Laurent Kuras * ^@ , Tilman Borggrefe ¹, and Roger D. Kornberg ¹

*Centre de Génétique Moléculaire, Centre National de la Recherche Scientifique, 91198 Gif-sur-Yvette, France;
¹ Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305

^@ To whom correspondence should be addressed. E-mail:  laurent.kuras@cgm.cnrs-gif.fr

The multiprotein Mediator complex has been shown to interact with gene-specific regulatory proteins and RNA polymerase II in vitro. Here, we use chromatin immunoprecipitation to analyze the recruitment of Mediator to GAL genes of yeast in vivo. We find that Mediator associates exclusively with transcriptionally active and not inactive GAL genes. This association maps to the upstream activating sequence, rather than the core promoter, and is independent of RNA polymerase II, general transcription factors, and core promoter sequences. These findings support the idea of Mediator as a primary conduit of regulatory information from enhancers to promoters in eukaryotic cells.

1. Ling J, Pi W, Yu X, Bengra C, Long Q, Jin H, Seyfang A, and Tuan D, "The ERV-9 LTR Enhancer is Not Blocked by the HS5 Insulator and Synthesizes Through the HS5 Site Non-Coding, Long RNAs that Regulate LTR Enhancer Function", Nucleic Acids Research, vol. 31, no. 15, pp. 4582-4596 (August 1, 2003).

2. Chung W-H, Craighead JL, Chang W-H, Ezeokonkwo C, Bareket-Samish A, Kornberg RD, and Asturias FJ,
"RNA Polymerase II/TFIIF Structure and Conserved Organization of the Initiation Complex", Molecular Cell, vol. 12, no. 4, pp. 1003-1013 (October, 2003).

3. Boeger H, Griesenbeck J, Strattan JS, and Kornberg RD, "Nucleosomes Unfold Completely at a Transcriptionally Active Promoter", Molecular Cell, vol 11, no. 6, pp. 1587-1598  (June, 2003).

4. Hovsepian JA, and Frenster JH, "RNA-Induced Melting of DNA during Selective Gene Transcription", Molec. Biol. Cell, vol. 13, supp. p. 239a (November, 2002).

5. Gottesfeld JM, and Barbas CF III, "RNA as a Transcriptional Activator", Chemistry and Biology, vol 10, no.7, pp. 584-585 (July, 2003).

6. Saha S, Ansari AZ, Jarell KA, and Ptashne M, "RNA Sequences that Work as Transcriptional Activating Regions", Nucleic Acid Research, vol. 31, no. 5, pp. 1565-1570 (March 1, 2003).

7. Buskirk AR, Kehayova PD, Landrigan A, and Liu DR, "In Vivo Evolution of an RNA-Based Transcriptional Activator", Chemistry and Biology, vol 10, no. 6, pp. 533-540 (June, 2003).

8. Lanz RB, Chua SS, Barron N, Söder BM, DeMayo F, and O'Malley BW, "Steroid Receptor RNA Activator Stimulates Proliferation as Well as Apoptosis In Vivo", Molecular and Cellular Biology, vol. 23, no. 20, pp. 7163-7176 (October, 2003).

9. Frenster JH, "Ultrastructural Probes of Active DNA Sites, and the RNA Activators of DNA",  Reviews and Slide Presentation.
 

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