Stable MCC binding to the APC/C is required for a functional spindle assembly checkpoint

EMBO Rep. 2014 Mar;15(3):264-72. doi: 10.1002/embr.201337496. Epub 2014 Jan 25.

Abstract

The spindle assembly checkpoint (SAC) delays progression into anaphase until all chromosomes have aligned on the metaphase plate by inhibiting Cdc20, the mitotic co-activator of the APC/C. Mad2 and BubR1 bind and inhibit Cdc20, thereby forming the mitotic checkpoint complex (MCC), which can bind stably to the APC/C. Whether MCC formation per se is sufficient for a functional SAC or MCC association with the APC/C is required remains unclear. Here, we analyze the role of two conserved motifs in Cdc20, IR and C-Box, in binding of the MCC to the APC/C. Mutants in both motifs assemble the MCC normally, but IR motif integrity is particularly important for stable binding to the APC/C. Cells expressing Cdc20 with a mutated IR motif have a compromised SAC, as uninhibited Cdc20 can compete with the MCC for APC/C binding and activate it. We thus show that stable MCC association with the APC/C is critical for a functional SAC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Anaphase-Promoting Complex-Cyclosome / metabolism*
  • Binding Sites
  • Cdc20 Proteins / chemistry
  • Cdc20 Proteins / genetics
  • Cdc20 Proteins / metabolism*
  • Conserved Sequence
  • HeLa Cells
  • Humans
  • M Phase Cell Cycle Checkpoints*
  • Mad2 Proteins / metabolism*
  • Protein Binding
  • Protein Serine-Threonine Kinases / metabolism*
  • Spindle Apparatus / metabolism*

Substances

  • Cdc20 Proteins
  • MAD2L1 protein, human
  • Mad2 Proteins
  • CDC20 protein, human
  • Anaphase-Promoting Complex-Cyclosome
  • BUB1 protein, human
  • Protein Serine-Threonine Kinases