Structural and functional conservation of vertebrate corticotropin- releasing factor genes

Evidence for a critical role for a conserved cyclic AMP response element

Meng Yao, Mary Stenzel-Poore, Robert J. Denver

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Corticotropin-releasing factor (CRF) plays a central role in neuroendocrine, autonomic, immune, and behavioral responses to stressors. We analyzed the proximal promoters of two Xenopus laevis CRF genes and found them to be remarkably conserved with mammalian CRF genes. We found several conserved cis elements in the frog CRF genes including a cAMP response element (CRE), activator protein 1 binding sites, and glucocorticoid response elements. Exposure to a physical stressor caused a rapid elevation in phosphorylated CRE binding protein (CREB; 20 min) and CRF (1 h) in the anterior preoptic area of juvenile frogs. CREB bound to the putative frog CREs in vitro, which was disrupted by point mutations introduced into the CRE. The frog proximal CRF promoters supported basal transcription in transfection assays, and forskolin caused robust transcriptional activation. Mutagenesis of the CRE or overexpression of a dominant-negative CREB reduced forskolin-induced promoter activation. Using electroporation-mediated gene transfer in tadpole brain, we show that the proximal CRF promoters support cAMP or stressor-dependent transcription in vivo, which was abolished by mutation of the CRE. Using chromatin immunoprecipitation, we found that CREB associated with the proximal frog CRF promoter in vivo in a stressor-dependent manner. These data provide strong support for the hypothesis that stressor-induced CRF gene activation in vivo depends on CREB binding to the CRE in the promoter. Our findings show that the basic regulatory elements of the CRF gene responsible for stressor-induced activation arose early in vertebrate evolution and have been maintained by strong positive selection.

Original languageEnglish (US)
Pages (from-to)2518-2531
Number of pages14
JournalEndocrinology
Volume148
Issue number5
DOIs
StatePublished - May 2007

Fingerprint

Corticotropin-Releasing Hormone
Response Elements
Cyclic AMP
Vertebrates
Anura
Genes
Colforsin
Transcriptional Activation
Cyclic AMP Response Element-Binding Protein
Preoptic Area
Electroporation
Chromatin Immunoprecipitation
Transcription Factor AP-1
Xenopus laevis
Point Mutation
Protein Binding
Mutagenesis
Glucocorticoids
Transfection
Larva

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

@article{423435ce936648f396b64be7d07ebcbf,
title = "Structural and functional conservation of vertebrate corticotropin- releasing factor genes: Evidence for a critical role for a conserved cyclic AMP response element",
abstract = "Corticotropin-releasing factor (CRF) plays a central role in neuroendocrine, autonomic, immune, and behavioral responses to stressors. We analyzed the proximal promoters of two Xenopus laevis CRF genes and found them to be remarkably conserved with mammalian CRF genes. We found several conserved cis elements in the frog CRF genes including a cAMP response element (CRE), activator protein 1 binding sites, and glucocorticoid response elements. Exposure to a physical stressor caused a rapid elevation in phosphorylated CRE binding protein (CREB; 20 min) and CRF (1 h) in the anterior preoptic area of juvenile frogs. CREB bound to the putative frog CREs in vitro, which was disrupted by point mutations introduced into the CRE. The frog proximal CRF promoters supported basal transcription in transfection assays, and forskolin caused robust transcriptional activation. Mutagenesis of the CRE or overexpression of a dominant-negative CREB reduced forskolin-induced promoter activation. Using electroporation-mediated gene transfer in tadpole brain, we show that the proximal CRF promoters support cAMP or stressor-dependent transcription in vivo, which was abolished by mutation of the CRE. Using chromatin immunoprecipitation, we found that CREB associated with the proximal frog CRF promoter in vivo in a stressor-dependent manner. These data provide strong support for the hypothesis that stressor-induced CRF gene activation in vivo depends on CREB binding to the CRE in the promoter. Our findings show that the basic regulatory elements of the CRF gene responsible for stressor-induced activation arose early in vertebrate evolution and have been maintained by strong positive selection.",
author = "Meng Yao and Mary Stenzel-Poore and Denver, {Robert J.}",
year = "2007",
month = "5",
doi = "10.1210/en.2006-1413",
language = "English (US)",
volume = "148",
pages = "2518--2531",
journal = "Endocrinology",
issn = "0013-7227",
publisher = "The Endocrine Society",
number = "5",

