Direct Interaction of the τ1 Transactivation Domain of the Human Glucocorticoid Receptor with the Basal Transcriptional Machinery

Iain J. McEwan, Anthony P.H. Wright, Karin Dahlman-Wright, Jan Carlstedt-Duke, Jan Åke Gustafsson

Research output: Contribution to journalArticle

55 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)399-407
Number of pages9
JournalMolecular and Cellular Biology
Volume13
Issue number1
DOIs
StatePublished - Jan 1993

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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