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Molecular and Cellular Biology, August 2007, p. 5575-5586, Vol. 27, No. 15
0270-7306/07/$08.00+0     doi:10.1128/MCB.00459-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Analysis of Transcriptional Activation at a Distance in Saccharomyces cerevisiae{triangledown}

Krista C. Dobi and Fred Winston*

Department of Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115

Received 16 March 2007/ Returned for modification 18 April 2007/ Accepted 16 May 2007

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.

* Corresponding author. Mailing address: Department of Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA 02115. Phone: (617) 432-7768. Fax: (617) 432-6506. E-mail: winston{at}genetics.med.harvard.edu

{triangledown} Published ahead of print on 25 May 2007.

Molecular and Cellular Biology, August 2007, p. 5575-5586, Vol. 27, No. 15
0270-7306/07/$08.00+0     doi:10.1128/MCB.00459-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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