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DNA breaks and chromosome pulverization from errors in mitosis

Nature volume 482, pages 53–58 (2012)Cite this article

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Abstract

The involvement of whole-chromosome aneuploidy in tumorigenesis is the subject of debate, in large part because of the lack of insight into underlying mechanisms. Here we identify a mechanism by which errors in mitotic chromosome segregation generate DNA breaks via the formation of structures called micronuclei. Whole-chromosome-containing micronuclei form when mitotic errors produce lagging chromosomes. We tracked the fate of newly generated micronuclei and found that they undergo defective and asynchronous DNA replication, resulting in DNA damage and often extensive fragmentation of the chromosome in the micronucleus. Micronuclei can persist in cells over several generations but the chromosome in the micronucleus can also be distributed to daughter nuclei. Thus, chromosome segregation errors potentially lead to mutations and chromosome rearrangements that can integrate into the genome. Pulverization of chromosomes in micronuclei may also be one explanation for ‘chromothripsis’ in cancer and developmental disorders, where isolated chromosomes or chromosome arms undergo massive local DNA breakage and rearrangement.

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Figure 1: Micronuclei from lagging chromosomes develop DNA breaks.
Figure 2: DNA breaks in a HAC targeted to a micronucleus.
Figure 3: DNA damage in micronuclei results from aberrant DNA replication.
Figure 4: Defective MCM2-7 complex recruitment, DNA damage response and nucleocytoplasmic transport in micronuclei.
Figure 5: The fate of chromosomes in micronuclei.

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Acknowledgements

We thank A. D’Andrea, M. E. McLaughlin, T. A. Rapoport, J. Walters and Pellman laboratory members for discussions and/or comments on the manuscript; L. Cameron for advice and help on microscopy; H. Li for irradiation of cells; M. Nitta and M. Hennessy for preliminary experiments; and V. Larionov, B. Stillman, J. Ellenberg, I. Mattaj, A. Miyawaki and T. Kuroda for reagents. D.P. was supported by the Howard Hughes Medical Institute and the NIH (GM083299); K.C. was a fellow of A*STAR Singapore; N.J.G. was a fellow of the Leukemia and Lymphoma Society; Y.P. and D.C. were funded by the NIH (1R01CA142698-01).

Author information

Authors and Affiliations

  1. Department of Pediatric Oncology, Dana-Farber Cancer Institute, 450 Brookline Avenue, Boston, Massachusetts 02115, USA,

    Karen Crasta, Neil J. Ganem, Regina Dagher, Alexandra B. Lantermann, Luigi Nezi & David Pellman

  2. Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, Massachusetts 02115, USA,

    Karen Crasta, Neil J. Ganem, Regina Dagher & David Pellman

  3. Howard Hughes Medical Institute, 450 Brookline Avenue, Boston, 02115, Massachusetts, USA

    Karen Crasta, Neil J. Ganem, Regina Dagher & David Pellman

  4. Belfer Institute for Applied Cancer Science, Dana-Farber Cancer Institute, 4 Blackfan Street, Boston, Massachusetts 02115, USA ,

    Elena V. Ivanova & Alexei Protopopov

  5. Department of Radiation Oncology, Dana-Farber Cancer Institute, 450 Brookline Avenue, Boston, Massachusetts 02115, USA,

    Yunfeng Pan & Dipanjan Chowdhury

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Contributions

D.P. conceived the project; K.C., N.J.G., R.D., A.B.L., D.C. and D.P. designed the experiments; D.P., K.C. and N.J.G. wrote the manuscript with edits from all authors; K.C. contributed Figs. 1–3, 4a, b and Supplementary Figs 2–8 with help from R.D. N.J.G. contributed Fig. 5j, Supplementary Fig. 12, Supplementary Table 1 and Supplementary Movies. R.D., E.V.I. and A.P. contributed Fig. 5a–i and Supplementary Figs 1 and 11; A.B.L. contributed Fig. 4c, d and Supplementary Figs 9 and 10; Y.P. and D.C. contributed Supplementary Fig. 3d; L.N. contributed Fig. 2a.

Corresponding author

Correspondence to David Pellman.

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Supplementary information

Supplementary Information

The file contains Supplementary Figures 1-12 with legends, Supplementary References, Supplementary Table 1 and legends for Supplementary Movies 1-4. (PDF 11345 kb)

Supplementary Movie 1

The movie shows live-cell imaging of a MN in a U2OS cell expressing H2B-mRFP -see Supplementary Information file for full legend. (MOV 6862 kb)

Supplementary Movie 2

The movie shows live-cell imaging of a MN in a U2OS cell expressing H2B-mRFP -see Supplementary Information file for full legend. (MOV 9785 kb)

Supplementary Movie 3

The movie shows live-cell imaging of a MN in a U2OS cell expressing H2B-Kaede (corresponds to the cell shown in Figure 4g, top row) - see Supplementary Information file for full legend. (MOV 16698 kb)

Supplementary Movie 4

The movie shows live-cell imaging of a MN in a U2OS cell expressing H2B-Kaede (cell is located in the bottom-left portion of the video and corresponds to the cell shown in Figure 4g, bottom row) - see Supplementary Information file for full legend. (MOV 7152 kb)

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Crasta, K., Ganem, N., Dagher, R. et al. DNA breaks and chromosome pulverization from errors in mitosis. Nature 482, 53–58 (2012). https://doi.org/10.1038/nature10802

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