Structural organization of the anaphase-promoting complex bound to the mitotic activator Slp1.
The anaphase-promoting complex/cyclosome (APC/C) is a conserved multisubunit E3 ubiquitin (Ub) ligase required to signal the degradation of key cell-cycle regulators. Using single particle cryo-electron microscopy (cryo-EM), we have determined a three-dimensional (3D) structure of the core APC/C from Schizosaccharomyces pombe bound to the APC/C activator Slp1/Cdc20. At the 27 A resolution of our density map, the APC/C is a triangular-shaped structure, approximately 19x17x15 nm in size, with a deep internal cavity and a prominent horn-like protrusion emanating from a lip of the cavity. Using antibody labeling and mutant analysis, we have localized 12 of 13 core APC/C components, as well as the position of the activator Slp1, enabling us to propose a structural model of APC/C organization. Comparison of the APC/C with another multiprotein E3 ligase, the SCF complex, uncovers remarkable structural similarities.
Ohi
Melanie D
MD
Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA.
Feoktistova
Anna
A
Ren
Liping
L
Yip
Calvin
C
Cheng
Yifan
Y
Chen
Jun-Song
JS
Yoon
Hyun-Joo
HJ
Wall
Joseph S
JS
Huang
Zhong
Z
Penczek
Pawel A
PA
Gould
Kathleen L
KL
Walz
Thomas
T
D064173:Anaphase-Promoting Complex-Cyclosome; D064199:Cdc20 Proteins; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000378:metabolism*; D002846:Chromatography, Affinity; D020285:Cryoelectron Microscopy; D015151:Immunoblotting; D008938:Mitosis / Q000502:physiology; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011487:Protein Conformation; D012568:Schizosaccharomyces / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry / Q000378:metabolism*; D043743:Ubiquitin-Protein Ligase Complexes / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D054875:Ubiquitination
D064199:Cdc20 Proteins; D018797:Cell Cycle Proteins; D029702:Schizosaccharomyces pombe Proteins; C577504:slp1 protein, S pombe; D043743:Ubiquitin-Protein Ligase Complexes; D064173:Anaphase-Promoting Complex-Cyclosome; D044767:Ubiquitin-Protein Ligases