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Structure and TBP binding of the Mediator head subcomplex Med8–Med18–Med20

Nature Structural & Molecular Biology volume 13, pages 895–901 (2006)Cite this article

Abstract

The Mediator head module stimulates basal RNA polymerase II (Pol II) transcription and enables transcriptional regulation. Here we show that the head subunits Med8, Med18 and Med20 form a subcomplex (Med8/18/20) with two submodules. The highly conserved N-terminal domain of Med8 forms one submodule that binds the TATA box–binding protein (TBP) in vitro and is essential in vivo. The second submodule consists of the C-terminal region of Med8 (Med8C), Med18 and Med20. X-ray analysis of this submodule reveals that Med18 and Med20 form related β-barrel folds. A conserved putative protein-interaction face on the Med8C/18/20 submodule includes sites altered by srb mutations, which counteract defects resulting from Pol II truncation. Our results and published data support a positive role of the Med8/18/20 subcomplex in initiation-complex formation and suggest that the Mediator head contains a multipartite TBP-binding site that can be modulated by transcriptional activators.

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Figure 1: Primary and secondary structure of Med8, Med18 and Med20.
Figure 2: Crystal structure of the Mediator head submodule Med8C/18/20.
Figure 3: Comparison of the Med18/20 heterodimer with similar known structures.
Figure 4: Modularity of Med8 in vivo.
Figure 5: TBP binding of Med8/18/20, Med8 and Med8N.
Figure 6: Med8/18/20 architecture and surface conservation.

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Acknowledgements

We thank S. Baumli for discussions, K. Armache for help with TFIIB preparation and other members of the Cramer laboratory for help. This work was supported by a European Molecular Biology Organization long-term fellowship to L.L., by grants of the Deutsche Forschungsgemeinschaft, the Sonderforschungsbereich SFB646 and the Fonds der chemischen Industrie to P.C. and K.S. and by the EU-grant 3D repertoire, contract no. LSHG-CT-2005-512028. Part of this work was performed at the Swiss Light Source at the Paul Scherrer Institute, Villigen, Switzerland. We thank C. Schulze-Briese and his team at the Swiss Light Source for help.

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  1. Department of Chemistry and Biochemistry, Gene Center Munich, Ludwig-Maximilians-Universität München, Feodor-Lynen-Str. 25, Munich, 81377, Germany

    Laurent Larivière, Sebastian Geiger, Sabine Hoeppner, Susanne Röther, Katja Sträßer & Patrick Cramer

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  1. Laurent Larivière
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Correspondence to Patrick Cramer.

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Supplementary information

Supplementary Fig. 1

Structure of the free Med18/20 heterodimer. (PDF 1508 kb)

Supplementary Fig. 2

Conserved intersubunit contacts within the Med8C/18/20 structure. (PDF 2278 kb)

Supplementary Fig. 3

Flexibility of the Med8C/18/20 trimer. (PDF 1562 kb)

Supplementary Table 1

Med18-Med20 contacts. (PDF 35 kb)

Supplementary Table 2

Med8-Med18 contacts. (PDF 30 kb)

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Larivière, L., Geiger, S., Hoeppner, S. et al. Structure and TBP binding of the Mediator head subcomplex Med8–Med18–Med20. Nat Struct Mol Biol 13, 895–901 (2006). https://doi.org/10.1038/nsmb1143

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