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Steroid hormones--Receptors; Gene regulatory networks; DNA; Chromatin


A cell-free system was established to study the role of steroid hormones In transcriptional control. The system consists of hormone-receptor-chromatin complex formation using partially purified receptor and chromatin. Micrococcal nuclease digestion of the complex released a 7S form of receptor. The absorbance at 260 nm also showed a peak in the 7S region of the sucrose gradient. DNAase I digestion caused the 7S receptor to shift to 2.8S form, while RNAase had no effect. When the receptor-chromatin complex was digested with DNAase I, different forms of receptors were observed, depending on the digestion time. Digestion of one minute produced a heterogeneous population of receptors ranging from 3S to 7S. Prolonged digestion caused a stepwise reduction In receptor size. These results suggest that 7S receptor contains an associated DNA fragment, and the transcriptional control mechanism involves binding of the steroid receptor with DNA of active chromatin.

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