TY  - JOUR
AU  - Popović, Natasa M
AU  - Ruždijić, Sabera
AU  - Kanazir, Dusan T.
AU  - Niciforović, Ana
AU  - Adžić, Miroslav
AU  - Paraskevopoulou, Elissavet
AU  - Pantelidou, Constantia
AU  - Radojcić, Marija B
AU  - Demonacos, Constantinos
AU  - Krstić-Demonacos, Marija
PY  - 2010
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1375
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
IS  - 6
VL  - 75
EP  - 465
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1375
ER  - 

@article{
author = "Popović, Natasa M and Ruždijić, Sabera and Kanazir, Dusan T. and Niciforović, Ana and Adžić, Miroslav and Paraskevopoulou, Elissavet and Pantelidou, Constantia and Radojcić, Marija B 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",
number = "6",
volume = "75",
pages = "465",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1375"
}

Popović, N. M., Ruždijić, S., Kanazir, D. T., Niciforović, A., Adžić, M., Paraskevopoulou, E., Pantelidou, C., Radojcić, M. B., Demonacos, C.,& Krstić-Demonacos, M.. (2010). Site-specific and dose-dependent effects of glucocorticoid receptor phosphorylation in yeast Saccharomyces cerevisiae. in Steroids, 75(6).
https://hdl.handle.net/21.15107/rcub_ibiss_1375

Popović NM, Ruždijić S, Kanazir DT, Niciforović A, Adžić M, Paraskevopoulou E, Pantelidou C, Radojcić MB, Demonacos C, Krstić-Demonacos M. Site-specific and dose-dependent effects of glucocorticoid receptor phosphorylation in yeast Saccharomyces cerevisiae. in Steroids. 2010;75(6):null-465.
https://hdl.handle.net/21.15107/rcub_ibiss_1375 .

Popović, Natasa M, Ruždijić, Sabera, Kanazir, Dusan T., Niciforović, Ana, Adžić, Miroslav, Paraskevopoulou, Elissavet, Pantelidou, Constantia, Radojcić, Marija B, 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),
https://hdl.handle.net/21.15107/rcub_ibiss_1375 .

TY  - JOUR
AU  - Vujčić, Miroslava T.
AU  - Ruždijić, Sabera
AU  - Terzić, Nataša A.
AU  - Ristić-Fira, Aleksandra M.
AU  - Kanazir, Dusan T.
PY  - 2005
PY  - 2005
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/551
AB  - Abstract: In order to contribute to the understanding of mechanisms by which regulatory proteins recognize genetic information stored in DNA, analyses of their interaction with specific nucleotides are usually performed. In this study, the electrophoretic mobility shift assay (EMSA) was applied to analyze the interaction of nuclear proteins from the liver of rats of different age i.e., young (3-month-old), middle- aged (12-month-old) and aged (24-month-old), with radioactively labelled synthetic oligonucleotide analogues, corresponding to GRE. The levels of GRE binding activity were assessed by quantitative densitometric scanning of the autoradiograms. The results showed statistically significant decreasing values of up to 78% and 49% in middle aged and old animals, respectively, compared to young animals (p < 0.05). The specificity of the nuclear proteins-GRE interaction was demonstrated by competition experiments with unlabelled GRE. In a supershift assay, using the antibody BuGR2, it was shown that the GR proteins present in nuclear extracts have a high affinity for the GRE probe. The stabilities of the protein-DNA complexes were analysed and it was concluded that they changed during ageing.
AB  - U cilju doprinosa razumevanju mehanizama pomoću kojih regulatorni proteini prepoznaju genetičku informaciju koju nosi DNK, analiziraju se njihove interakcije sa specifičnim nukleotidima. U ovom radu je metodom EMSA analizirana interakcija jedarnih proteina iz jetri pacova iz tri starosne grupe (mladi - 3 meseca, srednje doba – 12 meseci i stari – 24 meseca) sa sintetičkim, radioaktivno obeleženim, oligonukleotidnim analogom GRE. Nivo vezujuće aktivnosti GRE je određivan kvantitativno denzitometrijskom autoradiografijom. Rezultati su pokazali da postoji statistički značajan pad vrednosti GRE-vezujuće aktivnosti do 78% kod životinja srednjeg starosnog doba i do 49 % kod starih životinja, u poređenju sa vrednostima dobijenim za mlade životinje (p < 0.05). Specifičnost interakcije jedarnih proteina i GRE je određena eksperimentima kompeticije sa neobeleženim GRE. Korišćenjem antitela BuGR2 pokazano je da je glukokortikoidni receptor protein koji u jedarnom ekstraktu ima najveći afinitet za GRE probu. Analizirana je stabilnost kompleksa protein-DNK i zaključeno je da se menja tokom starenja.
T2  - Journal of the Serbian Chemical Society
T1  - Analiza interakcije jedarnog glukokortikoidnog receptora i DNK u jetri pacova tokom starenja
T1  - Analysis of nuclear glucocorticoid receptor-DNA interaction in aged rat liver
IS  - 5
VL  - 70
SP  - 705
EP  - 712
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_551
ER  - 

