The Critical cis-Acting Element Required for IMD2 Feedback Regulation by GDP Is a TATA Box Located 202 Nucleotides Upstream of the Transcription Start Site

Abstract

Guanylic nucleotides are essential cellular players, and the critical enzyme in their tightly regulated synthesis in Saccharomyces cerevisiae is encoded by the IMD2 gene. The transcription of IMD2 is subject to general repression by nutrient limitation through the cis nutrient-sensing element. It is also subject to specific feedback regulation by the end products of the guanylic nucleotide synthesis pathway. The critical cis element for this latter mechanism is the guanine response element (GRE), a TATAATA sequence which is located 202 nucleotides upstream of the transcription initiation site and which functions as the IMD2 TATA box. We show that the GRE functions in conjunction with a 52-nucleotide stretch near the transcription start site. This very unusual promoter structure ensures low, basal expression of IMD2 and the recruitment of TFIID to the GRE in response to guanylic nucleotide limitation.

ACKNOWLEDGMENTS

We thank Kevin Struhl for plasmids encoding wild-type or altered-specificity mutant TBP. We thank Steve Buratowski and Lisete Fernandes for helpful comments and Cécile Deluen and Eve Lenssen for critical reading of the manuscript.

This work was supported by grants from the CNRS (UMR5095) and ARC (4749) to B.D.-F., by a fellowship to M.E.-H from the Portuguese Government (FCT SFRH/BPD/5725/2001), and by a grant (3100-059199.99) from the Swiss National Science Foundation to M.A.C.