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Deficiency in TR4 nuclear receptor abrogates Gadd45a expression and increases cytotoxicity induced by ionizing radiation

Cellular & Molecular Biology Letters volume 17, pages 309–322 (2012)Cite this article

Abstract

The testicular receptor 4 (TR4) is a member of the nuclear receptor superfamily that controls various biological activities. A protective role of TR4 against oxidative stress has recently been discovered. We here examined the protective role of TR4 against ionizing radiation (IR) and found that small hairpin RNA mediated TR4 knockdown cells were highly sensitive to IR-induced cell death. IR exposure increased the expression of TR4 in scramble control small hairpin RNA expressing cells but not in TR4 knockdown cells. Examination of IR-responsive molecules found that the expression of Gadd45a, the growth arrest and DNA damage response gene, was dramatically decreased in Tr4 deficient (TR4KO) mice tissues and could not respond to IR stimulation in TR4KO mouse embryonic fibroblast cells. This TR4 regulation of GADD45A was at the transcriptional level. Promoter analysis identified four potential TR4 response elements located in intron 3 and exon 4 of the GADD45A gene. Reporter and chromatin immunoprecipitation (ChIP) assays provided evidence indicating that TR4 regulated the GADD45A expression through TR4 response elements located in intron 3 of the GADD45A gene. Together, we find that TR4 is essential in protecting cells from IR stress. Upon IR challenges, TR4 expression is increased, thereafter inducing GADD45A through transcriptional regulation. As GADD45A is directly involved in the DNA repair pathway, this suggests that TR4 senses genotoxic stress and up-regulates GADD45A expression to protect cells from IR-induced genotoxicity.

Abbreviations

ChIP:

chromatin immunoprecipitation

DR:

direct repeat

GADD45A:

growth arrest and DNA-damage-inducible, alpha

GADDLuc:

GADD45A gene controlled luciferase reporter

IgG:

immunoglobulin G

IR:

ionizing radiation

MEF:

mouse embryonic fibroblast

MTT:

3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide

SC:

cells expressing scramble control shRNA

shRNA:

small hairpin RNA

shTR4:

cell expressing shRNA targeting TR4

TR4:

testicular nuclear receptor 4

TR4KO:

TR4 knockout

TR4RE:

TR4 response element

UV:

ultraviolet light irradiation

WT:

wild type

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Authors and Affiliations

  1. George Whipple Lab for Cancer Research, Departments of Pathology, Urology, and Radiation Oncology, University of Rochester Medical Center, Rochester, NY, 14642, USA

    Shian-Jang Yan, Yi-Fen Lee, Huei-Ju Ting, Ning-Chun Liu, Su Liu, Shin-Jen Lin, Gonghui Li & Chawnshang Chang

  2. Department of Urology, Taipei Medical University, Taipei, 110, Taiwan

    Shauh-Der Yeh

  3. Sex Hormone Research Center, China Medical University/Hospital, Taichung, 404, Taiwan

    Chawnshang Chang

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  1. Shian-Jang Yan
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  2. Yi-Fen Lee
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Correspondence to Chawnshang Chang.

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These authors contributed equally to this paper.

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Yan, SJ., Lee, YF., Ting, HJ. et al. Deficiency in TR4 nuclear receptor abrogates Gadd45a expression and increases cytotoxicity induced by ionizing radiation. Cell Mol Biol Lett 17, 309–322 (2012). https://doi.org/10.2478/s11658-012-0012-9

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  • DOI: https://doi.org/10.2478/s11658-012-0012-9

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Cellular & Molecular Biology Letters

ISSN: 1689-1392

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