title: APC15 mediates CDC20 autoubiquitylation by APC/C(MCC) and disassembly of the mitotic checkpoint complex.
abstract: The anaphase-promoting complex/cyclosome (APC/C) bound to CDC20 (APC/C(CDC20)) initiates anaphase by ubiquitylating B-type cyclins and securin. During chromosome bi-orientation, CDC20 assembles with MAD2, BUBR1 and BUB3 into a mitotic checkpoint complex (MCC) that inhibits substrate recruitment to the APC/C. APC/C activation depends on MCC disassembly, which was proposed to require CDC20 autoubiquitylation. Here we characterize APC15, a human APC/C subunit related to yeast Mnd2. APC15 is located near APC/C's MCC binding site; it is required for APC/C-bound MCC (APC/C(MCC))-dependent CDC20 autoubiquitylation and degradation and for timely anaphase initiation but is dispensable for substrate ubiquitylation by APC/C(CDC20) and APC/C(CDH1). Our results support the model wherein MCC is continuously assembled and disassembled to enable rapid activation of APC/C(CDC20) and CDC20 autoubiquitylation promotes MCC disassembly. We propose that APC15 and Mnd2 negatively regulate APC/C coactivators and report generation of recombinant human APC/C.
authors.affiliation: Research Institute of Molecular Pathology, Vienna, Austria.
authors.forename: Kristina
authors.initials: K
authors.lastname: Uzunova
authors.forename: Billy T
authors.initials: BT
authors.lastname: Dye
authors.forename: Hannelore
authors.initials: H
authors.lastname: Schutz
authors.forename: Rene
authors.initials: R
authors.lastname: Ladurner
authors.forename: Georg
authors.initials: G
authors.lastname: Petzold
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Toyoda
authors.forename: Marc A
authors.initials: MA
authors.lastname: Jarvis
authors.forename: Nicholas G
authors.initials: NG
authors.lastname: Brown
authors.forename: Ina
authors.initials: I
authors.lastname: Poser
authors.forename: Maria
authors.initials: M
authors.lastname: Novatchkova
authors.forename: Karl
authors.initials: K
authors.lastname: Mechtler
authors.forename: Anthony A
authors.initials: AA
authors.lastname: Hyman
authors.forename: Holger
authors.initials: H
authors.lastname: Stark
authors.forename: Brenda A
authors.initials: BA
authors.lastname: Schulman
authors.forename: Jan-Michael
authors.initials: JM
authors.lastname: Peters
mesh_terms: D064173:Anaphase-Promoting Complex-Cyclosome; D002135:Calcium-Binding Proteins / Q000378:metabolism; D064199:Cdc20 Proteins; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D059566:M Phase Cell Cycle Checkpoints / Q000235:genetics / Q000502:physiology*; D064246:Mad2 Proteins; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D000075223:Poly-ADP-Ribose Binding Proteins; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D012097:Repressor Proteins / Q000378:metabolism; D059008:Time-Lapse Imaging; D043743:Ubiquitin-Protein Ligase Complexes / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases; D054875:Ubiquitination
chemical_list: C577350:ANAPC15 protein, human; C489444:BUB3 protein, human; D002135:Calcium-Binding Proteins; D064199:Cdc20 Proteins; D018797:Cell Cycle Proteins; C577697:MAD2L1 protein, human; D064246:Mad2 Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; D034741:RNA, Small Interfering; D012097:Repressor Proteins; C577488:CDC20 protein, human; D043743:Ubiquitin-Protein Ligase Complexes; D064173:Anaphase-Promoting Complex-Cyclosome; D044767:Ubiquitin-Protein Ligases; C584277:BUB1 protein, human; C090464:Bub1 spindle checkpoint protein; D017346:Protein Serine-Threonine Kinases