Reciprocal regulation of β1-adrenergic receptor gene transcription by Sp1 and early growth response gene 1

Induction of EGR-1 inhibits the expression of the β1-adrenergic receptor gene

Suleiman Bahouth, Michael J. Beauchamp, Kim N. Vu

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

The β1-adrenergic receptor (β1-AR) plays a key role in regulating heart rate and contractility in response to catecholamines. Our studies have focused on defining the factors that regulate the expression of β1-AR gene. We determined that a 65-base-pair (bp) region in the β1-AR promoter between bp -394 and bp -330 directs basal transcription. An element located between -377 and -365 can bind Sp1 and Sp3. In Drosophila melanogaster SL2 cells, Sp1 stimulated the expression of the β1-AR promoter, whereas Sp3 was unable to activate transcription. Site-directed mutagenesis indicated that an intact Sp1-binding site is essential for maintaining the activity of the basal promoter. In addition to binding Sp family members, the nucleotides between -381 and -367 can bind the zinc-finger transcription factor Egr-1. The Egr-1 and Sp1 binding sites are partially overlapping and their binding sequence is conserved among mammalian β1-AR genes. The induction of Egr-1 in rat neonatal ventricular myocytes with phorbol-12-myristate-13-acetate or in HeLa S3 cells by regulated expression of Egr-1 in a tetracycline-responsive promoter, suppressed expression from the β1-AR promoter. Overexpression of Sp1 in SK-N-MC cells increased β1-AR mRNA by 2.4-fold, whereas overexpression of Egr-1 reduced β1-AR mRNA by 40%. Coexpression of Egr-1 with Sp1 reduced Sp1-mediated up-regulation of β1-AR mRNA by 60%. Mutagenesis revealed that an intact Sp1-binding site is essential for observing transcriptional repression by Egr-1 and that Egr-1 suppressed the transcription of the β1-AR gene by competing with Sp1 for binding to their overlapping sites. These results reveal a novel physiologically relevant transcriptional mechanism for reciprocal regulation of β1-AR gene expression.

Original languageEnglish (US)
Pages (from-to)379-390
Number of pages12
JournalMolecular Pharmacology
Volume61
Issue number2
DOIs
StatePublished - Feb 11 2002

Fingerprint

Genetic Transcription
Adrenergic Receptors
Growth
Genes
Base Pairing
Binding Sites
Messenger RNA
Myocardial Contraction
Conserved Sequence
Zinc Fingers
Site-Directed Mutagenesis
Tetracycline
Drosophila melanogaster
HeLa Cells
Mutagenesis
Muscle Cells
Catecholamines
Acetates

All Science Journal Classification (ASJC) codes

  • Pharmacology

Cite this

@article{94c451f896b9437494209fd0fbb0bcf2,
title = "Reciprocal regulation of β1-adrenergic receptor gene transcription by Sp1 and early growth response gene 1: Induction of EGR-1 inhibits the expression of the β1-adrenergic receptor gene",
abstract = "The β1-adrenergic receptor (β1-AR) plays a key role in regulating heart rate and contractility in response to catecholamines. Our studies have focused on defining the factors that regulate the expression of β1-AR gene. We determined that a 65-base-pair (bp) region in the β1-AR promoter between bp -394 and bp -330 directs basal transcription. An element located between -377 and -365 can bind Sp1 and Sp3. In Drosophila melanogaster SL2 cells, Sp1 stimulated the expression of the β1-AR promoter, whereas Sp3 was unable to activate transcription. Site-directed mutagenesis indicated that an intact Sp1-binding site is essential for maintaining the activity of the basal promoter. In addition to binding Sp family members, the nucleotides between -381 and -367 can bind the zinc-finger transcription factor Egr-1. The Egr-1 and Sp1 binding sites are partially overlapping and their binding sequence is conserved among mammalian β1-AR genes. The induction of Egr-1 in rat neonatal ventricular myocytes with phorbol-12-myristate-13-acetate or in HeLa S3 cells by regulated expression of Egr-1 in a tetracycline-responsive promoter, suppressed expression from the β1-AR promoter. Overexpression of Sp1 in SK-N-MC cells increased β1-AR mRNA by 2.4-fold, whereas overexpression of Egr-1 reduced β1-AR mRNA by 40{\%}. Coexpression of Egr-1 with Sp1 reduced Sp1-mediated up-regulation of β1-AR mRNA by 60{\%}. Mutagenesis revealed that an intact Sp1-binding site is essential for observing transcriptional repression by Egr-1 and that Egr-1 suppressed the transcription of the β1-AR gene by competing with Sp1 for binding to their overlapping sites. These results reveal a novel physiologically relevant transcriptional mechanism for reciprocal regulation of β1-AR gene expression.",
author = "Suleiman Bahouth and Beauchamp, {Michael J.} and Vu, {Kim N.}",
year = "2002",
month = "2",
day = "11",
doi = "10.1124/mol.61.2.379",
language = "English (US)",
volume = "61",
pages = "379--390",
journal = "Molecular Pharmacology",
issn = "0026-895X",
publisher = "American Society for Pharmacology and Experimental Therapeutics",
number = "2",

