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 for the retinoic acid receptor (RAR), the estrogen receptor (ER), and the progesterone receptor (PR). 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. We have also used the fluorescent protein fusion technology to show that U.S. water supplies are heavily contaminated with chemicals that have glucocorticoid activity.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC010308-13
Application #
8348970
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
13
Fiscal Year
2011
Total Cost
$549,339
Indirect Cost
Name
National Cancer Institute Division of Basic Sciences
Department
Type
DUNS #
City
State
Country
Zip Code
Lynch, Caitlin; Sakamuru, Srilatha; Huang, Ruili et al. (2017) Identifying environmental chemicals as agonists of the androgen receptor by using a quantitative high-throughput screening platform. Toxicology 385:48-58
Presman, Diego M; Hager, Gordon L (2017) More than meets the dimer: What is the quaternary structure of the glucocorticoid receptor? Transcription 8:32-39
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
Voss, Ty C; Hager, Gordon L (2014) Dynamic regulation of transcriptional states by chromatin and transcription factors. Nat Rev Genet 15:69-81
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
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
Stavreva, Diana A; George, Anuja A; Klausmeyer, Paul et al. (2012) Prevalent glucocorticoid and androgen activity in US water sources. Sci Rep 2:937
Biddie, Simon C; John, Sam; Sabo, Pete J et al. (2011) Transcription factor AP1 potentiates chromatin accessibility and glucocorticoid receptor binding. Mol Cell 43:145-55
Sung, Myong-Hee; McNally, James G (2011) Live cell imaging and systems biology. Wiley Interdiscip Rev Syst Biol Med 3:167-82
Dull, Angie; Goncharova, Ekaterina; Hager, Gordon et al. (2010) Development of an image analysis screen for estrogen receptor alpha (ER?) ligands through measurement of nuclear translocation dynamics. J Steroid Biochem Mol Biol 122:341-51

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