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Molecular and Cellular Biology, January 2005, p. 114-123, Vol. 25, No. 1
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.1.114-123.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

The Saccharomyces cerevisiae Srb8-Srb11 Complex Functions with the SAGA Complex during Gal4-Activated Transcription

Erica Larschan and Fred Winston*

Department of Genetics, Harvard Medical School, Boston, Massachusetts

Received 26 July 2004/ Returned for modification 21 August 2004/ Accepted 30 September 2004

The Saccharomyces cerevisiae SAGA (Spt-Ada-Gcn5-acetyltransferase) complex functions as a coactivator during Gal4-activated transcription. A functional interaction between the SAGA component Spt3 and TATA-binding protein (TBP) is important for TBP binding at Gal4-activated promoters. To better understand the role of SAGA and other factors in Gal4-activated transcription, we selected for suppressors that bypass the requirement for SAGA. We obtained eight complementation groups and identified the genes corresponding to three of the groups as NHP10, HDA1, and SRB9. In contrast to the srb9 suppressor mutation that we identified, an srb9{Delta} mutation causes a strong defect in Gal4-activated transcription. Our studies have focused on this requirement for Srb9. Srb9 is part of the Srb8-Srb11 complex, associated with the Mediator coactivator. Srb8-Srb11 contains the Srb10 kinase, whose activity is important for GAL1 transcription. Our data suggest that Srb8-Srb11, including Srb10 kinase activity, is directly involved in Gal4 activation. By chromatin immunoprecipitation studies, Srb9 is present at the GAL1 promoter upon induction and facilitates the recruitment or stable association of TBP. Furthermore, the association of Srb9 with the GAL1 upstream activation sequence requires SAGA and specifically Spt3. Finally, Srb9 association also requires TBP. These results suggest that Srb8-Srb11 associates with the GAL1 promoter subsequent to SAGA binding, and that the binding of TBP and Srb8-Srb11 is interdependent.

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

Molecular and Cellular Biology, January 2005, p. 114-123, Vol. 25, No. 1
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.1.114-123.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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