Publication Date:
2011-02-11
Description:
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.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Schreiber, Anne -- Stengel, Florian -- Zhang, Ziguo -- Enchev, Radoslav I -- Kong, Eric H -- Morris, Edward P -- Robinson, Carol V -- da Fonseca, Paula C A -- Barford, David -- Cancer Research UK/United Kingdom -- England -- Nature. 2011 Feb 10;470(7333):227-32. doi: 10.1038/nature09756.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Section of Structural Biology, Institute of Cancer Research, Chester Beatty Laboratories, 237 Fulham Road, London, SW3 6JB, UK.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/21307936" target="_blank"〉PubMed〈/a〉
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/chemistry/metabolism/ultrastructure
;
Mass Spectrometry
;
Microscopy, Electron
;
Models, Molecular
;
Molecular Weight
;
Protein Binding
;
Protein Conformation
;
Protein Subunits/chemistry/isolation & purification/metabolism
;
Recombinant Proteins/chemistry/metabolism/ultrastructure
;
Saccharomyces cerevisiae/chemistry/genetics
;
Saccharomyces cerevisiae Proteins/chemistry/isolation &
;
purification/metabolism/ultrastructure
;
Scattering, Radiation
;
Schizosaccharomyces/chemistry
;
Structure-Activity Relationship
;
Substrate Specificity
;
Ubiquitin-Protein Ligase Complexes/*chemistry/*metabolism/ultrastructure
;
Ubiquitination
Print ISSN:
0028-0836
Electronic ISSN:
1476-4687
Topics:
Biology
,
Chemistry and Pharmacology
,
Medicine
,
Natural Sciences in General
,
Physics
