirus-2 major late promoter involves a specific factor.
    Nucleic Acids Res. 1984 Dec 11;12(23):8779-99 PMID: 6096806
  17. Duplications of a mutated simian virus 40 enhancer restore its activity.
    Nature. 1985 Feb 21-27;313(6004):711-4 PMID: 2983220
  18. Expression of hepatitis B virus surface and e antigen genes cloned in bovine papillomavirus vectors.
    Gene. 1984 Dec;32(3):357-68 PMID: 6099321
  19. A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus.
    Cell. 1985 Jun;41(2):521-30 PMID: 2985280
  20. Effect of the tripartite leader on synthesis of a non-viral protein in an adenovirus 5 recombinant.
    Nucleic Acids Res. 1985 Feb 11;13(3):841-57 PMID: 3839074
  21. Factors involved in specific transcription by human RNA polymerase II: analysis by a rapid and quantitative in vitro assay.
    Proc Natl Acad Sci U S A. 1985 Jul;82(13):4394-8 PMID: 3925456
  22. Transactivation of a bovine papilloma virus transcriptional regulatory element by the E2 gene product.
    Cell. 1985 Aug;42(1):183-91 PMID: 2990724
  23. Lines of BPV-transformed murine cells that constitutively express influenza virus hemagglutinin.
    EMBO J. 1985 Jan;4(1):91-103 PMID: 2990897
  24. Negative regulation of viral enhancers in undifferentiated embryonic stem cells.
    Cell. 1985 Sep;42(2):519-26 PMID: 2992802
  25. A new technique for the assay of infectivity of human adenovirus 5 DNA.
    Virology. 1973 Apr;52(2):456-67 PMID: 4705382
  26. Helper-independent transformation by unintegrated Harvey sarcoma virus DNA.
    J Virol. 1978 May;26(2):291-8 PMID: 26810
  27. Mapping temperature-sensitive and host-range mutations of adenovirus type 5 by marker rescue.
    Virology. 1978 Nov;91(1):39-50 PMID: 726266
  28. Nucleotide sequence analysis of the leader segments in a cloned copy of adenovirus 2 fiber mRNA.
    Cell. 1979 Apr;16(4):851-61 PMID: 455453
  29. Selective and accurate initiation of transcription at the Ad2 major late promotor in a soluble system dependent on purified RNA polymerase II and DNA.
    Cell. 1979 Oct;18(2):469-84 PMID: 498279
  30. In vitro tumorigenic transformation by a defined sub-genomic fragment of bovine papilloma virus DNA.
    Nature. 1980 Sep 4;287(5777):72-4 PMID: 6251381
  31. High-level expression of the bovine growth hormone gene in heterologous mammalian cells.
    Gene. 1985;38(1-3):111-8 PMID: 2998942
  32. Interaction of a gene-specific transcription factor with the adenovirus major late promoter upstream of the TATA box region.
    Cell. 1985 Nov;43(1):165-75 PMID: 4075392
  33. Nucleotide sequences of integrated Moloney sarcoma provirus long terminal repeats and their host and viral junctions.
    Proc Natl Acad Sci U S A. 1980 Jul;77(7):3937-41 PMID: 6254003
  34. Regulated expression of an extrachromosomal human beta-interferon gene in mouse cells.
    Proc Natl Acad Sci U S A. 1982 Aug;79(16):4897-901 PMID: 6956899
  35. Host-specific activation of transcription by tandem repeats from simian virus 40 and Moloney murine sarcoma virus.
    Proc Natl Acad Sci U S A. 1982 Nov;79(21):6453-7 PMID: 6292901
  36. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells.
    Mol Cell Biol. 1982 Sep;2(9):1044-51 PMID: 6960240
  37. Sequences upstream from the T-A-T-A box are required in vivo and in vitro for efficient transcription from the adenovirus serotype 2 major late promoter.
    Proc Natl Acad Sci U S A. 1982 Dec;79(23):7132-6 PMID: 6961401
  38. Transformation and replication in mouse cells of a bovine papillomavirus--pML2 plasmid vector that can be rescued in bacteria.
    Proc Natl Acad Sci U S A. 1982 Dec;79(23):7147-51 PMID: 6296820
  39. Interferon induces morphologic reversion with elimination of extrachromosomal viral genomes in bovine papillomavirus-transformed mouse cells.
    Proc Natl Acad Sci U S A. 1982 Dec;79(24):7914-8 PMID: 6185955
  40. Inducible expression of the human interferon beta 1 gene linked to a bovine papilloma virus DNA vector and maintained extrachromosomally in mouse cells.
    Mol Cell Biol. 1983 Feb;3(2):233-40 PMID: 6300659
  41. Regulation of a metallothionein-growth hormone hybrid gene in bovine papilloma virus.
    Proc Natl Acad Sci U S A. 1983 Jan;80(2):397-401 PMID: 6300840
  42. Translational control of SV40 T antigen expressed from the adenovirus late promoter.
    Cell. 1983 Jun;33(2):455-64 PMID: 6305512
  43. An RNA polymerase II transcription factor binds to an upstream element in the adenovirus major late promoter.
    Cell. 1985 Dec;43(2 Pt 1):439-48 PMID: 4075400
  44. SV40 enhancer activation during retinoic acid-induced differentiation of F9 embryonal carcinoma cells.
    EMBO J. 1985 Dec 30;4(13B):3831-7 PMID: 3004973
  45. The SV40 enhancer is composed of multiple functional elements that can compensate for one another.
    Cell. 1986 May 9;45(3):461-70 PMID: 3009027
  46. Physical state, expression and regulation of two glucocorticoid-controlled genes on bovine papilloma virus vectors.
    J Mol Biol. 1986 Feb 20;187(4):557-68 PMID: 3012094
  47. Control of adenovirus late promoter expression in two human cell lines.
    Mol Cell Biol. 1985 Sep;5(9):2433-42 PMID: 2942764
  48. Downstream sequences affect transcription initiation from the adenovirus major late promoter.
    Mol Cell Biol. 1986 Jul;6(7):2684-94 PMID: 3023944
  49. A tissue-specific transcription enhancer element is located in the major intron of a rearranged immunoglobulin heavy chain gene.
    Cell. 1983 Jul;33(3):717-28 PMID: 6409417
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/15.17.7137
Published
1987-09-11
Pages
7137-53
Language
English
Region
England
NLM ID
0411011
PMCID
PMC306198
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/15.17.7137 PMC Full Text View journal details
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