Receptors from Glucocorticoid-Sensitive Lymphoma Cells and Two Classes of Insensitive Clones: Physical and DNA-Binding Properties
- 1 October 1974
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 71 (10), 3901-3905
- https://doi.org/10.1073/pnas.71.10.3901
Abstract
Mouse lymphoma tissue culture cells (S49.1A) are normally killed by dexamethasone, a synthetic glucocorticoid hormone. Dexamethasone-resistant clones have been selected from this line, some of which retain the ability to specifically bind dexamethasone. Addition of [(3)H]dexamethasone to cultures, followed by cell fractionation, reveals that the nuclear transfer of hormone-receptor complexes in some of these variant clones is deficient (nt(-)), while others show increased nuclear transfer (nt(i)) relative to the parental line. Two independently selected members of each class have been studied here, in an effort to elucidate the molecular determinants involved in the receptor-nucleus interaction in vivo. The labeled receptors in cell-free extracts bind to DNA-cellulose, but only after previous incubation of the extract at 20 degrees , similar to the treatment required for cell-free interaction of receptors with nuclei. More importantly, the apparent DNA-binding affinity of the nt(-) receptors is lower than the wild type, whereas the nt(i) receptors bind DNA with an affinity higher than the parental molecules. The parallelism of nuclear and DNA binding, together with the observations that the receptors from the variants have sedimentation properties different from the wild-type cells, lead us to conclude that (i) these variants may contain altered receptor molecules and (ii) DNA is probably the primary nuclear binding site for steroid receptors in vivo.Keywords
This publication has 26 references indexed in Scilit:
- Role of DNA and Specific Cytoplasmic Receptors in Glucocorticoid ActionProceedings of the National Academy of Sciences of the United States of America, 1972
- Estrogen receptor in the rat uterus. Physiological forms and artifactsBiochemistry, 1972
- Glucocorticoid receptors: Relations between steroid binding and biological effectsJournal of Molecular Biology, 1972
- Mechanisms of Interaction of a Hormone—Receptor Complex with the Genome of a Eukaryotic Target CellNature, 1972
- A reconstituted cell-free system for the specific transfer of steroid–receptor complexes into nuclear chromatin isolated from rat ventral prostate glandBiochemical Journal, 1971
- Mutation rates in cells at different ploidy levelsJournal of Cellular Physiology, 1971
- Specific Cytoplasmic Glucocorticoid Hormone Receptors in Hepatoma Tissue Culture CellsProceedings of the National Academy of Sciences of the United States of America, 1971
- Progesterone-binding components of chick oviduct. II. Nuclear components.1971
- Differentiated functions expressed by cultured mouse lymphoma cells: I. Specificity and kinetics of cell responses to corticosteroidsExperimental Cell Research, 1970
- Some properties of a nuclear binding site of estradiolJournal of Molecular Biology, 1967