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ΔMEKK3:ER* activation induces a p38α/β2-dependent cell cycle arrest at the G2 checkpoint

Oncogene volume 21, pages 8089–8104 (2002)Cite this article

Abstract

Whilst many studies have examined the role of the MAP Kinases in regulating the G1→S transition, much less is known about the function of these pathways in regulating other cell cycle transitions. Stimulation of the conditional mutant ΔMEKK3:ER* in asynchronous hamster (CCl39) and rat (Rat-1) fibroblasts resulted in the strong activation of endogenous JNK and p38 but only a weak activation of ERK. Activation of ΔMEKK3:ER* inhibited cell proliferation through a combination of an initial G1 and G2 cell cycle arrest, followed by a delayed onset of apoptosis. When cells were synchronized in S phase with aphidicolin and then released, activation of ΔMEKK3:ER* resulted in the up-regulation of p21CIP1 and a pronounced inhibition of cyclin A/CDK2 and cyclin B1/CDK1 kinase activity. Analysis of mitotic figures indicated that cells failed to enter mitosis, arresting late in G2. ΔMEKK3:ER*-mediated CDK inhibition and G2 arrest did not absolutely require p21CIP1, since both events were observed in Rat-1 cells in which p21CIP1 is transcriptionally silenced due to promoter methylation. Rather, CDK inhibition was associated with a down-regulation of cyclin A and B1 expression. Finally, application of the p38 inhibitor SB203580 partially restored cyclin B associated kinase activity and allowed cells to proceed through mitosis into the next G1 phase, suggesting that activation of the p38α/β2 pathway can promote a G2 cell cycle arrest.

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Acknowledgements

We would like to thank Geoff Morgan for maintenance of the Babraham Institute Flow Cytometry Facility and for his technical advice, Adele Murrell for advice on counting mitotic indices and members of the Cook group for interesting discussions and comments. We are grateful to Tim Hunt for the generous provision of cyclin A and B antibodies, Jiri Lukas for Cdh1 antibodies, James Trzaskos (DuPont) for providing U0126 and Bert Vogelstein for providing WT and p53−/− HCT116 cells. This work was supported by Cancer Research UK (SP2458/0201), the BBSRC Science of Ageing Initiative (202/SAG10012) and a Competitive Strategic Grant from the BBSRC. CR Weston was funded by a MRC PhD studentship and SJ Cook is a Senior Cancer Research Fellow of Cancer Research UK.

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  1. Andrew P Garner

    Present address: Cancer and Infection Bioscience, AstraZeneca, Alderley Park, Macclesfield, SK10 4TG, UK

  2. Claire R Weston

    Present address: University of Massachusetts Medical School, II Biotech Park, 373 Plantation St., Suite 306, Worcester, 01605, MA, USA

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  1. Inositide Laboratory, Signalling Programme, The Babraham Institute, Babraham Hall, Cambridge, CB2 4AT, UK

    Andrew P Garner, Claire R Weston, Daniel E Todd, Kathryn Balmanno & Simon J Cook

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Garner, A., Weston, C., Todd, D. et al. ΔMEKK3:ER* activation induces a p38α/β2-dependent cell cycle arrest at the G2 checkpoint. Oncogene 21, 8089–8104 (2002). https://doi.org/10.1038/sj.onc.1206000

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