title: The Spindle Assembly Checkpoint Is Not Essential for Viability of Human Cells with Genetically Lowered APC/C Activity.
abstract: The anaphase-promoting complex/cyclosome (APC/C) and the spindle assembly checkpoint (SAC), which inhibits the APC/C, are essential determinants of mitotic timing and faithful division of genetic material. Activation of the APC/C is known to depend on two APC/C-interacting E2 ubiquitin-conjugating enzymes-UBE2C and UBE2S. We show that APC/C activity in human cells is tuned by the combinatorial use of three E2s, namely UBE2C, UBE2S, and UBE2D. Genetic deletion of UBE2C and UBE2S, individually or in combination, leads to discriminative reduction in APC/C function and sensitizes cells to UBE2D depletion. Reduction of APC/C activity results in loss of switch-like metaphase-to-anaphase transition and, strikingly, renders cells insensitive to chemical inhibition of MPS1 and genetic ablation of MAD2, both of which are essential for the SAC. These results provide insights into the regulation of APC/C activity and demonstrate that the essentiality of the SAC is imposed by the strength of the APC/C.
authors.affiliation: Proteomics Program, the Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen, Denmark.
authors.forename: Thomas
authors.initials: T
authors.lastname: Wild
authors.affiliation: Protein Signaling Program, the Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen, Denmark.
authors.forename: Marie Sofie Yoo
authors.initials: MS
authors.lastname: Larsen
authors.affiliation: Proteomics Program, the Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen, Denmark.
authors.forename: Takeo
authors.initials: T
authors.lastname: Narita
authors.affiliation: Protein Signaling Program, the Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen, Denmark.
authors.forename: Julie
authors.initials: J
authors.lastname: Schou
authors.affiliation: Protein Signaling Program, the Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen, Denmark. Electronic address: jakob.nilsson@cpr.ku.dk.
authors.forename: Jakob
authors.initials: J
authors.lastname: Nilsson
authors.affiliation: Proteomics Program, the Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Blegdamsvej 3B, 2200 Copenhagen, Denmark. Electronic address: chuna.choudhary@cpr.ku.dk.
authors.forename: Chunaram
authors.initials: C
authors.lastname: Choudhary
mesh_terms: D000705:Anaphase / Q000187:drug effects; D064173:Anaphase-Promoting Complex-Cyclosome / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D064113:CRISPR-Cas Systems; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D017353:Gene Deletion; D015870:Gene Expression; D045325:HCT116 Cells; D006801:Humans; D059566:M Phase Cell Cycle Checkpoints; D064246:Mad2 Proteins / Q000172:deficiency / Q000235:genetics*; D008677:Metaphase / Q000187:drug effects; D009025:Morpholines / Q000494:pharmacology; D015739:Nocodazole / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D011687:Purines / Q000494:pharmacology; D015398:Signal Transduction; D008941:Spindle Apparatus / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D044763:Ubiquitin-Conjugating Enzymes / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism
chemical_list: D018797:Cell Cycle Proteins; C577697:MAD2L1 protein, human; D064246:Mad2 Proteins; D009025:Morpholines; D011687:Purines; C105156:UBE2C protein, human; C097308:UBE2D1 protein, human; C544962:Ube2S protein, human; D044763:Ubiquitin-Conjugating Enzymes; D064173:Anaphase-Promoting Complex-Cyclosome; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; C075840:TTK protein, human; D015739:Nocodazole; C484369:2-(4-morpholinoanilino)-6-cyclohexylaminopurine