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Transcriptional activation independent of TFIIH kinase and the RNA polymerase II mediator in vivo

Nature volume 393, pages 389–392 (1998)Cite this article

Abstract

The carboxy-terminal domain (CTD) of the largest subunit of RNA polymerase II becomes multiply phosphorylated by protein kinases during early steps in the gene transcription cycle both in vivo1 and in vitro2. In yeast, the major CTD kinase is a subunit of the general transcription factor TFIIH, and is encoded by an essential gene, KIN28 (ref. 3). Although the CTD and its phosphorylation are important for transcription4,5,6, in vitro studies7,8 have challenged whether CTD phosphorylation is an absolutely required step. The general importance of CTD phosphorylation by Kin28 for transcription in yeast has been suggested because, for all genes tested, transcription is inhibited at the non-permissive temperature in temperature-sensitive kin28 mutants9,10. However, using such a mutant and a copper-inducible targeted destruction method, we show here that transcription of certain genes can be highly induced even when cells lack Kin28. We also show that transcription of these Kin28-independent genes is independent of Srb4 and Srb6, critical components of the CTD-associated transcriptional mediator complex11. These results indicate that there are at least two distinct pathways for transcriptional activation: one is dependent on Kin28 and the mediator complex, and the other is not.

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Figure 1: CUP1 gene activation is independent of TFIIH kinase, but dependent on TFIIH helicase activity.
Figure 2: The SSA4 gene is activated after depletion of Kin28.
Figure 3: Kin28-independent genes are also independent of mediator.

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Acknowledgements

We thank A. Bensaude, G. Faye, M. Cismowski, S. Reed, A. Greenleaf, S. Prakash and R. Young for strains; M. Cismowski for pSF-KIN28 plasmid; Z. Moqtaderi and K. Struhl for strains and plasmids used for generating copper knockout strains; S. Walker and M. Green for sequences of S1 probes; D. Thiele for advice on CUP1 induction; C. Wang for generating the Kin28 copper knockout strain; J.Roberts and E. Alani for critically reading the manuscript; and members of the Lis lab for criticism and encouragement. This work was supported by an NIH grant to J.T.L.

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  1. Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, 14853, New York, USA

    Dong-ki Lee & John T. Lis

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Correspondence to John T. Lis.

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Lee, Dk., Lis, J. Transcriptional activation independent of TFIIH kinase and the RNA polymerase II mediator in vivo. Nature 393, 389–392 (1998). https://doi.org/10.1038/30770

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