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

Mark D J Gurden, Andrew 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 journalArticle

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 (US)
Pages (from-to)321-330
Number of pages10
JournalJournal of Cell Science
Volume123
Issue number3
DOIs
StatePublished - Feb 1 2010
Externally publishedYes

    Fingerprint

Keywords

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

ASJC Scopus subject areas

  • Cell Biology

Cite this

Gurden, M. D. J., Holland, A., Van Zon, W., Tighe, A., Vergnolle, M. A., Andres, D. A., Spielmann, H. P., Malumbres, M., Wolthuis, R. M. F., Cleveland, D. W., & Taylor, S. S. (2010). Cdc20 is required for the post-anaphase, KEN-dependent degradation of centromere protein F. Journal of Cell Science, 123(3), 321-330. https://doi.org/10.1242/jcs.062075