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首页> 外文期刊>Molecular and Cellular Biology >Identification and Characterization of a Novel Component of the Human Minichromosome Maintenance Complex
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Identification and Characterization of a Novel Component of the Human Minichromosome Maintenance Complex

机译：人微染色体维持复合物新型成分的鉴定与表征

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Minichromosome maintenance (MCM) complex replicative helicase complexes play essential roles in DNA replication in all eukaryotes. Using a tandem affinity purification-tagging approach in human cells, we discovered a form of the MCM complex that contains a previously unstudied protein, MCM binding protein (MCM-BP). MCM-BP is conserved in multicellular eukaryotes and shares limited homology with MCM proteins. MCM-BP formed a complex with MCM3 to MCM7, which excluded MCM2; and, conversely, hexameric complexes of MCM2 to MCM7 lacked MCM-BP, indicating that MCM-BP can replace MCM2 in the MCM complex. MCM-BP-containing complexes exhibited increased stability under experimental conditions relative to those containing MCM2. MCM-BP also formed a complex with the MCM4/6/7 core helicase in vitro, but, unlike MCM2, did not inhibit this helicase activity. A proportion of MCM-BP bound to cellular chromatin in a cell cycle-dependent manner typical of MCM proteins, and, like other MCM subunits, preferentially associated with a cellular origin in G1 but not in S phase. In addition, down-regulation of MCM-BP decreased the association of MCM4 with chromatin, and the chromatin association of MCM-BP was at least partially dependent on MCM4 and cdc6. The results indicate that multicellular eukaryotes contain two types of hexameric MCM complexes with unique properties and functions.
机译：微型染色体维持（MCM）复合物解旋酶复合物在所有真核生物的DNA复制中都起着重要作用。在人细胞中使用串联亲和纯化标记方法，我们发现了一种MCM复合物，其中包含一种以前尚未研究的蛋白MCM结合蛋白（MCM-BP）。 MCM-BP在多细胞真核生物中是保守的，与MCM蛋白具有有限的同源性。 MCM-BP与MCM3至MCM7形成复合物，其中排除了MCM2;相反，MCM2至MCM7的六聚体缺乏MCM-BP，表明MCM-BP可以取代MCM复合物中的MCM2。相对于含有MCM2的复合物，含MCM-BP的复合物在实验条件下显示出更高的稳定性。 MCM-BP在体外还与MCM4 / 6/7核心解旋酶形成了复合物，但与MCM2不同，它不抑制这种解旋酶活性。一部分MCM-BP以MCM蛋白典型的细胞周期依赖性方式与细胞染色质结合，并且像其他MCM亚基一样，优先与G 1 中的细胞起源相关，但与S期无关。此外，MCM-BP的下调降低了MCM4与染色质的缔合，并且MCM-BP的染色质缔合至少部分取决于MCM4和cdc6。结果表明，多细胞真核生物包含两种类型的具有独特特性和功能的六聚体MCM复合物。

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