Analysis of nuclear glucocorticoid receptor-DNA interaction in aged rat liver
| Autor: | Miroslava Vujčić, M Aleksandra Ristic-Fira, D Sabera Ruzdijic, A Natasa Terzic, Dusan T. Kanazir |
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| Rok vydání: | 2005 |
| Předmět: |
liver
EMSA lcsh:Chemistry 03 medical and health sciences chemistry.chemical_compound Glucocorticoid receptor gre polycyclic compounds glucocorticoid receptor Nucleotide Electrophoretic mobility shift assay Nuclear protein 030304 developmental biology chemistry.chemical_classification 0303 health sciences biology Oligonucleotide 030302 biochemistry & molecular biology General Chemistry lcsh:QD1-999 Biochemistry chemistry Ageing ageing GRE biology.protein Antibody DNA emsa |
| Zdroj: | Journal of the Serbian Chemical Society Journal of the Serbian Chemical Society, Vol 70, Iss 5, Pp 705-712 (2005) |
| Popis: | 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. . |
| Databáze: | OpenAIRE |
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