Liver-specific expression of the mouse α-fetoprotein gene is mediated by cis-acting DNA elements

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Abstract

We have identified cis-acting regulatory elements in the 5' flanking region of the mouse α-fetoprotein (Afp) gene, using the expression of the bacterial gene for chloramphenicol acetyltransferase (CAT) in a transient expression assay. Tissue-specific enhancer activity was determined by transfection of mouse hepatoma (BWTG3) and fibroblast cells (C127, NIH 3T3) with various DNA fragments linked to the CAT gene. A 5.4-kilobase restriction fragment was shown to have characteristics typical of enhancers, including the ability to function independent of orientation and position and the ability to enhance transcription from a heterologous promoter. The enhancer activity was greatest in the hepatoma cells, which express Afp. By deletion analysis, it was demonstrated that enhancer activity is present in several subfragments, indicating the presence of more than one element in this fragment. An additional regulatory element within 950 base pairs of the Afp transcription initiation site has been identified and shown to confer tissue-specific expression on the CAT gene. This fragment, which lacks enhancer activity, contains the Afp promoter region and mediates the tissue-specific expression of the CAT gene when driven by nonspecific viral enhancers. We conclude from our studies that there are several types of regulatory elements in the 5' flanking region of the Afp gene that help mediate tissue-specific expression.

Original languageEnglish (US)
Pages (from-to)8196-8200
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume83
Issue number21
StatePublished - 1986

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Fetal Proteins
Chloramphenicol O-Acetyltransferase
5' Flanking Region
Liver
DNA
Genes
Hepatocellular Carcinoma
Bacterial Genes
NIH 3T3 Cells
Transcription Initiation Site
Genetic Promoter Regions
Base Pairing
Transfection
Fibroblasts
Gene Expression

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

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title = "Liver-specific expression of the mouse α-fetoprotein gene is mediated by cis-acting DNA elements",
abstract = "We have identified cis-acting regulatory elements in the 5' flanking region of the mouse α-fetoprotein (Afp) gene, using the expression of the bacterial gene for chloramphenicol acetyltransferase (CAT) in a transient expression assay. Tissue-specific enhancer activity was determined by transfection of mouse hepatoma (BWTG3) and fibroblast cells (C127, NIH 3T3) with various DNA fragments linked to the CAT gene. A 5.4-kilobase restriction fragment was shown to have characteristics typical of enhancers, including the ability to function independent of orientation and position and the ability to enhance transcription from a heterologous promoter. The enhancer activity was greatest in the hepatoma cells, which express Afp. By deletion analysis, it was demonstrated that enhancer activity is present in several subfragments, indicating the presence of more than one element in this fragment. An additional regulatory element within 950 base pairs of the Afp transcription initiation site has been identified and shown to confer tissue-specific expression on the CAT gene. This fragment, which lacks enhancer activity, contains the Afp promoter region and mediates the tissue-specific expression of the CAT gene when driven by nonspecific viral enhancers. We conclude from our studies that there are several types of regulatory elements in the 5' flanking region of the Afp gene that help mediate tissue-specific expression.",
author = "Steven Widen and John Papaconstantinou",
year = "1986",
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pages = "8196--8200",
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T1 - Liver-specific expression of the mouse α-fetoprotein gene is mediated by cis-acting DNA elements

AU - Widen, Steven

AU - Papaconstantinou, John

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N2 - We have identified cis-acting regulatory elements in the 5' flanking region of the mouse α-fetoprotein (Afp) gene, using the expression of the bacterial gene for chloramphenicol acetyltransferase (CAT) in a transient expression assay. Tissue-specific enhancer activity was determined by transfection of mouse hepatoma (BWTG3) and fibroblast cells (C127, NIH 3T3) with various DNA fragments linked to the CAT gene. A 5.4-kilobase restriction fragment was shown to have characteristics typical of enhancers, including the ability to function independent of orientation and position and the ability to enhance transcription from a heterologous promoter. The enhancer activity was greatest in the hepatoma cells, which express Afp. By deletion analysis, it was demonstrated that enhancer activity is present in several subfragments, indicating the presence of more than one element in this fragment. An additional regulatory element within 950 base pairs of the Afp transcription initiation site has been identified and shown to confer tissue-specific expression on the CAT gene. This fragment, which lacks enhancer activity, contains the Afp promoter region and mediates the tissue-specific expression of the CAT gene when driven by nonspecific viral enhancers. We conclude from our studies that there are several types of regulatory elements in the 5' flanking region of the Afp gene that help mediate tissue-specific expression.

AB - We have identified cis-acting regulatory elements in the 5' flanking region of the mouse α-fetoprotein (Afp) gene, using the expression of the bacterial gene for chloramphenicol acetyltransferase (CAT) in a transient expression assay. Tissue-specific enhancer activity was determined by transfection of mouse hepatoma (BWTG3) and fibroblast cells (C127, NIH 3T3) with various DNA fragments linked to the CAT gene. A 5.4-kilobase restriction fragment was shown to have characteristics typical of enhancers, including the ability to function independent of orientation and position and the ability to enhance transcription from a heterologous promoter. The enhancer activity was greatest in the hepatoma cells, which express Afp. By deletion analysis, it was demonstrated that enhancer activity is present in several subfragments, indicating the presence of more than one element in this fragment. An additional regulatory element within 950 base pairs of the Afp transcription initiation site has been identified and shown to confer tissue-specific expression on the CAT gene. This fragment, which lacks enhancer activity, contains the Afp promoter region and mediates the tissue-specific expression of the CAT gene when driven by nonspecific viral enhancers. We conclude from our studies that there are several types of regulatory elements in the 5' flanking region of the Afp gene that help mediate tissue-specific expression.

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