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PMID: 9207017 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Tetracycline-controlled transcription in eukaryotes: novel transactivators with graded transactivation potential.

Nucleic acids research ·Vol. 25 ·No. 14 ·1997-07-15 ·Pages 2723-9

Baron U, Gossen M, Bujard H

Abstract

Several tetracycline-controlled transactivators (tTA) were generated which differ in their activation potential by >3 orders of magnitude. The transactivators are fusions between the Tet repressor and minimal transcriptional activation domains derived from Herpes simplex virus protein 16 (VP16). By reducing the VP16 moiety of the previously described tTA to 12 amino acids, potential targets for interactions with various cellular transcription factors were eliminated, as were potential epitopes which may elicit a cellular immune response. When compared with the originally described tTA, these new transactivators are tolerated at higher intracellular concentrations. This will facilitate establishment of tet regulatory systems under a variety of conditions, but particularly when cell type-restricted tetracycline-controlled gene expression is to be achieved in transgenic organisms via homologous recombination.

MeSH Terms
Bacterial Proteins/genetics Carrier Proteins Eukaryotic Cells HeLa Cells Herpes Simplex Virus Protein Vmw65/genetics Humans Luciferases/metabolism Recombinant Fusion Proteins/genetics Repressor Proteins/genetics Tetracycline/pharmacology Trans-Activators/drug effects,metabolism Transcription, Genetic/drug effects Transcriptional Activation
Chemicals
Bacterial Proteins Carrier Proteins Herpes Simplex Virus Protein Vmw65 Recombinant Fusion Proteins Repressor Proteins Tet O resistance protein, Bacteria Trans-Activators tetracycline resistance-encoding transposon repressor protein Luciferases Tetracycline
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Baron U
ZMBH, Im Neuenheimer Feld 282, 69120 Heidelberg, Germany.
Gossen M
Bujard H
References (26)
26 references, click to expand
  1. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  2. Long-term control of erythropoietin secretion by doxycycline in mice transplanted with engineered primary myoblasts.
    Nat Med. 1997 Mar;3(3):299-305 PMID: 9055857
  3. A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus.
    Cell. 1985 Jun;41(2):521-30 PMID: 2985280
  4. Construction of a fusion gene that confers resistance against hygromycin B to mammalian cells in culture.
    Exp Cell Res. 1985 May;158(1):237-43 PMID: 2987009
  5. Dynamics of repressor-operator recognition: the Tn10-encoded tetracycline resistance control.
    Biochemistry. 1988 Feb 23;27(4):1094-104 PMID: 2835082
  6. Negative effect of the transcriptional activator GAL4.
    Nature. 1988 Aug 25;334(6184):721-4 PMID: 3412449
  7. Functional dissection of VP16, the trans-activator of herpes simplex virus immediate early gene expression.
    Genes Dev. 1988 Jun;2(6):718-29 PMID: 2843425
  8. Direct and selective binding of an acidic transcriptional activation domain to the TATA-box factor TFIID.
    Nature. 1990 Jun 28;345(6278):783-6 PMID: 2193231
  9. A novel mediator between activator proteins and the RNA polymerase II transcription apparatus.
    Cell. 1990 Jun 29;61(7):1209-15 PMID: 2163759
  10. Binding of general transcription factor TFIIB to an acidic activating region.
    Nature. 1991 Oct 10;353(6344):569-71 PMID: 1922364
  11. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters.
    Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5547-51 PMID: 1319065
  12. Genetic isolation of ADA2: a potential transcriptional adaptor required for function of certain acidic activation domains.
    Cell. 1992 Jul 24;70(2):251-65 PMID: 1638630
  13. Different activation domains stimulate transcription from remote ('enhancer') and proximal ('promoter') positions.
    EMBO J. 1992 Dec;11(13):4961-8 PMID: 1464321
  14. Estradiol-inducible squelching and cell growth arrest by a chimeric VP16-estrogen receptor expressed in Saccharomyces cerevisiae: suppression by an allele of PDR1.
    Mol Cell Biol. 1993 Jan;13(1):462-72 PMID: 8417344
  15. Mapping of a major surface-exposed site in herpes simplex virus protein Vmw65 to a region of direct interaction in a transcription complex assembly.
    J Virol. 1993 Feb;67(2):852-62 PMID: 8380468
  16. Pattern of aromatic and hydrophobic amino acids critical for one of two subdomains of the VP16 transcriptional activator.
    Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):883-7 PMID: 8381535
  17. Drosophila TAFII40 interacts with both a VP16 activation domain and the basal transcription factor TFIIB.
    Cell. 1993 Nov 5;75(3):519-30 PMID: 8221891
  18. Acceleration of the G1/S phase transition by expression of cyclins D1 and E with an inducible system.
    Mol Cell Biol. 1994 Mar;14(3):1669-79 PMID: 8114703
  19. Transcriptional activation by herpes simplex virus type 1 VP16 in vitro and its inhibition by oligopeptides.
    Mol Cell Biol. 1994 May;14(5):3484-93 PMID: 8164693
  20. A chimeric transactivator allows tetracycline-responsive gene expression in whole plants.
    Plant J. 1994 Apr;5(4):559-69 PMID: 8012406
  21. Binding of basal transcription factor TFIIH to the acidic activation domains of VP16 and p53.
    Mol Cell Biol. 1994 Oct;14(10):7013-24 PMID: 7935417
  22. Yeast ADA2 protein binds to the VP16 protein activation domain and activates transcription.
    Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11665-8 PMID: 7972120
  23. Transcriptional activation by tetracyclines in mammalian cells.
    Science. 1995 Jun 23;268(5218):1766-9 PMID: 7792603
  24. Identification of human proteins functionally conserved with the yeast putative adaptors ADA2 and GCN5.
    Mol Cell Biol. 1996 Feb;16(2):593-602 PMID: 8552087
  25. Doxycycline-mediated quantitative and tissue-specific control of gene expression in transgenic mice.
    Proc Natl Acad Sci U S A. 1996 Oct 1;93(20):10933-8 PMID: 8855286
  26. Comparison of initiation of protein synthesis in procaryotes, eucaryotes, and organelles.
    Microbiol Rev. 1983 Mar;47(1):1-45 PMID: 6343825
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/25.14.2723
Published
1997-07-15
Pages
2723-9
Language
English
Region
England
NLM ID
0411011
PMCID
PMC146828
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/25.14.2723 PMC Full Text View journal details
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