Mnd2 and Swm1 are core subunits of the Saccharomyces cerevisiae anaphase-promoting complex
Mnd2 and Swm1 are core subunits of the Saccharomyces cerevisiae anaphase-promoting complex
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DOI：
10.1074/jbc.m213109200
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发表时间：
2003-05-09
影响因子：
4.8
通讯作者：
Borchers, CH
中科院分区：
文献类型：
--
作者：
Hall, MC;Torres, MP;Borchers, CH
后期促进复合物（APC）是一种多亚基E3泛素连接酶，在真核细胞中调节中期-后期转换和退出有丝分裂。先前已经在来自芽殖酵母的APC中鉴定了11个亚基。我们已经确定了两个额外的亚基，Mnd 2和Swm 1，通过质谱。Mnd 2和Swm 1都被发现与来自单倍体酵母全细胞提取物的高度纯化的APC制剂特异性相关。此外，APC与表位标记的Mnd 2和Swm 1共纯化。这两种蛋白质存在于APC制剂从单倍体细胞停滞在G（1），S和M期，并从减数分裂二倍体细胞，表明它们是组成成分的复合物在整个酵母细胞周期。Mnd 2相互作用强烈，Cdc 23，Apc 5，Apc 1共表达时，在体外转录/翻译反应。Swm 1与Cdc 23和Apc 5也存在相互作用。先前的研究描述了MND 2和SWM 1突变的减数分裂缺陷。在这里，我们发现mnd 2Delta和swm 1Delta单倍体菌株表现出缓慢的生长和G（2）/M细胞的积累，与apc 9Delta或apc 10 Delta菌株中观察到的相当，并与APC缺陷一致。综上所述，这些结果证明Swm 1和Mnd 2是酵母APC的功能组分。
The anaphase-promoting complex (APC) is a multi-subunit E3 ubiquitin ligase that regulates the metaphase-anaphase transition and exit from mitosis in eukaryotic cells. Eleven subunits have been previously identified in APC from budding yeast. We have identified two additional subunits, Mnd2 and Swm1, by mass spectrometry. Both Mnd2 and Swm1 were found specifically associated with a highly purified preparation of APC from haploid yeast whole cell extract. Moreover, the APC co-purified with epitope-tagged Mnd2 and Swm1. Both proteins were present in APC preparations from haploid cells arrested in G(1), S, and M phases and from meiotic diploid cells, indicating that they are constitutive components of the complex throughout the yeast cell cycle. Mnd2 interacted strongly with Cdc23, Apc5, and Apc1 when coexpressed in an in vitro transcription/translation reaction. Swm1 also interacted with Cdc23 and Apc5 in this system. Previous studies described meiotic defects for mutations in MND2 and SWM1. Here, we show that mnd2Delta and swm1Delta haploid strains exhibit slow growth and accumulation of G(2)/M cells comparable with that seen in apc9Delta or apc10Delta strains and consistent with an APC defect. Taken together, these results demonstrate that Swm1 and Mnd2 are functional components of the yeast APC.