Functional analysis of the mouse alpha-fetoprotein enhancers and their subfragments in primary mouse hepatocyte cultures.
Identifieur interne : 004730 ( Main/Exploration ); précédent : 004729; suivant : 004731Functional analysis of the mouse alpha-fetoprotein enhancers and their subfragments in primary mouse hepatocyte cultures.
Auteurs : D E Zhang ; J P Rabek ; C C Hsieh ; C. Torres-Ramos ; J. PapaconstantinouSource :
- The Journal of biological chemistry [ 0021-9258 ] ; 1992.
Descripteurs français
- KwdFr :
- ADN, ARN messager (métabolisme), Alphafoetoprotéines (génétique), Animaux, Cellules cultivées, Chloramphenicol O-acetyltransferase (génétique), Chloramphenicol O-acetyltransferase (métabolisme), Données de séquences moléculaires, Encéphale (métabolisme), Facteurs de transcription (métabolisme), Foie (cytologie), Foie (métabolisme), Mutagenèse dirigée, Plasmides, Profilage d'ADN, Protéines de liaison à l'ADN (métabolisme), Protéines liant les séquences stimulatrices de type CCAAT, Protéines nucléaires (métabolisme), Rein (métabolisme), Souris, Séquence nucléotidique, Transfection, Éléments activateurs (génétique).
- MESH :
- cytologie : Foie.
- génétique : Alphafoetoprotéines, Chloramphenicol O-acetyltransferase.
- métabolisme : ARN messager, Chloramphenicol O-acetyltransferase, Encéphale, Facteurs de transcription, Foie, Protéines de liaison à l'ADN, Protéines nucléaires, Rein.
- ADN, Animaux, Cellules cultivées, Données de séquences moléculaires, Mutagenèse dirigée, Plasmides, Profilage d'ADN, Protéines liant les séquences stimulatrices de type CCAAT, Souris, Séquence nucléotidique, Transfection, Éléments activateurs (génétique).
English descriptors
- KwdEn :
- Animals, Base Sequence, Brain (metabolism), CCAAT-Enhancer-Binding Proteins, Cells, Cultured, Chloramphenicol O-Acetyltransferase (genetics), Chloramphenicol O-Acetyltransferase (metabolism), DNA, DNA Fingerprinting, DNA-Binding Proteins (metabolism), Enhancer Elements, Genetic, Kidney (metabolism), Liver (cytology), Liver (metabolism), Mice, Molecular Sequence Data, Mutagenesis, Site-Directed, Nuclear Proteins (metabolism), Plasmids, RNA, Messenger (metabolism), Transcription Factors (metabolism), Transfection, alpha-Fetoproteins (genetics).
- MESH :
- chemical , genetics : Chloramphenicol O-Acetyltransferase, alpha-Fetoproteins.
- chemical , metabolism : Chloramphenicol O-Acetyltransferase, DNA-Binding Proteins, Nuclear Proteins, RNA, Messenger, Transcription Factors.
- chemical : CCAAT-Enhancer-Binding Proteins, DNA.
- cytology : Liver.
- metabolism : Brain, Kidney, Liver.
- Animals, Base Sequence, Cells, Cultured, DNA Fingerprinting, Enhancer Elements, Genetic, Mice, Molecular Sequence Data, Mutagenesis, Site-Directed, Plasmids, Transfection.
