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Fission yeast Apc15 stabilises MCC-Cdc20-APC/C complexes ensuring efficient Cdc20 ubiquitination and checkpoint arrest

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http://linkinghub.elsevier.com/retrieve/pii/S0960982217302828
Original languageEnglish
JournalCurrent Biology
Early online date30 Mar 2017
DOIs
Publication statusPublished - 24 Apr 2017

Abstract

During mitosis cells must segregate the replicated copies of their genome to their daughter cells with extremely high fidelity. Segregation errors lead to an abnormal chromosome number (aneuploidy), which typically results in disease or cell death [1]. Chromosome segregation and anaphase onset are initiated through the action of the multi-subunit E3 ubiquitin ligase known as the anaphase-promoting complex or cyclosome (APC/C [2]). The APC/C is inhibited by the spindle checkpoint in the presence of kinetochore attachment defects [3, 4]. Here we demonstrate that two non-essential APC/C sub-units (Apc14 and Apc15) regulate association of spindle checkpoint proteins, in the form of the mitotic checkpoint complex (MCC), with the APC/C. apc14 mutants display increased MCC association with APC/C and are unable to silence the checkpoint efficiently. Conversely, apc15 mutants display reduced association between MCC and APC/C, are defective in poly-ubiquitination of Cdc20 and are checkpoint defective. In vitro reconstitution studies have shown that human MCC-APC/C can contain two molecules of Cdc20 [5-7]. Using a yeast strain expressing two Cdc20 genes with different epitope-tags, we show by co-immunoprecipitation that this is true in vivo. MCC binding to the second molecule of Cdc20 is mediated via the C-terminal KEN box in Mad3. Somewhat surprisingly, complexes containing both molecules of Cdc20 accumulate in apc15 cells and the implications of this observation are discussed.

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