Binding of the glucocorticoid receptor complex to the nucleosomal core in the P1798 mouse lymphosarcoma.
Binding of the glucocorticoid receptor complex to nucleosomes has been studied using the mouse P1798 lymphosarcoma. Cells were incubated with [3H]triamcinolone acetonide (TA), and nuclei prepared and digested with 3 different concentrations of micrococcal nuclease. After fractionation with EDTA and NaCl, it was observed that [3H]TA bound with similar specific radioactivity to mononucleosomes containing both core and linker DNA, of 183 +/- 5, and 168 +/- 4 base pair lengths, respectively, as well as to core size DNA, of 148 +/- 3 base pair length, suggesting that the glucocorticoid receptor bound to the core portion of the nucleosome. Steroid binding was found to be associated with regions of the nucleosome that were depleted in histone H1 and enriched in high mobility group (HMG) proteins 1 and 2; only negligible binding was noted in nucleosomes enriched in histone H1 and depleted in HMG proteins. In addition to binding to core nucleosomes, the glucocorticoid receptor complex was also shown to bind to a fraction sedimenting at 5-6 S on sucrose gradients characterized by subnucleosome and mononucleosome size DNA, as well as by core histones. While binding of the steroid receptor complex to linker regions of the nucleosome cannot be ruled out, this data would appear to present the first concrete evidence that glucocorticoid binding, at least in the P1798 lymphosarcoma, is to core nucleosomes. Some caution in interpretation of the results is indicated, however, on 2 points: (1) receptor redistribution during nuclease digestion cannot be ruled out; (2) only the binding of a small proportion of the steroid receptor complex may be physiologically relevant.
D000818:Animals; D001482:Base Composition; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D002843:Chromatin / Q000378:metabolism; D004247:DNA; D004492:Edetic Acid; D008228:Lymphoma, Non-Hodgkin / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008811:Mice, Inbred DBA; D008836:Micrococcal Nuclease / Q000378:metabolism; D009707:Nucleosomes / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D011987:Receptors, Steroid / Q000378:metabolism*; D012965:Sodium Chloride; D012995:Solubility; D014158:Transcription, Genetic; D014222:Triamcinolone Acetonide / Q000378:metabolism
D002843:Chromatin; D009707:Nucleosomes; D011506:Proteins; D011965:Receptors, Glucocorticoid; D011987:Receptors, Steroid; D012965:Sodium Chloride; D004247:DNA; D004492:Edetic Acid; D008836:Micrococcal Nuclease; D014222:Triamcinolone Acetonide