Efficient APC/C substrate degradation in cells undergoing mitotic exit depends on K11 ubiquitin linkages

Mol Biol Cell. 2015 Dec 1;26(24):4325-32. doi: 10.1091/mbc.E15-02-0102. Epub 2015 Oct 7.

Abstract

The ubiquitin proteasome system (UPS) directs programmed destruction of key cellular regulators via posttranslational modification of its targets with polyubiquitin chains. These commonly contain Lys-48 (K48)-directed ubiquitin linkages, but chains containing atypical Lys-11 (K11) linkages also target substrates to the proteasome--for example, to regulate cell cycle progression. The ubiquitin ligase called the anaphase-promoting complex/cyclosome (APC/C) controls mitotic exit. In higher eukaryotes, the APC/C works with the E2 enzyme UBE2S to assemble K11 linkages in cells released from mitotic arrest, and these are proposed to constitute an improved proteolytic signal during exit from mitosis. We tested this idea by correlating quantitative measures of in vivo K11-specific ubiquitination of individual substrates, including Aurora kinases, with their degradation kinetics tracked at the single-cell level. All anaphase substrates tested by this methodology are stabilized by depletion of K11 linkages via UBE2S knockdown, even if the same substrates are significantly modified with K48-linked polyubiquitin. Specific examination of substrates depending on the APC/C coactivator Cdh1 for their degradation revealed Cdh1-dependent enrichment of K11 chains on these substrates, whereas other ubiquitin linkages on the same substrates added during mitotic exit were Cdh1-independent. Therefore we show that K11 linkages provide the APC/C with a means to regulate the rate of substrate degradation in a coactivator-specified manner.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anaphase / physiology*
  • Anaphase-Promoting Complex-Cyclosome / metabolism*
  • Aurora Kinases / metabolism
  • Cell Cycle / physiology
  • Cell Line, Tumor
  • HeLa Cells
  • Humans
  • Lysine / metabolism
  • Proteasome Endopeptidase Complex / metabolism*
  • Protein Processing, Post-Translational
  • Proteolysis
  • Ubiquitin / metabolism*
  • Ubiquitin-Conjugating Enzymes / metabolism
  • Ubiquitination

Substances

  • Ubiquitin
  • Ubiquitin-Conjugating Enzymes
  • Anaphase-Promoting Complex-Cyclosome
  • Aurora Kinases
  • Proteasome Endopeptidase Complex
  • Lysine