Abstract
We have compared the activities of mouse alpha-fetoprotein (AFP) enhancers I, II, and III with their minimal enhancer fragments (Mers) I, II, and III and with the entire 7-kilobase pair enhancer domain by transient expression assay in primary fetal mouse liver cells. The level of expression directed by the AFP promoter [p(-1009)AFPcat] alone is stimulated at least 10-fold by the entire AFP enhancer domain (-1009 to -6983). Enhancer I can drive the level of chloramphenicol acetyltransferase activity equivalent to that of the entire enhancer domain, whereas the increase in activity by enhancers II and III is significantly lower (1.5-fold). MersI, II, and III all mediate a greater increase in activity than their corresponding enhancer regions. The increase with MerI is 16-fold. Using DNase I protection analyses we identified 3 protein-binding regions in MerI; site Ia binds liver and brain nuclear proteins; site Ib binds liver, kidney, and brain nuclear proteins as well as purified C/EBP; site Ic binds liver and kidney nuclear proteins. Site-specific mutation of Ia, Ib, or Ic showed a 10-25% reduction in chloramphenicol acetyltransferase expression; deletion of the C/EBP-binding site in Ib showed a 45% reduction in activity and mutation of all 3 sites (Ia, Ib, and Ic) resulted in a 75% reduction in activity. Our studies indicate no single trans-acting factor is absolutely essential for enhancer activity, and that the enhancer activity of MerI is mediated via a combinatorial and additive mechanism.
PubMed: 1375227
Affiliations:
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Le document en format XML
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<author><name sortKey="Zhang, D E" sort="Zhang, D E" uniqKey="Zhang D" first="D E" last="Zhang">D E Zhang</name>
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<author><name sortKey="Rabek, J P" sort="Rabek, J P" uniqKey="Rabek J" first="J P" last="Rabek">J P Rabek</name>
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<author><name sortKey="Hsieh, C C" sort="Hsieh, C C" uniqKey="Hsieh C" first="C C" last="Hsieh">C C Hsieh</name>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term>
<term>Base Sequence</term>
<term>Brain (metabolism)</term>
<term>CCAAT-Enhancer-Binding Proteins</term>
<term>Cells, Cultured</term>
<term>Chloramphenicol O-Acetyltransferase (genetics)</term>
<term>Chloramphenicol O-Acetyltransferase (metabolism)</term>
<term>DNA</term>
<term>DNA Fingerprinting</term>
<term>DNA-Binding Proteins (metabolism)</term>
<term>Enhancer Elements, Genetic</term>
<term>Kidney (metabolism)</term>
<term>Liver (cytology)</term>
<term>Liver (metabolism)</term>
<term>Mice</term>
<term>Molecular Sequence Data</term>
<term>Mutagenesis, Site-Directed</term>
<term>Nuclear Proteins (metabolism)</term>
<term>Plasmids</term>
<term>RNA, Messenger (metabolism)</term>
<term>Transcription Factors (metabolism)</term>
<term>Transfection</term>
<term>alpha-Fetoproteins (genetics)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>ADN</term>
<term>ARN messager (métabolisme)</term>
<term>Alphafoetoprotéines (génétique)</term>
<term>Animaux</term>
<term>Cellules cultivées</term>
<term>Chloramphenicol O-acetyltransferase (génétique)</term>
<term>Chloramphenicol O-acetyltransferase (métabolisme)</term>
<term>Données de séquences moléculaires</term>
<term>Encéphale (métabolisme)</term>
<term>Facteurs de transcription (métabolisme)</term>
<term>Foie (cytologie)</term>
<term>Foie (métabolisme)</term>
<term>Mutagenèse dirigée</term>
<term>Plasmides</term>
<term>Profilage d'ADN</term>
<term>Protéines de liaison à l'ADN (métabolisme)</term>
<term>Protéines liant les séquences stimulatrices de type CCAAT</term>
<term>Protéines nucléaires (métabolisme)</term>
<term>Rein (métabolisme)</term>
<term>Souris</term>
<term>Séquence nucléotidique</term>
<term>Transfection</term>
<term>Éléments activateurs (génétique)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Chloramphenicol O-Acetyltransferase</term>
<term>alpha-Fetoproteins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Chloramphenicol O-Acetyltransferase</term>
<term>DNA-Binding Proteins</term>
