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PMID: 8417339 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Direct interaction of the tau 1 transactivation domain of the human glucocorticoid receptor with the basal transcriptional machinery.

Molecular and cellular biology ·Vol. 13 ·No. 1 ·1993-01-00 ·Pages 399-407

McEwan IJ, Wright AP, Dahlman-Wright K, Carlstedt-Duke J, Gustafsson JA

Abstract

We have used a yeast (Saccharomyces cerevisiae) cell free transcription system to study protein-protein interactions involving the tau 1 transactivation domain of the human glucocorticoid receptor that are important for transcriptional transactivation by the receptor. Purified tau 1 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 tau 1. Recombinant yeast TATA-binding protein (yTFIID), although active in vitro, did not specifically reverse the inhibitory effect of tau 1. In addition, no specific interaction between tau 1 and yTFIID could be shown in vitro by affinity chromatography. Taken together, these results indicate that the tau 1 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 tau 1 was found.

MeSH Terms
Adenoviruses, Human/genetics Amino Acid Sequence Base Sequence Cell-Free System DNA-Binding Proteins/physiology Gene Expression Regulation Humans Molecular Sequence Data Promoter Regions, Genetic Receptors, Glucocorticoid/physiology Saccharomyces cerevisiae TATA-Box Binding Protein Transcription Factors/physiology Transcription, Genetic Transcriptional Activation
Chemicals
DNA-Binding Proteins Receptors, Glucocorticoid TATA-Box Binding Protein Transcription Factors
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
McEwan I J
Centre for Biotechnology, NOVUM, Huddinge University Hospital, Sweden.
Wright A P
Dahlman-Wright K
Carlstedt-Duke J
Gustafsson J A
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.13.1.399-407.1993
Published
1993-01-00
Pages
399-407
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC358920
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.13.1.399-407.1993 PMC Full Text View journal details
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