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PMID: 2179950 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Mutations that increase the activity of a transcriptional activator in yeast and mammalian cells.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 87 ·No. 6 ·1990-03-00 ·Pages 2127-31

Gill G, Sadowski I, Ptashne M

Abstract

Activating region I of GAL4 protein is a stretch of amino acids, positioned adjacent to the DNA-binding region, that activates transcription in yeast and, as we show here, in mammalian cells. Here we describe mutations located throughout a 65-amino acid region that increase the activation function of region I. Most of these mutations replace positively charged amino acids in the region with neutral ones, although we also describe substitutions at one position that do not alter the charge of the region. Mutations of region I that alter the activation function in yeast have similar effects on activation when assayed in mammalian cells. When individual mutations that raise the acidity of the activating region are recombined, the activities of the mutant proteins increase with increasing negative charge in both yeast and mammalian cells. These results extend and modify the correlation between acidity and activation and suggest that the requirements for a strong activating region are conserved in yeast and mammals.

MeSH Terms
Amino Acids Animals Cell Line Chromosome Deletion Cloning, Molecular DNA-Binding Proteins Fungal Proteins/genetics,metabolism Mutation Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins Transcription Factors/genetics
Chemicals
Amino Acids DNA-Binding Proteins Fungal Proteins GAL4 protein, S cerevisiae Saccharomyces cerevisiae Proteins Transcription Factors
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Gill G
Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, MA 02138.
Sadowski I
Ptashne M
References (38)
38 references, click to expand
  1. A single glutamic acid residue plays a key role in the transcriptional activation function of lambda repressor.
    Cell. 1989 Sep 22;58(6):1163-71 PMID: 2570642
  2. Transcription activation by the adenovirus E1a protein.
    Nature. 1989 Mar 2;338(6210):39-44 PMID: 2521923
  3. Transformation of intact yeast cells treated with alkali cations.
    J Bacteriol. 1983 Jan;153(1):163-8 PMID: 6336730
  4. A GAL10-CYC1 hybrid yeast promoter identifies the GAL4 regulatory region as an upstream site.
    Proc Natl Acad Sci U S A. 1982 Dec;79(23):7410-4 PMID: 6760197
  5. Repressor structure and the mechanism of positive control.
    Cell. 1983 Feb;32(2):319-25 PMID: 6218886
  6. Mapping of multiple phosphorylation sites within the structural and catalytic domains of the Fujinami avian sarcoma virus transforming protein.
    J Virol. 1983 Apr;46(1):29-41 PMID: 6298463
  7. Expression and regulation of Escherichia coli lacZ gene fusions in mammalian cells.
    J Mol Appl Genet. 1983;2(1):101-9 PMID: 6302193
  8. Primary structure of the Saccharomyces cerevisiae GAL4 gene.
    Mol Cell Biol. 1984 Feb;4(2):260-7 PMID: 6366516
  9. Saccharomyces cerevisiae GAL1-GAL10 divergent promoter region: location and function of the upstream activating sequence UASG.
    Mol Cell Biol. 1984 Nov;4(11):2467-78 PMID: 6392852
  10. Specific protein binding to far upstream activating sequences in polymerase II promoters.
    Proc Natl Acad Sci U S A. 1985 Jan;82(1):43-7 PMID: 3881758
  11. Rapid and efficient site-specific mutagenesis without phenotypic selection.
    Proc Natl Acad Sci U S A. 1985 Jan;82(2):488-92 PMID: 3881765
  12. Specific DNA binding of GAL4, a positive regulatory protein of yeast.
    Cell. 1985 Apr;40(4):767-74 PMID: 3886158
  13. A general method for saturation mutagenesis of cloned DNA fragments.
    Science. 1985 Jul 19;229(4710):242-7 PMID: 2990046
