Activator-Specific Requirement of Yeast Mediator Proteins for RNA Polymerase II Transcriptional Activation

Abstract

The multisubunit Mediator complex of Saccharomyces cerevisiae is required for most RNA polymerase II (Pol II) transcription. The Mediator complex is composed of two subcomplexes, the Rgr1 and Srb4 subcomplexes, which appear to function in the reception of activator signals and the subsequent modulation of Pol II activity, respectively. In order to determine the precise composition of the Mediator complex and to explore the specific role of each Mediator protein, our goal was to identify all of the Mediator components. To this end, we cloned three previously unidentified Mediator subunits, Med9/Cse2, Med10/Nut2, and Med11, and isolated mutant forms of each of them to analyze their transcriptional defects. Differential display and Northern analyses of mRNAs from wild-type and Mediator mutant cells demonstrated an activator-specific requirement for each Mediator subunit. Med9/Cse2 and Med10/Nut2 were required, respectively, for Bas1/Bas2- and Gcn4-mediated transcription of amino acid biosynthetic genes. Gal11 was required for Gal4- and Rap1-mediated transcriptional activation. Med11 was also required specifically for MFα1 transcription. On the other hand, Med6 was required for all of these transcriptional activation processes. These results suggest that distinct Mediator proteins in the Rgr1 subcomplex are required for activator-specific transcriptional activation and that the activation signals mediated by these Mediator proteins converge on Med6 (or the Srb4 subcomplex) to modulate Pol II activity.

ACKNOWLEDGMENTS

We thank Jin Mo Park and Juri Kim for technical help, Kelly LaMarco for careful reading of the manuscript, and R. Robinson, D. Kirby, K. Pierce, and E. Spooner of the Harvard Microchemistry Facility for their expertise and technical assistance. We also thank C. Gustafsson, R. Roeder, D. Reinberg, R. Kornberg, and I. Herskowitz for sharing information before publication. We give special thanks to C. Gustafsson, S. Bjoklund, and A. Hinnebusch for Mediator antibodies and Bas2-related plasmids.

This work was supported by grants from SBRI (B-96-004) and the Ministry of Health and Welfare, Republic of Korea (HMP-97-B-3-0030 of the 1997 Good Health R&D project), to Y.-J.K.