Abstract
The anaphase-promoting complex/cyclosome (APC/C) is a 22S ubiquitin ligase complex that initiates chromosome segregation and mitotic exit. We have used biochemical and electron microscopic analyses of Saccharomyces cerevisiae and human APC/C to address how the APC/C subunit Doc1 contributes to recruitment and processive ubiquitylation of APC/C substrates, and to understand how APC/C monomers interact to form a 36S dimeric form. We show that Doc1 interacts with Cdc27, Cdc16 and Apc1 and is located in the vicinity of the cullin-RING module Apc2-Apc11 in the inner cavity of the APC/C. Substrate proteins also bind in the inner cavity, in close proximity to Doc1 and the coactivator Cdh1, and induce conformational changes in Apc2-Apc11. Our results suggest that substrates are recruited to the APC/C by binding to a bipartite substrate receptor composed of a coactivator protein and Doc1.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Anaphase-Promoting Complex-Cyclosome
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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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Apc11 Subunit, Anaphase-Promoting Complex-Cyclosome
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Apc2 Subunit, Anaphase-Promoting Complex-Cyclosome
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Apc3 Subunit, Anaphase-Promoting Complex-Cyclosome
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Apc6 Subunit, Anaphase-Promoting Complex-Cyclosome
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Cadherins / metabolism*
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Cdh1 Proteins
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Cell Cycle Proteins / metabolism*
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Cytokines / metabolism*
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Humans
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Intracellular Signaling Peptides and Proteins
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Saccharomyces cerevisiae / enzymology*
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Saccharomyces cerevisiae Proteins / metabolism*
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Ubiquitin-Protein Ligase Complexes / chemistry
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Ubiquitin-Protein Ligase Complexes / metabolism*
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Ubiquitin-Protein Ligase Complexes / ultrastructure
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Ubiquitin-Protein Ligases / metabolism
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Ubiquitination
Substances
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ANAPC1 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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Apc6 Subunit, Anaphase-Promoting Complex-Cyclosome
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CDC16 protein, S cerevisiae
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CDC27 protein, S cerevisiae
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CDH1 protein, S cerevisiae
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CDH1 protein, human
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Cadherins
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Cdh1 Proteins
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Cell Cycle Proteins
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Cytokines
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DOC1 protein, S cerevisiae
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FILIP1L protein, human
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Intracellular Signaling Peptides and Proteins
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Saccharomyces cerevisiae Proteins
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Ubiquitin-Protein Ligase Complexes
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ANAPC11 protein, human
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APC2 protein, S cerevisiae
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Anaphase-Promoting Complex-Cyclosome
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Apc11 Subunit, Anaphase-Promoting Complex-Cyclosome
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Apc2 Subunit, Anaphase-Promoting Complex-Cyclosome
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Ubiquitin-Protein Ligases