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Title: Structure of an APC3–APC16 Complex: Insights into Assembly of the Anaphase-Promoting Complex/Cyclosome

Abstract

The anaphase-promoting complex/cyclosome (APC/C) is a massive E3 ligase that controls mitosis by catalyzing ubiquitination of key cell cycle regulatory proteins. The APC/C assembly contains two subcomplexes: the “Platform” centers around a cullin-RING-like E3 ligase catalytic core; the “Arc Lamp” is a hub that mediates transient association with regulators and ubiquitination substrates. The Arc Lamp contains the small subunits APC16, CDC26, and APC13, and tetratricopeptide repeat (TPR) proteins (APC7, APC3, APC6, and APC8) that homodimerize and stack with quasi-2-fold symmetry. Within the APC/C complex, APC3 serves as center for regulation. APC3's TPR motifs recruit substrate-binding coactivators, CDC20 and CDH1, via their C-terminal conserved Ile-Arg (IR) tail sequences. Human APC3 also binds APC16 and APC7 and contains a > 200-residue loop that is heavily phosphorylated during mitosis, although the basis for APC3 interactions and whether loop phosphorylation is required for ubiquitination are unclear. Here, we map the basis for human APC3 assembly with APC16 and APC7, report crystal structures of APC3Δloop alone and in complex with the C-terminal domain of APC16, and test roles of APC3's loop and IR tail binding surfaces in APC/C-catalyzed ubiquitination. The structures show how one APC16 binds asymmetrically to the symmetric APC3 dimer and, together withmore » biochemistry and prior data, explain how APC16 recruits APC7 to APC3, show how APC3's C-terminal domain is rearranged in the full APC/C assembly, and visualize residues in the IR tail binding cleft important for coactivator-dependent ubiquitination. Overall, the results provide insights into assembly, regulation, and interactions of TPR proteins and the APC/C.« less

Authors:
 [1];  [1];  [2];  [2];  [1];  [3];  [1];  [1];  [2];  [4]
  1. St. Jude Children's Research Hospital, Memphis, TN (United States). Dept. of Structural Biology
  2. Research Inst. of Molecular Pathology (IMP), Vienna (Austria). Vienna Biocenter (VBC)
  3. Howard Hughes Medical Inst., Memphis, TN (United States). St. Jude Children's Research Hospital
  4. St. Jude Children's Research Hospital, Memphis, TN (United States). Dept. of Structural Biology; Howard Hughes Medical Inst., Memphis, TN (United States). St. Jude Children's Research Hospital
Publication Date:
Research Org.:
Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
USDOE
OSTI Identifier:
1177429
Alternate Identifier(s):
OSTI ID: 1367766
Grant/Contract Number:  
AC02-06CH11357; P41 GM103403; R37GM065930; P30CA021765
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Journal of Molecular Biology
Additional Journal Information:
Journal Volume: 427; Journal Issue: 8; Journal ID: ISSN 0022-2836
Publisher:
Elsevier
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Yamaguchi, Masaya, Yu, Shanshan, Qiao, Renping, Weissmann, Florian, Miller, Darcie J., VanderLinden, Ryan, Brown, Nicholas G., Frye, Jeremiah J., Peters, Jan-Michael, and Schulman, Brenda A. Structure of an APC3–APC16 Complex: Insights into Assembly of the Anaphase-Promoting Complex/Cyclosome. United States: N. p., 2014. Web. doi:10.1016/j.jmb.2014.11.020.
Yamaguchi, Masaya, Yu, Shanshan, Qiao, Renping, Weissmann, Florian, Miller, Darcie J., VanderLinden, Ryan, Brown, Nicholas G., Frye, Jeremiah J., Peters, Jan-Michael, & Schulman, Brenda A. Structure of an APC3–APC16 Complex: Insights into Assembly of the Anaphase-Promoting Complex/Cyclosome. United States. https://doi.org/10.1016/j.jmb.2014.11.020
Yamaguchi, Masaya, Yu, Shanshan, Qiao, Renping, Weissmann, Florian, Miller, Darcie J., VanderLinden, Ryan, Brown, Nicholas G., Frye, Jeremiah J., Peters, Jan-Michael, and Schulman, Brenda A. 2014. "Structure of an APC3–APC16 Complex: Insights into Assembly of the Anaphase-Promoting Complex/Cyclosome". United States. https://doi.org/10.1016/j.jmb.2014.11.020. https://www.osti.gov/servlets/purl/1177429.
@article{osti_1177429,
title = {Structure of an APC3–APC16 Complex: Insights into Assembly of the Anaphase-Promoting Complex/Cyclosome},
author = {Yamaguchi, Masaya and Yu, Shanshan and Qiao, Renping and Weissmann, Florian and Miller, Darcie J. and VanderLinden, Ryan and Brown, Nicholas G. and Frye, Jeremiah J. and Peters, Jan-Michael and Schulman, Brenda A.},
abstractNote = {The anaphase-promoting complex/cyclosome (APC/C) is a massive E3 ligase that controls mitosis by catalyzing ubiquitination of key cell cycle regulatory proteins. The APC/C assembly contains two subcomplexes: the “Platform” centers around a cullin-RING-like E3 ligase catalytic core; the “Arc Lamp” is a hub that mediates transient association with regulators and ubiquitination substrates. The Arc Lamp contains the small subunits APC16, CDC26, and APC13, and tetratricopeptide repeat (TPR) proteins (APC7, APC3, APC6, and APC8) that homodimerize and stack with quasi-2-fold symmetry. Within the APC/C complex, APC3 serves as center for regulation. APC3's TPR motifs recruit substrate-binding coactivators, CDC20 and CDH1, via their C-terminal conserved Ile-Arg (IR) tail sequences. Human APC3 also binds APC16 and APC7 and contains a > 200-residue loop that is heavily phosphorylated during mitosis, although the basis for APC3 interactions and whether loop phosphorylation is required for ubiquitination are unclear. Here, we map the basis for human APC3 assembly with APC16 and APC7, report crystal structures of APC3Δloop alone and in complex with the C-terminal domain of APC16, and test roles of APC3's loop and IR tail binding surfaces in APC/C-catalyzed ubiquitination. The structures show how one APC16 binds asymmetrically to the symmetric APC3 dimer and, together with biochemistry and prior data, explain how APC16 recruits APC7 to APC3, show how APC3's C-terminal domain is rearranged in the full APC/C assembly, and visualize residues in the IR tail binding cleft important for coactivator-dependent ubiquitination. Overall, the results provide insights into assembly, regulation, and interactions of TPR proteins and the APC/C.},
doi = {10.1016/j.jmb.2014.11.020},
url = {https://www.osti.gov/biblio/1177429}, journal = {Journal of Molecular Biology},
issn = {0022-2836},
number = 8,
volume = 427,
place = {United States},
year = {Sat Dec 06 00:00:00 EST 2014},
month = {Sat Dec 06 00:00:00 EST 2014}
}

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Cited by: 28 works
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Works referencing / citing this record:

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