@article{
author = "Vujčić, Miroslava T. and Ruždijić, Sabera and Terzić, Nataša A. and Ristić-Fira, Aleksandra M. and Kanazir, Dusan T.",
year = "2005, 2005",
abstract = "Abstract: In order to contribute to the understanding of mechanisms by which regulatory proteins recognize genetic information stored in DNA, analyses of their interaction with specific nucleotides are usually performed. In this study, the electrophoretic mobility shift assay (EMSA) was applied to analyze the interaction of nuclear proteins from the liver of rats of different age i.e., young (3-month-old), middle- aged (12-month-old) and aged (24-month-old), with radioactively labelled synthetic oligonucleotide analogues, corresponding to GRE. The levels of GRE binding activity were assessed by quantitative densitometric scanning of the autoradiograms. The results showed statistically significant decreasing values of up to 78% and 49% in middle aged and old animals, respectively, compared to young animals (p < 0.05). The specificity of the nuclear proteins-GRE interaction was demonstrated by competition experiments with unlabelled GRE. In a supershift assay, using the antibody BuGR2, it was shown that the GR proteins present in nuclear extracts have a high affinity for the GRE probe. The stabilities of the protein-DNA complexes were analysed and it was concluded that they changed during ageing., U cilju doprinosa razumevanju mehanizama pomoću kojih regulatorni proteini prepoznaju genetičku informaciju koju nosi DNK, analiziraju se njihove interakcije sa specifičnim nukleotidima. U ovom radu je metodom EMSA analizirana interakcija jedarnih proteina iz jetri pacova iz tri starosne grupe (mladi - 3 meseca, srednje doba – 12 meseci i stari – 24 meseca) sa sintetičkim, radioaktivno obeleženim, oligonukleotidnim analogom GRE. Nivo vezujuće aktivnosti GRE je određivan kvantitativno denzitometrijskom autoradiografijom. Rezultati su pokazali da postoji statistički značajan pad vrednosti GRE-vezujuće aktivnosti do 78% kod životinja srednjeg starosnog doba i do 49 % kod starih životinja, u poređenju sa vrednostima dobijenim za mlade životinje (p < 0.05). Specifičnost interakcije jedarnih proteina i GRE je određena eksperimentima kompeticije sa neobeleženim GRE. Korišćenjem antitela BuGR2 pokazano je da je glukokortikoidni receptor protein koji u jedarnom ekstraktu ima najveći afinitet za GRE probu. Analizirana je stabilnost kompleksa protein-DNK i zaključeno je da se menja tokom starenja.",
journal = "Journal of the Serbian Chemical Society",
title = "Analiza interakcije jedarnog glukokortikoidnog receptora i DNK u jetri pacova tokom starenja, Analysis of nuclear glucocorticoid receptor-DNA interaction in aged rat liver",
number = "5",
volume = "70",
pages = "705-712",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_551"
}

Vujčić, M. T., Ruždijić, S., Terzić, N. A., Ristić-Fira, A. M.,& Kanazir, D. T.. (2005). Analiza interakcije jedarnog glukokortikoidnog receptora i DNK u jetri pacova tokom starenja. in Journal of the Serbian Chemical Society, 70(5), 705-712.
https://hdl.handle.net/21.15107/rcub_ibiss_551

Vujčić MT, Ruždijić S, Terzić NA, Ristić-Fira AM, Kanazir DT. Analiza interakcije jedarnog glukokortikoidnog receptora i DNK u jetri pacova tokom starenja. in Journal of the Serbian Chemical Society. 2005;70(5):705-712.
https://hdl.handle.net/21.15107/rcub_ibiss_551 .

