The properties of a nuclear acidic protein fraction that binds [6,7−3H]oestradiol-17β

Abstract

1. Additional evidence was obtained that the nuclear oestradiol-17β receptor is an acidic protein. Partial purification of the receptor protein was obtained by chromatography on hydroxyapatite and it contains protein-bound phosphate. 2. The nuclear ‘5s’ and cytoplasmic ‘9·5s’ and ‘5s’ receptors from uterus, dimethylbenzanthracene-induced mammary adenocarcinoma and kidney are precipitated together with bound oestradiol-17β by protamine sulphate. This common property suggests that the nuclear and cytoplasmic receptors are related to each other. 3. The properties of two acidic protein fractions from both liver and dimethylbenzanthracene-induced mammary adenocarcinoma are described. Fraction 1 contains two major components and fraction 2 contains one component, as judged from polyacrylamide-gel electrophoresis. Fraction 2 contains RNA and both fractions contain protein-bound phosphate. 4. These fractions form insoluble complexes with calf thymus histone, protamine sulphate and poly-l-lysine. The formation of these complexes is markedly affected by ionic strength and pH. Ionization of both the ∈-amino group of lysine and carboxyl group are involved. RNA and DNA do not appear to be involved. The interaction is not affected by EDTA or 1mm-Na⁺, -K⁺, -Ca²⁺, -Mg²⁺ or -Mn²⁺. Per unit weight, whole histone has 4–5 times as many binding sites for the acidic proteins as the latter have for the former. 5. No convincing evidence was obtained for DNA–acidic protein interaction, but, as judged from precipitation experiments, there was competition between DNA and acidic protein for histone. 6. Relatively large amounts of acidic protein partly relieved the histone inhibition of the template activity of DNA for Escherichia coli RNA polymerase (EC 2.7.7.6).