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Molecular and Cellular Biology, September 1998, p. 5364-5370, Vol. 18, No. 9
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Requirement for a Functional Interaction between Mediator Components Med6 and Srb4 in RNA Polymerase II Transcription

Young Chul Lee and Young-Joon Kim*

Laboratory of Molecular Genetics, Center for Molecular Medicine, Samsung Biomedical Research Institute, Kangnam-ku, Seoul 135-230, Korea

Received 3 April 1998/Returned for modification 25 May 1998/Accepted 9 June 1998

Regulated transcription of class II genes of the yeast Saccharomyces cerevisiae requires the diverse functions of mediator complex. In particular, MED6 is essential for activated transcription from many class II promoters, suggesting that it functions as a key player in the relay of activator signals to the basal transcription machinery. To identify the functional relationship between MED6 and other transcriptional regulators, we conducted a genetic screen to isolate a suppressor of a temperature-sensitive (ts) med6 mutation. We identified an SRB4 allele as a dominant and allele-specific suppressor of med6-ts. A single missense mutation in SRB4 can specifically suppress transcriptional defects caused by the med6 ts mutation, indicating a functional interaction between these two mediator subunits in the activation of transcription. Biochemical analysis of mediator subassembly revealed that mediator can be dissociated into two tightly associated subcomplexes. The Med6 and Srb4 proteins are contained in the same subcomplex together with other dominant Srb proteins, consistent with their functional relationship revealed by the genetic study. Our results suggest not only the existence of a specific interaction between Med6 and Srb4 but also the requirement of this interaction in transcriptional regulation of RNA polymerase II holoenzyme.

* Corresponding author. Mailing address: Laboratory of Molecular Genetics, Center for Molecular Medicine, Samsung Biomedical Research Institute, 50 Ilwon-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.

Molecular and Cellular Biology, September 1998, p. 5364-5370, Vol. 18, No. 9
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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