Characterization of the DOC1/APC10 subunit of the yeast and the human anaphase-promoting complex
Characterization of the DOC1/APC10 subunit of the yeast and the human anaphase-promoting complex
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DOI：
10.1074/jbc.274.20.14500
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发表时间：
1999-05-14
影响因子：
4.8
通讯作者：
Peters, JM
中科院分区：
文献类型：
--
作者：
Grossberger, R;Gieffers, C;Peters, JM
后期促进复合物/环体 (APC) 是一种泛素蛋白连接酶，其活性对于有丝分裂的进展至关重要。脊椎动物 APC 被认为由 8 个亚基组成，而在芽殖酵母中，已鉴定出几种其他 APC 相关蛋白，包括称为 Doc1 或 Apc10 的 33 kDa 蛋白。在这里，我们表明 Doc1/Apc10 是整个细胞周期中酵母 APC 的一个亚基。 Doc1/Apc10 的突变使 APC 失活，但不会破坏复合体的稳定性。Doc1/Apc10 的直系同源物（我们称为 APC10）与不同脊椎动物（包括人类和青蛙）的 APC 相关。对纯化的人 APC 成分的生化分级分离实验和质谱分析表明，APC10 是真正的 APC 亚基，其细胞水平或与 APC 的关联不受细胞周期调节。我们进一步在也包含 cullin 或 HECT 结构域的几个蛋白质序列中鉴定了 APC10 同源区域，我们建议将其称为 DOC 结构域。 Cullins 存在于包括 APC 在内的多种泛素化复合物中，而 HECT 结构域代表不同类型泛素蛋白连接酶的催化核心，因此 DOC 结构域对于几种类型的泛素蛋白连接酶催化的反应可能很重要。
The anaphase-promoting complex/cyclosome (APC) is a ubiquitin-protein ligase whose activity is essential for progression through mitosis. The vertebrate APC is thought to be composed of 8 subunits, whereas in budding yeast several additional APC-associated proteins have been identified, including a 33-kDa protein called Doc1 or Apc10, Here, we show that Doc1/Apc10 is a subunit of the yeast APC throughout the cell cycle. Mutation of Doc1/Apc10 inactivates the APC without destabilizing the complex, An ortholog of Doc1/Apc10, which we call APC10, is associated with the APC in different vertebrates, including humans and frogs. Biochemical fractionation experiments and mass spectrometric analysis of a component of the purified human APC show that APC10 is a genuine APC subunit whose cellular levels or association with the APC are not cell cycle-regulated, We have further identified an APC10 homology region, which we propose to call the DOC domain, in several protein sequences that also contain either cullin or HECT domains. Cullins are present in several ubiquitination complexes including the APC, whereas HECT domains represent the catalytic core of a different type of ubiquitin-protein ligase, DOC domains may therefore be important for reactions catalyzed by several types of ubiquitin-protein ligases.