Letter | Published:

A PP1–PP2A phosphatase relay controls mitotic progression

Nature volume 517, pages 9498 (01 January 2015) | Download Citation

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Abstract

The widespread reorganization of cellular architecture in mitosis is achieved through extensive protein phosphorylation, driven by the coordinated activation of a mitotic kinase network and repression of counteracting phosphatases. Phosphatase activity must subsequently be restored to promote mitotic exit. Although Cdc14 phosphatase drives this reversal in budding yeast, protein phosphatase 1 (PP1) and protein phosphatase 2A (PP2A) activities have each been independently linked to mitotic exit control in other eukaryotes1,2,3,4,5,6. Here we describe a mitotic phosphatase relay in which PP1 reactivation is required for the reactivation of both PP2A-B55 and PP2A-B56 to coordinate mitotic progression and exit in fission yeast. The staged recruitment of PP1 (the Dis2 isoform) to the regulatory subunits of the PP2A-B55 and PP2A-B56 (B55 also known as Pab1; B56 also known as Par1) holoenzymes sequentially activates each phosphatase. The pathway is blocked in early mitosis because the Cdk1–cyclin B kinase (Cdk1 also known as Cdc2) inhibits PP1 activity, but declining cyclin B levels later in mitosis permit PP1 to auto-reactivate1,7,8,9,10. PP1 first reactivates PP2A-B55; this enables PP2A-B55 in turn to promote the reactivation of PP2A-B56 by dephosphorylating a PP1-docking site in PP2A-B56, thereby promoting the recruitment of PP1. PP1 recruitment to human, mitotic PP2A-B56 holoenzymes and the sequences of these conserved PP1-docking motifs11,12 suggest that PP1 regulates PP2A-B55 and PP2A-B56 activities in a variety of signalling contexts throughout eukaryotes.

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Acknowledgements

This work was supported by Cancer Research UK (CRUK) grant numbers C147/A16406 and C29/A13678. We thank I. Alvarez-Tabares, K. Labib, K. Gull, M. Yanagida, V. Simanis, J. Bähler and K. Gould for reagents.

Author information

Author notes

    • Agnes Grallert
    •  & Elvan Boke

    These authors contributed equally to this work.

    • Elvan Boke

    Present address: Department of Systems Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.

Affiliations

  1. Cell Division Group, CRUK Manchester Institute, University of Manchester, Wilmslow Road, Manchester M20 4BX, UK

    • Agnes Grallert
    • , Elvan Boke
    • , Ben Hodgson
    •  & Iain M. Hagan
  2. The Gurdon Institute, Tennis Court Road, University of Cambridge, Cambridge, CB2 1QN, UK

    • Anja Hagting
    •  & Jonathon Pines
  3. Biological Mass Spectrometry, CRUK Manchester Institute, University of Manchester, Wilmslow Road, Manchester M20 4BX, UK

    • Yvonne Connolly
    • , John R. Griffiths
    •  & Duncan L. Smith

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Contributions

E.B. carried out the initial identification, validation and timing of PP1Dis2–B55Pab1/B56Par1 association, initial two-hybrid and co-immunoprecipitation assays, initial determination of specificity and timing of PP1Dis2.T316 phospho-recognition, and phospho-mapping of B56Par1 (with D.L.S. and Y.C.) and generated some mutant alleles. E.B. generated Fig. 4d; all other data, except for Fig. 4k and Extended Data Figs 5c, 9b–d, were generated and largely devised by A.G. Data for Extended Data Fig. 5c were generated by A.G., Y.C., D.L.S. and J.R.G. Molecular biology and background work for human two-hybrid assays and generation of human vectors was carried out by B.H. I.M.H. devised the work presented in Figs 1c, 2a, c, f and 4d with E.B., and devised the remainder of the work with A.G. A.H. and J.P. designed and executed the human work and gel filtration.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Iain M. Hagan.

Extended data

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    Supplementary Table 1

    This file contains the strains used in this study.

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DOI

https://doi.org/10.1038/nature14019

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