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The anaphase-promoting complex or cyclosome (APC/C) is an unusually large E3 ubiquitin ligase responsible for regulating defined cell cycle transitions. Information on how its 13 constituent proteins are assembled, and how they interact with co-activators, substrates and regulatory proteins is limited. Here, we describe a recombinant expression system that allows the reconstitution of holo APC/C and its sub-complexes that, when combined with electron microscopy, mass spectrometry and docking of crystallographic and homology-derived coordinates, provides a precise definition of the organization and structure of all essential APC/C subunits, resulting in a pseudo-atomic model for 70% of the APC/C. A lattice-like appearance of the APC/C is generated by multiple repeat motifs of most APC/C subunits. Three conserved tetratricopeptide repeat (TPR) subunits (Cdc16, Cdc23 and Cdc27) share related superhelical homo-dimeric architectures that assemble to generate a quasi-symmetrical structure. Our structure explains how this TPR sub-complex, together with additional scaffolding subunits (Apc1, Apc4 and Apc5), coordinate the juxtaposition of the catalytic and substrate recognition module (Apc2, Apc11 and Apc10 (also known as Doc1)), and TPR-phosphorylation sites, relative to co-activator, regulatory proteins and substrates.

Original publication

DOI

10.1038/nature09756

Type

Journal article

Journal

Nature

Publication Date

10/02/2011

Volume

470

Pages

227 - 232

Keywords

Amino Acid Motifs, Anaphase-Promoting Complex-Cyclosome, Animals, Apc2 Subunit, Anaphase-Promoting Complex-Cyclosome, Apc5 Subunit, Anaphase-Promoting Complex-Cyclosome, Apc8 Subunit, Anaphase-Promoting Complex-Cyclosome, Biocatalysis, Cell Line, Holoenzymes, Mass Spectrometry, Microscopy, Electron, Models, Molecular, Molecular Weight, Protein Binding, Protein Conformation, Protein Subunits, Recombinant Proteins, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Scattering, Radiation, Schizosaccharomyces, Structure-Activity Relationship, Substrate Specificity, Ubiquitin-Protein Ligase Complexes, Ubiquitination