}

TY - JOUR

T1 - Reciprocal regulation of β1-adrenergic receptor gene transcription by Sp1 and early growth response gene 1

T2 - Induction of EGR-1 inhibits the expression of the β1-adrenergic receptor gene

AU - Bahouth, Suleiman

AU - Beauchamp, Michael J.

AU - Vu, Kim N.

PY - 2002/2/11

Y1 - 2002/2/11

N2 - The β1-adrenergic receptor (β1-AR) plays a key role in regulating heart rate and contractility in response to catecholamines. Our studies have focused on defining the factors that regulate the expression of β1-AR gene. We determined that a 65-base-pair (bp) region in the β1-AR promoter between bp -394 and bp -330 directs basal transcription. An element located between -377 and -365 can bind Sp1 and Sp3. In Drosophila melanogaster SL2 cells, Sp1 stimulated the expression of the β1-AR promoter, whereas Sp3 was unable to activate transcription. Site-directed mutagenesis indicated that an intact Sp1-binding site is essential for maintaining the activity of the basal promoter. In addition to binding Sp family members, the nucleotides between -381 and -367 can bind the zinc-finger transcription factor Egr-1. The Egr-1 and Sp1 binding sites are partially overlapping and their binding sequence is conserved among mammalian β1-AR genes. The induction of Egr-1 in rat neonatal ventricular myocytes with phorbol-12-myristate-13-acetate or in HeLa S3 cells by regulated expression of Egr-1 in a tetracycline-responsive promoter, suppressed expression from the β1-AR promoter. Overexpression of Sp1 in SK-N-MC cells increased β1-AR mRNA by 2.4-fold, whereas overexpression of Egr-1 reduced β1-AR mRNA by 40%. Coexpression of Egr-1 with Sp1 reduced Sp1-mediated up-regulation of β1-AR mRNA by 60%. Mutagenesis revealed that an intact Sp1-binding site is essential for observing transcriptional repression by Egr-1 and that Egr-1 suppressed the transcription of the β1-AR gene by competing with Sp1 for binding to their overlapping sites. These results reveal a novel physiologically relevant transcriptional mechanism for reciprocal regulation of β1-AR gene expression.

AB - The β1-adrenergic receptor (β1-AR) plays a key role in regulating heart rate and contractility in response to catecholamines. Our studies have focused on defining the factors that regulate the expression of β1-AR gene. We determined that a 65-base-pair (bp) region in the β1-AR promoter between bp -394 and bp -330 directs basal transcription. An element located between -377 and -365 can bind Sp1 and Sp3. In Drosophila melanogaster SL2 cells, Sp1 stimulated the expression of the β1-AR promoter, whereas Sp3 was unable to activate transcription. Site-directed mutagenesis indicated that an intact Sp1-binding site is essential for maintaining the activity of the basal promoter. In addition to binding Sp family members, the nucleotides between -381 and -367 can bind the zinc-finger transcription factor Egr-1. The Egr-1 and Sp1 binding sites are partially overlapping and their binding sequence is conserved among mammalian β1-AR genes. The induction of Egr-1 in rat neonatal ventricular myocytes with phorbol-12-myristate-13-acetate or in HeLa S3 cells by regulated expression of Egr-1 in a tetracycline-responsive promoter, suppressed expression from the β1-AR promoter. Overexpression of Sp1 in SK-N-MC cells increased β1-AR mRNA by 2.4-fold, whereas overexpression of Egr-1 reduced β1-AR mRNA by 40%. Coexpression of Egr-1 with Sp1 reduced Sp1-mediated up-regulation of β1-AR mRNA by 60%. Mutagenesis revealed that an intact Sp1-binding site is essential for observing transcriptional repression by Egr-1 and that Egr-1 suppressed the transcription of the β1-AR gene by competing with Sp1 for binding to their overlapping sites. These results reveal a novel physiologically relevant transcriptional mechanism for reciprocal regulation of β1-AR gene expression.

UR - http://www.scopus.com/inward/record.url?scp=0036156038&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036156038&partnerID=8YFLogxK

U2 - 10.1124/mol.61.2.379

DO - 10.1124/mol.61.2.379

M3 - Article

VL - 61

SP - 379

EP - 390

JO - Molecular Pharmacology

JF - Molecular Pharmacology

SN - 0026-895X

IS - 2

ER -