Analysis of transcriptional activation at a distance in Saccharomyces cerevisiae.
Most fundamental aspects of transcription are conserved among eukaryotes. One striking difference between yeast Saccharomyces cerevisiae and metazoans, however, is the distance over which transcriptional activation occurs. In S. cerevisiae, upstream activation sequences (UASs) are generally located within a few hundred base pairs of a target gene, while in Drosophila and mammals, enhancers are often several kilobases away. To study the potential for long-distance activation in S. cerevisiae, we constructed and analyzed reporters in which the UAS-TATA distance varied. Our results show that UASs lose the ability to activate normal transcription as the UAS-TATA distance increases. Surprisingly, transcription does initiate, but proximally to the UAS, regardless of its location. To identify factors affecting long-distance activation, we screened for mutants allowing activation of a reporter when the UAS-TATA distance is 799 bp. These screens identified four loci, SIN4, SPT2, SPT10, and HTA1-HTB1, with sin4 mutations being the strongest. Our results strongly suggest that long-distance activation in S. cerevisiae is normally limited by Sin4 and other factors and that this constraint plays a role in ensuring UAS-core promoter specificity in the compact S. cerevisiae genome.
Dobi
Krista C
KC
Department of Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA 02115, USA.
Winston
Fred
F
D000426:Alcohol Dehydrogenase / Q000235:genetics; D015152:Blotting, Northern; D002843:Chromatin / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D017930:Genes, Reporter; D050505:Mutant Proteins / Q000302:isolation & purification; D009154:Mutation / Q000235:genetics; D009690:Nucleic Acid Conformation; D010641:Phenotype; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D016385:TATA Box / Q000235:genetics; D013728:Terminator Regions, Genetic / Q000235:genetics; D014158:Transcription, Genetic; D015533:Transcriptional Activation
D002843:Chromatin; D050505:Mutant Proteins; D029701:Saccharomyces cerevisiae Proteins; C516801:ADH1 protein, S cerevisiae; D000426:Alcohol Dehydrogenase