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  • Original Paper
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DNA-damaging agents cause inactivation of translational regulators linked to mTOR signalling

Oncogene volume 19, pages 3021–3031 (2000)Cite this article

Abstract

Treatment of cells with DNA-damaging agents, such as etoposide, can cause growth arrest or apoptosis. Treatment of Swiss 3T3 or RAT-1 cells with etoposide led to the dephosphorylation of both p70 S6 kinase and eukaryotic initiation factor (eIF) 4E-binding protein 1 (4E-BP1), resulting in decreased p70 S6 kinase activity and an increase in 4E-BP1 binding to eIF4E. These effects were not prevented by the general caspase inhibitor, Z-VAD.FMK. These findings indicate caspase-independent inhibition of signalling pathways that involve the mammalian target of rapamycin (mTOR). Similar effects were observed in response to two other DNA-damaging agents, cisplatin and mitomycin-C. These events preceded apoptosis, which was assessed by caspase-3 activity assays and FACS analysis. This shows that inhibition of mTOR signalling is not a consequence of apoptosis, although it may play a role in the events that precede cell death. 4E-BP1 was cleaved during apoptosis yielding a fragment that retained the ability to bind eIF4E. Cleavage of 4E-BP1 was inhibited by treatment of the cells with Z-VAD.FMK, indicating it is caspase-dependent. Insulin elicited full activation of p70 S6 kinase and phosphorylation of 4E-PB1 in etoposide-treated cells prior to the onset of apoptosis, but not during cell death. This suggests that mTOR signalling becomes irreversibly inhibited only after entry into apoptosis.

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Acknowledgements

We would like to thank Dr BA Spruce for the cell-lines, helpful discussions and critical reading of the manuscript, Dr U Weigand for technical assistance on the FACS analysis, and Dr M Kleijn for critical reading of the manuscript. We would also like to thank Dr G Krause (Wayne State University, MI, USA) and Prof RM Denton (University of Bristol) for supplying antibodies. This work was supported by the Wellcome Trust through a Studentship to AR Tee.

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  1. Department of Anatomy & Physiology, Medical Sciences Institute/Wellcome Trust Building Complex, University of Dundee, Dow Street, Dundee, DD1 5EH, UK

    Andrew R Tee & Christopher G Proud

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  1. Andrew R Tee
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  2. Christopher G Proud
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Tee, A., Proud, C. DNA-damaging agents cause inactivation of translational regulators linked to mTOR signalling. Oncogene 19, 3021–3031 (2000). https://doi.org/10.1038/sj.onc.1203622

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