Synergistic activation of a human promoter in vivo by transcription factor Sp1

Anderson, G M; Freytag, S O

doi:10.1128/MCB.11.4.1935

Title: Synergistic activation of a human promoter in vivo by transcription factor Sp1

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Abstract

Many eucaryotic promoters contain multiple binding sites for sequence-specific DNA-binding proteins. In some cases, these proteins have been shown to interact synergistically to activate transcription. In this study, the authors address the possibility that the transcription factor Sp1 can synergistically activate a native human promoter in a cellular context that closely resembles that of a single-copy gene. Using DNase I footprinting with affinity-purified Sp1, they show that the human argininosuccinate synthetase (AS) promoter contains three sites that bind Sp1 with different affinities. These binding sites were mutated to abolish Sp1 binding, individually and in all possible combinations, to generate a series of AS promoter-CAT constructs was then measured in stably transfected human RPMI 2650 cells lines. The results show that each of the three Sp1-binding sites contributes to full activation of the human AS promoter and the relative contribution of each site correlates well with its in vitro affinity for Sp1. They provide direct evidence that Sp1-binding sites in their native context in a human promoter can interact synergistically in vivo to activate transcription. The ability to activate transcription synergistically may be the reason that many cellular promoters have multiple Sp1-binding sites arranged in tandem and in close proximity.

Authors:

Anderson, G M ^[1]; Freytag, S O ^[2]

Henry Ford Hospital, Detroit, MI (United States)
University of Michigan, Ann Arbor (United States)

Publication Date:: Mon Apr 01 00:00:00 EST 1991

OSTI Identifier:: 6969204

Resource Type:: Journal Article

Journal Name:: Molecular and Cellular Biology; (United States)

Additional Journal Information:: Journal Volume: 11:4; Journal ID: ISSN 0270-7306

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; GENE REPRESSORS; CONFIGURATION INTERACTION; LIGASES; GENE REGULATION; TRANSCRIPTION FACTORS; DNA-ASE; RNA POLYMERASES; ENZYMES; ESTERASES; HYDROLASES; NUCLEOTIDYLTRANSFERASES; ORGANIC COMPOUNDS; PHOSPHODIESTERASES; PHOSPHORUS-GROUP TRANSFERASES; POLYMERASES; PROTEINS; TRANSFERASES; 550200* - Biochemistry

Citation Formats


                    Anderson, G M, and Freytag, S O. Synergistic activation of a human promoter in vivo by transcription factor Sp1.  United States: N. p., 1991. 
        Web.  doi:10.1128/MCB.11.4.1935.

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                    Anderson, G M, & Freytag, S O. Synergistic activation of a human promoter in vivo by transcription factor Sp1.  United States.  https://doi.org/10.1128/MCB.11.4.1935

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                    Anderson, G M, and Freytag, S O. 1991.  
        "Synergistic activation of a human promoter in vivo by transcription factor Sp1".  United States.  https://doi.org/10.1128/MCB.11.4.1935.

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@article{osti_6969204,

  title        = {Synergistic activation of a human promoter in vivo by transcription factor Sp1},

  author       = {Anderson, G M and Freytag, S O},

  abstractNote = {Many eucaryotic promoters contain multiple binding sites for sequence-specific DNA-binding proteins. In some cases, these proteins have been shown to interact synergistically to activate transcription. In this study, the authors address the possibility that the transcription factor Sp1 can synergistically activate a native human promoter in a cellular context that closely resembles that of a single-copy gene. Using DNase I footprinting with affinity-purified Sp1, they show that the human argininosuccinate synthetase (AS) promoter contains three sites that bind Sp1 with different affinities. These binding sites were mutated to abolish Sp1 binding, individually and in all possible combinations, to generate a series of AS promoter-CAT constructs was then measured in stably transfected human RPMI 2650 cells lines. The results show that each of the three Sp1-binding sites contributes to full activation of the human AS promoter and the relative contribution of each site correlates well with its in vitro affinity for Sp1. They provide direct evidence that Sp1-binding sites in their native context in a human promoter can interact synergistically in vivo to activate transcription. The ability to activate transcription synergistically may be the reason that many cellular promoters have multiple Sp1-binding sites arranged in tandem and in close proximity.},

  doi          = {10.1128/MCB.11.4.1935},

  url          = {https://www.osti.gov/biblio/6969204},
  journal      = {Molecular and Cellular Biology; (United States)},

  issn         = {0270-7306},

  number       = ,

  volume       = 11:4,

  place        = {United States},

  year         = {Mon Apr 01 00:00:00 EST 1991},

  month        = {Mon Apr 01 00:00:00 EST 1991}

}

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