  14. A eukaryotic transcriptional activator bearing the DNA specificity of a prokaryotic repressor.
    Cell. 1985 Dec;43(3 Pt 2):729-36 PMID: 3907859
  15. Separation of DNA binding from the transcription-activating function of a eukaryotic regulatory protein.
    Science. 1986 Feb 14;231(4739):699-704 PMID: 3080805
  16. Replacement of insulin receptor tyrosine residues 1162 and 1163 compromises insulin-stimulated kinase activity and uptake of 2-deoxyglucose.
    Cell. 1986 Jun 6;45(5):721-32 PMID: 3518947
  17. Single amino acid substitution, from Glu1025 to Asp, of the fps oncogenic protein causes temperature sensitivity in transformation and kinase activity.
    Virology. 1986 Nov;155(1):106-19 PMID: 2877522
  18. Deletion analysis of GAL4 defines two transcriptional activating segments.
    Cell. 1987 Mar 13;48(5):847-53 PMID: 3028647
  19. Promoters, activator proteins, and the mechanism of transcriptional initiation in yeast.
    Cell. 1987 May 8;49(3):295-7 PMID: 2882858
  20. A new class of yeast transcriptional activators.
    Cell. 1987 Oct 9;51(1):113-9 PMID: 3115591
  21. Mutants of GAL4 protein altered in an activation function.
    Cell. 1987 Oct 9;51(1):121-6 PMID: 3115592
  22. Transcription in yeast activated by a putative amphipathic alpha helix linked to a DNA binding unit.
    Nature. 1987 Dec 17-23;330(6149):670-2 PMID: 3317067
  23. The C-terminal domain of the largest subunit of RNA polymerase II of Saccharomyces cerevisiae, Drosophila melanogaster, and mammals: a conserved structure with an essential function.
    Mol Cell Biol. 1988 Jan;8(1):321-9 PMID: 3122024
  24. Cooperative DNA binding of the yeast transcriptional activator GAL4.
    Proc Natl Acad Sci U S A. 1988 Jan;85(2):382-6 PMID: 3124106
  25. GAL4 activates gene expression in mammalian cells.
    Cell. 1988 Jan 29;52(2):161-7 PMID: 2830021
  26. The yeast UASG is a transcriptional enhancer in human HeLa cells in the presence of the GAL4 trans-activator.
    Cell. 1988 Jan 29;52(2):169-78 PMID: 2830022
  27. GAL4 activates transcription in Drosophila.
    Nature. 1988 Apr 28;332(6167):853-6 PMID: 3128741
  28. Structural and functional characterization of the short acidic transcriptional activation region of yeast GCN4 protein.
    Nature. 1988 Jun 16;333(6174):635-40 PMID: 3287180
  29. Function of a yeast TATA element-binding protein in a mammalian transcription system.
    Nature. 1988 Jul 7;334(6177):37-42 PMID: 3290687
  30. A yeast activity can substitute for the HeLa cell TATA box factor.
    Nature. 1988 Jul 7;334(6177):77-80 PMID: 3290688
  31. Turning lambda Cro into a transcriptional activator.
    Cell. 1988 Jul 15;54(2):191-7 PMID: 2968842
  32. Yeast activators stimulate plant gene expression.
    Nature. 1988 Aug 18;334(6183):631-3 PMID: 3165494
  33. Mechanism of action of a yeast activator: direct effect of GAL4 derivatives on mammalian TFIID-promoter interactions.
    Cell. 1988 Aug 26;54(5):665-9 PMID: 3044608
  34. Negative effect of the transcriptional activator GAL4.
    Nature. 1988 Aug 25;334(6184):721-4 PMID: 3412449
  35. 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
  36. GAL4-VP16 is an unusually potent transcriptional activator.
    Nature. 1988 Oct 6;335(6190):563-4 PMID: 3047590
  37. How eukaryotic transcriptional activators work.
    Nature. 1988 Oct 20;335(6192):683-9 PMID: 3050531
  38. Mutant lambda phage repressor with a specific defect in its positive control function.
    Proc Natl Acad Sci U S A. 1982 Apr;79(7):2236-9 PMID: 6212933
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.87.6.2127
Published
1990-03-00
Pages
2127-31
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC53639
Subset
IM
Grants
NIGMS NIH HHS · GM32308 · United States
External Links
PubMed source DOI: 10.1073/pnas.87.6.2127 PMC Full Text View journal details
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