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Chromosome congression in the absence of kinetochore fibres

Nature Cell Biology volume 11, pages 832–838 (2009)Cite this article

Abstract

Proper chromosome congression (the process of aligning chromosomes on the spindle) contributes to accurate and faithful chromosome segregation. It is widely accepted that congression requires kinetochore fibres (K-fibres), microtubule bundles that extend from the kinetochores to spindle poles1,2. Here, we demonstrate that chromosomes in human cells co-depleted of HSET (human kinesin-14)3,4 and hNuf2 (human Ndc80/Hec1-complex component)5 can congress to the metaphase plate in the absence of K-fibres. However, the chromosomes are not stably maintained at the metaphase plate under these conditions. Chromosome congression in HSET + hNuf2 co-depleted cells required the plus-end directed motor CENP-E (centromere protein E; kinesin-7 family member)6, which has been implicated in the gliding of mono-oriented kinetochores alongside adjacent K-fibres7. Thus, proper end-on attachment of kinetochores to microtubules is not necessary for chromosome congression. Instead, our data support the idea that congression allows unattached chromosomes to move to the middle of the spindle where they have a higher probability of establishing connections with both spindle poles. These bi-oriented connections are also used to maintain stable chromosome alignment at the spindle equator.

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Figure 1: Chromosomes can align in cells lacking K-fibres.
Figure 2: Kinetochores laterally bind stabilized microtubule bundles in the absence of K-fibres.
Figure 3: In the absence of K-fibres, kinetochores congress with alternating kinetochores leading.
Figure 4: Chromosome congression in cells with disrupted K-fibres is dependent on CENP-E.
Figure 5: K-fibres are needed to maintain chromosome alignment.

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Acknowledgements

We thank S. Ems-McClung and J. Powers for discussions and critical comments on the manuscript, G. Fang for HURP antibodies, D. Compton for HSET constructs, B. Sullivan for the GFP–CENP A construct and T. Yen for the CENP-E antibody. S.C. is grateful to members of the Marine Biological Laboratory Physiology course for instruction in imaging and many thoughtful discussions. The Indiana University Bloomington Light Microscopy Imaging Center and Wadsworth Center's electron microscopy Core facility provided microscopy resources. This work was supported by NIH grants (GM59618) to C.E.W. and (GM59363) A.K; C.B.O. was supported by a Kirschstein-National Research Service Award post-doctoral fellowship (GM077911).

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Authors and Affiliations

  1. Biochemistry Program, Indiana University, Myers Hall 262, 915 East 3rd Street, Bloomington, 47405, IN, USA

    Shang Cai

  2. Medical Sciences Program, Indiana University, Myers Hall 262, 915 East 3rd Street, Bloomington, 47405, IN, USA

    Claire E. Walczak

  3. Division of Molecular Medicine, New York State Department of Health, Wadsworth Center, Albany, 12201, NY, USA

    Christopher B. O'Connell & Alexey Khodjakov

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Contributions

S.C. performed most of the experiments, analysed the data and prepared the initial draft of the manuscript; C.O. performed electron microscopy analyses and C.E.W. and A.K. in conjunction with S.C. and C.O. designed the experiments, interpreted the data, edited the manuscript and prepared the final data for publication.

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Correspondence to Claire E. Walczak.

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Cai, S., O'Connell, C., Khodjakov, A. et al. Chromosome congression in the absence of kinetochore fibres. Nat Cell Biol 11, 832–838 (2009). https://doi.org/10.1038/ncb1890

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