Identification of a Retinoid Receptor Response Element in the Human Aldehyde Dehydrogenase-2 Promoter

Jane Pinaire, Wan Yin Chou, Michael Morton, Min You, Yan Zeng, Won Kyoo Cho, Andrea Galli, Lynn Everett, Heather Breen, Natividad Dumaual, Jennifer R. Smith, David Crabb

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Background: The human aldehyde dehydrogenase-2 promoter contains sites that bind members of the nuclear receptor family, and one (designated FP330-3′) is predicted to bind retinoic acid receptors. Methods: Binding of retinoid receptors to the FP330-3′ oligonucleotide duplex and point mutations thereof was assayed using electrophoretic mobility shift assays. The function of the promoter element was determined in transfection assays. Results: Heterodimers of retinoic acid receptor (RAR)α, β, and γ with retinoid X receptor (RXR)α bound the FP330-3′ site. Mutagenesis of the FP330-3′ site suggested that either the upstream DR-5 or downstream DR-1 could mediate binding of RAR/RXR. FP330-3′ oligonucleotide duplexes were not bound by in vitro translated RXR homodimers but weakly competed with a synthetic DR-1 oligonucleotide duplex for binding by RXR. A reporter construct carrying four copies of the FP330-3′ element was induced by cotransfection of rat hepatoma cells with a construct encoding RARa, when the RAR-specific ligand AM580 was present. Each of the three RXR isoforms α, β, and γ stimulated the expression of reporter constructs containing the FP330-3′ sites in a 9-cis retinoic acid-dependent fashion in cells in culture. This was confirmed in the case of RXRα using the RXR-specific ligand methoprene. Conclusion: The human aldehyde dehydrogenase-2 promoter contains a retinoid response element, which may contribute to regulation of the gene.

Original languageEnglish
Pages (from-to)1860-1866
Number of pages7
JournalAlcoholism: Clinical and Experimental Research
Volume27
Issue number12
DOIs
StatePublished - Dec 2003

Fingerprint

Retinoid X Receptors
Aldehyde Dehydrogenase
Retinoids
Response Elements
Retinoic Acid Receptors
Oligonucleotides
Assays
Methoprene
Ligands
Electrophoretic mobility
Mutagenesis
Electrophoretic Mobility Shift Assay
Cytoplasmic and Nuclear Receptors
Nuclear Family
Point Mutation
Transfection
Rats
Hepatocellular Carcinoma
Protein Isoforms
Cell Culture Techniques

Keywords

  • Aldehyde Dehydrogenase
  • Liver
  • Receptor
  • Retinoic Acid
  • Transcription
  • Vitamin A

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Toxicology

Cite this

Identification of a Retinoid Receptor Response Element in the Human Aldehyde Dehydrogenase-2 Promoter. / Pinaire, Jane; Chou, Wan Yin; Morton, Michael; You, Min; Zeng, Yan; Cho, Won Kyoo; Galli, Andrea; Everett, Lynn; Breen, Heather; Dumaual, Natividad; Smith, Jennifer R.; Crabb, David.

In: Alcoholism: Clinical and Experimental Research, Vol. 27, No. 12, 12.2003, p. 1860-1866.

Research output: Contribution to journalArticle

Pinaire, J, Chou, WY, Morton, M, You, M, Zeng, Y, Cho, WK, Galli, A, Everett, L, Breen, H, Dumaual, N, Smith, JR & Crabb, D 2003, 'Identification of a Retinoid Receptor Response Element in the Human Aldehyde Dehydrogenase-2 Promoter', Alcoholism: Clinical and Experimental Research, vol. 27, no. 12, pp. 1860-1866. https://doi.org/10.1097/01.ALC.0000100941.86227.4F
Pinaire, Jane ; Chou, Wan Yin ; Morton, Michael ; You, Min ; Zeng, Yan ; Cho, Won Kyoo ; Galli, Andrea ; Everett, Lynn ; Breen, Heather ; Dumaual, Natividad ; Smith, Jennifer R. ; Crabb, David. / Identification of a Retinoid Receptor Response Element in the Human Aldehyde Dehydrogenase-2 Promoter. In: Alcoholism: Clinical and Experimental Research. 2003 ; Vol. 27, No. 12. pp. 1860-1866.
@article{497d39c6c5934b618c13e419c96ac038,
title = "Identification of a Retinoid Receptor Response Element in the Human Aldehyde Dehydrogenase-2 Promoter",
abstract = "Background: The human aldehyde dehydrogenase-2 promoter contains sites that bind members of the nuclear receptor family, and one (designated FP330-3′) is predicted to bind retinoic acid receptors. Methods: Binding of retinoid receptors to the FP330-3′ oligonucleotide duplex and point mutations thereof was assayed using electrophoretic mobility shift assays. The function of the promoter element was determined in transfection assays. Results: Heterodimers of retinoic acid receptor (RAR)α, β, and γ with retinoid X receptor (RXR)α bound the FP330-3′ site. Mutagenesis of the FP330-3′ site suggested that either the upstream DR-5 or downstream DR-1 could mediate binding of RAR/RXR. FP330-3′ oligonucleotide duplexes were not bound by in vitro translated RXR homodimers but weakly competed with a synthetic DR-1 oligonucleotide duplex for binding by RXR. A reporter construct carrying four copies of the FP330-3′ element was induced by cotransfection of rat hepatoma cells with a construct encoding RARa, when the RAR-specific ligand AM580 was present. Each of the three RXR isoforms α, β, and γ stimulated the expression of reporter constructs containing the FP330-3′ sites in a 9-cis retinoic acid-dependent fashion in cells in culture. This was confirmed in the case of RXRα using the RXR-specific ligand methoprene. Conclusion: The human aldehyde dehydrogenase-2 promoter contains a retinoid response element, which may contribute to regulation of the gene.",
keywords = "Aldehyde Dehydrogenase, Liver, Receptor, Retinoic Acid, Transcription, Vitamin A",
author = "Jane Pinaire and Chou, {Wan Yin} and Michael Morton and Min You and Yan Zeng and Cho, {Won Kyoo} and Andrea Galli and Lynn Everett and Heather Breen and Natividad Dumaual and Smith, {Jennifer R.} and David Crabb",
year = "2003",
month = "12",
doi = "10.1097/01.ALC.0000100941.86227.4F",
language = "English",
volume = "27",
pages = "1860--1866",
journal = "Alcoholism: Clinical and Experimental Research",
issn = "0145-6008",
publisher = "Wiley-Blackwell",
number = "12",

