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.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
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)
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).
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).
Rickert, P. et al. Cyclin C/CDK8 is a novel CTD kinase associated with RNA polymerase II. Oncogene 12, 2631–2640 (1996).
Liao, S.-M. et al. A kinase–cyclin pair in the RNA polymerase II holoenzyme. Nature 374, 193–196 (1995).
Maldonado, E. et al. A human RNA polymerase II complex with SRB and DNA-repair proteins. Nature 381, 86– 89 (1996).
Hengarther, C. J. et al. Temporal regulation of RNA polymerase II by Srb10 and Kin28 cyclin-dependent kinases. Mol. Cell 2, 45 –53 (1998).
Carlson, M. Genetics of transcriptional regulation in yeast: connections to the RNA polymerase II CTD. Annu. Rev. Cell. Dev. Biol. 13, 1–23 (1997).
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).
Gu, W. et al. A novel human Srb/Med-containing cofactor complex, SMCC, involved in transcription regulation. Mol. Cell 3, 97–108 (1999).
Boyer, T. G. et al. Mammalian Srb/Mediator complex is targeted by adenovirus E1A protein. Nature 399, 276– 279 (1999).
Hampsey, M. & Reinberg, D. RNA polymerase II as a control for multiple coactivator complexes. Curr. Opin. Genet. Dev. 9, 132–139 (1999).
Andersen, G. et al. The structure of cyclin H: common mode of kinase activation and specific features. EMBO J. 16, 958– 967 (1997).
Rachez, C. et al. Ligand-dependent transcription activation by nuclear receptors requires the DRIP complex. Nature 398, 824 –828 (1999).
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).
Naar, A. M. et al. Composite coactivator ARC mediates chromatin-directed transcriptional activation. Nature 398, 828– 832 (1999).
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).
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).
Gould, K. L. in Protein Kinases 149–176 (IRL Press, New York, 1994).
Kazmi, M. A. et al. High-level inducible expression of visual pigments in transfected cells. Biotechniques 21, 304– 311 (1996).
Hauer, C. A. et al. Epstein-virus episome-based promoter function in human myeloid cells. Nucleic Acids Res. 17, 989– 2003 (1989).
Akoulitchev, S. & Reinberg, D. The molecular mechanism of mitotic inhibition of TFIIH is mediated by phosphorylation of CDK7. Genes Dev. 12, 3541– 3550 (1998).
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).
Harper, J. W. & Elledge, S. J. The role of Cdk7 in CAK function, a retro-retrospective. Genes Dev. 12, 285 –289 (1998).
Orphanides, G., Lagrange, T. & Reinberg, D. The general transcription factors of RNA polymerase II. Genes Dev. 10, 2657– 2683 (1996).
Kong, M. et al. Cyclin F regulates the nuclear localization of cyclin B1 through a cyclin–cyclin interaction. EMBO J. 19, 1378–1388 (2000).
LeRoy, G., Drapkin, R., Weis, L. & Reinberg, D. Immunoaffinity purification of the human multisubunit transcription factor IIH. J. Biol. Chem. 273, 7134–7140 (1998).
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).
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).
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.
Author information
Authors and Affiliations
Corresponding author
Supplementary information
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/35024111
This article is cited by
-
Hit discovery of potential CDK8 inhibitors and analysis of amino acid mutations for cancer therapy through computer-aided drug discovery
BMC Chemistry (2024)
-
Cyclin-dependent kinase 7 (CDK7) inhibitors as a novel therapeutic strategy for different molecular types of breast cancer
British Journal of Cancer (2024)
-
Cyclins and cyclin-dependent kinases: from biology to tumorigenesis and therapeutic opportunities
Journal of Cancer Research and Clinical Oncology (2023)
-
MED12 is overexpressed in glioblastoma patients and serves as an oncogene by targeting the VDR/BCL6/p53 axis
Cellular and Molecular Life Sciences (2022)
-
CDK7 inhibitors as anticancer drugs
Cancer and Metastasis Reviews (2020)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.