Modulation of DNA binding of glucocorticoid receptors

Glucocorticoid receptors of several rodent and human cell lines were subjected to mild proteolysis with several proteases. A hormone binding fragment of Mr approximately 40,000 was generated which had increased affinity for DNA as revealed by DNA-cellulose chromatography. It behaved similar to the truncated nti ('increased nuclear transfer') receptor of mutant mouse lymphoma cells. These data led to the view that wild-type receptors of Mr approximately 94,000 contain in addition to the functional domains for hormone binding and interaction with DNA a third domain ('modulation domain') which is essential for biological activity. Monoclonal antibodies against wild-type receptors were used in DNA binding experiments and increased affinity for DNA was observed. The data suggest that reacting the receptor with antibody leads to functional elimination of the modulation domain as if it were cleaved off by mild proteolysis. Antibody treatment neither caused nor inhibited receptor activation to a DNA binding form.