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Abnormal mitosis triggers p53-dependent cell cycle arrest in human tetraploid cells

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Abstract

Erroneously arising tetraploid mammalian cells are chromosomally instable and may facilitate cell transformation. An increasing body of evidence shows that the propagation of mammalian tetraploid cells is limited by a p53-dependent arrest. The trigger of this arrest has not been identified so far. Here we show by live cell imaging of tetraploid cells generated by an induced cytokinesis failure that most tetraploids arrest and die in a p53-dependent manner after the first tetraploid mitosis. Furthermore, we found that the main trigger is a mitotic defect, in particular, chromosome missegregation during bipolar mitosis or spindle multipolarity. Both a transient multipolar spindle followed by efficient clustering in anaphase as well as a multipolar spindle followed by multipolar mitosis inhibited subsequent proliferation to a similar degree. We found that the tetraploid cells did not accumulate double-strand breaks that could cause the cell cycle arrest after tetraploid mitosis. In contrast, tetraploid cells showed increased levels of oxidative DNA damage coinciding with the p53 activation. To further elucidate the pathways involved in the proliferation control of tetraploid cells, we knocked down specific kinases that had been previously linked to the cell cycle arrest and p53 phosphorylation. Our results suggest that the checkpoint kinase ATM phosphorylates p53 in tetraploid cells after abnormal mitosis and thus contributes to proliferation control of human aberrantly arising tetraploids.

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Acknowledgments

We are thankful to Steven Taylor for providing the RPE1 cells stably transfected with H2B–GFP. We thank Tom Gaitanos, Steven Bergink, and Sabine Elowe for their helpful comments on the manuscript. We are thankful to Aline Sewo Pires de Campos, Susanne Gutmann, Daniela Fellner, and Miriam Penka for technical support.

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Authors and Affiliations

  1. Max Planck Institute of Biochemistry, Am Klopferspitz 18, 82152, Martinsried, Germany

    Christian Kuffer, Anastasia Yurievna Kuznetsova & Zuzana Storchová

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  1. Christian Kuffer
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  2. Anastasia Yurievna Kuznetsova
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  3. Zuzana Storchová
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Correspondence to Zuzana Storchová.

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Communicated by Erich Nigg

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Kuffer, C., Kuznetsova, A.Y. & Storchová, Z. Abnormal mitosis triggers p53-dependent cell cycle arrest in human tetraploid cells. Chromosoma 122, 305–318 (2013). https://doi.org/10.1007/s00412-013-0414-0

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