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Calcium triggers exit from meiosis II by targeting the APC/C inhibitor XErp1 for degradation

Nature volume 437, pages 1048–1052 (2005)Cite this article

Abstract

Vertebrate eggs awaiting fertilization are arrested at metaphase of meiosis II by a biochemical activity termed cytostatic factor (CSF)1,2. This activity inhibits the anaphase-promoting complex/cyclosome (APC/C), a ubiquitin ligase that triggers anaphase onset and mitotic/meiotic exit by targeting securin and M-phase cyclins for destruction3,4,5. On fertilization a transient rise in free intracellular calcium6 causes release from CSF arrest and thus APC/C activation. Although it has previously been shown that calcium induces the release of APC/C from CSF inhibition through calmodulin-dependent protein kinase II (CaMKII)7,8, the relevant substrates of this kinase have not been identified. Recently, we characterized XErp1 (Emi2), an inhibitor of the APC/C and key component of CSF activity in Xenopus egg extract9. Here we show that calcium-activated CaMKII triggers exit from meiosis II by sensitizing the APC/C inhibitor XErp1 for polo-like kinase 1 (Plx1)-dependent degradation. Phosphorylation of XErp1 by CaMKII leads to the recruitment of Plx1 that in turn triggers the destruction of XErp1 by phosphorylating a site known to serve as a phosphorylation-dependent degradation signal. These results provide a molecular explanation for how the fertilization-induced calcium increase triggers exit from meiosis II.

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Figure 1: XErp1 is a substrate of CaMKII in vitro.
Figure 2: CaMKII converts XErp1 into an efficient Plx1 substrate.
Figure 3: CaMKII activity is essential for the degradation of XErp1 in Xenopus egg extract.
Figure 4: Model of how calcium triggers release from CSF arrest.

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Acknowledgements

We thank O. Stemmann and F. Barr for reagents and discussions during this work. We are grateful to I. Gorr and D. Boos for advice on Δ90 experiments, S. Hümmer for continuous support and R. Neef for assistance with the far-western analyses.

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Author notes

  1. Nadine R. Rauh and Andreas Schmidt: *These authors contributed equally to this work

Authors and Affiliations

  1. Chemical Biology, Independent Research Group,

    Nadine R. Rauh, Andreas Schmidt, Jenny Bormann & Thomas U. Mayer

  2. Department of Cell Biology, Max Planck Institute of Biochemistry, Am Klopferspitz 18, D-82152, Martinsried, Germany

    Erich A. Nigg

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Correspondence to Thomas U. Mayer.

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Supplementary information

Supplementary Figure S1

Excess IVT-XErp1WT or -XErp1T195A blocks calcium-induced CSF release demonstrating that both proteins are active and folded properly. The stability of 35S-labelled IVT-securin and the morphology of the chromatin was used to monitor the cell cycle stage of the extract. (PDF 547 kb)

Supplementary Figure S2

XErp1 is degraded in Xenopus δ90 extracts possesing CaMKII activity. (PDF 436 kb)

Supplementary Figure S3

CaMKII triggers Plx1-dependent degradation of XErp1. Endogenous XErp1 is rapidly degraded in CSF extract supplemented with constitutively active CaMKII or calcium. CaMKII induced degradation of XErp1 depends on Plx1 activity as indicated by the fact that a dominant negative acting form of Plx1 (PBD) prevents CaMKII triggered XErp1 destruction. (PDF 327 kb)

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Rauh, N., Schmidt, A., Bormann, J. et al. Calcium triggers exit from meiosis II by targeting the APC/C inhibitor XErp1 for degradation. Nature 437, 1048–1052 (2005). https://doi.org/10.1038/nature04093

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