Purification and functional inactivation of the fission yeast MCMMCM-BP complex
Purification and functional inactivation of the fission yeast MCMMCM-BP complex
复制标题
DOI：
10.1016/j.febslet.2011.10.033
复制
发表时间：
2011-12-15
期刊：
影响因子：
3.5
通讯作者：
MacNeill, Stuart A.
中科院分区：
文献类型：
--
作者：
Li, Juan-juan;Schnick, Jasmin;MacNeill, Stuart A.
MCM（微型染色体维持）复合物是真核复制解旋酶的核心，由六种蛋白质组成：Mcm2-Mcm7。在人类中，该复合物的一种变体形式是 MCM2 被 MCM-BP 蛋白取代。最近的结果表明，裂殖酵母中存在类似的复合物，在 DNA 复制和细胞周期进展中发挥重要作用。在这里，我们描述了裂殖酵母 MCMMcb1 复合物的纯化和亚基组成。使用新生成的温度敏感等位基因，我们表明MCMMcb1功能的丧失会导致DNA损伤的积累、检查点激活和细胞周期停滞，并为MCMMcb1在减数分裂中的作用提供证据。蛋白质相互作用的结构化摘要：Mcb1通过pull down与Mcm4物理相互作用（查看相互作用）Mcb1通过pull down与Mcm5物理相互作用（查看相互作用）Mcb1通过pull down与Mcm6物理相互作用（查看相互作用）Mcm4 和 Mcb1 通过双分子荧光互补进行物理相互作用（查看相互作用）Mcb1 通过串联亲和纯化与 Mcm3、Mcm4、Mcm5、Mcm6 和 Mcm7 进行物理相互作用（查看相互作用）Mcb1 通过下拉与 Mcm7 进行物理相互作用（查看相互作用）(C) 2011 年欧洲生化协会联合会。由 Elsevier B.V 出版。保留所有权利。
The MCM (mini-chromosome maintenance) complex is the core of the eukaryotic replicative helicase and comprises six proteins, Mcm2-Mcm7. In humans, a variant form of the complex has Mcm2 replaced by the MCM-BP protein. Recent results suggest that a similar complex exists in fission yeast with an essential role in DNA replication and cell cycle progression. Here, we describe the purification and subunit composition of the fission yeast MCMMcb1 complex. Using newly generated temperature-sensitive alleles, we show that loss of MCMMcb1 function leads to accumulation of DNA damage, checkpoint activation and cell cycle arrest, and provide evidence for a role for MCMMcb1 in meiosis.Structured summary of protein interactions:Mcb1 physically interacts with Mcm4 by pull down (View interaction)Mcb1 physically interacts with Mcm5 by pull down (View interaction)Mcb1 physically interacts with Mcm6 by pull down (View interaction)Mcm4 and Mcb1 physically interact by bimolecular fluorescence complementation (View interaction)Mcb1 physically interacts with Mcm3, Mcm4, Mcm5, Mcm6 and Mcm7 by tandem affinity purification (View interaction)Mcb1 physically interacts with Mcm7 by pull down (View interaction) (C) 2011 Federation of European Biochemical Societies. Published by Elsevier B. V. All rights reserved.