Site-specific and dose-dependent effects of glucocorticoid receptor phosphorylation in yeast Saccharomyces cerevisiae
Authors
Popović, Nataša M.Ruždijić, Sabera
Kanazir, Dušan T.
Nićiforović, Ana
Adžić, Miroslav
Paraskevopoulou, Elissavet
Pantelidou, Constantia
Radojčić, Marija
Demonacos, Constantinos
Krstić-Demonacos, Marija
Article (Published version)
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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. Crown Copyright (C) 2010 Published by Elsevier Inc. All rights reserved.
Keywords:
Transcription / Glucocorticoid receptor / Phosphorylation / YeastSource:
Steroids, 2010, 75, 6, 457-465Funding / projects:
- Ministry of Science and Technological Development of Serbia [03E24, 143042B, 143044B], Serbian Academy of Sciences and Arts, Wellcome Trust [069024]
DOI: 10.1016/j.steroids.2010.03.001
ISSN: 0039-128X
PubMed: 20223255
WoS: 000277670100011
Scopus: 2-s2.0-77950537809
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VinčaTY - JOUR AU - Popović, Nataša M. AU - Ruždijić, Sabera AU - Kanazir, Dušan T. AU - Nićiforović, Ana AU - Adžić, Miroslav AU - Paraskevopoulou, Elissavet AU - Pantelidou, Constantia AU - Radojčić, Marija AU - Demonacos, Constantinos AU - Krstić-Demonacos, Marija PY - 2010 UR - https://vinar.vin.bg.ac.rs/handle/123456789/3999 AB - 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. Crown Copyright (C) 2010 Published by Elsevier Inc. All rights reserved. T2 - Steroids T1 - Site-specific and dose-dependent effects of glucocorticoid receptor phosphorylation in yeast Saccharomyces cerevisiae VL - 75 IS - 6 SP - 457 EP - 465 DO - 10.1016/j.steroids.2010.03.001 ER -
@article{ author = "Popović, Nataša M. and Ruždijić, Sabera and Kanazir, Dušan T. and Nićiforović, Ana and Adžić, Miroslav and Paraskevopoulou, Elissavet and Pantelidou, Constantia and Radojčić, Marija and Demonacos, Constantinos and Krstić-Demonacos, Marija", year = "2010", abstract = "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. Crown Copyright (C) 2010 Published by Elsevier Inc. All rights reserved.", journal = "Steroids", title = "Site-specific and dose-dependent effects of glucocorticoid receptor phosphorylation in yeast Saccharomyces cerevisiae", volume = "75", number = "6", pages = "457-465", doi = "10.1016/j.steroids.2010.03.001" }
Popović, N. M., Ruždijić, S., Kanazir, D. T., Nićiforović, A., Adžić, M., Paraskevopoulou, E., Pantelidou, C., Radojčić, M., Demonacos, C.,& Krstić-Demonacos, M.. (2010). Site-specific and dose-dependent effects of glucocorticoid receptor phosphorylation in yeast Saccharomyces cerevisiae. in Steroids, 75(6), 457-465. https://doi.org/10.1016/j.steroids.2010.03.001
Popović NM, Ruždijić S, Kanazir DT, Nićiforović A, Adžić M, Paraskevopoulou E, Pantelidou C, Radojčić M, Demonacos C, Krstić-Demonacos M. Site-specific and dose-dependent effects of glucocorticoid receptor phosphorylation in yeast Saccharomyces cerevisiae. in Steroids. 2010;75(6):457-465. doi:10.1016/j.steroids.2010.03.001 .
Popović, Nataša M., Ruždijić, Sabera, Kanazir, Dušan T., Nićiforović, Ana, Adžić, Miroslav, Paraskevopoulou, Elissavet, Pantelidou, Constantia, Radojčić, Marija, Demonacos, Constantinos, Krstić-Demonacos, Marija, "Site-specific and dose-dependent effects of glucocorticoid receptor phosphorylation in yeast Saccharomyces cerevisiae" in Steroids, 75, no. 6 (2010):457-465, https://doi.org/10.1016/j.steroids.2010.03.001 . .