Abstract
The anaphase-promoting complex (APC/C) is a multimeric RING E3 ubiquitin ligase that controls chromosome segregation and mitotic exit. Its regulation by coactivator subunits, phosphorylation, the mitotic checkpoint complex and interphase early mitotic inhibitor 1 (Emi1) ensures the correct order and timing of distinct cell-cycle transitions. Here we use cryo-electron microscopy to determine atomic structures of APC/C-coactivator complexes with either Emi1 or a UbcH10-ubiquitin conjugate. These structures define the architecture of all APC/C subunits, the position of the catalytic module and explain how Emi1 mediates inhibition of the two E2s UbcH10 and Ube2S. Definition of Cdh1 interactions with the APC/C indicates how they are antagonized by Cdh1 phosphorylation. The structure of the APC/C with UbcH10-ubiquitin reveals insights into the initiating ubiquitination reaction. Our results provide a quantitative framework for the design of future experiments to investigate APC/C functions in vivo.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Anaphase-Promoting Complex-Cyclosome / chemistry
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Anaphase-Promoting Complex-Cyclosome / metabolism*
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Anaphase-Promoting Complex-Cyclosome / ultrastructure*
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Antigens, CD
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Apc1 Subunit, Anaphase-Promoting Complex-Cyclosome / chemistry
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Apc1 Subunit, Anaphase-Promoting Complex-Cyclosome / metabolism
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Apc1 Subunit, Anaphase-Promoting Complex-Cyclosome / ultrastructure
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Apc10 Subunit, Anaphase-Promoting Complex-Cyclosome / chemistry
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Apc10 Subunit, Anaphase-Promoting Complex-Cyclosome / metabolism
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Apc10 Subunit, Anaphase-Promoting Complex-Cyclosome / ultrastructure
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Apc11 Subunit, Anaphase-Promoting Complex-Cyclosome / chemistry
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Apc11 Subunit, Anaphase-Promoting Complex-Cyclosome / metabolism
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Apc3 Subunit, Anaphase-Promoting Complex-Cyclosome / chemistry
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Apc3 Subunit, Anaphase-Promoting Complex-Cyclosome / metabolism
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Apc8 Subunit, Anaphase-Promoting Complex-Cyclosome / chemistry
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Apc8 Subunit, Anaphase-Promoting Complex-Cyclosome / metabolism
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Apc8 Subunit, Anaphase-Promoting Complex-Cyclosome / ultrastructure
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Cadherins / chemistry
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Cadherins / metabolism
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Cadherins / ultrastructure
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Catalytic Domain
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Cell Cycle Proteins / chemistry
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Cell Cycle Proteins / metabolism
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Cell Cycle Proteins / ultrastructure
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Cryoelectron Microscopy
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Cytoskeletal Proteins / chemistry
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Cytoskeletal Proteins / metabolism
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F-Box Proteins / chemistry
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F-Box Proteins / metabolism
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F-Box Proteins / ultrastructure
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Humans
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Lysine / metabolism
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Models, Molecular
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Phosphorylation
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Protein Binding
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Protein Subunits / chemistry
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Protein Subunits / metabolism
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Structure-Activity Relationship
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Substrate Specificity
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Ubiquitin / chemistry
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Ubiquitin / metabolism
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Ubiquitin / ultrastructure
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Ubiquitin-Conjugating Enzymes / chemistry
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Ubiquitin-Conjugating Enzymes / metabolism
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Ubiquitin-Conjugating Enzymes / ultrastructure
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Ubiquitination*
Substances
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ANAPC1 protein, human
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ANAPC10 protein, human
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APC2 protein, human
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Antigens, CD
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Apc1 Subunit, Anaphase-Promoting Complex-Cyclosome
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Apc10 Subunit, Anaphase-Promoting Complex-Cyclosome
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Apc3 Subunit, Anaphase-Promoting Complex-Cyclosome
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Apc8 Subunit, Anaphase-Promoting Complex-Cyclosome
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CDC23 protein, human
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CDC27 protein, human
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CDH1 protein, human
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Cadherins
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Cell Cycle Proteins
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Cytoskeletal Proteins
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F-Box Proteins
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FBXO5 protein, human
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Protein Subunits
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Ubiquitin
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UBE2C protein, human
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Ube2S protein, human
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Ubiquitin-Conjugating Enzymes
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ANAPC11 protein, human
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Anaphase-Promoting Complex-Cyclosome
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Apc11 Subunit, Anaphase-Promoting Complex-Cyclosome
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Lysine