Department of Physiological Chemistry and Cancer Genomics Centre, UMC Utrecht, Utrecht, The Netherlands.
J Cell Sci. 2010 May 15;123(Pt 10):1623-33. doi: 10.1242/jcs.061549. Epub 2010 Apr 14.
Error-free chromosome segregation depends on timely activation of the multi-subunit E3 ubiquitin ligase APC/C. Activation of the APC/C initiates chromosome segregation and mitotic exit by targeting critical cell-cycle regulators for destruction. The APC/C is the principle target of the mitotic checkpoint, which prevents segregation while chromosomes are unattached to spindle microtubules. We now report the identification and characterization of APC16, a conserved subunit of the APC/C. APC16 was found in association with tandem-affinity-purified mitotic checkpoint complex protein complexes. APC16 is a bona fide subunit of human APC/C: it is present in APC/C complexes throughout the cell cycle, the phenotype of APC16-depleted cells copies depletion of other APC/C subunits, and APC16 is important for APC/C activity towards mitotic substrates. APC16 sequence homologues can be identified in metazoans, but not fungi, by four conserved primary sequence stretches. We provide evidence that the C. elegans gene K10D2.4 and the D. rerio gene zgc:110659 are functional equivalents of human APC16. Our findings show that APC/C is composed of previously undescribed subunits, and raise the question of why metazoan APC/C is molecularly different from unicellular APC/C.
染色体分离无误依赖于多亚基 E3 泛素连接酶 APC/C 的适时激活。APC/C 的激活通过靶向关键细胞周期调节剂进行破坏来启动染色体分离和有丝分裂退出。APC/C 是有丝分裂检查点的主要靶标,该检查点可防止染色体在未与纺锤体微管连接时进行分离。我们现在报告 APC16 的鉴定和特征,APC16 是 APC/C 的保守亚基。APC16 与串联亲和纯化的有丝分裂检查点复合物蛋白复合物相关联。APC16 是人类 APC/C 的真正亚基:它存在于整个细胞周期的 APC/C 复合物中,APC16 耗尽细胞的表型复制了其他 APC/C 亚基的耗尽,并且 APC16 对 APC/C 对有丝分裂底物的活性很重要。通过四个保守的主要序列延伸,可以在后生动物中但在真菌中识别 APC16 序列同源物。我们提供的证据表明,秀丽隐杆线虫基因 K10D2.4 和斑马鱼基因 zgc:110659 是人类 APC16 的功能等同物。我们的研究结果表明 APC/C 由以前未描述的亚基组成,并提出后生动物 APC/C 在分子上与单细胞 APC/C 不同的问题。