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| Status |
Public on Apr 07, 2017 |
| Title |
MDFIC alteration of the GR transcriptome in COS-1 cells depends on S211 phosphorylation of GR |
| Platform organism |
Homo sapiens |
| Sample organism |
Chlorocebus aethiops |
| Experiment type |
Expression profiling by array
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| Summary |
Glucocorticoids are primary stress hormones that regulate many physiological processes, and synthetic derivatives of these molecules and are widely used in the clinic. The cellular response to glucocorticoids is remarkably diverse; however, the molecular factors that govern tissue specificity are poorly understood. The actions of glucocorticoids are mediated by the glucocorticoid receptor (GR). To discover new proteins that interact with GR and modulate its function, we performed a yeast 2 hybrid assay using as bait the hinge region of GR. The MyoD family inhibitor domain-containing (MDFIC) protein was identified as a binding partner for GR. Knockdown of MDFIC in A549 cells alters the GR transcriptome. Overexpression of MDFIC with GR in COS-1 cells also modulates the GR transcriptome by expanding the number of genes regulated in response to glucocorticoid treatment. Our findings in A549 cells suggest that MDFIC alters the gene regulatory profile of GR by modulating receptor phosphorylation at several residues, including S211. To further investigate the molecular link between MDFIC-mediated effects on GR phosphorylation at S211 and alterations in the GR transcriptome, we performed a genome-wide microarray in COS-1 cells that were transfected with the GR phosphorylation mutant S211A or S211A and MDFIC. The transfected cells were treated with vehicle or the synthetic glucocorticoid Dexamethasone (Dex) for 6 hours. The ability of MDFIC to expand the GR transcriptome was attenuated in cells expressing S211A mutant.
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| Overall design |
COS-1 cells were transfected with the GR phosphorylation mutant S211A or S211A and MDFIC. The day after transfection, medium was replaced with new medium supplemented with 10% charcoal-stripped FBS. Forty-eight hours post-transfection, cells were stimulated with either vehicle (control) or 100nM Dex for 6 hours. Total RNA was harvested for microarray analysis using the RNeasy Mini Kit and RNase-Free DNase Kit (Qiagen) from 3 biological replicates from each group.
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| Contributor(s) |
Oakley RH, Cidlowski JA |
| Citation missing |
Has this study been published? Please login to update or notify GEO. |
| Submission date |
Jan 21, 2017 |
| Last update date |
Feb 22, 2018 |
| Contact name |
NIEHS Microarray Core |
| E-mail(s) |
microarray@niehs.nih.gov, liuliw@niehs.nih.gov
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| Organization name |
NIEHS
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| Department |
DIR
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| Lab |
Microarray Core
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| Street address |
111 T.W. Alexander Drive
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| City |
RTP |
| State/province |
NC |
| ZIP/Postal code |
27709 |
| Country |
USA |
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| Platforms (1) |
| GPL4133 |
Agilent-014850 Whole Human Genome Microarray 4x44K G4112F (Feature Number version) |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA362784 |
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