Dual inhibition of sister chromatid separation at metaphase

Titelangaben

Stemmann, Olaf ; Zou, Hui ; Gerber, Scott A. ; Gygi, Steven P. ; Kirschner, Marc W.:
Dual inhibition of sister chromatid separation at metaphase.
In: Cell. Bd. 107 (2001) Heft 6 . - S. 715-726.
ISSN 1097-4172
DOI: https://doi.org/10.1016/S0092-8674(01)00603-1

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Abstract

Separation of sister chromatids in anaphase is mediated by separase, an endopeptidase that cleaves the chromosomal cohesin SCC1. Separase is inhibited by securin, which is degraded at the metaphase-anaphase transition. Using Xenopus egg extracts, we demonstrate that high CDC2 activity inhibits anaphase but not securin degradation. We show that separase is kept inactive under these conditions by a mechanism independent of binding to securin. Mutation of a single phosphorylation site on separase relieves the inhibition and rescues chromatid separation in extracts with high CDC2 activity. Using quantitative mass spectrometry, we show that, in intact cells, there is complete phosphorylation of this site in metaphase and significant dephosphorylation in anaphase. We propose that separase activation at the metaphase-anaphase transition requires the removal of both securin and an inhibitory phosphate.