Abstract
Treatment of Swiss 3T3 cells with staurosporine resulted in dephosphorylation of two proteins which play key roles in regulating mRNA translation. This occurred before the execution of apoptosis, assessed by caspase-3 activity. These translation regulators are p70 S6 kinase, which phosphorylates ribosomal protein S6, and eukaryotic initiation factor (eIF) 4E binding protein 1 (4E-BP1), which both lie downstream of the mammalian target of rapamycin (mTOR). This resulted in decreased p70 S6 kinase activity, dephosphorylation of ribosomal protein S6, increased binding of 4E-BP1 to eIF4E and a concomitant decrease in eIF4F complexes. Our data show that staurosporine impairs mTOR signalling in vivo but that this not due to direct inhibition of mTOR or to inhibition of protein kinase C. It is becoming clear that agents which cause apoptosis inactivate mTOR signalling as a common early response prior to the execution of apoptosis, i.e., before caspase activation. Cell Death and Differentiation (2001) 8, 841–849
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Abbreviations
- BIM:
-
bisindolylmaleimide
- eIF:
-
eukaryotic initiation factor
- 4E-BP1:
-
eukaryotic initiation factor 4E-binding protein 1
- HEK 293:
-
human embryonic kidney 293
- MEK:
-
MAP kinase/Erk kinase
- m7GTP:
-
7-methylguanosine triphosphate
- mTOR:
-
mammalian target of rapamycin
- p70 S6 kinase:
-
70 kDa protein kinase that phosphorylates ribosomal protein S6
- PKC:
-
protein kinase C
- PKR:
-
double-stranded RNA-dependent protein kinase
- Z-VAD.FMK:
-
Z-Val-Ala-Asp.fluoromethylketone
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Acknowledgements
We would like to thank Dr. BA Spruce for the cell-lines and helpful discussions, Dr. G Krause (Wayne State University, MI, USA) and Prof. RM Denton (University of Bristol) for supplying antibodies. We would also like to thank Dr. P Shepherd (University College, London) for providing the FLAG-tagged mTOR construct, Dr. P Scott (University of Glasgow) for supplying the FKBP12 recombinant protein, Dr. DR Alessi (University of Dundee) for providing the ribosomal protein S6 phospho-specific antibody and Dr. TP Herbert (University of Dundee) for critical reading of the manuscript. This work was supported by the Wellcome Trust through the award of a Prize Studentship to AR Tee.
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Tee, A., Proud, C. Staurosporine inhibits phosphorylation of translational regulators linked to mTOR. Cell Death Differ 8, 841–849 (2001). https://doi.org/10.1038/sj.cdd.4400876
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DOI: https://doi.org/10.1038/sj.cdd.4400876
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