Vujčić, Miroslava T., Ruždijić, Sabera, Terzić, Nataša A., Ristić-Fira, Aleksandra M., Kanazir, Dusan T., "Analiza interakcije jedarnog glukokortikoidnog receptora i DNK u jetri pacova tokom starenja" in Journal of the Serbian Chemical Society, 70, no. 5 (2005):705-712,
https://hdl.handle.net/21.15107/rcub_ibiss_551 .

TY  - JOUR
AU  - Đorđević-Marković, R
AU  - Radić, O
AU  - Jelić, V
AU  - Radojcić, M
AU  - Rapić-Otrin, V
AU  - Ruždijić, Sabera
AU  - Krstić-Demonacos, Marija
AU  - Kanazir, Selma
AU  - Kanazir, Dusan T.
PY  - 1999
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/1840
AB  - The role of the glucocorticoid receptor (GR) in senescence was studied in rats of increasing age. Statistically significant changes in the number of GRs from rat liver were detected, whereas the affinity for the ligand triamcinolone acetonide (TA) did not change with increasing age, and was in the range of 1-2 nM. In all cases the number of receptors was lower in rats treated with hormone in vivo relative to untreated animals. In addition, we have found changes in GR activation, as measured by the binding to DNA cellulose in the mentioned age groups. Furthermore, expression of the glucocorticoid hormone (GH)-inducible gene, tyrosine amino transferase (TAT) also showed age-related alterations. We conclude that receptor function shows oscillatory changes during ageing. In addition, response to GH generally declines towards the older age. This. specific periodicity in functional characteristics of the GR may reconcile conflicting results about the receptor number and properties during the ageing process, and marks particular age at which individual organism shows the highest or the lowest sensitivity to the actions of GH. (C) 1999 Elsevier Science Inc. All rights reserved.
T2  - Experimental Gerontology
T1  - Glucocorticoid receptors in ageing rats
IS  - 8
VL  - 34
EP  - 982
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_1840
ER  - 

@article{
author = "Đorđević-Marković, R and Radić, O and Jelić, V and Radojcić, M and Rapić-Otrin, V and Ruždijić, Sabera and Krstić-Demonacos, Marija and Kanazir, Selma and Kanazir, Dusan T.",
year = "1999",
abstract = "The role of the glucocorticoid receptor (GR) in senescence was studied in rats of increasing age. Statistically significant changes in the number of GRs from rat liver were detected, whereas the affinity for the ligand triamcinolone acetonide (TA) did not change with increasing age, and was in the range of 1-2 nM. In all cases the number of receptors was lower in rats treated with hormone in vivo relative to untreated animals. In addition, we have found changes in GR activation, as measured by the binding to DNA cellulose in the mentioned age groups. Furthermore, expression of the glucocorticoid hormone (GH)-inducible gene, tyrosine amino transferase (TAT) also showed age-related alterations. We conclude that receptor function shows oscillatory changes during ageing. In addition, response to GH generally declines towards the older age. This. specific periodicity in functional characteristics of the GR may reconcile conflicting results about the receptor number and properties during the ageing process, and marks particular age at which individual organism shows the highest or the lowest sensitivity to the actions of GH. (C) 1999 Elsevier Science Inc. All rights reserved.",
journal = "Experimental Gerontology",
title = "Glucocorticoid receptors in ageing rats",
number = "8",
volume = "34",
pages = "982",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_1840"
}

Đorđević-Marković, R., Radić, O., Jelić, V., Radojcić, M., Rapić-Otrin, V., Ruždijić, S., Krstić-Demonacos, M., Kanazir, S.,& Kanazir, D. T.. (1999). Glucocorticoid receptors in ageing rats. in Experimental Gerontology, 34(8).
https://hdl.handle.net/21.15107/rcub_ibiss_1840

Đorđević-Marković R, Radić O, Jelić V, Radojcić M, Rapić-Otrin V, Ruždijić S, Krstić-Demonacos M, Kanazir S, Kanazir DT. Glucocorticoid receptors in ageing rats. in Experimental Gerontology. 1999;34(8):null-982.
https://hdl.handle.net/21.15107/rcub_ibiss_1840 .

Đorđević-Marković, R, Radić, O, Jelić, V, Radojcić, M, Rapić-Otrin, V, Ruždijić, Sabera, Krstić-Demonacos, Marija, Kanazir, Selma, Kanazir, Dusan T., "Glucocorticoid receptors in ageing rats" in Experimental Gerontology, 34, no. 8 (1999),
https://hdl.handle.net/21.15107/rcub_ibiss_1840 .