The ATF/CREB transcription factor-binding site in the polymerase β promoter mediates the positive effect of N-methyl-N′-nitro-N-nitrosoguanidine on transcription

Padmini S. Kedar, Steven Widen, Ella Englander, Albert J. Fornace, Samuel H. Wilson

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42 Citations (Scopus)

Abstract

DNA polymerase β(pol β) is a constitutively expressed DNA repair enzyme in vertebrate cells. Yet, it had been shown previously that the pol β mRNA level increases in Chinese hamster ovary (CHO) cells within 4 h after treatment with several monofunctional DNA damaging agents, notably, N-methyl-N′-nitro-N-nitrosoguanidine (MNNG). Herein we report that a transfected pol β promoter fusion gene is activated by MNNG treatment of CHO cells; mRNA from the transfected gene is ≈ 10-fold higher in treated cells than in untreated cells 16 h after treatment. This activation is mediated through the decamicleotide palindromic element GTGACGTCAC at positions -49 to -40 in the "TATA-less" core promoter. This clement, which is similar to the ATF/CREB transcription factor-binding site in a number of mammalian genes, forms the center of a strong protein-binding site for CHO cell nuclear extract proteins. Mutated pol β promoter fusion genes lacking the element fail to bind protein at this site and fail to respond to MNNG treatment of cells.

Original languageEnglish (US)
Pages (from-to)3729-3733
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume88
Issue number9
StatePublished - 1991
Externally publishedYes

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Activating Transcription Factors
Methylnitronitrosoguanidine
Binding Sites
Cricetulus
Nitrosoguanidines
Ovary
Gene Fusion
DNA Repair Enzymes
Messenger RNA
DNA-Directed DNA Polymerase
Nuclear Proteins
Cell Extracts
Protein Binding
Genes
Vertebrates
DNA

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

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title = "The ATF/CREB transcription factor-binding site in the polymerase β promoter mediates the positive effect of N-methyl-N′-nitro-N-nitrosoguanidine on transcription",
abstract = "DNA polymerase β(pol β) is a constitutively expressed DNA repair enzyme in vertebrate cells. Yet, it had been shown previously that the pol β mRNA level increases in Chinese hamster ovary (CHO) cells within 4 h after treatment with several monofunctional DNA damaging agents, notably, N-methyl-N′-nitro-N-nitrosoguanidine (MNNG). Herein we report that a transfected pol β promoter fusion gene is activated by MNNG treatment of CHO cells; mRNA from the transfected gene is ≈ 10-fold higher in treated cells than in untreated cells 16 h after treatment. This activation is mediated through the decamicleotide palindromic element GTGACGTCAC at positions -49 to -40 in the {"}TATA-less{"} core promoter. This clement, which is similar to the ATF/CREB transcription factor-binding site in a number of mammalian genes, forms the center of a strong protein-binding site for CHO cell nuclear extract proteins. Mutated pol β promoter fusion genes lacking the element fail to bind protein at this site and fail to respond to MNNG treatment of cells.",
author = "Kedar, {Padmini S.} and Steven Widen and Ella Englander and Fornace, {Albert J.} and Wilson, {Samuel H.}",
year = "1991",
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T1 - The ATF/CREB transcription factor-binding site in the polymerase β promoter mediates the positive effect of N-methyl-N′-nitro-N-nitrosoguanidine on transcription

AU - Kedar, Padmini S.

AU - Widen, Steven

AU - Englander, Ella

AU - Fornace, Albert J.

AU - Wilson, Samuel H.

PY - 1991

Y1 - 1991

N2 - DNA polymerase β(pol β) is a constitutively expressed DNA repair enzyme in vertebrate cells. Yet, it had been shown previously that the pol β mRNA level increases in Chinese hamster ovary (CHO) cells within 4 h after treatment with several monofunctional DNA damaging agents, notably, N-methyl-N′-nitro-N-nitrosoguanidine (MNNG). Herein we report that a transfected pol β promoter fusion gene is activated by MNNG treatment of CHO cells; mRNA from the transfected gene is ≈ 10-fold higher in treated cells than in untreated cells 16 h after treatment. This activation is mediated through the decamicleotide palindromic element GTGACGTCAC at positions -49 to -40 in the "TATA-less" core promoter. This clement, which is similar to the ATF/CREB transcription factor-binding site in a number of mammalian genes, forms the center of a strong protein-binding site for CHO cell nuclear extract proteins. Mutated pol β promoter fusion genes lacking the element fail to bind protein at this site and fail to respond to MNNG treatment of cells.

AB - DNA polymerase β(pol β) is a constitutively expressed DNA repair enzyme in vertebrate cells. Yet, it had been shown previously that the pol β mRNA level increases in Chinese hamster ovary (CHO) cells within 4 h after treatment with several monofunctional DNA damaging agents, notably, N-methyl-N′-nitro-N-nitrosoguanidine (MNNG). Herein we report that a transfected pol β promoter fusion gene is activated by MNNG treatment of CHO cells; mRNA from the transfected gene is ≈ 10-fold higher in treated cells than in untreated cells 16 h after treatment. This activation is mediated through the decamicleotide palindromic element GTGACGTCAC at positions -49 to -40 in the "TATA-less" core promoter. This clement, which is similar to the ATF/CREB transcription factor-binding site in a number of mammalian genes, forms the center of a strong protein-binding site for CHO cell nuclear extract proteins. Mutated pol β promoter fusion genes lacking the element fail to bind protein at this site and fail to respond to MNNG treatment of cells.

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