Identification of a retinoic acid response domain involved in the activation of the β1-adrenergic receptor gene by retinoic acid in F9 teratocarcinoma cells

Suleiman Bahouth, Michael J. Beauchamp, Edwards Park

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Abstract

The density of β1-adrenergic receptors (β1-AR) is up-regulated upon differentiation of embryonic F9 teratocarcinoma cells by retinoic acid (RA) to the primitive endodermal phenotype. To identify the domains involved in RA-mediated activation of β1-AR gene transcription, three kb of 5'-flanking sequence of the β1-AR gene were ligated to a luciferase reporter gene and transiently transfected into F9 cells that were pre-exposed to 100 nM RA for 2 days. By generating deletions in the β1-AR promoter, a region between -125 and -100 was found to mediate a 3-fold induction in cells exposed to RA for an additional 2 days. Through site directed. mutagenesis of this region, it was determined that the RA responsive element (RARE) was organized as a direct repeat separated by 5 nucleotides in which the 5'-most AGGTCG half-site was between nucleotides -106 and -101 and the 3' most AGGTCA half-site was between nucleotides -117 and -112. The RA receptor α (RARα) isoform bound to the oligomer representing the sequences between -125 and -100 as a heterodimer complex with the retinoid X receptor or (RXRα). In a separate study, it was determined that the nucleotides between -125 and -100 are involved in thyroid hormone-mediated activation of the β1-AR gene in ventricular myocytes. Therefore, transcriptional activation of the β1-AR gene by thyroid hormone or RA involves a single binding site in the promoter.

Original languageEnglish (US)
Pages (from-to)215-225
Number of pages11
JournalBiochemical Pharmacology
Volume55
Issue number2
DOIs
StatePublished - Jan 15 1998

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Teratocarcinoma
Tretinoin
Adrenergic Receptors
Genes
Chemical activation
Nucleotides
Thyroid Hormones
Retinoid X Receptors
Mutagenesis
Retinoic Acid Receptors
Nucleic Acid Repetitive Sequences
5' Flanking Region
Transcription
Luciferases
Reporter Genes
Oligomers
Genetic Promoter Regions
Muscle Cells
Transcriptional Activation
Protein Isoforms

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Pharmacology

Cite this

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title = "Identification of a retinoic acid response domain involved in the activation of the β1-adrenergic receptor gene by retinoic acid in F9 teratocarcinoma cells",
abstract = "The density of β1-adrenergic receptors (β1-AR) is up-regulated upon differentiation of embryonic F9 teratocarcinoma cells by retinoic acid (RA) to the primitive endodermal phenotype. To identify the domains involved in RA-mediated activation of β1-AR gene transcription, three kb of 5'-flanking sequence of the β1-AR gene were ligated to a luciferase reporter gene and transiently transfected into F9 cells that were pre-exposed to 100 nM RA for 2 days. By generating deletions in the β1-AR promoter, a region between -125 and -100 was found to mediate a 3-fold induction in cells exposed to RA for an additional 2 days. Through site directed. mutagenesis of this region, it was determined that the RA responsive element (RARE) was organized as a direct repeat separated by 5 nucleotides in which the 5'-most AGGTCG half-site was between nucleotides -106 and -101 and the 3' most AGGTCA half-site was between nucleotides -117 and -112. The RA receptor α (RARα) isoform bound to the oligomer representing the sequences between -125 and -100 as a heterodimer complex with the retinoid X receptor or (RXRα). In a separate study, it was determined that the nucleotides between -125 and -100 are involved in thyroid hormone-mediated activation of the β1-AR gene in ventricular myocytes. Therefore, transcriptional activation of the β1-AR gene by thyroid hormone or RA involves a single binding site in the promoter.",
author = "Suleiman Bahouth and Beauchamp, {Michael J.} and Edwards Park",
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N2 - The density of β1-adrenergic receptors (β1-AR) is up-regulated upon differentiation of embryonic F9 teratocarcinoma cells by retinoic acid (RA) to the primitive endodermal phenotype. To identify the domains involved in RA-mediated activation of β1-AR gene transcription, three kb of 5'-flanking sequence of the β1-AR gene were ligated to a luciferase reporter gene and transiently transfected into F9 cells that were pre-exposed to 100 nM RA for 2 days. By generating deletions in the β1-AR promoter, a region between -125 and -100 was found to mediate a 3-fold induction in cells exposed to RA for an additional 2 days. Through site directed. mutagenesis of this region, it was determined that the RA responsive element (RARE) was organized as a direct repeat separated by 5 nucleotides in which the 5'-most AGGTCG half-site was between nucleotides -106 and -101 and the 3' most AGGTCA half-site was between nucleotides -117 and -112. The RA receptor α (RARα) isoform bound to the oligomer representing the sequences between -125 and -100 as a heterodimer complex with the retinoid X receptor or (RXRα). In a separate study, it was determined that the nucleotides between -125 and -100 are involved in thyroid hormone-mediated activation of the β1-AR gene in ventricular myocytes. Therefore, transcriptional activation of the β1-AR gene by thyroid hormone or RA involves a single binding site in the promoter.

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