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Recruiting a microtubule-binding complex to DNA directs chromosome segregation in budding yeast

Nature Cell Biology volume 11, pages 11161120 (2009) | Download Citation

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Abstract

Accurate chromosome segregation depends on the kinetochore, which is the complex of proteins that link microtubules to centromeric DNA1. The kinetochore of the budding yeast Saccharomyces cerevisiae consists of more than 80 proteins assembled on a 125-bp region of DNA1. We studied the assembly and function of kinetochore components by fusing individual kinetochore proteins to the lactose repressor (LacI) and testing their ability to improve segregation of a plasmid carrying tandem repeats of the lactose operator (LacO). Targeting Ask1, a member of the Dam1–DASH microtubule-binding complex, creates a synthetic kinetochore that performs many functions of a natural kinetochore: it can replace an endogenous kinetochore on a chromosome, bi-orient sister kinetochores at metaphase during the mitotic cycle, segregate sister chromatids, and repair errors in chromosome attachment. We show the synthetic kinetochore functions do not depend on the DNA-binding components of the natural kinetochore but do require other kinetochore proteins. We conclude that tethering a single kinetochore protein to DNA triggers assembly of the complex structure that directs mitotic chromosome segregation.

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Acknowledgements

We thank Greg Lang, Joana Gonçalves-Sá, John Koschwanez, Gregg Wildenberg and Dai Tsuchiya for technical advice; Charles Asbury, Ted Salmon, Steve Elledge, Ajit Joglekar, Frank Solomon, Amy Rowat and members of the Murray lab for critical reading of the manuscript; and Steve Elledge, John Kilmartin and Sue Biggins for strains. This work was supported by a US National Institutes of Health (NIH) National Research Service Award fellowship to S.L. and an NIH grant to A.W.M. (GM043987).

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Author notes

    • Soni Lacefield

    Current Address: Department of Biology, Indiana University, 1001 E 3rd St, Bloomington, Indiana 47405, USA.

Affiliations

  1. Department of Molecular and Cellular Biology, Harvard University, 52 Oxford St, Cambridge, Massachusetts 02138, USA.

    • Soni Lacefield
    • , Derek T. C. Lau
    •  & Andrew W. Murray

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Contributions

S.L. and A.W.M. conceived and designed the experiments. S.L. and D.T.L. performed and analysed the experiments. S.L. and A.W.M. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Andrew W. Murray.

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DOI

https://doi.org/10.1038/ncb1925

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