Analysis of nuclear glucocorticoid receptor-DNA interaction in aged rat liver
Апстракт
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 LT 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 show...n 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.
Кључне речи:
ageing / liver / glucocorticoid receptor / GRE / EMSAИзвор:
Journal of the Serbian Chemical Society, 2005, 70, 5, 705-712
DOI: 10.2298/JSC0505705V
ISSN: 0352-5139
WoS: 000230375900004
Scopus: 2-s2.0-31544483056
Колекције
Институција/група
VinčaTY - JOUR AU - Vujčić, Miroslava T. AU - Terzić, N AU - Ristić-Fira, Aleksandra AU - Kanazir, Selma AU - Ruždijić, Sabera PY - 2005 UR - https://vinar.vin.bg.ac.rs/handle/123456789/2906 AB - 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 LT 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. T2 - Journal of the Serbian Chemical Society T1 - Analysis of nuclear glucocorticoid receptor-DNA interaction in aged rat liver VL - 70 IS - 5 SP - 705 EP - 712 DO - 10.2298/JSC0505705V ER -
@article{ author = "Vujčić, Miroslava T. and Terzić, N and Ristić-Fira, Aleksandra and Kanazir, Selma and Ruždijić, Sabera", year = "2005", 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 LT 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.", journal = "Journal of the Serbian Chemical Society", title = "Analysis of nuclear glucocorticoid receptor-DNA interaction in aged rat liver", volume = "70", number = "5", pages = "705-712", doi = "10.2298/JSC0505705V" }
Vujčić, M. T., Terzić, N., Ristić-Fira, A., Kanazir, S.,& Ruždijić, S.. (2005). Analysis of nuclear glucocorticoid receptor-DNA interaction in aged rat liver. in Journal of the Serbian Chemical Society, 70(5), 705-712. https://doi.org/10.2298/JSC0505705V
Vujčić MT, Terzić N, Ristić-Fira A, Kanazir S, Ruždijić S. Analysis of nuclear glucocorticoid receptor-DNA interaction in aged rat liver. in Journal of the Serbian Chemical Society. 2005;70(5):705-712. doi:10.2298/JSC0505705V .
Vujčić, Miroslava T., Terzić, N, Ristić-Fira, Aleksandra, Kanazir, Selma, Ruždijić, Sabera, "Analysis of nuclear glucocorticoid receptor-DNA interaction in aged rat liver" in Journal of the Serbian Chemical Society, 70, no. 5 (2005):705-712, https://doi.org/10.2298/JSC0505705V . .