DNA-binding and non-DNA-binding forms of the transformed glucocorticoid receptor.
In this work we have probed the mechanism responsible for two non-DNA-binding states of the mouse glucocorticoid receptor. In the first case, transformed receptors were treated with hydrogen peroxide. It is known that oxidizing agents promote the formation of disulfide bonds in the glucocorticoid receptor, but it has not been determined what domains are involved in any disulfide bond formation that leads to inactivation of DNA-binding activity. We show here that hydrogen peroxide inhibits DNA-binding by the 15-kDa tryptic fragment containing the DNA-binding fingers with the same concentration dependency as it inhibits DNA-binding by the uncleaved receptor. This suggests that all of the effect of peroxide is on sulfhydryl groups within the zinc fingers. After dissociation (transformation) of cytosolic heteromeric glucocorticoid receptor complexes, only a portion (40-60%) of the dissociated receptors can bind to DNA-cellulose. We show that the 15-kDa tryptic fragment derived from the portion of transformed receptors that do not bind to DNA is itself competent at DNA-binding.
Hutchison
K A
KA
Department of Pharmacology, University of Michigan Medical School, Ann Arbor 48109.
Matić
G
G
Czar
M J
MJ
Pratt
W B
WB
D000818:Animals; D002460:Cell Line; D003600:Cytosol / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D003907:Dexamethasone / Q000378:metabolism / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D015151:Immunoblotting; D051379:Mice; D008970:Molecular Weight; D010449:Peptide Mapping; D011965:Receptors, Glucocorticoid / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D014221:Triamcinolone / Q000494:pharmacology
D004268:DNA-Binding Proteins; D011965:Receptors, Glucocorticoid; D014221:Triamcinolone; D003907:Dexamethasone; D004247:DNA; D006861:Hydrogen Peroxide