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  • Original Paper
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Cell type-specific responses of human cells to inhibition of replication licensing

Oncogene volume 21, pages 6624–6632 (2002)Cite this article

Abstract

Replication origins are ‘licensed' for a single initiation event by loading Mcm2-7 complexes during late mitosis and G1. Licensing is blocked at other cell cycle stages by the activity of cyclin-dependent kinases and a small protein called geminin. Here, we describe the effects of over-expressing a non-degradable form of geminin in various cell lines. Geminin expression reduced the quantity of Mcm2 bound to chromatin and blocked cell proliferation. U2OS (p53+/Rb+) cells showed an early S phase arrest with high cyclin E and undetectable cyclin A levels, consistent with the activation of an intra-S checkpoint. Saos2 (p53-/Rb-) cells showed an accumulation of cells in late S and G2/M with approximately normal levels of cyclin A, consistent with loss of this intra-S phase checkpoint. Geminin also induced apoptosis in both these cell lines. In contrast, IMR90 primary fibroblasts over-expressing geminin arrested in G1 with reduced cyclin E levels and no detectable apoptosis. A ‘licensing checkpoint’ may therefore act in primary cells to prevent passage into S phase in the absence of sufficient origin licensing. These results suggest that inhibition of the licensing system may cause cancer-specific cell killing and therefore represent a novel anti-cancer target.

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Acknowledgements

We thank Neil Perkins, Pedro Jares, Pamela Robertson, Jean Christophe Bourdon and Marcus Achison for help at various stages of this project. This work is the subject of UK patent application 0209508.1. This work was supported by a British Commonwealth scholarship to S Shreeram and Cancer Research UK grant SP2385/0101.

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  1. Wellcome Trust Biocentre, University of Dundee, Dow Street, Dundee DD1 5EH, UK

    S Shreeram & J Julian Blow

  2. Department of Surgery and Molecular Oncology, University of Dundee, Ninewells Hospital and Medical School, Dundee, DD1 9SY, UK

    S Shreeram, Alison Sparks & David P Lane

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  1. S Shreeram
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Correspondence to J Julian Blow.

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Shreeram, S., Sparks, A., Lane, D. et al. Cell type-specific responses of human cells to inhibition of replication licensing. Oncogene 21, 6624–6632 (2002). https://doi.org/10.1038/sj.onc.1205910

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