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K-fibre minus ends are stabilized by a RanGTP-dependent mechanism essential for functional spindle assembly

Nature Cell Biology volume 13, pages 1406–1414 (2011)Cite this article

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Abstract

Chromosome segregation requires the formation of K-fibres, microtubule bundles that attach sister kinetochores to spindle poles. Most K-fibre microtubules originate around the chromosomes through a non-centrosomal RanGTP-dependent pathway and become oriented with the plus ends attached to the kinetochore and the minus ends focused at the spindle poles. The capture and stabilization of microtubule plus ends at the kinetochore has been extensively studied but very little is known on how their minus-end dynamics are controlled. Here we show that MCRS1 is a RanGTP-regulated factor essential for non-centrosomal microtubule assembly. MCRS1 localizes to the minus ends of chromosomal microtubules and K-fibres, where it protects them from depolymerization. Our data reveal the existence of a mechanism that stabilizes the minus ends of chromosomal microtubules and K-fibres, and is essential for the assembly of a functional bipolar spindle.

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Figure 1: MCRS1 localizes to K-fibres and chromosomal microtubules.
Figure 2: MCRS1 is important for chromosomal microtubule assembly.
Figure 3: MCRS1 silencing induces spindle instability.
Figure 4: MCRS1 is involved in K-fibre minus-end stability.
Figure 5: Xenopus MCRS1 is an importin- β-binding protein included in a RanGTP-dependent microtubule-stabilizing complex.
Figure 6: MCRS1 counteracts MCAK activity in vitro and in vivo.

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Acknowledgements

We thank S. Gross for statistical analysis of data; M. Mendoza, J. Solon, T. Surrey and the Vernos laboratory’s members for helpful discussions and critical comments on the manuscript; P. Meraldi for the gift of the H2B–eGFP/α-tubulin–mRFP and H2B–RFP/PA-GFP–tubulin HeLa cell lines and anti-Mad1 antibodies, A. Khodjakov for the gift of the GFP–centrin-1 HeLa cell line and S. Rybina and E. Karsenti for the gift of purified MCAK and anti-MCAK antibodies. M. Sanz, N. Mallol, V. Beltran-Sastre and L. Ávila provided technical assistance. We thank the Advanced Light Microscopy Unit of the CRG for microscopy support. This work was funded by grants from the Spanish MEC and MICINN BFU2006-04694, BFU2009-10202 and CSD2006-00023.

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  1. Cell and Developmental Biology Program, Centre for Genomic Regulation (CRG), Universitat Pompeu Fabra, 08003 Barcelona, Spain

    Sylvain Meunier & Isabelle Vernos

  2. Institució Catalana de Recerca i Estudis Avançats (ICREA), 08010 Barcelona, Spain

    Isabelle Vernos

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I.V. and S.M. designed the experiments. S.M. carried out all the experiments and analysed the data. I.V. and S.M. wrote the manuscript.

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Correspondence to Isabelle Vernos.

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Meunier, S., Vernos, I. K-fibre minus ends are stabilized by a RanGTP-dependent mechanism essential for functional spindle assembly. Nat Cell Biol 13, 1406–1414 (2011). https://doi.org/10.1038/ncb2372

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