ADR1-Mediated Transcriptional Activation Requires the Presence of an Intact TFIID Complex

Abstract

ABSTRACT The yeast transcriptional activator ADR1, which is required for ADH2 and other genes’ expression, contains four transactivation domains (TADs). While previous studies have shown that these TADs act through GCN5 and ADA2, and presumably TFIIB, other factors are likely to be involved in ADR1 function. In this study, we addressed the question of whether TFIID is also required for ADR1 action. In vitro binding studies indicated that TADI of ADR1 was able to retain TAF II 90 from yeast extracts and TADII could retain TBP and TAF II 130/145. TADIV, however, was capable of retaining multiple TAF II s, suggesting that TADIV was binding TFIID from yeast whole-cell extracts. The ability of TADIV truncation derivatives to interact with TFIID correlated with their transcription activation potential in vivo. In addition, the ability of LexA-ADR1-TADIV to activate transcription in vivo was compromised by a mutation in TAF II 130/145. ADR1 was found to associate in vivo with TFIID in that immunoprecipitation of either TAF II 90 or TBP from yeast whole-cell extracts specifically coimmunoprecipitated ADR1. Most importantly, depletion of TAF II 90 from yeast cells dramatically reduced ADH2 derepression. These results indicate that ADR1 physically associates with TFIID and that its ability to activate transcription requires an intact TFIID complex.