Abstract

We created fusion chimeras between GR and other nuclear receptors that manifest ligand dependent cytoplasmic/nuclear translocation for the chimeric receptor in response to the heterologous ligand. The approach has been successfuly developed both for the retinoic acid receptor (RAR) and the estrogen receptor (ER). We developed cell lines which express fluorescently tagged versions of the receptor chimeras. The ER-GR chimeras have been utilized to screen chemical libraries, and identify new compounds with ER stimulating activity. Thus, the concept that chimeric GR fusions can be used to identify new ligands has been confirmed in a large scale screening protocol. We now believe that this approach can be generalized to many members of the nuclear receptor super-family, opening a new avenue for ligand discovery.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC010027-15
Application #
7965200
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
15
Fiscal Year
2009
Total Cost
$270,388
Indirect Cost

  Institution

Name
National Cancer Institute Division of Basic Sciences
Department
Type
DUNS #
City
State
Country
Zip Code

  Publications

Oh, Kyu-Seon; Gottschalk, Rachel A; Lounsbury, Nicolas W et al. (2018) Dual Roles for Ikaros in Regulation of Macrophage Chromatin State and Inflammatory Gene Expression. J Immunol 201:757-771
Presman, Diego M; Ball, David A; Paakinaho, Ville et al. (2017) Quantifying transcription factor binding dynamics at the single-molecule level in live cells. Methods 123:76-88
Oh, Kyu-Seon; Patel, Heta; Gottschalk, Rachel A et al. (2017) Anti-Inflammatory Chromatinscape Suggests Alternative Mechanisms of Glucocorticoid Receptor Action. Immunity 47:298-309.e5
Stavreva, Diana A; Varticovski, Lyuba; Levkova, Ludmila et al. (2016) Novel cell-based assay for detection of thyroid receptor beta-interacting environmental contaminants. Toxicology 368-369:69-79
Stavreva, Diana A; Hager, Gordon L (2015) Chromatin structure and gene regulation: a dynamic view of enhancer function. Nucleus 6:442-8
Dull, Angie B; George, Anuja A; Goncharova, Ekaterina I et al. (2014) Identification of compounds by high-content screening that induce cytoplasmic to nuclear localization of a fluorescent estrogen receptor ? chimera and exhibit agonist or antagonist activity in vitro. J Biomol Screen 19:242-52
Guertin, Michael J; Zhang, Xuesen; Coonrod, Scott A et al. (2014) Transient estrogen receptor binding and p300 redistribution support a squelching mechanism for estradiol-repressed genes. Mol Endocrinol 28:1522-33
Miranda, Tina B; Voss, Ty C; Hager, Gordon L (2013) High-throughput fluorescence-based screen to identify factors involved in nuclear receptor recruitment to response elements. Methods Mol Biol 1042:3-12
Aguilar-Arnal, Lorena; Hakim, Ofir; Patel, Vishal R et al. (2013) Cycles in spatial and temporal chromosomal organization driven by the circadian clock. Nat Struct Mol Biol 20:1206-13
Kieffer-Kwon, Kyong-Rim; Tang, Zhonghui; Mathe, Ewy et al. (2013) Interactome maps of mouse gene regulatory domains reveal basic principles of transcriptional regulation. Cell 155:1507-20

Showing the most recent 10 out of 27 publications