Role of ubiquitylation of components of mitotic checkpoint complex in their dissociation from anaphase-promoting complex/cyclosome

Proc Natl Acad Sci U S A. 2018 Feb 20;115(8):1777-1782. doi: 10.1073/pnas.1720312115. Epub 2018 Feb 5.


The mitotic checkpoint system ensures the fidelity of chromosome segregation in mitosis by preventing premature initiation of anaphase until correct bipolar attachment of chromosomes to the mitotic spindle is reached. It promotes the assembly of a mitotic checkpoint complex (MCC), composed of BubR1, Bub3, Cdc20, and Mad2, which inhibits the activity of the anaphase-promoting complex/cyclosome (APC/C) ubiquitin ligase. When the checkpoint is satisfied, anaphase is initiated by the disassembly of MCC. Previous studies indicated that the dissociation of APC/C-bound MCC requires ubiquitylation and suggested that the target of ubiquitylation is the Cdc20 component of MCC. However, it remained unknown how ubiquitylation causes the release of MCC from APC/C and its disassembly and whether ubiquitylation of additional proteins is involved in this process. We find that ubiquitylation causes the dissociation of BubR1 from Cdc20 in MCC and suggest that this may lead to the release of MCC components from APC/C. BubR1 in MCC is ubiquitylated by APC/C, although to a lesser degree than Cdc20. The extent of BubR1 ubiquitylation was markedly increased in recombinant MCC that contained a lysine-less mutant of Cdc20. Mutation of lysine residues to arginines in the N-terminal region of BubR1 partially inhibited its ubiquitylation and slowed down the release of MCC from APC/C, provided that Cdc20 ubiquitylation was also blocked. It is suggested that ubiquitylation of both Cdc20 and BubR1 may be involved in their dissociation from each other and in the release of MCC components from APC/C.

Keywords: cell cycle; protein degradation; ubiquitin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anaphase*
  • Anaphase-Promoting Complex-Cyclosome / genetics
  • Anaphase-Promoting Complex-Cyclosome / metabolism*
  • Cdc20 Proteins / genetics
  • Cdc20 Proteins / metabolism
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism
  • HeLa Cells
  • Humans
  • Mad2 Proteins / genetics
  • Mad2 Proteins / metabolism
  • Mitosis
  • Poly-ADP-Ribose Binding Proteins / genetics
  • Poly-ADP-Ribose Binding Proteins / metabolism
  • Protein Binding
  • Spindle Apparatus / genetics
  • Spindle Apparatus / metabolism
  • Ubiquitination


  • BUB3 protein, human
  • Cdc20 Proteins
  • Cell Cycle Proteins
  • Mad2 Proteins
  • Poly-ADP-Ribose Binding Proteins
  • Anaphase-Promoting Complex-Cyclosome