Identification of pathways regulating cell size and cell-cycle progression by RNAi (original) (raw)

Nature volume 439, pages 1009–1013 (2006)Cite this article

Abstract

Many high-throughput loss-of-function analyses of the eukaryotic cell cycle have relied on the unicellular yeast species Saccharomyces cerevisiae and Schizosaccharomyces pombe. In multicellular organisms, however, additional control mechanisms regulate the cell cycle to specify the size of the organism and its constituent organs1. To identify such genes, here we analysed the effect of the loss of function of 70% of Drosophila genes (including 90% of genes conserved in human) on cell-cycle progression of S2 cells using flow cytometry. To address redundancy, we also targeted genes involved in protein phosphorylation simultaneously with their homologues. We identify genes that control cell size, cytokinesis, cell death and/or apoptosis, and the G1 and G2/M phases of the cell cycle. Classification of the genes into pathways by unsupervised hierarchical clustering on the basis of these phenotypes shows that, in addition to classical regulatory mechanisms such as Myc/Max, Cyclin/Cdk and E2F, cell-cycle progression in S2 cells is controlled by vesicular and nuclear transport proteins, COP9 signalosome activity and four extracellular-signal-regulated pathways (Wnt, p38βMAPK, FRAP/TOR and JAK/STAT). In addition, by simultaneously analysing several phenotypes, we identify a translational regulator, eIF-3p66, that specifically affects the Cyclin/Cdk pathway activity.

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Acknowledgements

We thank the Berkeley Drosophila Genome Project, L. Lum and P. A. Beachy for constructs and reagents; T. Mäkelä, M. Laiho, M. Bonke and P. Ojala for critical review of the manuscript, and R. Medema for discussion. This work was supported by the Centre of Excellence in Translational Genome-Scale Biology of the Academy of Finland, Biocentrum Helsinki, University of Helsinki, Sigrid Jusélius Foundation, Finnish Cultural Foundation, Maud Kuistila Foundation and Finnish Cancer Research Organizations.

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  1. Mikael Björklund and Minna Taipale: *These authors contributed equally to this work

Authors and Affiliations

  1. Molecular and Cancer Biology Program, University of Helsinki, PO Box 63 , Haartmaninkatu 8, FI-00014, Finland
    Mikael Björklund, Minna Taipale, Markku Varjosalo & Jussi Taipale
  2. High Throughput Center, Biomedicum Helsinki, University of Helsinki, PO Box 63, Haartmaninkatu 8, FI-00014, Finland
    Juhani Lahdenperä & Jussi Taipale
  3. Department of Molecular Medicine, National Public Health Institute, FI-00251, Helsinki, Finland
    Mikael Björklund, Minna Taipale, Markku Varjosalo, Juha Saharinen & Jussi Taipale
  4. Biomedicum Bioinformatics Unit, Biomedicum, FI-00251, Helsinki, Finland
    Juha Saharinen

Authors

  1. Mikael Björklund
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  2. Minna Taipale
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  3. Markku Varjosalo
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  4. Juha Saharinen
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  5. Juhani Lahdenperä
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  6. Jussi Taipale
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Corresponding authors

Correspondence toMinna Taipale or Jussi Taipale.

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Björklund, M., Taipale, M., Varjosalo, M. et al. Identification of pathways regulating cell size and cell-cycle progression by RNAi.Nature 439, 1009–1013 (2006). https://doi.org/10.1038/nature04469

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Editorial Summary

Cell cycle paths

Recent advances in our understanding of control of the eukaryotic cell cycle have relied on loss-of-function analyses of unicellular yeasts. But in multicellular organisms there are additional control mechanisms that regulate cell-cycle progression in the G1 phase, and specify the size of the organism and its constituent organs. An RNA interference (RNAi) screening for pathways regulating cell size and cell cycle in Drosophila has now identified genes that specifically control cell size, cytokinesis, and cell death/apoptosis. The panel of genes involved in cell-cycle control will serve as a basis for a systems biology approach to analysis of the metazoan cell cycle.