- Open access
- Published:
Chromatin immunoprecipitation assay detects er recruitment to gene specific promoters in uterus
Biological Procedures Online volume 8, pages 69–76 (2006)
Abstract
Chromatin immunoprecipitation (ChIP) technique allows detection of proteins that bind to chromatin. While this technique has been applied extensively in cell-based studies, its tissue-based application remains poorly explored. We are specifically interested in examining estrogen-dependent transcriptional mechanism in respect of recruitment of estrogen receptor-alpha (ERα), a ligand-activated transcription factor, to uterine gene promoters in mice. Recent genearray studies, utilizing ERα knock-out vs. wild-type mice, have revealed that estrogen regulates numerous uterine genes temporally and most importantly via ERα during the phase-II response, including three well characterized genes viz., lactoferrin (Ltf), progesterone receptor (Pgr) and cyclinD1 (Ccnd1). Here, utilizing systematic ChIP studies, we demonstrate endogenous recruitment of ERα to above uterine gene promoters following estradiol-17β (E2) injection in mice.
Abbreviations
- ChIP:
-
chromatin immunoprecipitation
- ERα:
-
estrogen receptor-alpha
- Ltf :
-
lactoferrin
- Pgr :
-
progesterone receptor
- Ccnd1:
-
cyclin D1
- IP:
-
immunoprecipitation
References
Hearnes JM, Mays DJ, Schavolt KL, Tang L, Jiang X, Pietenpol JA. Chromatin immunoprecipitation-based screen to identify functional genomic binding sites for sequence-specific transactivators. Mol Cell Biol 2005; 25:10148–10158.
Couse JF, Korach KS. Estrogen receptor null mice: what have we learned and where will they lead us? Endo Rev 1999; 20(3):358–417.
Tsai MJ, O’Malley BW. Molecular mechanisms of action of steroid/thyroid receptor superfamily members. Ann Rev Biochem 1994; 63:451–486.
Ray S, Hou X, Zhou H-E, Wang H, Das SK. Bip is a molecular link between the phase-I and phase-II estrogenic responses in uterus. Mol Endocrinol 2006; 20(8):1825–1837.
Hewitt SC, Deroo BJ, Hansen K, Collins J, Grissom S, Afshari CA, Korach KS. Estrogen receptordependent genomic responses in the uterus mirror the biphasic physiological response to estrogen. Mol Endocrinol 2003; 17:2070–2083.
Das SK, Taylor JA, Korach KS, Paria BC, Dey SK, Lubahn DB. Estrogenic responses in estrogen receptor-α deficient mice reveal a novel estrogen signaling pathway. Proc Natl Acad Sci USA 1997; 94:12786–12791.
Das SK, Tan J, Raja S, Halder J, Paria BC, Dey SK. Estrogen targets genes involved in protein processing, calcium homeostasis and Wnt signaling in the mouse uterus independent of estrogen receptor-α and - β. J Biol Chem 2000; 275:28834–28842.
Hou X, Tan Y, Li M, Dey SK, Das SK. Canonical Wnt Signaling Is Critical to Estrogen Mediated Uterine Growth. Mol Endocrinol 2004; 18:3035–3049.
Tan J, Paria BC, Dey SK, Das SK. Differential uterine expression of estrogen and progesterone receptors correlates with uterine preparation for implantation and decidualization in the mouse. Endocrinology 1999; 140:5310–5321.
Krege JH, Hodgin JB, Couse JF, Enmark E, Warner M, Mahler JF, Sar M, Korach KS, Gustafsson JA, Smithies O. Generation and reproductive phenotypes of mice lacking estrogen receptor β. Proc Natl Acad Sci USA 1998; 95:15677–15682.
Klein-Hitpass L, Ryffel GU, Heitlinger E, Cato AC. A 13 bp palindrome is a functional estrogen responsive element and interacts specifically with estrogen receptor. Nucleic Acids Res 1988; 16:647–663.
Sukovich DA, Mukherjee R, Benfield PA. A novel, celltype-specific mechanism for estrogen receptormediated gene activation in the absence of an estrogen-responsive element. Mol Cell Biol 1994; 14:7134–7143.
O’Lone R, Frith MC, Karlsson EK, Hansen U. Genomic targets of nuclear estrogen receptors. Mol Endocrinol 2004; 18:1859–1875.
Webb P, Nguyen P, Valentine C, Lopez GN, Kwok GR, McInerney E, Katzenellenbogen BS, Enmark E, Gustafsson JA, Nilsson S, Kushner PJ. The estrogen receptor enhances AP-1 activity by two distinct mechanisms with different requirements for receptor transactivation functions. Mol Endocrinol 1999; 13:1672–1685.
Das SK, Tan J, Johnson DC, Dey SK. Differential spatiotemporal regulation of lactoferrin and progesterone receptor genes in the mouse uterus by primary estrogen, catechol estrogen, and xenoestrogen. Endocrinology 1998; 139: 2905–2915.
Tong W, Pollard JW. Progesterone inhibits estrogeninduced cyclin D1 and cdk4 nuclear translocation, cyclin E- and cyclin A-cdk2 kinase activation, and cell proliferation in uterine epithelial cells in mice. Mol Cell Biol 1999; 19:2251–2264
Liu Y, Teng CT. Estrogen response module of the mouse lactoferrin gene contains overlapping chicken ovalbumin upstream promoter transcription factor and estrogen receptor-binding elements. Mol Endocrinol 1992; 6:355–364.
Planas-Silva MD, Shang Y, Donaher JL, Brown M, Weinberg RA. AIB1 enhances estrogen dependent induction of cyclin D1 expression. Cancer Res 2001; 61:3858–3862.
Petz LN, Ziegler YS, Schultz JR, Kim H, Kemper JK, Nardulli AM. Differential regulation of the human progesterone receptor gene through an estrogen response element half site and Sp1 sites. J Steroid Biochem Mol Biol 2004; 88:113–122.
Kazi AA, Jones JM, Koos RD. Chromatin immunoprecipitation analysis of gene expression in the rat uterus in vivo: estrogen-induced recruitment of both estrogen receptor alpha and hypoxiainducible factor 1 to the vascular endothelial growth factor promoter. Mol Endocrinol 2005; 19:2006–2019.
Rahman MA, Li M, Li P, Wang H, Dey SK, Das SK. Hoxa-10 deficiency alters region-specific gene expression and perturbs differentiation of natural killer cells during decidualization. Dev Biol 2006; 290:105–117.
Orlando V. Mapping chromosomal proteins in vivo by formaldehyde-crosslinked-chromatin immunoprecipitation. Trends Biochem Sci 2000; 25:99–104.
Yahata T, Shao W, Endoh H, Hur J, Coser KR, Sun H, Ueda Y, Kato S, Isselbacher KJ, Brown M, Shioda T. Selective coactivation of estrogen-dependent transcription by CITED1 CBP/p300-binding protein. Genes Dev 2001; 15:2598–2612.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
Open Access This article is published under license to BioMed Central Ltd. This is an Open Access article is distributed under the terms of the Creative Commons Attribution License ( https://creativecommons.org/licenses/by/2.0 ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
About this article
Cite this article
Ray, S., Das, S.K. Chromatin immunoprecipitation assay detects er recruitment to gene specific promoters in uterus. Biol. Proced. Online 8, 69–76 (2006). https://doi.org/10.1251/bpo120
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1251/bpo120