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

J Cell Sci. 2010 Feb 1;123(Pt 3):321-30. doi: 10.1242/jcs.062075. Epub 2010 Jan 5.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anaphase-Promoting Complex-Cyclosome
  • Animals
  • Antigens, CD
  • Cadherins / genetics
  • Cadherins / metabolism
  • Cdc20 Proteins
  • Cdh1 Proteins
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism*
  • Cell Line, Tumor
  • Cells, Cultured
  • Chromosomal Proteins, Non-Histone / metabolism*
  • Humans
  • Kinetochores / metabolism
  • Mice
  • Microfilament Proteins / metabolism*
  • Microscopy
  • Prenylation
  • RNA Interference
  • Ubiquitin-Protein Ligase Complexes / metabolism

Substances

  • Antigens, CD
  • CDH1 protein, human
  • Cadherins
  • Cdc20 Proteins
  • Cdc20 protein, mouse
  • Cdh1 Proteins
  • Cell Cycle Proteins
  • Chromosomal Proteins, Non-Histone
  • Fzr1 protein, mouse
  • Microfilament Proteins
  • centromere protein E
  • centromere protein F
  • CDC20 protein, human
  • Ubiquitin-Protein Ligase Complexes
  • Anaphase-Promoting Complex-Cyclosome