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Molecular and Cellular Biology, February 1999, p. 979-988, Vol. 19, No. 2
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

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

Sang Jun Han,1 Young Chul Lee,1,dagger Byung Soo Gim,1 Gi-Hyuck Ryu,1 Soon Jung Park,1,Dagger William S. Lane,2 and Young-Joon Kim1,*

Center for Molecular Medicine, Samsung Biomedical Research Institute, Sungkyunkwan University College of Medicine, Seoul 135-230, Korea,1 and Harvard Microchemistry Facility, Harvard University, Cambridge, Massachusetts 021382

Received 15 September 1998/Returned for modification 26 October 1998/Accepted 3 November 1998

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 MFalpha 1 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.

* Corresponding author. Mailing address: Center for Molecular Medicine, Samsung Biomedical Research Institute, Sungkyunkwan University College of Medicine, 50 IIwon-dong, Kangnam-ku, Seoul 135-230, Korea. Phone: 82-2-3410-3638. Fax: 82-2-3410-3649. E-mail: yjkim{at}smc.samsung.co.kr.

dagger Present address: Center for Ligand and Transcription, Chonnam National University, Kwangju 500-757, Korea.

Dagger Present address: Department of Parasitology, College of Medicine, Yonsei University, Seoul 120-752, Korea.

Molecular and Cellular Biology, February 1999, p. 979-988, Vol. 19, No. 2
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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