Analysis of nuclear glucocorticoid receptor-DNA interaction in aged rat liver
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 a...
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| Format: | Article |
| Language: | English |
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Serbian Chemical Society
2005-01-01
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| Series: | Journal of the Serbian Chemical Society |
| Subjects: | |
| Online Access: | http://www.doiserbia.nb.rs/img/doi/0352-5139/2005/0352-51390505705V.pdf |
| _version_ | 1818293167682748416 |
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| author | Vujčić Miroslava Terzić Nataša A. Ristić-Fira Aleksandra M. Kanazir Dušan Ruždijić Sabera D. |
| author_facet | Vujčić Miroslava Terzić Nataša A. Ristić-Fira Aleksandra M. Kanazir Dušan Ruždijić Sabera D. |
| author_sort | Vujčić Miroslava |
| collection | DOAJ |
| description | 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. . |
| first_indexed | 2024-12-13T03:11:33Z |
| format | Article |
| id | doaj.art-28c938b05c444aa3b4d0ebfe47d482d1 |
| institution | Directory Open Access Journal |
| issn | 0352-5139 1820-7421 |
| language | English |
| last_indexed | 2024-12-13T03:11:33Z |
| publishDate | 2005-01-01 |
| publisher | Serbian Chemical Society |
| record_format | Article |
| series | Journal of the Serbian Chemical Society |
| spelling | doaj.art-28c938b05c444aa3b4d0ebfe47d482d12022-12-22T00:01:35ZengSerbian Chemical SocietyJournal of the Serbian Chemical Society0352-51391820-74212005-01-0170570571210.2298/JSC0505705V0352-51390505705VAnalysis of nuclear glucocorticoid receptor-DNA interaction in aged rat liverVujčić Miroslava0Terzić Nataša A.1Ristić-Fira Aleksandra M.2Kanazir Dušan3Ruždijić Sabera D.4Institute of Chemistry, Technology and Metallurgy, Department of Chemistry, Belgrade'Vinča' Institute of Nuclear Sciences, Belgrade'Vinča' Institute of Nuclear Sciences, BelgradeSerbian Academy of Sciences and Arts, BelgradeInstitute for Biological Research 'Siniša Stanković', BelgradeAbstract: 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. .http://www.doiserbia.nb.rs/img/doi/0352-5139/2005/0352-51390505705V.pdfageingliverglucocorticoid receptorgreemsa. |
| spellingShingle | Vujčić Miroslava Terzić Nataša A. Ristić-Fira Aleksandra M. Kanazir Dušan Ruždijić Sabera D. Analysis of nuclear glucocorticoid receptor-DNA interaction in aged rat liver Journal of the Serbian Chemical Society ageing liver glucocorticoid receptor gre emsa. |
| title | Analysis of nuclear glucocorticoid receptor-DNA interaction in aged rat liver |
| title_full | Analysis of nuclear glucocorticoid receptor-DNA interaction in aged rat liver |
| title_fullStr | Analysis of nuclear glucocorticoid receptor-DNA interaction in aged rat liver |
| title_full_unstemmed | Analysis of nuclear glucocorticoid receptor-DNA interaction in aged rat liver |
| title_short | Analysis of nuclear glucocorticoid receptor-DNA interaction in aged rat liver |
| title_sort | analysis of nuclear glucocorticoid receptor dna interaction in aged rat liver |
| topic | ageing liver glucocorticoid receptor gre emsa. |
| url | http://www.doiserbia.nb.rs/img/doi/0352-5139/2005/0352-51390505705V.pdf |
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