Localization of the coactivator Cdh1 and the cullin subunit Apc2 in a cryo-electron microscopy model of vertebrate APC/C

被引:80
作者
Dube, P
Herzog, F
Gieffers, C
Sander, B
Riedel, D
Müller, SA
Engel, A
Peters, JM
Stark, H
机构
[1] Res Inst Mol Pathol, A-1030 Vienna, Austria
[2] Max Planck Inst Biophys Chem, D-37077 Gottingen, Germany
[3] Univ Basel, Biozentrum, Maurice E Muller Inst, CH-4056 Basel, Switzerland
基金
奥地利科学基金会;
关键词
D O I
10.1016/j.molcel.2005.11.008
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The anaphase-promoting complex/cyclosome (APC/C) is a ubiquitin ligase with essential functions in mitosis, meiosis, and G1 phase of the cell cycle. APC/C recognizes substrates via coactivator proteins such as Cdh1, and bound substrates are ubiquitinated by E2 enzymes that interact with a hetero-dimer of the RING subunit Apc11 and the cullin Apc2. We have obtained three-dimensional (3D) models of human and Xenopus APC/C by angular reconstitution and random conical tilt (RCT) analyses of negatively stained cryo-electron microscopy (cryo-EM) preparations, have determined the masses of these particles by scanning transmission electron microscopy (STEM), and have mapped the locations of Cdh1 and Apc2. These proteins are located on the same side of the asymmetric APC/C, implying that this is where substrates are ubiquitinated. We have further identified a large flexible domain in APC/C that adopts a different orientation upon Cdh1 binding. Cdh1 may thus activate APC/C both by recruiting substrates and by inducing conformational changes.
引用
收藏
页码:867 / 879
页数:13
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