}

TY - JOUR

T1 - Structural and functional conservation of vertebrate corticotropin- releasing factor genes

T2 - Evidence for a critical role for a conserved cyclic AMP response element

AU - Yao, Meng

AU - Stenzel-Poore, Mary

AU - Denver, Robert J.

PY - 2007/5

Y1 - 2007/5

N2 - Corticotropin-releasing factor (CRF) plays a central role in neuroendocrine, autonomic, immune, and behavioral responses to stressors. We analyzed the proximal promoters of two Xenopus laevis CRF genes and found them to be remarkably conserved with mammalian CRF genes. We found several conserved cis elements in the frog CRF genes including a cAMP response element (CRE), activator protein 1 binding sites, and glucocorticoid response elements. Exposure to a physical stressor caused a rapid elevation in phosphorylated CRE binding protein (CREB; 20 min) and CRF (1 h) in the anterior preoptic area of juvenile frogs. CREB bound to the putative frog CREs in vitro, which was disrupted by point mutations introduced into the CRE. The frog proximal CRF promoters supported basal transcription in transfection assays, and forskolin caused robust transcriptional activation. Mutagenesis of the CRE or overexpression of a dominant-negative CREB reduced forskolin-induced promoter activation. Using electroporation-mediated gene transfer in tadpole brain, we show that the proximal CRF promoters support cAMP or stressor-dependent transcription in vivo, which was abolished by mutation of the CRE. Using chromatin immunoprecipitation, we found that CREB associated with the proximal frog CRF promoter in vivo in a stressor-dependent manner. These data provide strong support for the hypothesis that stressor-induced CRF gene activation in vivo depends on CREB binding to the CRE in the promoter. Our findings show that the basic regulatory elements of the CRF gene responsible for stressor-induced activation arose early in vertebrate evolution and have been maintained by strong positive selection.

AB - Corticotropin-releasing factor (CRF) plays a central role in neuroendocrine, autonomic, immune, and behavioral responses to stressors. We analyzed the proximal promoters of two Xenopus laevis CRF genes and found them to be remarkably conserved with mammalian CRF genes. We found several conserved cis elements in the frog CRF genes including a cAMP response element (CRE), activator protein 1 binding sites, and glucocorticoid response elements. Exposure to a physical stressor caused a rapid elevation in phosphorylated CRE binding protein (CREB; 20 min) and CRF (1 h) in the anterior preoptic area of juvenile frogs. CREB bound to the putative frog CREs in vitro, which was disrupted by point mutations introduced into the CRE. The frog proximal CRF promoters supported basal transcription in transfection assays, and forskolin caused robust transcriptional activation. Mutagenesis of the CRE or overexpression of a dominant-negative CREB reduced forskolin-induced promoter activation. Using electroporation-mediated gene transfer in tadpole brain, we show that the proximal CRF promoters support cAMP or stressor-dependent transcription in vivo, which was abolished by mutation of the CRE. Using chromatin immunoprecipitation, we found that CREB associated with the proximal frog CRF promoter in vivo in a stressor-dependent manner. These data provide strong support for the hypothesis that stressor-induced CRF gene activation in vivo depends on CREB binding to the CRE in the promoter. Our findings show that the basic regulatory elements of the CRF gene responsible for stressor-induced activation arose early in vertebrate evolution and have been maintained by strong positive selection.

UR - http://www.scopus.com/inward/record.url?scp=34249782869&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34249782869&partnerID=8YFLogxK

U2 - 10.1210/en.2006-1413

DO - 10.1210/en.2006-1413

M3 - Article

VL - 148

SP - 2518

EP - 2531

JO - Endocrinology

JF - Endocrinology

SN - 0013-7227

IS - 5

ER -