Ubiquitin chain elongation requires E3-dependent tracking of the emerging conjugate

Mol Cell. 2014 Oct 23;56(2):232-245. doi: 10.1016/j.molcel.2014.09.010. Epub 2014 Oct 9.

Abstract

Protein modification with ubiquitin chains is an essential signaling event catalyzed by E3 ubiquitin ligases. Most human E3s contain a signature RING domain that recruits a ubiquitin-charged E2 and a separate domain for substrate recognition. How RING-E3s can build polymeric ubiquitin chains while binding substrates and E2s at defined interfaces remains poorly understood. Here, we show that the RING-E3 APC/C catalyzes chain elongation by strongly increasing the affinity of its E2 for the distal acceptor ubiquitin in a growing conjugate. This function of the APC/C requires its coactivator as well as conserved residues of the E2 and ubiquitin. APC/C's ability to track the tip of an emerging conjugate is required for APC/C-substrate degradation and accurate cell division. Our results suggest that RING-E3s tether the distal ubiquitin of a growing chain in proximity to the active site of their E2s, allowing them to assemble polymeric conjugates without altering their binding to substrate or E2.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apc11 Subunit, Anaphase-Promoting Complex-Cyclosome / metabolism*
  • Apc2 Subunit, Anaphase-Promoting Complex-Cyclosome / metabolism*
  • Catalytic Domain
  • Cdc20 Proteins / genetics
  • Cdc20 Proteins / metabolism*
  • Cell Cycle Checkpoints / genetics
  • Cell Line, Tumor
  • Enzyme Activation
  • HeLa Cells
  • Humans
  • Peptide Biosynthesis, Nucleic Acid-Independent / genetics*
  • Protein Binding
  • Protein Structure, Tertiary
  • RNA Interference
  • RNA, Small Interfering
  • Ubiquitin / biosynthesis
  • Ubiquitin / genetics*
  • Ubiquitin-Conjugating Enzymes / metabolism*
  • Ubiquitination

Substances

  • Cdc20 Proteins
  • RNA, Small Interfering
  • Ubiquitin
  • Ube2S protein, human
  • Ubiquitin-Conjugating Enzymes
  • ANAPC11 protein, human
  • Apc11 Subunit, Anaphase-Promoting Complex-Cyclosome
  • Apc2 Subunit, Anaphase-Promoting Complex-Cyclosome