Dual RING E3 Architectures Regulate Multiubiquitination and Ubiquitin Chain Elongation by APC/C

  • Cell. 2016 Jun 2;165(6):1440-1453. doi: 10.1016/j.cell.2016.05.037.
Nicholas G Brown  1 ,  Ryan VanderLinden  2 ,  Edmond R Watson  1 ,  Florian Weissmann  3 ,  Alban Ordureau  4 ,  Kuen-Phon Wu  1 ,  Wei Zhang  5 ,  Shanshan Yu  1 ,  Peter Y Mercredi  1 ,  Joseph S Harrison  6 ,  Iain F Davidson  3 ,  Renping Qiao  3 ,  Ying Lu  7 ,  Prakash Dube  8 ,  Michael R Brunner  1 ,  Christy R R Grace  1 ,  Darcie J Miller  1 ,  David Haselbach  8 ,  Marc A Jarvis  3 ,  Masaya Yamaguchi  1 ,  David Yanishevski  1 ,  Georg Petzold  3 ,  Sachdev S Sidhu  5 ,  Brian Kuhlman  6 ,  Marc W Kirschner  7 ,  J Wade Harper  4 ,  Jan-Michael Peters  9 ,  Holger Stark  10 ,  Brenda A Schulman  11
Affiliations
  • 1. Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
  • 2. Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA; Howard Hughes Medical Institute, Memphis, TN 38105, USA.
  • 3. Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC), 1030 Vienna, Austria.
  • 4. Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA.
  • 5. Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 160 College Street, Toronto, Ontario M5S3E1, Canada.
  • 6. Department of Biochemistry and Biophysics and Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
  • 7. Department of Systems Biology, Harvard Medical School, Boston, MA, 02115, USA.
  • 8. Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany.
  • 9. Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC), 1030 Vienna, Austria. Electronic address: [email protected].
  • 10. Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany. Electronic address: [email protected].
  • 11. Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA; Howard Hughes Medical Institute, Memphis, TN 38105, USA. Electronic address: [email protected].
PMID: 27259151 DOI: 10.1016/j.cell.2016.05.037
Abstract

Protein ubiquitination involves E1, E2, and E3 trienzyme cascades. E2 and RING E3 Enzymes often collaborate to first prime a substrate with a single ubiquitin (UB) and then achieve different forms of polyubiquitination: multiubiquitination of several sites and elongation of linkage-specific UB chains. Here, cryo-EM and biochemistry show that the human E3 anaphase-promoting complex/cyclosome (APC/C) and its two partner E2s, UBE2C (aka UBCH10) and UBE2S, adopt specialized catalytic architectures for these two distinct forms of polyubiquitination. The APC/C RING constrains UBE2C proximal to a substrate and simultaneously binds a substrate-linked UB to drive processive multiubiquitination. Alternatively, during UB chain elongation, the RING does not bind UBE2S but rather lures an evolving substrate-linked UB to UBE2S positioned through a cullin interaction to generate a Lys11-linked chain. Our findings define mechanisms of APC/C regulation, and establish principles by which specialized E3-E2-substrate-UB architectures control different forms of polyubiquitination.