}

TY - JOUR

T1 - Identification of a Retinoid Receptor Response Element in the Human Aldehyde Dehydrogenase-2 Promoter

AU - Pinaire, Jane

AU - Chou, Wan Yin

AU - Morton, Michael

AU - You, Min

AU - Zeng, Yan

AU - Cho, Won Kyoo

AU - Galli, Andrea

AU - Everett, Lynn

AU - Breen, Heather

AU - Dumaual, Natividad

AU - Smith, Jennifer R.

AU - Crabb, David

PY - 2003/12

Y1 - 2003/12

N2 - Background: The human aldehyde dehydrogenase-2 promoter contains sites that bind members of the nuclear receptor family, and one (designated FP330-3′) is predicted to bind retinoic acid receptors. Methods: Binding of retinoid receptors to the FP330-3′ oligonucleotide duplex and point mutations thereof was assayed using electrophoretic mobility shift assays. The function of the promoter element was determined in transfection assays. Results: Heterodimers of retinoic acid receptor (RAR)α, β, and γ with retinoid X receptor (RXR)α bound the FP330-3′ site. Mutagenesis of the FP330-3′ site suggested that either the upstream DR-5 or downstream DR-1 could mediate binding of RAR/RXR. FP330-3′ oligonucleotide duplexes were not bound by in vitro translated RXR homodimers but weakly competed with a synthetic DR-1 oligonucleotide duplex for binding by RXR. A reporter construct carrying four copies of the FP330-3′ element was induced by cotransfection of rat hepatoma cells with a construct encoding RARa, when the RAR-specific ligand AM580 was present. Each of the three RXR isoforms α, β, and γ stimulated the expression of reporter constructs containing the FP330-3′ sites in a 9-cis retinoic acid-dependent fashion in cells in culture. This was confirmed in the case of RXRα using the RXR-specific ligand methoprene. Conclusion: The human aldehyde dehydrogenase-2 promoter contains a retinoid response element, which may contribute to regulation of the gene.

AB - Background: The human aldehyde dehydrogenase-2 promoter contains sites that bind members of the nuclear receptor family, and one (designated FP330-3′) is predicted to bind retinoic acid receptors. Methods: Binding of retinoid receptors to the FP330-3′ oligonucleotide duplex and point mutations thereof was assayed using electrophoretic mobility shift assays. The function of the promoter element was determined in transfection assays. Results: Heterodimers of retinoic acid receptor (RAR)α, β, and γ with retinoid X receptor (RXR)α bound the FP330-3′ site. Mutagenesis of the FP330-3′ site suggested that either the upstream DR-5 or downstream DR-1 could mediate binding of RAR/RXR. FP330-3′ oligonucleotide duplexes were not bound by in vitro translated RXR homodimers but weakly competed with a synthetic DR-1 oligonucleotide duplex for binding by RXR. A reporter construct carrying four copies of the FP330-3′ element was induced by cotransfection of rat hepatoma cells with a construct encoding RARa, when the RAR-specific ligand AM580 was present. Each of the three RXR isoforms α, β, and γ stimulated the expression of reporter constructs containing the FP330-3′ sites in a 9-cis retinoic acid-dependent fashion in cells in culture. This was confirmed in the case of RXRα using the RXR-specific ligand methoprene. Conclusion: The human aldehyde dehydrogenase-2 promoter contains a retinoid response element, which may contribute to regulation of the gene.

KW - Aldehyde Dehydrogenase

KW - Liver

KW - Receptor

KW - Retinoic Acid

KW - Transcription

KW - Vitamin A

UR - http://www.scopus.com/inward/record.url?scp=0345733800&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0345733800&partnerID=8YFLogxK

U2 - 10.1097/01.ALC.0000100941.86227.4F

DO - 10.1097/01.ALC.0000100941.86227.4F

M3 - Article

C2 - 14691372

AN - SCOPUS:0345733800

VL - 27

SP - 1860

EP - 1866

JO - Alcoholism: Clinical and Experimental Research

JF - Alcoholism: Clinical and Experimental Research

SN - 0145-6008

IS - 12

ER -