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Structures of APC/C(Cdh1) with substrates identify Cdh1 and Apc10 as the D-box co-receptor.

The ubiquitylation of cell-cycle regulatory proteins by the large multimeric anaphase-promoting complex (APC/C) controls sister chromatid segregation and the exit from mitosis. Selection of APC/C targets is achieved through recognition of destruction motifs, predominantly the destruction (D)-box and KEN (Lys-Glu-Asn)-box. Although this process is known to involve a co-activator protein (either Cdc20 or Cdh1) together with core APC/C subunits, the structural basis for substrate recognition and ubiquitylation is not understood. Here we investigate budding yeast APC/C using single-particle electron microscopy and determine a cryo-electron microscopy map of APC/C in complex with the Cdh1 co-activator protein (APC/C(Cdh1)) bound to a D-box peptide at ∼10 Å resolution. We find that a combined catalytic and substrate-recognition module is located within the central cavity of the APC/C assembled from Cdh1, Apc10--a core APC/C subunit previously implicated in substrate recognition--and the cullin domain of Apc2. Cdh1 and Apc10, identified from difference maps, create a co-receptor for the D-box following repositioning of Cdh1 towards Apc10. Using NMR spectroscopy we demonstrate specific D-box-Apc10 interactions, consistent with a role for Apc10 in directly contributing towards D-box recognition by the APC/C(Cdh1) complex. Our results rationalize the contribution of both co-activator and core APC/C subunits to D-box recognition and provide a structural framework for understanding mechanisms of substrate recognition and catalysis by the APC/C.

Pubmed ID: 21107322

Authors

  • da Fonseca PC
  • Kong EH
  • Zhang Z
  • Schreiber A
  • Williams MA
  • Morris EP
  • Barford D

Journal

Nature

Publication Data

February 10, 2011

Associated Grants

  • Agency: Cancer Research UK, Id: A7403
  • Agency: Cancer Research UK, Id: A8022
  • Agency: Cancer Research UK, Id:

Mesh Terms

  • Amino Acid Motifs
  • Anaphase-Promoting Complex-Cyclosome
  • Apc10 Subunit, Anaphase-Promoting Complex-Cyclosome
  • Apc2 Subunit, Anaphase-Promoting Complex-Cyclosome
  • Biocatalysis
  • Cdh1 Proteins
  • Cell Cycle Proteins
  • Cryoelectron Microscopy
  • Models, Molecular
  • Nuclear Magnetic Resonance, Biomolecular
  • Peptides
  • Protein Binding
  • Protein Conformation
  • Saccharomyces cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Substrate Specificity
  • Ubiquitin-Protein Ligase Complexes
  • Ubiquitination