Characterization of the DOC1/APC10 subunit of the yeast and the human anaphase-promoting complex

J Biol Chem. 1999 May 14;274(20):14500-7. doi: 10.1074/jbc.274.20.14500.

Abstract

The anaphase-promoting complex/cyclosome (APC) is a ubiquitin-protein ligase whose activity is essential for progression through mitosis. The vertebrate APC is thought to be composed of 8 subunits, whereas in budding yeast several additional APC-associated proteins have been identified, including a 33-kDa protein called Doc1 or Apc10. Here, we show that Doc1/Apc10 is a subunit of the yeast APC throughout the cell cycle. Mutation of Doc1/Apc10 inactivates the APC without destabilizing the complex. An ortholog of Doc1/Apc10, which we call APC10, is associated with the APC in different vertebrates, including humans and frogs. Biochemical fractionation experiments and mass spectrometric analysis of a component of the purified human APC show that APC10 is a genuine APC subunit whose cellular levels or association with the APC are not cell cycle-regulated. We have further identified an APC10 homology region, which we propose to call the DOC domain, in several protein sequences that also contain either cullin or HECT domains. Cullins are present in several ubiquitination complexes including the APC, whereas HECT domains represent the catalytic core of a different type of ubiquitin-protein ligase. DOC domains may therefore be important for reactions catalyzed by several types of ubiquitin-protein ligases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Anaphase*
  • Anaphase-Promoting Complex-Cyclosome
  • Apc10 Subunit, Anaphase-Promoting Complex-Cyclosome
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism*
  • Codon, Terminator
  • Fungal Proteins / metabolism*
  • Humans
  • Ligases / genetics
  • Ligases / metabolism*
  • Macromolecular Substances
  • Mass Spectrometry
  • Molecular Sequence Data
  • Saccharomyces cerevisiae Proteins*
  • Sequence Alignment
  • Ubiquitin-Protein Ligase Complexes*
  • Ubiquitin-Protein Ligases

Substances

  • Apc10 Subunit, Anaphase-Promoting Complex-Cyclosome
  • Cell Cycle Proteins
  • Codon, Terminator
  • DOC1 protein, S cerevisiae
  • Fungal Proteins
  • Macromolecular Substances
  • Saccharomyces cerevisiae Proteins
  • Ubiquitin-Protein Ligase Complexes
  • Anaphase-Promoting Complex-Cyclosome
  • Ubiquitin-Protein Ligases
  • Ligases

Associated data

  • GENBANK/AF132794