5G04

Structure of the human APC-Cdc20-Hsl1 complex

5G04 の概要

• エントリーDOI: 10.2210/pdb5g04/pdb
• 関連するPDBエントリー: 5G05
• EMDBエントリー: 3385
• 分子名称: ANAPHASE-PROMOTING COMPLEX SUBUNIT 1, ANAPHASE-PROMOTING COMPLEX SUBUNIT 10, ANAPHASE-PROMOTING COMPLEX SUBUNIT 13, ... (17 entities in total)
• 機能のキーワード: cell cycle, phosphorylation, mitosis, ubiquitination
• 由来する生物種: HOMO SAPIENS (HUMAN); 詳細
• 細胞内の位置: Nucleus : Q9BS18 P30260 Q8NHZ8; Cytoplasm, cytoskeleton, microtubule organizing center, centrosome : Q12834 Q13042; Bud neck : P34244; Cytoplasm : Q9NYG5 Q96DE5
• タンパク質・核酸の鎖数: 21
• 化学式量合計: 1250917.04

構造登録者

Zhang, S.,Chang, L.,Alfieri, C.,Zhang, Z.,Yang, J.,Maslen, S.,Skehel, M.,Barford, D. (登録日: 2016-03-16, 公開日: 2016-05-25, 最終更新日: 2024-05-08)

主引用文献

Zhang, S.,Chang, L.,Alfieri, C.,Zhang, Z.,Yang, J.,Maslen, S.,Skehel, M.,Barford, D.
Molecular Mechanism of Apc/C Activation by Mitotic Phosphorylation.
Nature, 533:260-, 2016

PubMed Abstract: In eukaryotes, the anaphase-promoting complex (APC/C, also known as the cyclosome) regulates the ubiquitin-dependent proteolysis of specific cell-cycle proteins to coordinate chromosome segregation in mitosis and entry into the G1 phase. The catalytic activity of the APC/C and its ability to specify the destruction of particular proteins at different phases of the cell cycle are controlled by its interaction with two structurally related coactivator subunits, Cdc20 and Cdh1. Coactivators recognize substrate degrons, and enhance the affinity of the APC/C for its cognate E2 (refs 4-6). During mitosis, cyclin-dependent kinase (Cdk) and polo-like kinase (Plk) control Cdc20- and Cdh1-mediated activation of the APC/C. Hyperphosphorylation of APC/C subunits, notably Apc1 and Apc3, is required for Cdc20 to activate the APC/C, whereas phosphorylation of Cdh1 prevents its association with the APC/C. Since both coactivators associate with the APC/C through their common C-box and Ile-Arg tail motifs, the mechanism underlying this differential regulation is unclear, as is the role of specific APC/C phosphorylation sites. Here, using cryo-electron microscopy and biochemical analysis, we define the molecular basis of how phosphorylation of human APC/C allows for its control by Cdc20. An auto-inhibitory segment of Apc1 acts as a molecular switch that in apo unphosphorylated APC/C interacts with the C-box binding site and obstructs engagement of Cdc20. Phosphorylation of the auto-inhibitory segment displaces it from the C-box-binding site. Efficient phosphorylation of the auto-inhibitory segment, and thus relief of auto-inhibition, requires the recruitment of Cdk-cyclin in complex with a Cdk regulatory subunit (Cks) to a hyperphosphorylated loop of Apc3. We also find that the small-molecule inhibitor, tosyl-l-arginine methyl ester, preferentially suppresses APC/C(Cdc20) rather than APC/C(Cdh1), and interacts with the binding sites of both the C-box and Ile-Arg tail motifs. Our results reveal the mechanism for the regulation of mitotic APC/C by phosphorylation and provide a rationale for the development of selective inhibitors of this state.

PubMed: 27120157
DOI: 10.1038/NATURE17973

実験手法

ELECTRON MICROSCOPY (3.9 Å)