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TFIIH is negatively regulated by cdk8-containing mediator complexes

Nature volume 407, pages 102–106 (2000)Cite this article

Abstract

The mammalian cyclin-dependent kinase 8 (cdk8)1 gene has been linked with a subset of acute lymphoblastic leukaemias2, and its corresponding protein has been functionally implicated in regulation of transcription3,4. Mammalian cdk8 and cyclin C, and their respective yeast homologues, Srb10 and Srb11, are components of the RNA polymerase II holoenzyme complex5,6 where they function as a protein kinase that phosphorylates the carboxy-terminal domain (CTD) of the largest subunit of RNA polymerase II (ref. 7). The yeast SRB10 and SRB11 genes have been implicated in the negative regulation of transcription8. The cdk8/cyclin C protein complex is also found in a number of mammalian Mediator-like protein complexes3,5,9,10,11,12, which repress activated transcription independently of the CTD in vitro 9,10. Here we show that cdk8/cyclin C can regulate transcription by targeting the cdk7/cyclin H subunits of the general transcription initiation factor IIH (TFIIH). cdk8 phosphorylates mammalian cyclin H in the vicinity of its functionally unique amino-terminal and carboxy-terminal α-helical domains13. This phosphorylation represses both the ability of TFIIH to activate transcription and its CTD kinase activity. In addition, mimicking cdk8 phosphorylation of cyclin H in vivo has a dominant-negative effect on cell growth. Our results link the Mediator complex and the basal transcription machinery by a regulatory pathway involving two cyclin-dependent kinases. This pathway appears to be unique to higher organisms.

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Figure 1: TFIIH mediates cdk8-dependent repression by NAT.
Figure 2: Properties of the Mediator-like complex, devoid of cdk8.
Figure 3: cdk8/cyclin C phosphorylates the cyclin H subunit of TFIIH.
Figure 4: Phosphorylation of cyclin H by cdk8 at Ser 5 and Ser 304 in vitro.
Figure 5: Regulatory phosphorylation of cyclin H in vivo.

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References

  1. Tassan, J.-P. et al. Identification of human cyclin-dependent kinase 8, a putative protein kinase partner for cyclin C. Proc. Natl Acad. Sci. USA 97, 8871–8875 ( 1995)

    Article  ADS  Google Scholar 

  2. Li, H. et al. Molecular cloning and chromosomal localization of the human cyclin C (CCNC) and cyclin E (CCNE) genes: deletion of the CCNC gene in human tumors. Genomics 32, 253–259 (1996).

    Article  CAS  Google Scholar 

  3. Gold, M. O. et al. Viral transactivators E1A and VP16 interact with a large complex that is associated with CTD kinase activity and contains CDK8. Nucleic Acids Res. 24, 3771–3777 (1996).

    Article  CAS  Google Scholar 

  4. Rickert, P. et al. Cyclin C/CDK8 is a novel CTD kinase associated with RNA polymerase II. Oncogene 12, 2631–2640 (1996).

    CAS  Google Scholar 

  5. Liao, S.-M. et al. A kinase–cyclin pair in the RNA polymerase II holoenzyme. Nature 374, 193–196 (1995).

    Article  ADS  CAS  Google Scholar 

  6. Maldonado, E. et al. A human RNA polymerase II complex with SRB and DNA-repair proteins. Nature 381, 86– 89 (1996).

    Article  ADS  CAS  Google Scholar 

  7. Hengarther, C. J. et al. Temporal regulation of RNA polymerase II by Srb10 and Kin28 cyclin-dependent kinases. Mol. Cell 2, 45 –53 (1998).

    Google Scholar 

  8. Carlson, M. Genetics of transcriptional regulation in yeast: connections to the RNA polymerase II CTD. Annu. Rev. Cell. Dev. Biol. 13, 1–23 (1997).

    Article  CAS  Google Scholar 

  9. Sun, X. et al. NAT a human complex containing Srb polypeptides that functions as a negative regulator of activated transcription. Mol. Cell 2, 213–222 (1998).

    Article  CAS  Google Scholar 

  10. Gu, W. et al. A novel human Srb/Med-containing cofactor complex, SMCC, involved in transcription regulation. Mol. Cell 3, 97–108 (1999).

    Article  CAS  Google Scholar 

  11. Boyer, T. G. et al. Mammalian Srb/Mediator complex is targeted by adenovirus E1A protein. Nature 399, 276– 279 (1999).

    Article  ADS  CAS  Google Scholar 

  12. Hampsey, M. & Reinberg, D. RNA polymerase II as a control for multiple coactivator complexes. Curr. Opin. Genet. Dev. 9, 132–139 (1999).

