Mechanisms of core promoter recognition by general transcription factor TFIID

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TFIID, the largest GTF composed of TBP and 14 TAFs, plays a critical role in transcription from TATA-less promoters. In metazoans, several core promoter elements (CE) other than the TATA element are thought to be recognition sites for TFIID. However, it is unclear whether functionally homologous elements also exist in TATA-less promoters in yeast.Here we develop a novel high-throughput screening system to measure gene expression in yeast by using VTC1 as a reporter gene. This method allows us to isolate several active and novel CE by screening a randomized oligonucleotide pool for the CYC1 promoter. The TAF N-terminal domain (TAND) of the TAF1 protein plays a significant role in transcriptional activation. TAND comprises two subdomains, TAND1 and TAND2, which interact with the concave and convex surfaces of TBP, respectively. In collaboration with other groups, we determine the crystal structure of the TAND-TBP complex that provides a novel insight into transcriptional activation.