Letter | Published:

Distinct kinetics of serine and threonine dephosphorylation are essential for mitosis

Nature Cell Biology volume 19, pages 14331440 (2017) | Download Citation

Abstract

Protein phosphatase 2A (PP2A) in complex with B55 regulatory subunits reverses cyclin-dependent kinase 1 (Cdk1) phosphorylations at mitotic exit1,2,3,4,5. Interestingly, threonine and serine residues phosphorylated by Cdk1 display distinct phosphorylation dynamics, but the biological significance remains unexplored. Here we demonstrate that the phosphothreonine preference of PP2A–B55 provides an essential regulatory element of mitotic exit. To allow rapid activation of the anaphase-promoting complex/cyclosome (APC/C) co-activator Cdc20, inhibitory phosphorylation sites are conserved as threonines while serine substitutions delay dephosphorylation and Cdc20 activation. Conversely, to ensure timely activation of the interphase APC/C co-activator Cdh1, inhibitory phosphorylation sites are conserved as serines, and threonine substitutions result in premature Cdh1 activation. Furthermore, rapid translocation of the chromosomal passenger complex to the central spindle is prevented by mutation of a single phosphorylated threonine to serine in inner centromere protein (INCENP), leading to failure of cytokinesis. Altogether, the findings of our work reveal that the inherent residue preference of a protein phosphatase can provide temporal regulation in biological processes.

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Acknowledgements

We thank T. Mayer and A. Heim (Universitat Konstanz, Germany), J. Pines (ICR, United Kingdom), D. Hermida Aponte and C. Lukas (Novo Nordisk Foundation Center for Protein Research, Denmark) for reagents and discussion, and the CPR protein production facility and, in particular, G. Cazzamali, M. Williamson, A. L. L. Vala and H. Koc in the eukaryotic and prokaryotic expression teams for purifying Cdc20 proteins. The Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, is supported financially by the Novo Nordisk Foundation (grant agreement NNF14CC0001). In addition, this work was supported by grants from the Danish Cancer Society (R72-A4351-13-S2 and R124-A7827-15-S2), a grant from the Danish Council for Independent Research (DFF-4183-00388) and a grant from the Novo Nordisk Foundation (NNF16OC0022394) to J.N.

Author information

Author notes

    • Emil P. T. Hertz
    • , Dimitriya H. Garvanska
    •  & Thomas Kruse

    These authors contributed equally to this work.

Affiliations

  1. The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen, Denmark

    • Jamin B. Hein
    • , Emil P. T. Hertz
    • , Dimitriya H. Garvanska
    • , Thomas Kruse
    •  & Jakob Nilsson

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Contributions

All authors contributed to project planning and data analysis of their respective experiments and J.N. coordinated the work. J.B.H. performed all analysis of Cdc20 and Cdh1, except in vitro APC/C ubiquitylation assays performed by D.H.G. E.P.T.H. performed Michaelis–Menten kinetics analysis of PP2A–B55 and all in vitro kinase and phosphatase assays and phosphorylation of Arpp19. T.K. performed in vivo analysis of INCENP. J.B.H. and D.H.G. performed analysis of phosphoantibodies. All authors contributed to the writing of the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Jakob Nilsson.

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DOI

https://doi.org/10.1038/ncb3634

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