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Genome-wide genetic analysis of polyploidy in yeast

Nature volume 443, pages 541547 (05 October 2006) | Download Citation

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Abstract

Polyploidy, increased sets of chromosomes, occurs during development, cellular stress, disease and evolution. Despite its prevalence, little is known about the physiological alterations that accompany polyploidy. We previously described ‘ploidy-specific lethality’, where a gene deletion that is not lethal in haploid or diploid budding yeast causes lethality in triploids or tetraploids. Here we report a genome-wide screen to identify ploidy-specific lethal functions. Only 39 out of 3,740 mutations screened exhibited ploidy-specific lethality. Almost all of these mutations affect genomic stability by impairing homologous recombination, sister chromatid cohesion, or mitotic spindle function. We uncovered defects in wild-type tetraploids predicted by the screen, and identified mechanisms by which tetraploidization affects genomic stability. We show that tetraploids have a high incidence of syntelic/monopolar kinetochore attachments to the spindle pole. We suggest that this defect can be explained by mismatches in the ability to scale the size of the spindle pole body, spindle and kinetochores. Thus, geometric constraints may have profound effects on genome stability; the phenomenon described here may be relevant in a variety of biological contexts, including disease states such as cancer.

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Acknowledgements

We are grateful to many colleagues from the yeast community for providing the reagents. We thank A. Amon, G. Fink, J. Haber, R. Rothstein, M. McLaughlin, M. Raschle and members of the Pellman laboratory for discussions; G. Fink, S. Elledge, J. Haber, A. Van Oudenaarden, M. McLaughlin, R. Rothstein and J. Walter for comments on the manuscript; C. Glavin for Supplementary Fig. 9; and M. Lenburg for guidance on the analysis of the expression profile data. D.P. was supported by an NIH grant and a gift from the G. Harold and Leila Y. Mathers Foundation. The Boulder Laboratory for 3D Electron Microscopy of Cells is supported by an NIH grant to J. R. McIntosh.

Author information

Author notes

    • Amanda Breneman
    • , Jessica Cande
    •  & Joshua Dunn

    *These authors contributed equally to this work

Affiliations

  1. Department of Pediatric Oncology, Dana-Farber Cancer Institute,

    • Zuzana Storchová
    • , Amanda Breneman
    • , Jessica Cande
    • , Joshua Dunn
    •  & David Pellman
  2. Department of Physics, Harvard University, and

    • Kendra Burbank
  3. Department of Pediatric Hematology/Oncology, Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA

    • David Pellman
  4. The Boulder Laboratory for 3D Electron Microscopy of Cells, University of Colorado, Boulder, Colorado 80309, USA

    • Eileen O'Toole

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Competing interests

The transcriptional profiling data are available at MIAMExpress database (http://www.ebi.ac.uk/miamexpress) under accession number E-MEXP-822. Reprints and permissions information is available at www.nature.com/reprints. The authors declare no competing financial interests.

Corresponding author

Correspondence to David Pellman.

Supplementary information

PDF files

  1. 1.

    Supplementary Notes

    This file provides detailed descriptions of the genome-wide strategy for tetraploid formation, the plasmid shuffle strategy for tetraploid formation, Nuf2-GFP fluorescence intensity measurements, high-voltage EM tomography of yeast and description and explanation of expression profiling analysis. This file also contains Supplementary Figures 1–9 and Supplementary Tables 1–3.

Excel files

  1. 1.

    Supplementary Data 1

    Complete results of the genome-wide screen for genes specifically required in yeast tetraploid cells.

  2. 2.

    Supplementary Data 2

    Complete results of the expression profiling.

Videos

  1. 1.

    Supplementary Movie

    3D reconstruction of a tetraploid forming spindle.

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DOI

https://doi.org/10.1038/nature05178

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