Transcriptional co-operativity between distant retinoic acid response elements in regulation of Cyp26A1 inducibility

Olivier Loudig, Glenn A. MacLean, Naomi L. Dore, Luong Luu, Martin Petkovich

Research output: Contribution to journalArticle

94 Citations (Scopus)

Abstract

Cyp26A1 encodes an RA (retinoic acid)-catabolizing CYP (cytochrome P450) protein that plays a critical role in regulating RA distribution in vivo. Cyp26A1 expression is inducible by RA, and the locus has previously been shown to contain a RARE (RA response element), R1, within the minimal promoter [Loudig, Babichuk, White, Abu-Abed, Mueller and Petkovich (2000) Mol. Endocrinol. 14, 1483-1497]. In the present study, we report the identification of a second functional RARE (R2) located 2.0 kb upstream of the Cyp26A1 transcriptional start site. Constructs containing murine sequences encompassing both R1 and R2 showed that these elements work together to generate higher transcriptional activity upon treatment with RA than those containing R1 alone. Inclusion of R2 also dramatically enhanced the sensitivity of reporter constructs to RA, as even treatment with 10-8 M RA resulted in a 5-fold induction of reporter activity. Mutational analysis identified R2 as the functional element responsible for the increased RA inducibility of promoter constructs. The element was shown to bind RARγ (RA receptor γ)/RXRα (retinoid X receptor α) heterodimers in vitro, and inclusion of nuclear receptors in transfections boosted the transcriptional response. A construct containing both R1 and R2 was used to generate a stable luciferase reporter cell line that can be used as a tool to identify factors regulating Cyp26A1 expression. The analysis of R1 and R2 has led to the proposal that the two elements work synergistically to provide a maximal response to RA and that R2 is an upstream enhancer.

Original languageEnglish (US)
Pages (from-to)241-248
Number of pages8
JournalBiochemical Journal
Volume392
Issue number1
DOIs
StatePublished - Nov 15 2005
Externally publishedYes

Fingerprint

Response Elements
Tretinoin
Retinoid X Receptors
Retinoic Acid Receptors
Cytoplasmic and Nuclear Receptors
Luciferases
Cytochrome P-450 Enzyme System
Transfection
Cells
Cell Line

Keywords

  • Cyp26A1
  • Embryonic development
  • Nuclear receptor
  • Retinoic acid
  • Retinoic acid response element (RARE)
  • Transcriptional regulation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Transcriptional co-operativity between distant retinoic acid response elements in regulation of Cyp26A1 inducibility. / Loudig, Olivier; MacLean, Glenn A.; Dore, Naomi L.; Luu, Luong; Petkovich, Martin.

In: Biochemical Journal, Vol. 392, No. 1, 15.11.2005, p. 241-248.

Research output: Contribution to journalArticle

Loudig, Olivier ; MacLean, Glenn A. ; Dore, Naomi L. ; Luu, Luong ; Petkovich, Martin. / Transcriptional co-operativity between distant retinoic acid response elements in regulation of Cyp26A1 inducibility. In: Biochemical Journal. 2005 ; Vol. 392, No. 1. pp. 241-248.
@article{885bc1c2d65a4f2387dda25bae64b25d,
title = "Transcriptional co-operativity between distant retinoic acid response elements in regulation of Cyp26A1 inducibility",
abstract = "Cyp26A1 encodes an RA (retinoic acid)-catabolizing CYP (cytochrome P450) protein that plays a critical role in regulating RA distribution in vivo. Cyp26A1 expression is inducible by RA, and the locus has previously been shown to contain a RARE (RA response element), R1, within the minimal promoter [Loudig, Babichuk, White, Abu-Abed, Mueller and Petkovich (2000) Mol. Endocrinol. 14, 1483-1497]. In the present study, we report the identification of a second functional RARE (R2) located 2.0 kb upstream of the Cyp26A1 transcriptional start site. Constructs containing murine sequences encompassing both R1 and R2 showed that these elements work together to generate higher transcriptional activity upon treatment with RA than those containing R1 alone. Inclusion of R2 also dramatically enhanced the sensitivity of reporter constructs to RA, as even treatment with 10-8 M RA resulted in a 5-fold induction of reporter activity. Mutational analysis identified R2 as the functional element responsible for the increased RA inducibility of promoter constructs. The element was shown to bind RARγ (RA receptor γ)/RXRα (retinoid X receptor α) heterodimers in vitro, and inclusion of nuclear receptors in transfections boosted the transcriptional response. A construct containing both R1 and R2 was used to generate a stable luciferase reporter cell line that can be used as a tool to identify factors regulating Cyp26A1 expression. The analysis of R1 and R2 has led to the proposal that the two elements work synergistically to provide a maximal response to RA and that R2 is an upstream enhancer.",
keywords = "Cyp26A1, Embryonic development, Nuclear receptor, Retinoic acid, Retinoic acid response element (RARE), Transcriptional regulation",
author = "Olivier Loudig and MacLean, {Glenn A.} and Dore, {Naomi L.} and Luong Luu and Martin Petkovich",
year = "2005",
month = "11",
day = "15",
doi = "10.1042/BJ20050874",
language = "English (US)",
volume = "392",
pages = "241--248",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "1",

