Abstract
The p16INK4a cyclin-dependent kinase inhibitor has a key role in establishing stable G1 cell-cycle arrest through activating the retinoblastoma (Rb) tumour suppressor protein pRb1,2,3,4,5 in cellular senescence. Here, we show that the p16INK4a /Rb-pathway also cooperates with mitogenic signals to induce elevated intracellular levels of reactive oxygen species (ROS), thereby activating protein kinase Cδ (PKCδ) in human senescent cells. Importantly, once activated by ROS, PKCδ promotes further generation of ROS, thus establishing a positive feedback loop to sustain ROS–PKCδ signalling6,7,8. Sustained activation of ROS–PKCδ signalling irreversibly blocks cytokinesis, at least partly through reducing the level of WARTS (also known as LATS1), a mitotic exit network (MEN) kinase required for cytokinesis9,10,11, in human senescent cells. This irreversible cytokinetic block is likely to act as a second barrier to cellular immortalization ensuring stable cell-cycle arrest in human senescent cells. These results uncover an unexpected role for the p16INK4a–Rb pathway and provide a new insight into how senescent cell-cycle arrest is enforced in human cells.
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Acknowledgements
We thank D. Mann, G. Peters, J. Campisi and U. Kikkawa for helpful comments on the manuscript; R. Agami for providing p16 knockdown construct; D. Beach and G. Hannon for providing pMarX retrovirus vector; W. Jiang and R. Fukunaga for providing anti-PRC1 antibody; and D. Kufe and S. Ohno for PKCδ cDNA. We also thank A. Hirao for his useful suggestion in analysis of intracellular ROS. This work was supported by grants from Ministry of Education, Science, Sports and Technology of Japan, Ministry of Health, Labour and Welfare of Japan, the Uehara Memorial Foundation, the Sumitomo Foundation, the Nakatomi Foundation, the Sagawa Foundation for Promotion of Cancer Research, the Inamori Foundation and Sankyo Foundation of Life Science (E.H.). A.T. is supported by the Japan Society for the Promotion of Science.
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Takahashi, A., Ohtani, N., Yamakoshi, K. et al. Mitogenic signalling and the p16INK4a–Rb pathway cooperate to enforce irreversible cellular senescence. Nat Cell Biol 8, 1291–1297 (2006). https://doi.org/10.1038/ncb1491
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DOI: https://doi.org/10.1038/ncb1491
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