Saccharomyces cerevisiae Ssd1p promotes CLN2 expression by binding to the 5'-untranslated region of CLN2 mRNA

Abstract

In Saccharomyces cerevisiae, TFIID, which is composed of TATA-binding protein (TBP) and a set of TBP-associated factors (TAFs), mediates the transcription of most class II genes. Previous studies have shown that CLN2 expression was significantly reduced by taf1-ts2, but not by taf1-N568Δ, although both mutations display similar temperature-sensitive growth phenotypes and transcriptional defects in other genes. Here, we show that the reduced expression of CLN2 is not because of differences in taf1 alleles in the previous experiments but because of allelic differences at the SSD1 locus in the host strains. Specifically, ssd1-d reduces CLN2 expression when combined with taf1. Ssd1p expressed from SSD1-V, but not from ssd1-d, stabilizes a subpopulation of CLN2 mRNA in wild-type and taf1-N568Δ strains and facilitates the continuous transcription of CLN2 after heat shock in the taf1-N568Δ strain. Reporter assays show that both activities appear to depend on the 5'-untranslated region of CLN2 mRNA and that Ssd1p binds to this region via its amino- and carboxy-terminal domains. Based on these observations, we propose a model for the action of Ssd1p and discuss its biologic role.