}

TY - JOUR

T1 - Transcriptional co-operativity between distant retinoic acid response elements in regulation of Cyp26A1 inducibility

AU - Loudig, Olivier

AU - MacLean, Glenn A.

AU - Dore, Naomi L.

AU - Luu, Luong

AU - Petkovich, Martin

PY - 2005/11/15

Y1 - 2005/11/15

N2 - Cyp26A1 encodes an RA (retinoic acid)-catabolizing CYP (cytochrome P450) protein that plays a critical role in regulating RA distribution in vivo. Cyp26A1 expression is inducible by RA, and the locus has previously been shown to contain a RARE (RA response element), R1, within the minimal promoter [Loudig, Babichuk, White, Abu-Abed, Mueller and Petkovich (2000) Mol. Endocrinol. 14, 1483-1497]. In the present study, we report the identification of a second functional RARE (R2) located 2.0 kb upstream of the Cyp26A1 transcriptional start site. Constructs containing murine sequences encompassing both R1 and R2 showed that these elements work together to generate higher transcriptional activity upon treatment with RA than those containing R1 alone. Inclusion of R2 also dramatically enhanced the sensitivity of reporter constructs to RA, as even treatment with 10-8 M RA resulted in a 5-fold induction of reporter activity. Mutational analysis identified R2 as the functional element responsible for the increased RA inducibility of promoter constructs. The element was shown to bind RARγ (RA receptor γ)/RXRα (retinoid X receptor α) heterodimers in vitro, and inclusion of nuclear receptors in transfections boosted the transcriptional response. A construct containing both R1 and R2 was used to generate a stable luciferase reporter cell line that can be used as a tool to identify factors regulating Cyp26A1 expression. The analysis of R1 and R2 has led to the proposal that the two elements work synergistically to provide a maximal response to RA and that R2 is an upstream enhancer.

AB - Cyp26A1 encodes an RA (retinoic acid)-catabolizing CYP (cytochrome P450) protein that plays a critical role in regulating RA distribution in vivo. Cyp26A1 expression is inducible by RA, and the locus has previously been shown to contain a RARE (RA response element), R1, within the minimal promoter [Loudig, Babichuk, White, Abu-Abed, Mueller and Petkovich (2000) Mol. Endocrinol. 14, 1483-1497]. In the present study, we report the identification of a second functional RARE (R2) located 2.0 kb upstream of the Cyp26A1 transcriptional start site. Constructs containing murine sequences encompassing both R1 and R2 showed that these elements work together to generate higher transcriptional activity upon treatment with RA than those containing R1 alone. Inclusion of R2 also dramatically enhanced the sensitivity of reporter constructs to RA, as even treatment with 10-8 M RA resulted in a 5-fold induction of reporter activity. Mutational analysis identified R2 as the functional element responsible for the increased RA inducibility of promoter constructs. The element was shown to bind RARγ (RA receptor γ)/RXRα (retinoid X receptor α) heterodimers in vitro, and inclusion of nuclear receptors in transfections boosted the transcriptional response. A construct containing both R1 and R2 was used to generate a stable luciferase reporter cell line that can be used as a tool to identify factors regulating Cyp26A1 expression. The analysis of R1 and R2 has led to the proposal that the two elements work synergistically to provide a maximal response to RA and that R2 is an upstream enhancer.

KW - Cyp26A1

KW - Embryonic development

KW - Nuclear receptor

KW - Retinoic acid

KW - Retinoic acid response element (RARE)

KW - Transcriptional regulation

UR - http://www.scopus.com/inward/record.url?scp=28044448712&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=28044448712&partnerID=8YFLogxK

U2 - 10.1042/BJ20050874

DO - 10.1042/BJ20050874

M3 - Article

VL - 392

SP - 241

EP - 248

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 1

ER -