Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Xpd/Ercc2 regulates CAK activity and mitotic progression

Nature volume 424, pages 228–232 (2003)Cite this article

Abstract

General transcription factor IIH (TFIIH) consists of nine subunits: cyclin-dependent kinase 7 (Cdk7), cyclin H and MAT1 (forming the Cdk-activating-kinase or CAK complex), the two helicases Xpb/Hay and Xpd, and p34, p44, p52 and p62 (refs 1–3). As the kinase subunit of TFIIH, Cdk7 participates in basal transcription by phosphorylating the carboxy-terminal domain of the largest subunit of RNA polymerase II1,4,5. As part of CAK, Cdk7 also phosphorylates other Cdks, an essential step for their activation6,7,8,9. Here we show that the Drosophila TFIIH component Xpd negatively regulates the cell cycle function of Cdk7, the CAK activity. Excess Xpd titrates CAK activity, resulting in decreased Cdk T-loop phosphorylation, mitotic defects and lethality, whereas a decrease in Xpd results in increased CAK activity and cell proliferation. Moreover, Xpd is downregulated at the beginning of mitosis when Cdk1, a cell cycle target of Cdk7, is most active. Downregulation of Xpd thus seems to contribute to the upregulation of mitotic CAK activity and to regulate mitotic progression positively. Simultaneously, the downregulation of Xpd might be a major mechanism of mitotic silencing of basal transcription.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Cell-cycle dependence of Xpd.
Figure 2: Reduced CAK activity and viability in embryos overexpressing xpd.
Figure 3: Cellular effects of xpd overexpression.
Figure 4: Decreased Xpd concentration is correlated with increased CAK activity and cell proliferation in S2 cells.

Similar content being viewed by others

References

  1. Frit, P., Bergmann, E. & Egly, J. M. Transcription factor IIH: A key player in the cellular response to DNA damage. Biochimie 81, 27–38 (1999)

    Article  CAS  Google Scholar 

  2. Egly, J. M. The 14th Datta Lecture. TFIIH: From transcription to clinic. FEBS Lett. 498, 124–128 (2001)

    Article  CAS  Google Scholar 

  3. Austin, R. J. & Biggin, M. D. Purification of the Drosophila RNA polymerase II general transcription factors. Proc. Natl Acad. Sci. USA 93, 5788–5792 (1996)

    Article  CAS  Google Scholar 

  4. Akoulitchev, S., Makela, T. P., Weinberg, R. A. & Reinberg, D. Requirement for TFIIH kinase activity in transcription by RNA polymerase II. Nature 377, 557–560 (1995)

    Article  CAS  Google Scholar 

  5. Makela, T. P. et al. A kinase-deficient transcription factor TFIIH is functional in basal and activated transcription. Proc. Natl Acad. Sci. USA 92, 5174–5178 (1995)

    Article  CAS  Google Scholar 

  6. Nigg, E. A. Cyclin-dependent protein kinases: Key regulators of the eukaryotic cell cycle. BioEssays 17, 471–480 (1995)

    Article  CAS  Google Scholar 

  7. Morgan, D. O. Cyclin-dependent kinases: Engines, clocks, and microprocessors. Annu. Rev. Cell Dev. Biol. 13, 261–291 (1997)

    Article  CAS  Google Scholar 

  8. Morgan, D. O. Principles of CDK regulation. Nature 374, 131–134 (1995)

    Article  CAS  Google Scholar 

  9. Larochelle, S., Pandur, J., Fisher, R. P., Salz, H. K. & Suter, B. Cdk7 is essential for mitosis and for in vivo Cdk-activating kinase activity. Genes Dev. 12, 370–381 (1998)

    Article  CAS  Google Scholar 

  10. Fisher, R. P. & Morgan, D. O. CAK in TFIIH: Crucial connection or confounding coincidence? Biochim. Biophys. Acta 1288, O7–O10 (1996)

    PubMed  Google Scholar 

  11. Foe, V. E., Odekk, G. M. & Edgar, B. A. in The Development of Drosophila melanogaster (eds Bate, M. & Martinez-Arias, A.) 149–300 (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1993)

