Abstract
Xenopus RINGO/Speedy (XRINGO) is a potent inducer of oocyte meiotic maturation that can directly activate Cdk1 and Cdk2. Here, we show that endogenous XRINGO protein accumulates transiently during meiosis I entry and then is downregulated. This tight regulation of XRINGO expression is the consequence of two interconnected mechanisms: processing and degradation. XRINGO processing involves recognition of at least three distinct phosphorylated recognition motifs by the SCFβTrCP ubiquitin ligase, followed by proteasome-mediated limited degradation, resulting in an amino-terminal XRINGO fragment. XRINGO processing is directly stimulated by several kinases, including protein kinase A and glycogen synthase kinase-3β, and may contribute to the maintenance of G2 arrest. On the other hand, XRINGO degradation after meiosis I is mediated by the ubiquitin ligase Siah-2, which probably requires phosphorylation of XRINGO on Ser 243 and may be important for the omission of S phase at the meiosis-I–meiosis-II transition in Xenopus oocytes.
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Acknowledgements
We thank J. Moore and T. Hunt (Cancer Research UK Clare Hall Laboratories, South Mimms, UK) for providing advice and reagents to perform DNA replication assays in cyclin-E-depleted extracts; E. Pogge von Strandmann (University of Cologne, Germany) for sending us the Xenopus Siah-2 cDNA; and O. Coux (CNRS-CRBM, France) for providing E1 enzymes, helpful suggestions and for critically reading the manuscript. A.R.N. acknowledges the grant BFU2004-03566 from the Ministerio de Educacion y Ciencia of Spain.
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G.J.G. performed all the experiments presented in this manuscript; most of the work was done during his PhD at the EMBL. A.V. critically contributed to the characterization of the PKA phosphorylation sites on XRINGO. I.F. and G.S. generated some XRINGO mutants and the 9260 and 9261 rabbit antisera. A.C., T.L. and Z.R. provided essential tools to study the regulation of XRINGO by SCFβTrCP and Siah-2. A.R.N. participated in the design of the experiments and the interpretation of the results and wrote the manuscript together with G.J.G.
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Gutierrez, G., Vögtlin, A., Castro, A. et al. Meiotic regulation of the CDK activator RINGO/Speedy by ubiquitin-proteasome-mediated processing and degradation. Nat Cell Biol 8, 1084–1094 (2006). https://doi.org/10.1038/ncb1472
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DOI: https://doi.org/10.1038/ncb1472
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