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Mitogenic signalling and the p16INK4a–Rb pathway cooperate to enforce irreversible cellular senescence

Nature Cell Biology volume 8, pages 1291–1297 (2006)Cite this article

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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Figure 1: Mitogenic signals cooperate with pRb and/or p53 to induce irreversible cell-cycle arrest.
Figure 2: Involvement of ROS–PKCδ signalling in irreversible cell-cycle arrest.
Figure 3: Downregulation of WARTS by PKCδ.
Figure 4: p16INK4a–Rb pathway elicits ROS–PKCδ signalling in primary human diploid fibroblasts.
Figure 5: Irreversibility of replicative senescence.

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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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Authors and Affiliations

  1. Institute for Genome Research, University of Tokushima, Tokushima, 770-8503, Japan

    Akiko Takahashi, Naoko Ohtani, Kimi Yamakoshi & Eiji Hara

  2. Graduate School of Medical Science, Kumamoto University, Kumamoto, 860-8556, Japan

    Shin-ichi Iida & Hideyuki Saya

  3. Hiroshima University School of Medicine, Hiroshima, 734-8551, Japan

    Hidetoshi Tahara

  4. Graduate School of Medicine, Tohoku University, Sendai, 980-8575, Japan

    Keiko Nakayama

  5. Medical Institute of Bioregulation, Kyushu University, Fukuoka, 812-8582, Japan

    Keiichi I. Nakayama

  6. Faculty of Pharmaceutical Sciences, Hiroshima International University, Kure, 737-0112, Japan

    Toshinori Ide

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Correspondence to Eiji Hara.

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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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