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Site-specific and dose-dependent effects of glucocorticoid receptor phosphorylation in yeast Saccharomyces cerevisiae
The glucocorticoid receptor (GR) signal transduction and transcriptional regulation are efficiently recapitulated when GR is expressed in Saccharomyces cerevisiae. In this report we demonstrate that the in vivo GR phosphorylation pattern, hormone dependency and interdependency of phosphorylation eve...
Autores principales: | , , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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Elsevier
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2880793/ https://www.ncbi.nlm.nih.gov/pubmed/20223255 http://dx.doi.org/10.1016/j.steroids.2010.03.001 |
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author | Popovic, Natasa Ruzdijic, Sabera Kanazir, Dusan T. Niciforovic, Ana Adzic, Miroslav Paraskevopoulou, Elissavet Pantelidou, Constantia Radojcic, Marija Demonacos, Constantinos Krstic-Demonacos, Marija |
author_facet | Popovic, Natasa Ruzdijic, Sabera Kanazir, Dusan T. Niciforovic, Ana Adzic, Miroslav Paraskevopoulou, Elissavet Pantelidou, Constantia Radojcic, Marija Demonacos, Constantinos Krstic-Demonacos, Marija |
author_sort | Popovic, Natasa |
collection | PubMed |
description | The glucocorticoid receptor (GR) signal transduction and transcriptional regulation are efficiently recapitulated when GR is expressed in Saccharomyces cerevisiae. In this report we demonstrate that the in vivo GR phosphorylation pattern, hormone dependency and interdependency of phosphorylation events were similar in yeast and mammalian cells. GR phosphorylation at S246 exhibited inhibitory effect on S224 and S232 phosphorylation, suggesting the conservation of molecular mechanisms that control this interdependence between yeast and mammalian cells. To assess the effects of GR phosphorylation the mutated GR derivatives T171A, S224A, S232A, S246A were overexpressed and their transcriptional activity was analysed. These receptor derivatives displayed significant hormone inducible transcription when overexpressed in S. cerevisiae. We have established an inducible methionine expression system, which allows the close regulation of the receptor protein levels to analyse the dependence of GR function on its phosphorylation and protein abundance. Using this system we observed that GR S246A mutation increased its activity across all of the GR concentrations tested. The activity of the S224A and S246A mutants was mostly independent of GR protein levels, whereas the WT, T171A and S232A mediated transcription diminished with declining GR protein levels. Our results suggest that GR phosphorylation at specific residues affects its transcriptional functions in a site selective manner and these effects were directly linked to GR dosage. |
format | Text |
id | pubmed-2880793 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-28807932010-07-09 Site-specific and dose-dependent effects of glucocorticoid receptor phosphorylation in yeast Saccharomyces cerevisiae Popovic, Natasa Ruzdijic, Sabera Kanazir, Dusan T. Niciforovic, Ana Adzic, Miroslav Paraskevopoulou, Elissavet Pantelidou, Constantia Radojcic, Marija Demonacos, Constantinos Krstic-Demonacos, Marija Steroids Article The glucocorticoid receptor (GR) signal transduction and transcriptional regulation are efficiently recapitulated when GR is expressed in Saccharomyces cerevisiae. In this report we demonstrate that the in vivo GR phosphorylation pattern, hormone dependency and interdependency of phosphorylation events were similar in yeast and mammalian cells. GR phosphorylation at S246 exhibited inhibitory effect on S224 and S232 phosphorylation, suggesting the conservation of molecular mechanisms that control this interdependence between yeast and mammalian cells. To assess the effects of GR phosphorylation the mutated GR derivatives T171A, S224A, S232A, S246A were overexpressed and their transcriptional activity was analysed. These receptor derivatives displayed significant hormone inducible transcription when overexpressed in S. cerevisiae. We have established an inducible methionine expression system, which allows the close regulation of the receptor protein levels to analyse the dependence of GR function on its phosphorylation and protein abundance. Using this system we observed that GR S246A mutation increased its activity across all of the GR concentrations tested. The activity of the S224A and S246A mutants was mostly independent of GR protein levels, whereas the WT, T171A and S232A mediated transcription diminished with declining GR protein levels. Our results suggest that GR phosphorylation at specific residues affects its transcriptional functions in a site selective manner and these effects were directly linked to GR dosage. Elsevier 2010-06 /pmc/articles/PMC2880793/ /pubmed/20223255 http://dx.doi.org/10.1016/j.steroids.2010.03.001 Text en © 2010 Elsevier Inc. https://creativecommons.org/licenses/by/4.0/ Open Access under CC BY 4.0 (https://creativecommons.org/licenses/by/4.0/) license |
spellingShingle | Article Popovic, Natasa Ruzdijic, Sabera Kanazir, Dusan T. Niciforovic, Ana Adzic, Miroslav Paraskevopoulou, Elissavet Pantelidou, Constantia Radojcic, Marija Demonacos, Constantinos Krstic-Demonacos, Marija Site-specific and dose-dependent effects of glucocorticoid receptor phosphorylation in yeast Saccharomyces cerevisiae |
title | Site-specific and dose-dependent effects of glucocorticoid receptor phosphorylation in yeast Saccharomyces cerevisiae |
title_full | Site-specific and dose-dependent effects of glucocorticoid receptor phosphorylation in yeast Saccharomyces cerevisiae |
title_fullStr | Site-specific and dose-dependent effects of glucocorticoid receptor phosphorylation in yeast Saccharomyces cerevisiae |
title_full_unstemmed | Site-specific and dose-dependent effects of glucocorticoid receptor phosphorylation in yeast Saccharomyces cerevisiae |
title_short | Site-specific and dose-dependent effects of glucocorticoid receptor phosphorylation in yeast Saccharomyces cerevisiae |
title_sort | site-specific and dose-dependent effects of glucocorticoid receptor phosphorylation in yeast saccharomyces cerevisiae |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2880793/ https://www.ncbi.nlm.nih.gov/pubmed/20223255 http://dx.doi.org/10.1016/j.steroids.2010.03.001 |
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