    Google Scholar 

  12. Edgar, B. A., Sprenger, F., Duronio, R. J., Leopold, P. & O'Farrell, P. H. Distinct molecular mechanism regulate cell cycle timing at successive stages of Drosophila embryogenesis. Genes Dev. 8, 440–452 (1994)

    Article  CAS  Google Scholar 

  13. Sandrock, B. & Egly, J. M. A yeast four-hybrid system identifies Cdk-activating kinase as a regulator of the XPD helicase, a subunit of transcription factor IIH. J. Biol. Chem. 276, 35328–35333 (2001)

    Article  CAS  Google Scholar 

  14. Wallenfang, M. R. & Seydoux, G. cdk-7 Is required for mRNA transcription and cell cycle progression in Caenorhabditis elegans embryos. Proc. Natl Acad. Sci. USA 99, 5527–5532 (2002)

    Article  CAS  Google Scholar 

  15. Clemens, J. C. et al. Use of double-stranded RNA interference in Drosophila cell lines to dissect signal transduction pathways. Proc. Natl Acad. Sci. USA 97, 6499–6503 (2000)

    Article  CAS  Google Scholar 

  16. Gottesfeld, J. M. & Forbes, D. J. Mitotic repression of the transcriptional machinery. Trends Biochem. Sci. 22, 197–202 (1997)

    Article  CAS  Google Scholar 

  17. Long, J. J., Leresche, A., Kriwacki, R. W. & Gottesfeld, J. M. Repression of TFIIH transcriptional activity and TFIIH-associated cdk7 kinase activity at mitosis. Mol. Cell. Biol. 18, 1467–1476 (1998)

    Article  CAS  Google Scholar 

  18. 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 

  19. Larochelle, S. et al. T-loop phosphorylation stabilizes the CDK7-cyclin H-MAT1 complex in vivo and regulates its CTD kinase activity. EMBO J. 20, 3749–3759 (2001)

    Article  CAS  Google Scholar 

  20. Foe, V. E. & Alberts, B. M. Reversible chromosome condensation induced in Drosophila embryos by anoxia: visualization of interphase nuclear organization. J. Cell Biol. 100, 1623–1636 (1985)

    Article  CAS  Google Scholar 

  21. Maldonado-Codina, G., Llamazares, S. & Glover, D. M. Heat shock results in cell cycle delay and synchronisation of mitotic domains in cellularised Drosophila melanogaster embryos. J. Cell Sci. 105, 711–720 (1993)

    PubMed  Google Scholar 

Download references

Acknowledgements

We thank M. Zurita for hsp–xpd transgenic flies and the xpd cDNA clone; M. Fuller for anti-(Xpb/Hay) antibody, hsp–xpb/hay transgenic flies and xpb/hay cDNA clone; T. Schüpbach, W. J. Mackay and B.-Z. Shilo for fly strains; R. Tanguay for providing S2 cells; M. Miron, O. Johnstone and M. Lindor for technical assistance with the cell culture; B. Hu for microinjection of the hsp–xpdC construct; A. Tsang for use of the confocal microscope; A. Spurmanis for technical assistance; P. Lasko for the use of the Axioplan microscope; R. Roy for use of the PhosphorImager; and R. Roy, N. Mashrouha and M. Cavey for their critical reading of the manuscript. This work was supported by the National Cancer Institute of Canada with funds from the Canadian Cancer Society. B.S. was a research scientist of the National Cancer Institute of Canada supported by funds from the Canadian Cancer Society and is now a Canadian Institute of Health Research investigator. J.C. was a recipient of a studentship from the Fonds pour la formation de chercheurs et l'aide à la recherche (FCAR Quebec).

Author information

Author notes

  1. Stéphane Larochelle

    Present address: Cell Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York, 10021, USA

Authors and Affiliations

  1. Department of Biology, McGill University, PQ, H3A 1B1, Montreal, Canada

    Jian Chen, Xiaoming Li & Beat Suter

Authors
  1. Jian Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Stéphane Larochelle
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiaoming Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Beat Suter
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Beat Suter.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chen, J., Larochelle, S., Li, X. et al. Xpd/Ercc2 regulates CAK activity and mitotic progression. Nature 424, 228–232 (2003). https://doi.org/10.1038/nature01746

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature01746

This article is cited by

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.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing