Protein Phosphatase 2A and Separase Form a Complex Regulated by Separase Autocleavage

J Biol Chem. 2007 Aug 24;282(34):24623-32. doi: 10.1074/jbc.M702545200. Epub 2007 Jun 29.

Abstract

The onset of anaphase is triggered by the activation of a site-specific protease called separase. Separase cleaves the chromosomal cohesins holding the duplicated sister chromatids together, allowing sisters to simultaneously separate and segregate to opposite ends of the cell before division. Activated separase cleaves not only cohesin, but also itself; however, the biological significance of separase self-cleavage has remained elusive. Before anaphase, separase is inhibited by at least two mechanisms. The first involves the binding of securin, whereas the second requires the phosphorylation-dependent binding of cyclin-dependent kinase 1 (Cdk1)/cyclin B1. Because securin and Cdk1/cyclin B1 interact with separase in a mutually exclusive manner, the degradation of both these inhibitors plays an important role in activating separase at anaphase. Here we identify a new separase interacting partner, a specific subtype of the heterotrimeric protein phosphatase 2A (PP2A). PP2A associates with separase through the B' (B56) regulatory subunit and does so independently of securin and cyclin B1 binding. The association of PP2A with separase requires a 55-amino acid domain closely juxtaposed to separase autocleavage sites. Strikingly, mutation of these cleavage sites increases PP2A binding, suggesting that separase cleavage disrupts the interaction of PP2A with separase. Furthermore, expression of a non-cleavable separase, but not a non-cleavable mutant that cannot bind PP2A, causes a premature loss of centromeric cohesion. Together these observations provide a new mechanistic insight into a physiological function for separase self-cleavage.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Catalysis
  • Cell Cycle Proteins / metabolism*
  • Cell Line
  • Cloning, Molecular
  • Endopeptidases / metabolism*
  • Gene Expression Regulation, Enzymologic*
  • HeLa Cells
  • Humans
  • Mitosis
  • Models, Biological
  • Phenotype
  • Phosphoprotein Phosphatases / metabolism*
  • Phosphorylation
  • Protein Binding
  • Protein Phosphatase 2
  • Protein Structure, Tertiary
  • Separase
  • Transfection

Substances

  • Cell Cycle Proteins
  • Phosphoprotein Phosphatases
  • Protein Phosphatase 2
  • Endopeptidases
  • ESPL1 protein, human
  • Separase