TY - JOUR
T1 - Direct Interaction of the τ1 Transactivation Domain of the Human Glucocorticoid Receptor with the Basal Transcriptional Machinery
AU - McEwan, Iain J.
AU - Wright, Anthony P.H.
AU - Dahlman-Wright, Karin
AU - Carlstedt-Duke, Jan
AU - Gustafsson, Jan Åke
N1 - Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 1993/1
Y1 - 1993/1
N2 - We have used a yeast (Saccharomyces cerevisiae) cell free transcription system to study protein-protein interactions involving the τ1 transactivation domain of the human glucocorticoid receptor that are important for transcriptional transactivation by the receptor. Purified τ1 specifically inhibited transcription from a basal promoter derived from the CYC1 gene and from the adenovirus 2 major late core promoter in a concentration-dependent manner. This inhibition or squelching was correlated with the transactivation activity of τ1. Recombinant yeast TATA-binding protein (yTFIID), although active in vitro, did not specifically reverse the inhibitory effect of τ1. In addition, no specific interaction between τ1 and yTFIID could be shown in vitro by affinity chromatography. Taken together, these results indicate that the τ1 transactivation domain of the human glucocorticoid receptor interacts directly with the general transcriptional apparatus through some target protein(s) that is distinct from the TATA-binding factor. Furthermore, this assay can be used to identify interacting factors, since after phosphocellulose chromatography of a whole-cell yeast extract, a fraction that contained an activity which selectively counteracted the squelching effect of τ1 was found.
AB - We have used a yeast (Saccharomyces cerevisiae) cell free transcription system to study protein-protein interactions involving the τ1 transactivation domain of the human glucocorticoid receptor that are important for transcriptional transactivation by the receptor. Purified τ1 specifically inhibited transcription from a basal promoter derived from the CYC1 gene and from the adenovirus 2 major late core promoter in a concentration-dependent manner. This inhibition or squelching was correlated with the transactivation activity of τ1. Recombinant yeast TATA-binding protein (yTFIID), although active in vitro, did not specifically reverse the inhibitory effect of τ1. In addition, no specific interaction between τ1 and yTFIID could be shown in vitro by affinity chromatography. Taken together, these results indicate that the τ1 transactivation domain of the human glucocorticoid receptor interacts directly with the general transcriptional apparatus through some target protein(s) that is distinct from the TATA-binding factor. Furthermore, this assay can be used to identify interacting factors, since after phosphocellulose chromatography of a whole-cell yeast extract, a fraction that contained an activity which selectively counteracted the squelching effect of τ1 was found.
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U2 - 10.1128/mcb.13.1.399
DO - 10.1128/mcb.13.1.399
M3 - Article
C2 - 8417339
AN - SCOPUS:0027529830
VL - 13
SP - 399
EP - 407
JO - Molecular and Cellular Biology
JF - Molecular and Cellular Biology
SN - 0270-7306
IS - 1
ER -