Receptor accessory factor enhances specific DNA binding of androgen and glucocorticoid receptors.
Protein-protein interactions are common among transcriptional activators and may have important consequences for gene regulation. Using the mobility shift assay, we have identified a factor that enhances specific DNA binding of truncated rat androgen (AR) and glucocorticoid (GR) receptors by 25- and 6-fold, respectively, through the formation of heteromeric complexes. This factor, designated receptor accessory factor, or RAF, also potentiates DNA binding of full-length human GR. RAF is temperature and trypsin sensitive and is present in a variety of cultured mammalian cells. By gel filtration RAF has a predicted molecular mass of 130 kDa. RAF enhancement of AR-DNA binding is optimal with androgen response element DNA. RAF appears to interact directly with AR because 1) deoxycholate, which interferes with protein-protein but not protein-DNA interactions, prevents RAF.AR.DNA complex formation, 2) RAF activity is recovered from an androgen response element DNA affinity column only in the presence of AR, and 3) RAF increases the size of an AR.DNA complex by gel filtration. Mutagenesis of truncated AR fragments indicates that a region in the NH2-terminal domain is required for RAF to enhance AR-DNA binding. The interaction of RAF with AR and GR suggests that RAF might influence the ability of these nuclear receptors to activate transcription.
Kupfer
S R
SR
Department of Pediatrics, University of North Carolina School of Medicine, Chapel Hill 27599.
Marschke
K B
KB
Wilson
E M
EM
French
F S
FS
D000818:Animals; D001483:Base Sequence; D001685:Biological Factors / Q000378:metabolism*; D002460:Cell Line; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D009036:Moths; D011485:Protein Binding; D011817:Rabbits; D051381:Rats; D011944:Receptors, Androgen / Q000378:metabolism*; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D013696:Temperature
D001685:Biological Factors; D011944:Receptors, Androgen; D011965:Receptors, Glucocorticoid; D004247:DNA