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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.

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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.

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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).

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  1. Stéphane Larochelle

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

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

    Jian Chen, Xiaoming Li & Beat Suter

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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

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