Cdc20 is required for the post-anaphase, KEN-dependent degradation of centromere protein F

Mark D J Gurden, Andrew J. Holland, Wouter Van Zon, Anthony Tighe, Mailys A. Vergnolle, Douglas A. Andres, H. Peter Spielmann, Marcos Malumbres, Rob M F Wolthuis, Don W. Cleveland, Stephen S. Taylor

    Research output: Contribution to journalArticlepeer-review

    Abstract

    Progression through mitosis and cytokinesis requires the sequential proteolysis of several cell-cycle regulators. This proteolysis is mediated by the ubiquitin-proteasome system, with the E3 ligase being the anaphase-promoting complex, also known as the cyclosome (APC/C). The APC/C is regulated by two activators, namely Cdc20 and Cdh1. The current view is that prior to anaphase, the APC/C is activated by Cdc20, but that following anaphase, APC/C switches to Cdh1-dependent activation. However, here we present an analysis of the kinetochore protein Cenp-F that is inconsistent with this notion. Although it has long been appreciated that Cenp-F is degraded sometime during or after mitosis, exactly when and how has not been clear. Here we show that degradation of Cenp-F initiates about six minutes after anaphase, and that this is dependent on a C-terminal KEN-box. Although these two observations are consistent with Cenp-F being a substrate of Cdh1-activated APC/C, Cenp-F is degraded normally in Cdh1-null cells. By contrast, RNAi-mediated repression of APC/C subunits or Cdc20 does inhibit Cenp-F degradation. These findings therefore suggest that the APC/C does not simply 'switch' upon anaphase onset; rather, our observations indicate that Cdc20 also contributes to post-anaphase activation of the APC/C. We also show that the post-anaphase, KEN-box-dependent degradation of Cenp-F requires it to be farnesylated, a post-translational modification usually linked to membrane association. Because so many of the behaviours of Cenp-F are farnesylation-dependent, we suggest that this modification plays a more global role in Cenp-F function.
    Original languageEnglish
    Pages (from-to)321-330
    Number of pages9
    JournalJournal of Cell Science
    Volume123
    Issue number3
    DOIs
    Publication statusPublished - 1 Feb 2010

    Keywords

    • APC/C
    • Cdc20
    • Cenp-E
    • Cenp-F
    • Farnesylation
    • Kinetochore

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