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Nucleic Acids Research Advance Access originally published online on February 29, 2008
Nucleic Acids Research 2008 36(8):2489-2504; doi:10.1093/nar/gkn070
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Nucleic Acids Research, 2008, Vol. 36, No. 8 2489-2504
© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Molecular Biology

Two conserved modules of Schizosaccharomyces pombe Mediator regulate distinct cellular pathways

Tomas Linder1, Nina N. Rasmussen2, Camilla O. Samuelsen2, Emmanouella Chatzidaki1, Vera Baraznenok1, Jenny Beve1, Peter Henriksen2, Claes M. Gustafsson1 and Steen Holmberg2,*

1Division of Metabolic Diseases, Karolinska Institutet, Novum, SE-141 86 Stockholm, Sweden and 2Department of Molecular Biology, Copenhagen Biocenter, University of Copenhagen, Ole Måloes Vej 5, DK-2200 Copenhagen N, Denmark

*To whom correspondence should be addressed. Tel: +45 35322119; Fax: +45 3532128; Email: gensteen{at}bio.ku.dk Correspondence may also be addressed to Tomas Linder. Tel: +1 415 476 3068; Fax: +1 415 514 9736; Email: tlinder{at}cmp.ucsf.edu

Received January 28, 2008. Revised February 5, 2008. Accepted February 5, 2008.

Mediator is an evolutionary conserved coregulator complex required for transcription of almost all RNA polymerase II-dependent genes. The Schizosaccharomyces pombe Mediator consists of two dissociable components—a core complex organized into a head and middle domain as well as the Cdk8 regulatory subcomplex. In this work we describe a functional characterization of the S. pombe Mediator. We report the identification of the S. pombe Med20 head subunit and the isolation of ts alleles of the core head subunit encoding med17+. Biochemical analysis of med8ts, med17ts, {Delta}med18, {Delta}med20 and {Delta}med27 alleles revealed a stepwise head domain molecular architecture. Phenotypical analysis of Cdk8 and head module alleles including expression profiling classified the Mediator mutant alleles into one of two groups. Cdk8 module mutants flocculate due to overexpression of adhesive cell-surface proteins. Head domain-associated mutants display a hyphal growth phenotype due to defective expression of factors required for cell separation regulated by transcription factor Ace2. Comparison with Saccharomyces cerevisiae Mediator expression data reveals that these functionally distinct modules are conserved between S. pombe and S. cerevisiae.

Present address: Tomas Linder, Department of Cellular and Molecular Pharmacology, University of California San Francisco, 1700 4th Street, PO Box 25430, Byers Hall 4308A, San Francisco, CA 94158-25430, USA

Nina N. Rasmussen, Evolva A/S, Bülowsvej 25, DK-1870 Frederiksberg C, Denmark


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