Differential sensitivity of chicken progesterone receptor forms to sulfhydryl reactive reagents.
DNA binding of chick progesterone receptor B form (PRB) has been examined and compared to that of the A form (PRA). We found that the elution profiles of the two receptors overlap on DNA-cellulose columns. Both PRA or PRB could bind to plasmid DNA equivalently as assayed by sedimentation velocity studies. However, DNA-binding activity of the two receptor forms showed differential sensitivity to reducing agents and to sulfhydryl (SH) reactive reagents. Reducing agents stabilized DNA-binding activity of PRA more efficiently than they stabilized PRB. Moreover, removal of reducing agents from receptor preparations caused preferential loss of DNA binding by PRB compared to the PRA. DNA-binding activity of PRA was readily destroyed by sulfhydryl modifying reagents such as N-ethylmaleimide and iodoacetamide while PRB was 3-4 times less sensitive to these reagents. We conclude the DNA-binding activity of PRB is less stable due to altered accessibility of SH groups despite the amino acid sequence identity of the DNA-binding domains of PRA and PRB.
Peleg
S
S
Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030.
Schrader
W T
WT
O'Malley
B W
BW
D000818:Animals; D015153:Blotting, Western; D002645:Chickens; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005033:Ethylmaleimide / Q000494:pharmacology; D007460:Iodoacetamide / Q000494:pharmacology; D010957:Plasmids; D011980:Receptors, Progesterone / Q000187:drug effects / Q000378:metabolism*; D012995:Solubility; D013438:Sulfhydryl Compounds / Q000494:pharmacology; D013439:Sulfhydryl Reagents / Q000494:pharmacology*; D014461:Ultracentrifugation
D004268:DNA-Binding Proteins; D011980:Receptors, Progesterone; D013438:Sulfhydryl Compounds; D013439:Sulfhydryl Reagents; D004247:DNA; D005033:Ethylmaleimide; D007460:Iodoacetamide