Impact of glucocorticoid receptor density on ligand- independent dimerization, cooperative ligand-binding and basal priming of transactivation : a cell culture model
| dc.contributor.author | Robertson, Steven | |
| dc.contributor.author | Rohwer, Johann M. | |
| dc.contributor.author | Hapgood, Janet P | |
| dc.contributor.author | Louw, Ann | |
| dc.date.accessioned | 2013-06-18T07:12:52Z | |
| dc.date.available | 2013-06-18T07:12:52Z | |
| dc.date.issued | 2013-05 | |
| dc.description | The original publication is available at www.plosone.org | en_ZA |
| dc.description | Publication of this article was funded by the Stellenbosch University Open Access Fund. | |
| dc.description.abstract | Glucocorticoid receptor (GR) levels vary between tissues and individuals and are altered by physiological and pharmacological effectors. However, the effects and implications of differences in GR concentration have not been fully elucidated. Using three statistically different GR concentrations in transiently transfected COS-1 cells, we demonstrate, using co-immunoprecipitation (CoIP) and fluorescent resonance energy transfer (FRET), that high levels of wild type GR (wtGR), but not of dimerization deficient GR (GRdim), display ligand-independent dimerization. Whole-cell saturation ligand-binding experiments furthermore establish that positive cooperative ligand-binding, with a concomitant increased ligand-binding affinity, is facilitated by ligand-independent dimerization at high concentrations of wtGR, but not GRdim. The down-stream consequences of ligand-independent dimerization at high concentrations of wtGR, but not GRdim, are shown to include basal priming of the system as witnessed by ligand-independent transactivation of both a GRE-containing promoterreporter and the endogenous glucocorticoid (GC)-responsive gene, GILZ, as well as ligand-independent loading of GR onto the GILZ promoter. Pursuant to the basal priming of the system, addition of ligand results in a significantly greater modulation of transactivation potency than would be expected solely from the increase in ligand-binding affinity. Thus ligand-independent dimerization of the GR at high concentrations primes the system, through ligand-independent DNA loading and transactivation, which together with positive cooperative ligand-binding increases the potency of GR agonists and shifts the bio-character of partial GR agonists. Clearly GR-levels are a major factor in determining the sensitivity to GCs and a critical factor regulating transcriptional programs. | en_ZA |
| dc.description.version | Publishers' Version | en_ZA |
| dc.format.extent | 17 p. : ill. | |
| dc.identifier.citation | Robertson, S. Rohwer, J. M. Hapgood, J. P. & Louw, A. 2013. Impact of glucocorticoid receptor density on ligand-independent dimerization, cooperative ligand-binding and basal priming of transactivation: a cell culture model. PLoS ONE, 8(5): e64831. doi:10.1371/journal.pone.0064831. | en_ZA |
| dc.identifier.issn | 1932-6203 (online) | |
| dc.identifier.other | doi:10.1371/journal.pone.0064831 | |
| dc.identifier.uri | http://hdl.handle.net/10019.1/80931 | |
| dc.language.iso | en_ZA | en_ZA |
| dc.publisher | Public Library of Science -- PLoS | en_ZA |
| dc.rights.holder | Authors retain copyright | en_ZA |
| dc.subject | Glucocorticoids -- Receptors | en_ZA |
| dc.subject | Inflammatory diseases | en_ZA |
| dc.subject | Ligand binding (Biochemistry) | en_ZA |
| dc.subject | DNA | en_ZA |
| dc.title | Impact of glucocorticoid receptor density on ligand- independent dimerization, cooperative ligand-binding and basal priming of transactivation : a cell culture model | en_ZA |
| dc.type | Article | en_ZA |