1. Cell Biology

A FRET-based study reveals site-specific regulation of spindle position checkpoint proteins at yeast centrosomes

  1. Yuliya Gryaznova
  2. Ayse Koca Caydasi
  3. Gabriele Malengo
  4. Victor Sourjik
  5. Gislene Pereira  Is a corresponding author
  1. German Cancer Research Centre, Germany
  2. Centre for Molecular Biology, Germany
https://doi.org/10.7554/eLife.14029
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  1. Yuliya Gryaznova
  2. Ayse Koca Caydasi
  3. Gabriele Malengo
  4. Victor Sourjik
  5. Gislene Pereira
(2016)
A FRET-based study reveals site-specific regulation of spindle position checkpoint proteins at yeast centrosomes
eLife 5:e14029.
https://doi.org/10.7554/eLife.14029
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Abstract

The spindle position checkpoint (SPOC) is a spindle pole body (SPB, equivalent of mammalian centrosome) associated surveillance mechanism that halts mitotic exit upon spindle mis-orientation. Here, we monitored the interaction between SPB proteins and the SPOC component Bfa1 by FRET microscopy. We show that Bfa1 binds to the scaffold-protein Nud1 and the γ-tubulin receptor Spc72. Spindle misalignment specifically disrupts Bfa1-Spc72 interaction by a mechanism that requires the 14-3-3-family protein Bmh1 and the MARK/PAR-kinase Kin4. Dissociation of Bfa1 from Spc72 prevents the inhibitory phosphorylation of Bfa1 by the polo-like kinase Cdc5. We propose Spc72 as a regulatory hub that coordinates the activity of Kin4 and Cdc5 towards Bfa1. In addition, analysis of spc72∆ cells shows that a mitotic-exit-promoting dominant signal, which is triggered upon elongation of the spindle into the bud, overrides the SPOC. Our data reinforce the importance of daughter-cell-associated factors and centrosome-based regulations in mitotic exit and SPOC control.

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Author details

  1. Yuliya Gryaznova

    DKFZ-ZMBH Alliance, German Cancer Research Centre, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.

  2. Ayse Koca Caydasi

    DKFZ-ZMBH Alliance, German Cancer Research Centre, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.

  3. Gabriele Malengo

    DKFZ-ZMBH Alliance, Centre for Molecular Biology, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.

  4. Victor Sourjik

    DKFZ-ZMBH Alliance, Centre for Molecular Biology, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.

  5. Gislene Pereira

    DKFZ-ZMBH Alliance, German Cancer Research Centre, Heidelberg, Germany
    For correspondence
    gislene.pereira@cos.uni-heidelberg.de
    Competing interests
    The authors declare that no competing interests exist.

Copyright

© 2016, Gryaznova et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Yuliya Gryaznova
  2. Ayse Koca Caydasi
  3. Gabriele Malengo
  4. Victor Sourjik
  5. Gislene Pereira
(2016)
A FRET-based study reveals site-specific regulation of spindle position checkpoint proteins at yeast centrosomes
eLife 5:e14029.
https://doi.org/10.7554/eLife.14029

Share this article

https://doi.org/10.7554/eLife.14029

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    1. Cell Biology

    Spatial signals link exit from mitosis to spindle position

    Jill Elaine Falk, Dai Tsuchiya ... Angelika Amon
    Research Article Updated

    In budding yeast, if the spindle becomes mispositioned, cells prevent exit from mitosis by inhibiting the mitotic exit network (MEN). The MEN is a signaling cascade that localizes to spindle pole bodies (SPBs) and activates the phosphatase Cdc14. There are two competing models that explain MEN regulation by spindle position. In the 'zone model', exit from mitosis occurs when a MEN-bearing SPB enters the bud. The 'cMT-bud neck model' posits that cytoplasmic microtubule (cMT)-bud neck interactions prevent MEN activity. Here we find that 1) eliminating cMT– bud neck interactions does not trigger exit from mitosis and 2) loss of these interactions does not precede Cdc14 activation. Furthermore, using binucleate cells, we show that exit from mitosis occurs when one SPB enters the bud despite the presence of a mispositioned spindle. We conclude that exit from mitosis is triggered by a correctly positioned spindle rather than inhibited by improper spindle position.