    Article  CAS  Google Scholar 

  13. Andersen, G. et al. The structure of cyclin H: common mode of kinase activation and specific features. EMBO J. 16, 958– 967 (1997).

    Article  CAS  Google Scholar 

  14. Rachez, C. et al. Ligand-dependent transcription activation by nuclear receptors requires the DRIP complex. Nature 398, 824 –828 (1999).

    Article  ADS  CAS  Google Scholar 

  15. Ryu, S., Zhou, S., Ladurner, A. G. & Tjian, R. The transcriptional cofactor complex CRSP is required for activity of the enhancer-binding protein Sp1. Nature 397, 446–450 (1999).

    Article  ADS  CAS  Google Scholar 

  16. Naar, A. M. et al. Composite coactivator ARC mediates chromatin-directed transcriptional activation. Nature 398, 828– 832 (1999).

    Article  ADS  CAS  Google Scholar 

  17. Ge, H., Zhao, Y., Chait, B. T. & Roeder, R. G. Phosphorylation negatively regulates the function of PC4. Proc. Natl Acad. Sci. USA 91, 12691–12695 ( 1994).

    Article  ADS  CAS  Google Scholar 

  18. Kim, Y. -J. et al. A multiprotein mediator of transcriptional activation and its interaction with the C-terminal repeat domain of RNA polymerase II. Cell 77, 599–608 (1994).

    Article  CAS  Google Scholar 

  19. Gould, K. L. in Protein Kinases 149–176 (IRL Press, New York, 1994).

    Google Scholar 

  20. Kazmi, M. A. et al. High-level inducible expression of visual pigments in transfected cells. Biotechniques 21, 304– 311 (1996).

    Article  CAS  Google Scholar 

  21. Hauer, C. A. et al. Epstein-virus episome-based promoter function in human myeloid cells. Nucleic Acids Res. 17, 989– 2003 (1989).

    ADS  Google Scholar 

  22. Akoulitchev, S. & Reinberg, D. The molecular mechanism of mitotic inhibition of TFIIH is mediated by phosphorylation of CDK7. Genes Dev. 12, 3541– 3550 (1998).

    Article  CAS  Google Scholar 

  23. Long, J. J. et al. Repression of TFIIH transcriptional activity and TFIIH-associated cdk7 kinase activity at mitosis. Mol. Cell. Biol. 18 , 1467–1476 (1998).

    Article  CAS  Google Scholar 

  24. Harper, J. W. & Elledge, S. J. The role of Cdk7 in CAK function, a retro-retrospective. Genes Dev. 12, 285 –289 (1998).

    Article  CAS  Google Scholar 

  25. Orphanides, G., Lagrange, T. & Reinberg, D. The general transcription factors of RNA polymerase II. Genes Dev. 10, 2657– 2683 (1996).

    Article  CAS  Google Scholar 

  26. Kong, M. et al. Cyclin F regulates the nuclear localization of cyclin B1 through a cyclin–cyclin interaction. EMBO J. 19, 1378–1388 (2000).

    Article  CAS  Google Scholar 

  27. LeRoy, G., Drapkin, R., Weis, L. & Reinberg, D. Immunoaffinity purification of the human multisubunit transcription factor IIH. J. Biol. Chem. 273, 7134–7140 (1998).

    Article  CAS  Google Scholar 

  28. Desai, D., Wessling, H., Fisher, R. & Morgan, D. Effects of phosphorylation by CAK on cyclin binding by CDC2 and CDK2. Mol. Cell. Biol. 15, 345–350 (1995).

    Article  CAS  Google Scholar 

  29. Parvin, J. D. & Sharp, P. A. DNA topology and a minimal set of basal factors for transcription by RNA polymerase II. Cell 73, 533–540 (1993).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank M. Tyckocinsky for pMEP4β; R. Roeder for the cdk8–Flag cell line; E. Lees for cDNAs encoding mammalian cdk8 and cyclin C; R. Young and P. Cook for communicating unpublished results; T. Kim for affinity purified TFIIH; A. Berk for anti-hSur2 antibodies; and L. Freedman for anti-DRIP150 antibodies. This work was supported by grants from NIH and the Howard Hughes Medical Institute to D.R.

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  1. Howard Hughes Medical Institute Division of Nucleic Acids Enzymology, Department of Biochemistry, Robert Wood Johnson Medical School, Piscataway, 08854, New Jersey, USA

    Sasha Akoulitchev, Sergei Chuikov & Danny Reinberg

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  1. Sasha Akoulitchev
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Correspondence to Danny Reinberg.

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Akoulitchev, S., Chuikov, S. & Reinberg, D. TFIIH is negatively regulated by cdk8-containing mediator complexes. Nature 407, 102–106 (2000). https://doi.org/10.1038/35024111

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