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Glucocorticoid receptor profiling in U2OS cells expressing GRalpha or GRgamma isoform

ABSTRACT: Glucocorticoid receptor profiling in U2OS cells expressing GRalpha or GRgamma isoform

PROVIDER: PRJEB6695 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR560462.fastq.gz	Fastqsanger.gz
	ERR560463.fastq.gz	Fastqsanger.gz
	ERR560464.fastq.gz	Fastqsanger.gz

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Glucocorticoid receptor profiling in U2OS cells expressing GRalpha or GRgamma isoform

Project description:To determine whether insertion of a single amino acid had an effect on Glucocorticoid receptor (GR) occupancy, we performed ChIP-seq of GRalpha and GRgamma in the same U2OS osteosarcoma cell lines, upon glucocorticoid treatment. These assays are also part of submission E-MTAB-2955

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Project description:To identify the sequences responsible for recruitment of Glucocorticoid receptor (GR) to individual loci, we performed ChIP-seq and ChIP-exo that combines chromatin immunoprecipitation with an exonuclease digestion step. We performed these experiments in three cell lines : IMR90 (ATTC:CCL-186), U2OS osteosarcoma cell lines, K562 (ATCC:CCL243), upon glucocorticoid treatment. The U2OS assays are the same as those in E-MTAB-2731.

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SUMOylation Regulates the Anti-Proliferative Gene Signature Programs of Glucocorticoid Receptor (U2Os cell line)

Project description:In addition to the glucocorticoids, the glucocorticoid receptor (GR) is regulated by post-translational modifications, including SUMOylation. We have analyzed how SUMOylation influences the activity of endogenous GR target genes and the receptor chromatin binding by using isogenic HEK293 cells expressing wild-type GR (wtGR) or SUMOylation-defective GR (GR3KR). Gene expression profiling revealed that both dexamethasone up- and down-regulated genes are affected by the GR sumoylation and that the affected genes are significantly associated with pathways of cellular proliferation and survival. The GR3KR-expressing cells proliferated more rapidly and their anti-proliferative response to dexamethasone was less pronounced than in the wtGR-expressing cells. ChIP-seq analyses indicated that the SUMOylation modulates the chromatin occupancy of GR on several loci associated with cellular growth in a fashion which parallels with their differential dexamethasone-regulated expression between the two cell lines. Moreover, genome-wide SUMO-2/3 marks, which were generally associated with active chromatin, showed markedly higher overlap with the wtGR cistrome than with the GR3KR cistrome. In sum, our results indicate that the SUMOylation does not simply repress the GR activity, but regulates the activity of the receptor in a target locus selective fashion, playing an important role in controlling the GR activity on genes influencing cell growth. Total RNA isolated from U2Os cell lines stably expressing either wild-type GR (wtGR) or SUMOylation-defective GR (GR3KR) treated with either vehicle (EtOH) or 100 nM of dexamethasone (dex) for 6h.

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