<term>Nuclear Proteins</term>
<term>RNA, Messenger</term>
<term>Transcription Factors</term>
</keywords>
<keywords scheme="MESH" type="chemical" xml:lang="en"><term>CCAAT-Enhancer-Binding Proteins</term>
<term>DNA</term>
</keywords>
<keywords scheme="MESH" qualifier="cytologie" xml:lang="fr"><term>Foie</term>
</keywords>
<keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>Liver</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Alphafoetoprotéines</term>
<term>Chloramphenicol O-acetyltransferase</term>
</keywords>
<keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Brain</term>
<term>Kidney</term>
<term>Liver</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>ARN messager</term>
<term>Chloramphenicol O-acetyltransferase</term>
<term>Encéphale</term>
<term>Facteurs de transcription</term>
<term>Foie</term>
<term>Protéines de liaison à l'ADN</term>
<term>Protéines nucléaires</term>
<term>Rein</term>
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<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Base Sequence</term>
<term>Cells, Cultured</term>
<term>DNA Fingerprinting</term>
<term>Enhancer Elements, Genetic</term>
<term>Mice</term>
<term>Molecular Sequence Data</term>
<term>Mutagenesis, Site-Directed</term>
<term>Plasmids</term>
<term>Transfection</term>
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<keywords scheme="MESH" xml:lang="fr"><term>ADN</term>
<term>Animaux</term>
<term>Cellules cultivées</term>
<term>Données de séquences moléculaires</term>
<term>Mutagenèse dirigée</term>
<term>Plasmides</term>
<term>Profilage d'ADN</term>
<term>Protéines liant les séquences stimulatrices de type CCAAT</term>
<term>Souris</term>
<term>Séquence nucléotidique</term>
<term>Transfection</term>
<term>Éléments activateurs (génétique)</term>
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<front><div type="abstract" xml:lang="en">We have compared the activities of mouse alpha-fetoprotein (AFP) enhancers I, II, and III with their minimal enhancer fragments (Mers) I, II, and III and with the entire 7-kilobase pair enhancer domain by transient expression assay in primary fetal mouse liver cells. The level of expression directed by the AFP promoter [p(-1009)AFPcat] alone is stimulated at least 10-fold by the entire AFP enhancer domain (-1009 to -6983). Enhancer I can drive the level of chloramphenicol acetyltransferase activity equivalent to that of the entire enhancer domain, whereas the increase in activity by enhancers II and III is significantly lower (1.5-fold). MersI, II, and III all mediate a greater increase in activity than their corresponding enhancer regions. The increase with MerI is 16-fold. Using DNase I protection analyses we identified 3 protein-binding regions in MerI; site Ia binds liver and brain nuclear proteins; site Ib binds liver, kidney, and brain nuclear proteins as well as purified C/EBP; site Ic binds liver and kidney nuclear proteins. Site-specific mutation of Ia, Ib, or Ic showed a 10-25% reduction in chloramphenicol acetyltransferase expression; deletion of the C/EBP-binding site in Ib showed a 45% reduction in activity and mutation of all 3 sites (Ia, Ib, and Ic) resulted in a 75% reduction in activity. Our studies indicate no single trans-acting factor is absolutely essential for enhancer activity, and that the enhancer activity of MerI is mediated via a combinatorial and additive mechanism.</div>
</front>
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<tree><noCountry><name sortKey="Hsieh, C C" sort="Hsieh, C C" uniqKey="Hsieh C" first="C C" last="Hsieh">C C Hsieh</name>
<name sortKey="Papaconstantinou, J" sort="Papaconstantinou, J" uniqKey="Papaconstantinou J" first="J" last="Papaconstantinou">J. Papaconstantinou</name>
<name sortKey="Rabek, J P" sort="Rabek, J P" uniqKey="Rabek J" first="J P" last="Rabek">J P Rabek</name>
<name sortKey="Torres Ramos, C" sort="Torres Ramos, C" uniqKey="Torres Ramos C" first="C" last="Torres-Ramos">C. Torres-Ramos</name>
<name sortKey="Zhang, D E" sort="Zhang, D E" uniqKey="Zhang D" first="D E" last="Zhang">D E Zhang</name>
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