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Genome-wide survey of protein kinases required for cell cycle progression

Nature volume 432, pages 980–987 (2004)Cite this article

Abstract

Cycles of protein phosphorylation are fundamental in regulating the progression of the eukaryotic cell through its division cycle. Here we test the complement of Drosophila protein kinases (kinome) for cell cycle functions after gene silencing by RNA-mediated interference. We observed cell cycle dysfunction upon downregulation of 80 out of 228 protein kinases, including most kinases that are known to regulate the division cycle. We find new enzymes with cell cycle functions; some of these have family members already known to phosphorylate microtubules, actin or their associated proteins. Additionally, depletion of several signalling kinases leads to specific mitotic aberrations, suggesting novel roles for familiar enzymes. The survey reveals the inter-digitation of systems that monitor cellular physiology, cell size, cellular stress and signalling processes with the basic cell cycle regulatory machinery.

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Figure 1: Cell cycle progression after RNAi of protein kinases.
Figure 2: Examples of mitotic phenotypes seen following downregulation by RNAi of selected protein kinases.
Figure 3: Quantitative analysis of mitotic RNAi phenotypes.
Figure 4: New cell cycle roles for Gwl, Fray and Pvr kinases.

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Acknowledgements

We thank Cancer Research UK and BBSRC for supporting work in the CR-UK Cell Cycle Genetics Research Group. The BBSRC support was part of a LINK Programme in which the Department of Trade and Industry also provide support for Cyclacel Ltd. An NIH grant supported work in R.W.C.'s group and JSPS funded S.Y. We thank N. B. Carmo, C. Malone and N. Miller for help with experiments. We are grateful to J. C. Pezzullo for his expertise with excel statistical macros and to F. Scaerou, C. Midgley and J. Pereira-Leal for discussions. We also thank T. Hunt, J. Maller and our colleagues, particularly A. Carpenter, M. Segal and M. Savoian for their comments on the manuscript.

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Authors and Affiliations

  1. Department of Genetics, Cancer Research UK Cell Cycle Genetics Research Group, University of Cambridge, Downing Street, CB2 3EH, Cambridge, UK

    M. Bettencourt-Dias, R. Giet, R. Sinka, A. Mazumdar, W. G. Lock, F. Balloux, P. J. Zafiropoulos & D. M. Glover

  2. CNRS UMR 6061 Université de Rennes 1, Equipe Labellisée Ligue Nationale Contre le Cancer, Groupe Cycle Cellulaire, IFR97, 2 avenue du Pr Léon Bernard, 35043, Rennes, France

    R. Giet

  3. Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, Chicago, Illinois, 60208, USA

    S. Yamaguchi, S. Winter & R. W. Carthew

  4. Polgen Division, Cyclacel Ltd, Babraham Science Park, Babraham, Cambridge, CB4 2AT, UK

    M. Cooper, D. Jones, L. Frenz & D. M. Glover

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  1. M. Bettencourt-Dias
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Correspondence to M. Bettencourt-Dias or D. M. Glover.

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Supplementary information

Supplementary Data

This file contains Supplementary Materials and Methods, Supplementary Tables 1-3, and Supplementary Figs 1 and 2. (PDF 460 kb)

Supplementary Table 4

Includes datasheet used to calculate Mitotic Parameters (before normalization) and instructions on how to use this datasheet. (XLS 264 kb)

Supplementary Table 5

Complete list of Drosophila protein kinases showing cell cycle phenotypes, their putative orthologues and functional information from literature and databases. Shown in alphabetic order and with links to Inparanoid database (for orthologies) and FlyBase (a Database of the Drosophila Genome). (DOC 524 kb)

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Bettencourt-Dias, M., Giet, R., Sinka, R. et al. Genome-wide survey of protein kinases required for cell cycle progression. Nature 432, 980–987 (2004). https://doi.org/10.1038/nature03160

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