Interactions between Mcm10p and other replication factors are required for proper initiation and elongation of chromosomal DNA replication in Saccharomyces cerevisiae
Interactions between Mcm10p and other replication factors are required for proper initiation and elongation of chromosomal DNA replication in Saccharomyces cerevisiae
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DOI：
10.1046/j.1365-2443.2000.00387.x
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发表时间：
2000-12-01
期刊：
影响因子：
2.1
通讯作者：
Sugino, A
中科院分区：
文献类型：
--
作者：
Kawasaki, Y;Hiraga, S;Sugino, A
背景：MCM 10是酿酒酵母染色体DNA复制启动所必需的。以前的工作表明，Mcm 10 p与Mcm 2 -7蛋白复合物相互作用，可能是作为复制许可因子。此外，Mcm 10 p是必需的在原点激活和拆卸的prereplicative复杂，这使得顺利通过的复制fork.Results：我们表明，一个MCM 10突变导致DNA合成的缓慢进展和染色体的完整性损失在S期，并防止进入有丝分裂，尽管明显完成染色体DNA复制在nonpermissive温度。此外，Mcm 10 p在遗传上与起源识别复合物（ORC）和各种复制延伸因子（包括DNA聚合酶epsilon和delta的亚基）相互作用。Mcm 10 p是一种丰富的蛋白质（每个单倍体细胞约4 × 10（4）拷贝），在细胞周期的所有阶段几乎完全定位于染色质和/或核基质组分中。结论：Mcm 10 p与Mcm 2 -7蛋白一样，是复制前染色质的重要组成部分，在染色体DNA复制的启动过程中与ORC共同起作用;此外，Mcm 10 p在DNA复制的延伸过程中起重要作用。
Background: MCM10 is essential for the initiation of chromosomal DNA replication in Saccharomyces cerevisiae. Previous work showed that Mcm10p interacts with the Mcm2-7 protein complex that may be functioning as the replication-licensing factor. In addition, Mcm10p is required during origin activation and disassembly of the prereplicative complex, which allows smooth passage of replication forks.Results: We show that an mcm10 mutation causes a slow progression of DNA synthesis and a loss of chromosome integrity during the S phase and prevents entry into mitosis, despite apparent completion of chromosomal DNA replication at nonpermissive temperatures. Furthermore, Mcm10p interacts genetically with the origin recognition complex (ORC) and various replication elongation factors, including a subunit of DNA polymerases epsilon and delta. Mcm10p is an abundant protein (approximately 4 x 10(4) copies per haploid cell) that is almost exclusively localized in the chromatin and/or nuclear matrix fractions during all phases of the cell cycle. When it is visualized by the chromosome-spreading method followed by immunostaining, Mcm10p forms punctate foci on chromatin throughout the cell cycle and these foci mostly overlap with those of Orc1p, a component of ORC.Conclusions: These results suggest that Mcm10p, like the Mcm2-7 proteins, is a critical component of the prereplication chromatin and acts together with ORC during the initiation of chromosomal DNA replication; in addition, Mcm10p plays an important role during the elongation of DNA replication.