Multiple domains of fission yeast Cdc19p (MCM2) are required for its association with the core MCM complex.
Multiple domains of fission yeast Cdc19p (MCM2) are required for its association with the core MCM complex.
复制标题
裂殖酵母 Cdc19p (MCM2) 的多个结构域需要与核心 MCM 复合物结合。
DOI：
10.1091/mbc.9.7.1833
复制
发表时间：
1998
影响因子：
3.3
通讯作者：
Forsburg,SL
中科院分区：
文献类型：
--
作者：
Sherman,DA;Pasion,SG;Forsburg,SL
MCM蛋白家族的成员是形成六元蛋白复合物的真核生物DNA复制必需因子。在这项研究中，我们使用四个MCM蛋白的抗体，调查的结构和要求，在体内形成的裂变酵母MCM复合物，特别是关于Cdc19p（MCM2）。凝胶过滤分析表明，MCM蛋白复合物是不稳定的，可以分解成亚复合物。使用免疫共沉淀，我们发现，Mis5p（MCM6）和Cdc21p（MCM4）是紧密相关的核心复合物与Cdc19p松散协会彼此。Cdc19 p与核心的组装依赖于Cdc21 p。有趣的是，有没有在Cdc19p的MCM配合物通过细胞周期的明显变化。使用面板的Cdc19p突变体，我们发现，多个域的Cdc19p所需的MCM结合。这些研究表明裂殖酵母中的MCM复合物具有不同的亚结构，这可能与功能有关。
The members of the MCM protein family are essential eukaryotic DNA replication factors that form a six-member protein complex. In this study, we use antibodies to four MCM proteins to investigate the structure of and requirements for the formation of fission yeast MCM complexes in vivo, with particular regard to Cdc19p (MCM2). Gel filtration analysis shows that the MCM protein complexes are unstable and can be broken down to subcomplexes. Using coimmunoprecipitation, we find that Mis5p (MCM6) and Cdc21p (MCM4) are tightly associated with one another in a core complex with which Cdc19p loosely associates. Assembly of Cdc19p with the core depends upon Cdc21p. Interestingly, there is no obvious change in Cdc19p-containing MCM complexes through the cell cycle. Using a panel of Cdc19p mutants, we find that multiple domains of Cdc19p are required for MCM binding. These studies indicate that MCM complexes in fission yeast have distinct substructures, which may be relevant for function.